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Hi I dont know if this technique is the best but it certainly works for us. Depending on what part of the beetle you want to examine, we stick and glue with silver dag, an entomological pin (for the small beetles) or a normal pin (for larger specimens)to an area of the specimen which is not going to be observed. From here we place the specimen and pin in a stub like vice. This allows you to gold coat the beetle and place in the SEM chamber with minimal handling. Ok yes the pin can be seen under the SEM. The idea is the mount the specimen in such away that the pin will not obscure in anyway the views that are wanted. ie if dorsal and front view are wanted then the pin would be placed in the ventral side at an angle less than 90 degrees sloping backwards towards the posterior end. This will give a greater angle to work with when observing the frontal position.
I know this is a brief explanation, however if you want to ask any questions please ring me on 02 9320 6198
Hope this helps
Sue Lindsay
SEM Lab Australian Museum
I would like to know if there is any new technique about SEM and beetles, what is the best way to mount the beetle.
Keep care and be of good cheer.
Regards
Vratislav Richard Eugene Maria John Baptiste of Bejsak (Bayshark)-Collorado-Mansfeld
Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
Temporally home address: 32 Girrawheen Ave. Kiama NSW 2533 Australia e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
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BEGIN:VCARD VERSION:2.1 N:Bejsak-Collorado-Mansfeld;Vratislav Richard;Eugene Maria John Baptist FN:Vratislav Richard Eugene Maria John Baptist Bejsak-Collorado-Mansfeld ORG:Bayshark Research Laboratory TITLE:director NOTE;ENCODING=QUOTED-PRINTABLE:Marketing and Coaching=0D=0A=0D=0ATenebrionidae Orbis and higher taxonomy TEL;WORK;VOICE:(+61 2) 9319 6380 TEL;CELL;VOICE:(+61 414) 540 465 ADR;WORK:;;29 Edward Street;Darlington, SYDNEY;;NSW 2008;Australie LABEL;WORK;ENCODING=QUOTED-PRINTABLE:29 Edward Street=0D=0ADarlington, SYDNEY NSW 2008=0D=0AAustralie ADR;HOME;ENCODING=QUOTED-PRINTABLE:;;(temporaly address):=0D=0A32 Girrawheen Ave;KIAMA;;NSW 2533;AUSTRALIA LABEL;HOME;ENCODING=QUOTED-PRINTABLE:(temporaly address):=0D=0A32 Girrawheen Ave=0D=0AKIAMA NSW 2533=0D=0AAUSTRAL= IA URL: URL:http://www.coleoptera.org EMAIL;INTERNET:ricardo-at-login.cz EMAIL;PREF;INTERNET:vratislav-at-bigfoot.com EMAIL;INTERNET:ricardo-at-ans.com.au REV:19981231T063007Z END:VCARD --IMA.Boundary.638414519--
} Hi } I dont know if this technique is the best but it certainly works for us. } Depending on what part of the beetle you want to examine, we stick and glue } with silver dag, an entomological pin (for the small beetles) or a normal } pin (for larger specimens)to an area of the specimen which is not going to } be observed. From here we place the specimen and pin in a stub like vice. } This allows you to gold coat the beetle and place in the SEM chamber with } minimal handling. } Ok yes the pin can be seen under the SEM.
Paint the pin with dag before mounting and if you get your contrast right the pin dissapear into the bagground!!!
Just a usefull hint. Mr. S H Coetzee Tell: (011) 716 2419 Electron Microscope Unit Fax: (011) 339 3407 Private bag X3 E-mail: Stephan-at-gecko.biol.wits.ac.za Wits Johannesburg 2050
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I highly reccomend MOC (Microscopical Optical Consulting) to service your microtomes. This is a small, independant company in Cottage Valley, NY. They do all of our's every year - Reichart, Leica, Sorvall, etc. Their # is 914-268-6450. Good luck! Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
A Research Fellow is required in the Department of Materials, University of Leeds, for a fixed period of two years, to work on an electron microscope study of the mechanism of graphitisation in steels. Applicants should have, or be near completing, a PhD in Metallurgy, Materials Science or a related discipline. Experience in transmission electron microscopy would be advantageous.
Salary: Research IA within the range stlg15,735- stlg19,197 p.a. according to qualifications and relevant experience.
Application forms and further particulars may be obtained from Professor D V Edmonds, Department of Materials, University of Leeds, Leeds, LS2 9JT, UK. tel: 0113 233 2348, fax: 0113 233 2384.
Job ref: 062-066-004-027.
Closing date: 29 January 1999.
Towards Equal Opportunities
Professor David V Edmonds Department of Materials School of Process, Environmental and Materials Engineering University of Leeds Leeds LS2 9JT United Kingdom
-- Jane M. Woodruff Polysciences Inc Phone: 215-343-6484 400 Valley Rd. 800-523-2575 Warrington, PA 18976 Fax:215-343-0214 E-mail jwoodruf-at-polysciences.com
Project MICRO is MSA's middle school educational outreach program. In his capacity as MSA webmaster Nestor gave MICRO a wonderful Christmas present; a major expansion of its web page. The new URL is http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html There is a LOT of microscopy education info there, including the quotations that listserver readers contributed last year. Please visit the site, and send me your comments.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I am a vendor, and we developed and sell "Entomounts", which are basically specimen mounts with the entomology pins already in them. They are provided as a convenience, and are so reasonably priced that I don't care much one way or the other whether we sell a bunch of them or not {grin} .
Happy New Year! Steven Slap
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
I have a position open for an EM TECHNICIAN, SENIOR, open. The job entails negative staining and thin sectioning of clinical specimens for viral examination. The salary is $26K plus good health benefits. Send resume to me below, or email for questions.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We have a researcher who is interested in imaging some molecules (proteins, primarily) in an aqeuous medium in order to determine if any conformational changes are occurring. Originally, this work was being done using x-ray microscopy; however, since there are only a very few such microscopes in the world, I suggested cryo TEM.
We are now looking for central facilities where high resolution cryo-TEM work might be done.
Thank you.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I fear that my copy of Microscopy News (I think that is correct) which had a procedure for the preparation of snowflakes my have hit the recycling bin without my knowledge or consent.
Can anyone forward a copy of the procedure to me? I would greatly appreciate it.
Thanks.
Alan Stone ASTON Metallurgical Services as-at-mcs.com
Maria Lucia Ribeiro Caldas wrote: ============================================= Does anyone have a protocol for the resin SPI-PON 812 (Sigma). ============================================== This product originally came from our firm, SPI Supplies. It is our SPI-Pon 812 epoxy resin, it is one of the components of our "Epon replacement" kit, and is described on our website given below.
We have not yet put up on our website the "Use Instructions" for this product, but if you send me your FAX number, I will make sure that a complete set is FAXed off to you right away. SPI-Pon 812 resin is "plug in" compatible with all known protocols based on the original Epon 812 protocols , so you could continue using whatever protocol(s) you have found successful in the past.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
------------------------------------------------------------------- FOR THOSE PLANNING TO SUBMIT A PAPER TO THE MICROSCOPY AND MICROANALYSIS MEETING.
This year information about the authors of Papers submitted to the M&M '99 meeting in Portland, OR should be submitted electronically. Unfortunately, the URL for submission was inadvertently left out of the "Call for Papers" booklet. The URL is:
The abstract itself still should be mailed to Microscopy & Microanalysis "99 Meeting Management (as indicated in the Call for Papers).
Additional information about the meeting and a link to the data entry page are available at:
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If you need a copy of the "Call for Papers" contact M&M '99 meeting management toll free at 877-672-6271 or toll call at 708-361-6045. Fax number: 708-361-6166.
Hello everybody, I would like to get some information on TEM diffraction pattern analysis. Specifically;
1) What software is available for analysis of diffraction patterns (both ring patterns and spot patterns)? What kind of accuracy can be obtained - are we getting close to the accuracy of X-ray diffractometry yet, or are there fundamental reasons such as lens aberrations, smaller Bragg angles, and accuracy of measurement which mean that we'll never get there?
2) What are the typical procedures people use for, say, measuring camera length or identifying unknown phases using diffraction?
I will make a summary of replies and distribute it to anyone who is interested.
Many thanks in advance, and a Happy New Year to all,
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK e-mail richard.beanland-at-gecm.com
Make a 1% solution of Formvar in methylene chloride (chloroform will work too). Chill the solution and some glass slides (leave them outside with a cover to keep the snow off). Dip a toothpick in the Formvar solution and place drops on a slide, then catch snowflakes on a piece of black velvet or something similar. With a toothpick wetted with the solution, touch a snowflake ever so slightly and it will cling so you can transfer it to a drop on the slide. Cover the slide and let the solvent evaporate (this happens very quickly). Take the slide inside and the snowflake will melt, leaving the Formvar replica. (To preserve the most delicate structure, leave the slide outside longer to let the water sublimate).
This is a great treatment for cabin fever. Happy New Year.
I've been using a variation of the "ento pin" for years that provides a little more flexibility than a rigid pin. Cut a long, thin triangular piece of thick aluminum foil (like that in a weighing dish - in a pinch you can use aluminum or copper tape), bend the base at a 90 degree angle, and stick it to the stub with carbon paint or your favorite conductive adhesive. Mount the insect on the point with conductive adhesive and coat. After coating, re-paint the stub surface and pin with carbon paint to darken the background. The mount is flexible enough to make fine adjustments to the positioning
of the insect (to get an exactly lateral view, or to hide the pin or whatever) and can be bent 90 degrees in any direction to get dorsal, ventral or other views. This works a lot better than trying to tilt the stage, as in most scopes you lose the ability to move in the X or Y direction at high tilts. Plus the background remains darker if you don't have to tilt.
Hope this helps
JME
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Lucy, I'm not familiar with that specific mounting media but if it is for permanent slides (similar to Cargill melt mounts), then place the container (if glass) on a hot plate and slowly heat it up until the crystals go back into solution. You may need to repeat this if recrystallization occurs between uses. Hope this is helpful. Mike Bucker Consolidated Labs of Va Feed Microscopy Unit
} } } maria lucia ribeiro caldas {caldasml-at-amcham.com.br} 01/04 6:29 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all
I have an old 100g bottle of Ultramount II (Ladd) which is completely crystallized. I would like to know if there any solution for this.
maria lucia ribeiro caldas wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Dear all } } I have an old } 100g bottle of } Ultramount II } (Ladd) which is } completely } crystallized. I } would like to } know if there } any solution } for this. } } Thanks } } Lucy
Dear Lucy,
It would depend. If there is some solvent left ther might be a chance to save it. If you wish to contact me directly and give me a liittle more detail I can advise you further. On your other post, SPI-812 is a replacement for the old Epon 812 as is our LX-112 and similar products sold be other supply houses. SPI-812 I believe probably is a trademark product from SPI, but I could supply you with the protocols from our product if you wish.
Thanks,
Dr. Charles Duvic Chemist -- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
Is anyone using or has anyone used Improvision's OpenLab software? We have had it in for a demo and are curious as to how it performs in "real life", how is the tech support, etc. Any comments (positive or negative) will be appreciated.
I have acquired 2 sets of human blood smears - one stained with Wright's stain and one with cresyl violet to show reticulocytes. I would like to incorporate them into my histology class student slide sets. I have coverslipped the smears but there is still a little stained area outside some of the coverslipped area. Are there any safety issues with stained smears or can one assume that any potential viral material would be killed by the staining step. I am unfamilair with the exact staining procedure but thought that there is ethanol in the stain. any comments from knowledge histotechs. Thanks in advance. Tom Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Date sent: Tue, 5 Jan 1999 09:55:29 -0500 (EST) } From: Tamara Howard {howard-at-cshl.org} To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com} We have been using openlab here for about 1 year now. It is being used for both time lapse imaging and low light GFP imaging. We are happy with the software. We have had no real problems. The only problems that we did run into were generally users (my self included) getting a little confused. The whole package is very complex and complete, depending on which modules you have and does require a little time to learn. I did find the automator, a module for automating repetitive or long protracted tasks, could become slightly cluttered, again more dependant on how the user was laying out the tasks. We did run into a couple of problems, or tasks that made us scratch our heads. When we phoned their UK tech support line, there was always someone able to help and they resolved all the problems that we had. When the line was busy they always phoned back. These remarks all relate to the earlier version of open lab, Open lab 2 only arrived with us just before christmas so I have not had long to play with it, but so far it appears to be as good as before. They also appear to be setting up internet user groups and an improved web site.
If you need any more info please give me a call.
Peter
Peter McHardy Technology Services Manager, Beatson Institute, Glasgow University, Garscube Estate, Bearsden, Glasgow G61 1BD Tel 0141 330 4818 Fax 0141 330 4127 http://www.beatson.gla.ac.uk/pmh
I am trying to embed L. dispar spermathecae for TEM, however I'm having = a terrible time dissecting / fixing the organs without losing the sperm = contained within them. Does anyone have any suggestions? ( I have = tried fixing the organs before dissecting them from the insect - this has not worked well )
Thanks,
Laura K. Garvey University of Connecticut Dept. of Molecular and Cell Biology=20 U-131, Beach Hall Storrs, CT 06269=20
DEPARTMENTS OF PHYSICS & ASTRONOMY AND CHEMISTRY=20
POST-DOCTORAL RESEARCH ASSISTANT =20
RA1A =A315,537 - =A323,651=20
A post-doctoral position is available for up to 24 months to work on an EPSRC funded project, "The use of XANES and ELNES for the characterisation of stabilised zirconia". The project is a collaboration between Glasgow University, The Queen's University, Belfast, MEL Chemicals Ltd and Johnson Matthey Ltd. The part of the project associated with this post involves modelling the near edge fine structure present on the edges observed in x-ray absorption spectroscopy and electron energy loss spectroscopy. Experience with first principles band structure calculations is essential and a background in the theoretical interpretation of spectroscopic techniques such as ELNES and XANES would be highly desirable, as would a knowledge of many-body physics. The post will be based in Glasgow but will involve extended periods at The Queens University working with Professor Finnis and the Atomic Simulations Group. =20
Further information is available at http://www.ssp.gla.ac.uk/ or from Professor Alan Craven, Department of Physics and Astronomy, University of Glasgow, Glasgow G12 8QQ. (Tel 0141 330 5892, FAX 0141 330 4464, a.craven-at-physics.gla.ac.uk) to whom applications, including a CV and the names of two referees, should be sent. Closing date - 12 January 1999.
------------------------------------------------------------------------ Dr Dave McComb Lecturer in Materials Chemistry Department of Chemistry University of Glasgow Glasgow G12 8QQ UK
} } Hello everybody, } I would like to get some information on TEM diffraction pattern } analysis. Specifically; } } 1) What software is available for analysis of diffraction patterns (both ring } patterns and spot patterns)?
One package is Desktop Microscopist. I have some FORTRAN code (written for a UNIX platform) which is helpful for indexing spot patterns to a known structure (If you are interested, you may contact me). I am sure there are others, too. A place to look would be the Sincris site at
http://www.lmcp.jussieu.fr/sincris/logiciel/
} What kind of accuracy can be obtained - are we } getting close to the accuracy of X-ray diffractometry yet, or are there } fundamental reasons such as lens aberrations, smaller Bragg angles, and } accuracy of measurement which mean that we'll never get there?
Because of lens hysteresis, it's not possible on a standard TEM to calibrate the camera constant L*(lambda) to an accuracy of greater than about 2 percent. A way to get more accuracy is to use an internal standard. For a good discussion of the problems, see Hirsch, Howie, Nicholson, Pashley and Whelan, Electron Microscopy of Thin Crystals, end of chapter 1.
In spite of the camera length inaccuracy, the RELATIVE spacings for two spots isn't affected. Therefore, it is potentially limited only by measurement inaccuracies (how precisely can we locate the center of the spot), and by the relrods (see Hirsch et al).
I believe that a spherical-type distortion of the pattern occurs if you use the beam convergence (condenser lens) to compensate for poor focus of the diffraction pattern. I've never read a good discussion of this (anyone know of one?) Also, ring patterns can be distorted from circular by astigmatism effects (in any post-specimen lens). These optical factors would also limit the accuracy of relative d-spacings, though to some extent one may be able to correct for them.
One place where electrons have a significant advantage over x-rays is with respect to noise. The interaction of electrons with matter is strong, so in very short experimental times good statistics can be obtained from miniscule sample volumse. Electron diffraction therefore has potential for structure refinements (e.g. recent work by Jansen and Zandbergen, L. D. Marks et al for surface diffraction). Also, the photographic film or CCD is a 2D detector (rare in XRD), so you can win big time in reducing noise. This can be used in studies of amorphous materials, via circumferential averaging of the patterns. However, strictly speaking, both dynamical and inelastic effects have to be considered in quantitatively interpreting the data.
} } 2) What are the typical procedures people use for, say, measuring camera length } or identifying unknown phases using diffraction?
Again, you can measure camera length as accurately as you want. Turn the scope off and on and it may differ by a couple of percent.
} } I will make a summary of replies and distribute it to anyone who is interested.
Thanks, I would be interested in hearing.
Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
mailto:wharton.sinkler-at-anlw.anl.gov
} } Many thanks in advance, and a Happy New Year to all, } } Richard Beanland } GMMT Ltd., } Caswell, } Towcester, } Northants NN12 8EQ } UK } e-mail richard.beanland-at-gecm.com } } }
Workshop on Quantitative Image Analysis May 20-22 and May 24-26, 1999, North Carolina State University, Raleigh, North Carolina, USA June 14-16, 1999, Danish Technological Institute, Taastrup, Denmark
This highly regarded hands-on course taught by Dr. John Russ and other expert faculty has been presented annually for more than 15 years. It deals with all phases of quantitative and computer-assisted imaging from acquisition and processing through measurement and stereological interpretation. Attendees receive The Image Processing Handbook plus a CD-ROM containing images, algorithms (Photoshop-compatible for Mac and Windows) and an extensive tutorial. The course is appropriate for professionals scientists, technicians and administrators using these techniques for research. Attendees typically come from materials science, geology, biological and medical sciences, pharmaceuticals, food science, industrial quality control, remote sensing, and other disciplines.
For detailed information and registration contact Cindy Allen, Dept. of Continuing and Professional Education, N. C. State University, Raleigh, NC 27695-7401, 919-515-8171, fax 919-515-7614, email: cindy_allen-at-ncsu.edu
On-line information is available at http://members.aol.com/IPCourse/
Dear listservers,=20 This topic was discussed awhile ago. Hopefully, some new developments = are now here to solve the problem easily. We are looking for a dedicated = software package for scheduling, in this case, instrument use = incorporated into our webpage. We have several microscopes, = workstations, microtomes, etc. and would like to have calenders or = something similar for each, so users could sign up in advance using the = web. It should be possible to assign different levels of privilege to = each user. Once an entry is made it could only be changed by an = administrator other than the original user. We like would like to then = to automatically transfer this information into a billing database, = either Access or FileMaker Pro based. Is something like what I have = described commercially available?
Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
I don't have the URL immediately at hand, but you might try the Filemaker website. They have additional templates besides those shipping with Filemaker. there are also links to Filemaker consultants who have free templates or who may be able to advise you regarding feasibility.
Regards, Glen
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
On Wed, 6 Jan 1999, hank p adams wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear listservers, } This topic was discussed awhile ago. Hopefully, some new developments are now here to solve the problem easily. We are looking for a dedicated software package for scheduling, in this case, instrument use incorporated into our webpage. We have several microscopes, workstations, microtomes, etc. and would like to have calenders or something similar for each, so users could sign up in advance using the web. It should be possible to assign different levels of privilege to each user. Once an entry is made it could only be changed by an administrator other than the original user. We like would like to then to automatically transfer this information into a billing database, either Access or FileMaker Pro based. Is something like what I have described commercially available? } } Hank Adams } Cell Biology } Integrated Microscopy Core } Baylor College of Medicine } One Baylor Plaza } Houston, Tx 77030 } } }
} I would like to get some information on TEM diffraction } pattern } analysis. Specifically; } } 1) What software is available for analysis of diffraction patterns (both ring } patterns and spot patterns)?
There is an operation in the SPIDER image processing program which will refine a lattice and determine the background-subtracted intensities. I have written a procedure which subtracts the circu- larly symmetric background, which improves the subsequent linear background subtraction. I have also written a routine (both in SPIDER and as a stand-alone) to determine the center and radius of a ring from up to 20 points. I can send you the source code for the stand-alone version.
} What kind of accuracy can be obtained - are we } getting close to the accuracy of X-ray diffractometry yet, or are there } fundamental reasons such as lens aberrations, smaller Bragg angles, and } accuracy of measurement which mean that we'll never get there? } When I selected 20 points from each of 13 rings on a gold pattern (5 from each quadrant), I got r/d* = 402.79+-0.49 pixels. The range of values was 401.89 to 403.71. The precision was close to 0.1%. Furthermore, there seemed to be no pattern of systematic variation, except that the larger rings, which were less intense and more diffuse, gave somewhat larger errors.
} 2) What are the typical procedures people use for, say, measuring camera } length } The best procedure, if it can be done, is to evaporate gold, or another standard, onto the crystal whose lattice constants are to be measured. This way one gets the lattice points and the standard on the same negative. I scan my negatives on a Perkin-Elmer micro- densitometer using a 10 mu x 10 mu window. For lattice constant measurement, I do not interpolate the file, but for intensity measure- ments, I reduce by a factor of 5 by averaging a 5 x 5 array for each pixel in the reduced image. This reduction does not produce errors in the background-subtracted intensities.
} I will make a summary of replies and distribute it to anyone who is } interested. } Thanx. I am on both listservers, so only those responses to you directly, rather than to a list, would be required.
} Many thanks in advance, and a Happy New Year to all, } And to you. Yours, Bill Tivol
We've had abit of a clean up here at RMIT and have some goodies to give away. We have: Two boxes Tungsten filaments, Item no. A050 purchased from AGAR these were used in an old ETEC. One H.V. supply for the ETEC, I've been told that when the ETEC was decommissioned it still worked!!!
Yes! they are old items, but perhaps someone has a creative use for them. If you want them you can contact me on the details below. Email has the best chance of getting me.
Richard, One major source of inaccuracy in the measurement of electron diffraction patterns lies in the rather large variation of camera length with the specimen height. If you use a double-tilt stage, the specimen height changes with tilt (only one tilt axis is eucentric). It is possible to calibrate the camera length against objective current when the specimen is in focus, but, as pointed out by Wharton Sinkler, switching the microscope off and on will undo your work. The height change with tilt can be obviated by using a rotation-tilt stage instead, but it can drive you nuts, especially if the crystal is not at the centre of the grid.
As was suggested, the best method is to use an internal standard as this compensates for both camera-length changes and projector astigmatism. It also enables the operator to estimate d-spacings on screen in order to decide whether the crystal is in a sensible orientation, and one can very quickly tell whether a pattern is rectangular or oblique.
I hope this helps, Eric ---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
} } Hi all. } } We are currently setting up a cryo-vitrification unit for TEM sample } analysis. Research into what is the best } cryogen to use for vitrification of our sample type gave the answer of } liquid ethane. } } Does anybody have any advice as to what materials to use or to NOT use } with liquid ethane (for material } incompatibility reasons - chemically and physically (i.e. extreme } temperatures))? Is copper okay with } liquid ethane? I've vaguely read somewhere that some groups use a copper } coil (in liquid nitrogen) to condense } their ethane. Any other confirmations? } } A small flexible length of tubing will also be needed to join the copper } (?) tubing to the cylinder regulator } (so we can move the coil in and out of the liquid N2 easily). Apparently } natural rubber is NOT good with ethane. } } Any other pearls of wisdom out there? } } Thanks in advance. I'll summarise my replies. } } Terri } } ------------------------------------- } Ms Terri Soar, PhD student, } University of South Australia } Email: terri-at-drage.com.au {mailto:Terri-at-drage.com.au} } ------------------------------------- }
Electron microscopy specialist needed immediately. Prepare ultra thin sections and photograph them using JEOL1200EX. Darkroom, Adobe Illustrator/Photoshop. Salary commensurate with experience.
Mail, e-mail, or FAX resume and two letters of recommendation to:
Dr. Peter Sterling 123 Anatomy-Chemistry BLDG Department of Neuroscience University of Pennsylvania Philadelphia, PA 19104-6058
I have used Formvar for over 50 years now and never known exactly what it is. In the back of my mind I seem to recall hearing that it is polyvinylformal, but I'm not certain that is correct. I have just consulted two polymer scientists in our department, and neither one of them knows what it is, nor could they find a chemical formula for it in their reference books.
If anyone knows what it is, I'd like to know too.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Philippe-Andr=E9 Buffat wrote: } =20 } I would be strongly interested in softwares able to handle digital } diffraction patterns. In particular a soft to flatten the background is } needed. From my experience, I can see on negatives faint rings or spots= , } but under the same condition it is impossible to see them on a computer } screen without increasing the contrast but then the range between black= and } white covers only a small part of densities in the whole pattern and th= e } observable area between the white center and the black periphery become= s } unacceptably small. The use of color coding of the intensities improves } only a little bit the visible range. An other soft to determine } automatically the spot position or the ring position by a refinement me= thod } or deconvolution of gaussian/lorentzian (or else) curves would also be } welcome.
Dear Philippe, In my old paper (Ultramicroscopy (1982) 9:117-130) I discuss=20 a technique of subtracting circularly-symmetric background, and give=20 a reference to an even older paper by Fraser et al. (Appl. Cryst.=20 (1977) 10:64- ). I have since written a procedure using the opera- tions in the image-processing program SPIDER (I'm sure the operations are also part of other programs), and I'd be happy to send you a copy. Briefly, the procedure refines a lattice, masks out the spots, replaces them with a local average background, makes a 2-D rotational average image, and subtracts this from the original pattern. The remaining=20 background is linear enough so that the usual form of background sub- traction will work. This process cannot be used for ring patterns, although a modification could work. Not only does this circularly- symmetric subtraction improve the visibility on a screen, it also increases the accuracy with which the intensities can be measured. As one can surmise, SPIDER has an operation to determine intensities and refine a lattice. It can also find the center and radius of a ring, and I have a stand-alone version of this. This program works by one choosing from 3 to 20 points on the ring, using the first 3 to make an initial guess, then least-squares-fitting the points (if there are more than 3). I'd be happy to send you the=20 FORTRAN code for the stand-alone ring program. Yours, Bill Tivol
Wharton, Nice summary, I would like to add a practical hint to those who may be new to ED. You can use an extrinsic standard such as a gold film. The improtant point is maintaining the same conditions for the standard and sample. To accomplish this the lens currents should be duplicated. An easy way to accomplish this is to focus the sample with the Z position while maintaining the lens currents from the standard. When using extrinsic standards the potential for a Z position mismatch and subsequent change in the objective lens current is the most likely error in camera length. Russ
-----Original Message----- } From: Wharton Sinkler [mailto:wharton.sinkler-at-anlw.anl.gov] Sent: Tuesday, January 05, 1999 2:07 PM To: Richard Beanland +44 1327 356363; Microscopy Listserver; lemas Listserver
} } Hello everybody, } I would like to get some information on TEM diffraction pattern } analysis. Specifically; } } 1) What software is available for analysis of diffraction patterns (both ring } patterns and spot patterns)?
One package is Desktop Microscopist. I have some FORTRAN code (written for a UNIX platform) which is helpful for indexing spot patterns to a known structure (If you are interested, you may contact me). I am sure there are others, too. A place to look would be the Sincris site at
http://www.lmcp.jussieu.fr/sincris/logiciel/
} What kind of accuracy can be obtained - are we } getting close to the accuracy of X-ray diffractometry yet, or are there } fundamental reasons such as lens aberrations, smaller Bragg angles, and } accuracy of measurement which mean that we'll never get there?
Because of lens hysteresis, it's not possible on a standard TEM to calibrate the camera constant L*(lambda) to an accuracy of greater than about 2 percent. A way to get more accuracy is to use an internal standard. For a good discussion of the problems, see Hirsch, Howie, Nicholson, Pashley and Whelan, Electron Microscopy of Thin Crystals, end of chapter 1.
In spite of the camera length inaccuracy, the RELATIVE spacings for two spots isn't affected. Therefore, it is potentially limited only by measurement inaccuracies (how precisely can we locate the center of the spot), and by the relrods (see Hirsch et al).
I believe that a spherical-type distortion of the pattern occurs if you use the beam convergence (condenser lens) to compensate for poor focus of the diffraction pattern. I've never read a good discussion of this (anyone know of one?) Also, ring patterns can be distorted from circular by astigmatism effects (in any post-specimen lens). These optical factors would also limit the accuracy of relative d-spacings, though to some extent one may be able to correct for them.
One place where electrons have a significant advantage over x-rays is with respect to noise. The interaction of electrons with matter is strong, so in very short experimental times good statistics can be obtained from miniscule sample volumse. Electron diffraction therefore has potential for structure refinements (e.g. recent work by Jansen and Zandbergen, L. D. Marks et al for surface diffraction). Also, the photographic film or CCD is a 2D detector (rare in XRD), so you can win big time in reducing noise. This can be used in studies of amorphous materials, via circumferential averaging of the patterns. However, strictly speaking, both dynamical and inelastic effects have to be considered in quantitatively interpreting the data.
} } 2) What are the typical procedures people use for, say, measuring camera length } or identifying unknown phases using diffraction?
Again, you can measure camera length as accurately as you want. Turn the scope off and on and it may differ by a couple of percent.
} } I will make a summary of replies and distribute it to anyone who is interested.
Thanks, I would be interested in hearing.
Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
mailto:wharton.sinkler-at-anlw.anl.gov
} } Many thanks in advance, and a Happy New Year to all, } } Richard Beanland } GMMT Ltd., } Caswell, } Towcester, } Northants NN12 8EQ } UK } e-mail richard.beanland-at-gecm.com } } }
You wrote: } } } Does anybody have any advice as to what materials to use or to NOT use } } with liquid ethane (for material } } incompatibility reasons - chemically and physically (i.e. extreme } } temperatures))? Is copper okay with } } liquid ethane? I've vaguely read somewhere that some groups use a copper } } coil (in liquid nitrogen) to condense } } their ethane. Any other confirmations? } } Both Cu and Al are compatable with LEt. The best scheme might be to pass the N2 vapor at ~80 K through the tube to liquify, but not freeze, the Et. We use an Al cup cooled by LN2, and there are problems with the Et solidifying.
} } A small flexible length of tubing will also be needed to join the copper } } (?) tubing to the cylinder regulator } } (so we can move the coil in and out of the liquid N2 easily). Apparently } } natural rubber is NOT good with ethane.
Tygon is also not good. Teflon tubing retains its strength and flexibility at 77 K, so I reccommend it. Good luck. Yours, Bill Tivol
I have done some more research on the matter of the composition of Formvar. In the book 'Techniques for Electron Microscopy' D. H. Kay, Ed., Blackwell Scientific, 1965 I find a statement indicating that Formvar is Polyvinyl Formal (p. 60) In the book 'Polymer Chemistry' by M. P. Stevens, Oxford Univ. Press, 1990, p.302, I find that the reaction of vinyl alcohol with butyl aldehyde produces a polymer called polyvinyl butyral. By analogy, if vinyl alcohol were reacted with formaldehyde (HCHO) one might assume it would produce polyvinyl formal. If this is so, AND IT IS ONLY A GUESS, then by analogy the chemical formula for the repeating unit in the polymer chain might be:
CH2 / \ -[CH2-CH CH]- | | O O \ / CH2
I hope this formula survives the process of being transmitted across the internet. This word processer is not ideal for writing organic chemical formulas.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Wil Bigelow wrote: } } I have used Formvar for over 50 years now and never known exactly what it is. In the back of my mind I seem to recall hearing that it is } polyvinylformal, but I'm not certain that is correct. I have just } consulted two polymer scientists in our department, and neither one of them knows what it is, nor could they find a chemical formula for it in their reference books.
Will et al:
According to the free sample, yes, I said free sample, I got from Monsanto, Formvar is a "polymer from polyvinyl alcohol and formaldehyde as as copolymer with polyvinyl acetate". If that is not enough information you could call Monsanto in St. Louis. I believe that it was originally developed to coat copper wire. Note that are several different types of Formvar. I think the type us EM folks use is 15/95 but I could be wrong.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Wilbur C. Bigelow wrote: =================================================== I have used Formvar for over 50 years now and never known exactly what it is . In the back of my mind I seem to recall hearing that it is polyvinylformal, but I'm not certain that is correct. I have just consulted two polymer scientists in our department, and neither one of them knows what it is, nor could they find a chemical formula for it in their reference books.
If anyone knows what it is, I'd like to know too. ================================================= You are correct, it is generically, polyvinyl formal, and the term "Formvar" is a trade name, originally registered (if my memory is correct) by Monsanto Chemical Company in St. Louis.
Disclaimer: SPI is a supplier of Formvar resin for use in electron microscopy.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
You will find all the information in my book "low Temperature Microscopy and Analysis" Plenum Press New York 1992
Patrick Echlin University of CambridheOn Wed, 6 Jan 1999, Mail Delivery
Subsystem wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } } Hi all. } } } } We are currently setting up a cryo-vitrification unit for TEM sample } } analysis. Research into what is the best } } cryogen to use for vitrification of our sample type gave the answer of } } liquid ethane. } } } } Does anybody have any advice as to what materials to use or to NOT use } } with liquid ethane (for material } } incompatibility reasons - chemically and physically (i.e. extreme } } temperatures))? Is copper okay with } } liquid ethane? I've vaguely read somewhere that some groups use a copper } } coil (in liquid nitrogen) to condense } } their ethane. Any other confirmations? } } } } A small flexible length of tubing will also be needed to join the copper } } (?) tubing to the cylinder regulator } } (so we can move the coil in and out of the liquid N2 easily). Apparently } } natural rubber is NOT good with ethane. } } } } Any other pearls of wisdom out there? } } } } Thanks in advance. I'll summarise my replies. } } } } Terri } } } } ------------------------------------- } } Ms Terri Soar, PhD student, } } University of South Australia } } Email: terri-at-drage.com.au {mailto:Terri-at-drage.com.au} } } ------------------------------------- } } } } } }
We use Tygon brand tubing connected to a copper coil when liquifying gaseous ethane. The tubing is connected to the ethane tank regulator at one end and the copper coil at the other. The copper coil is placed in a beaker in a styrofoam box in a fume hood. The ethane is turned on and liquid nitrogen is poured around the coil. As the copper and ethane start to get cold, the ethane will start to liquify and collect in the beaker. It should take approximately 2-3 minutes to start to liquify and 5-7 minutes to collect 2-300 ml.
You need to be extremely careful with liquid ethane. Besides the obvious (it's extremely cold and will produce severe burns rather quickly), it is volatile if it comes in contact with liquid nitrogen. So when you have a beaker of liquid ethane immersed in a box of liquid nitrogen, the potential for injury cannot be overstated. Hand (and forearm) as well as eye protection are essential.
We've been using liquid ethane for years and I even have photos of the setup in our lab. If you'd like, I could send you a copy of these. Feel free to write or call.
Hope this helps....
Russ
Russell W. Desnoyer Senior Research Technologist Cleveland Clinic Foundation Department of Molecular Cardiology 9500 Euclid Avenue Cleveland, Ohio 44195 Ph: (216) 444-4673 Fax: (216) 445-6062 E-mail: desnoyr-at-cesmtp.ccf.org
I need some info on binary alloys. Typically Aluminium, Titanium and Carbon Nitrides, Borides, etc. These are typically grown as thin films using cathodic arc deposition. What I need is some general info: What research has been done in the past? What research is currently happening? Where is the research is likely to go? What applications do these materials have as thin films and as bulk samples?
I have come across this several times, and there is no simple solution that I am aware of. I believe the reason has a lot to do with how the eye/brain interprets images. We do a good job of excluding noise, and we can often see patterns when it is very difficult to quantitatively measure them on a computer. In a sense we do a type of Maximum Entropy analysis -- we have a prior model of what is in the image and find features that fit this model, for instance weak spots or rings. (Of course, this also means that sometimes we find things that are not there.)
The best method that I am aware of is to combine a rank filter (good at reducing shot noise), some sort of high-pass filter to remove only the low frequencies (reducing the background) and pasting togethor images at different contrast levels to prevent the high intensity regions from dominating. At least for a picture this often works, although you have to play a lot with the kernel sizes. Quantitation is very hard. You have to set up a model (Maximum Entropy, Maximum Likelihood, Least-Squares) and perform a numerical fit. Sometimes Least-Squares works; I have never tried Maximum Entropy which should do better.
Philippe-Andr=E9 Buffat wrote: } =20 } I would be strongly interested in softwares able to handle digital } diffraction patterns. In particular a soft to flatten the background is } needed. From my experience, I can see on negatives faint rings or spots, } but under the same condition it is impossible to see them on a computer } screen without increasing the contrast but then the range between black a= nd } white covers only a small part of densities in the whole pattern and the } observable area between the white center and the black periphery becomes } unacceptably small. The use of color coding of the intensities improves } only a little bit the visible range. An other soft to determine } automatically the spot position or the ring position by a refinement meth= od } or deconvolution of gaussian/lorentzian (or else) curves would also be } welcome.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Formvar is indeed used to coat copper wire. My wife works for an overhead transformer manufacturing firm and they buy the stuff by the barrels for coating the wire (not just copper). It is a different grade and formulation, otherwise I would've been tempted to never buy the stuff again after purchasing a 55 gal drum of it...
On Wed, 06 Jan 1999 14:53:02 -0800 Geoff McAuliffe {mcauliff-at-UMDNJ.EDU} wrote: .. I got from } Monsanto, Formvar is a "polymer from polyvinyl alcohol and formaldehyde } as as copolymer with polyvinyl acetate". If that is not enough } information you could call Monsanto in St. Louis. I believe that it was } originally developed to coat copper wire. Note that are several } different types of Formvar.
I have some filters with particles of differing size but fixed density distributed over the surface. I would like to quantify and compare the "uniformity" of the particle mass distribution on each filter by SEM. My software estimates the volume of each particle using the min and max diameters and assuming an oblate spheroid. Is there a statistically sound recipe for using the observed variation in Vf - where Vf is the particle volume per field measured over many randomly selected fields - to quantify the uniformity of the sample's mass distribution and compare one sample to another? If two samples have different particle loadings, does one use different field areas on the two samples to get the same average Vf for both? The recipe should also include some confidence factor related to the fraction of filter area analyzed.
Thanks for your suggestions.
Bob Willis ManTech Environmental Technology, Inc Research Triangle Park, NC
For background and previous research, try "Phase Diagrams of Binary Titanium Alloys", edited by J. L. Murray. There may also be similar books (published by ASM) for aluminium and carbon. If not, the phase diagram compilation by T. B. Massalski would be a place to start, as well as "Journal of Phase Equilibria" (formerly "Bulletin of Alloy Phase Diagrams" ).
Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
mailto:wharton.sinkler-at-anlw.anl.gov
---------- } From: "George Theodossiou" {GEORGE-at-bunyip.ph.rmit.edu.au} } To: microscopy-at-sparc5.microscopy.com } Subject: Binary Alloys } Date: Thu, Jan 7, 1999, 10:43 AM }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One of our researchers has mouse embryos that express gfp-linked material. These embryos were preserved and maintained in PBS plus freshly made 4% p formaldehyde and 0.5% Tween 20. Initially, his control samples were pink in color without fluorescence. After 3 months, his control samples have started to turn green under fluorescent light. In so far as these are controls for his gfp expression samples, this fluorescence is not helpful.
Any idea what is causing this change in color? I recommended he should store his samples long-term in PBS plus 0.5% formaldehyde in PBS. Anyone have any better suggestions?
Also, those of you who work with gfp.....Could he treat his embryos, say with ammonium chloride, to try to get rid of his unwanted background fluorescence? Can he do an equivalent treatment of his controls and experimentals without damaging the gfp fluorescence he has previously observed?
thanks in advance
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-8759 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Chairman, Educational Outreach subcommittee promoting remote access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
You should look up the proceedings from the International Conference on Metallurgical Coatings and Thin Films for the past several years. The proceedings are two volumes and the papers are full papers. One volume is printed in Thin Solid Films and the other in Surface and Coatings Technology. You can find out more about this year's conference at the following web site: http://www.vacuum.org/icmctf/icmctf.html
Incidentally, I am a session chair for the "Microstructural, Microanalytical and Imaging Characterization" session in the "Coating and Thin Film Characterization" symposium.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: George Theodossiou To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hi all
I need some info on binary alloys. Typically Aluminium, Titanium and Carbon Nitrides, Borides, etc. These are typically grown as thin films using cathodic arc deposition. What I need is some general info: What research has been done in the past? What research is currently happening? Where is the research is likely to go? What applications do these materials have as thin films and as bulk samples?
i've been reading principals of heat treatment of steel by krauss and in the references of chapter 5 reference 4 lists a paper by S.Chattopadhyay and C.M. Sellars titled Quantitative measurements of pearlite spheroidization,Metallography,vol10,1977 pg 89-105
does anyone have this paper? if you do could you please fax it to me at 716-684-9433
or could you tell me what Metallography is ,is it a magizne?
Sounds like you may want to check with John Russ about many of these issues. I thought he was right there in your backyard. But I will offer a couple thoughts.
I think you would be better use the same field size for all of your measurements. Besides, Vf should have the same mean value no matter how large the field. The larger the field you measure, the less variation in field-specific measurements between fields. Supposing I find a standard deviation of 4% for some measurement for a single field (determined by measureing multiple fields). Then, if I measured a field 5 times as wide or measured 25 fields and averaged the results, the standard deviation on that measurement would be sqrt(25) less or 0.8%. Either way, 25 times more area was analyzed. So, you could make some adjustments to match up your measurements even if you did use different areas.
Regarding measuring the variation in Vf and compare samples - our work involves the measuring of void distributions. We are routinely measuring 20 frames per sample and calculating the variation in Af, but that is to have a measure of confidence in our Af measurements. It gives us some insight into the nature of our samples. A lower variability normally translates into a smaller average particle size, but it can also indicate some things about uniformity of distribution for the smae feature size between samples.
As to the statistical test for determining when the variation is significantly different between two populations with identical means, I will leave that to those that are better versed in statistics than I am at this moment. There must be one out there.
Hoping this helps. Warren
At 08:51 AM 1/7/99 -0600, you wrote: } I have some filters with particles of differing size but fixed density } distributed over the surface. I would like to quantify and compare the } "uniformity" of the particle mass distribution on each filter by SEM. My } software estimates the volume of each particle using the min and max } diameters and assuming an oblate spheroid. Is there a statistically } sound recipe for using the observed variation in Vf - where Vf is the } particle volume per field measured over many randomly selected fields - } to quantify the uniformity of the sample's mass distribution and compare } one sample to another? If two samples have different particle loadings, } does one use different field areas on the two samples to get the same } average Vf for both? The recipe should also include some confidence } factor related to the fraction of filter area analyzed. } } Bob Willis } ManTech Environmental Technology, Inc } Research Triangle Park, NC
For some reason, Philippe-Andre Buffat's posting to the listserver = never showed up in my mail. I wonder if this is common--getting partial = conversations.
Seeing faint features on TEM negatives and not on the digitized images = sounds as if it has more to do with limitations of the digitization process used = than it does with visualization. 8 bits probably just isn't enough integers to = preserve the faint image detail and keep the black and white extremes at the = same time. If the data is not preserved in the digital images, no amount of fancy = filtering is going to recover it. I'd suggest a careful look at what's going on = in the digitization process you're using. If your scanner allows spreading = the film density data over 12 or 14 bits instead of the usual 8, you may be able = to extract the fine detail information by postprocessing high-bit images = to level backgrounds, adjust tones, and filter. The unsharp mask filter in = Photoshop sometimes does a good job at enhancing faint details that are otherwise = lost or blurred in the scanning process.
Larry Thomas Mechanical and Materials Engineering Washington State University Pullman, WA=20
---------- From: L. D. Marks Sent: Thursday, January 7, 1999 12:59 PM To: Microscopy List Subject: Re: TEM; diffraction pattern analysis
The Microscopy ListServer -- Sponsor: The Microscopy Society of = America=20 To Subscribe/Unsubscribe -- Send Email to = ListServer-at-MSA.Microscopy.Com On-Line Help = http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html = -----------------------------------------------------------------------.=
I have come across this several times, and there is no simple solution that I am aware of. I believe the reason has a lot to do with how the eye/brain interprets images. We do a good job of excluding noise, and we can often see patterns when it is very difficult to quantitatively measure them on a computer. In a sense we do a type of Maximum = Entropy analysis -- we have a prior model of what is in the image and find features that fit this model, for instance weak spots or rings. (Of course, this also means that sometimes we find things that are not there.)
The best method that I am aware of is to combine a rank filter (good at reducing shot noise), some sort of high-pass filter to remove only the low frequencies (reducing the background) and pasting togethor images at different contrast levels to prevent the high intensity regions from dominating. At least for a picture this often works, although you have to play a lot with the kernel sizes. Quantitation is very hard. You have to set up a model (Maximum Entropy, Maximum Likelihood, Least-Squares) and perform a numerical fit. Sometimes Least-Squares works; I have never tried Maximum Entropy which should do better.
Philippe-Andr=E9 Buffat wrote: } =20 } I would be strongly interested in softwares able to handle digital } diffraction patterns. In particular a soft to flatten the background is } needed. From my experience, I can see on negatives faint rings or spots, } but under the same condition it is impossible to see them on a computer } screen without increasing the contrast but then the range between black and } white covers only a small part of densities in the whole pattern and the } observable area between the white center and the black periphery becomes } unacceptably small. The use of color coding of the intensities improves } only a little bit the visible range. An other soft to determine } automatically the spot position or the ring position by a refinement method } or deconvolution of gaussian/lorentzian (or else) curves would also = be } welcome.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Greetings, I need assistance with flat embedding of rat cerebellum (IEM-DAB), that is 100 um thick and 1.5 x 0.5 cm dimensions. Basically we are worried about tissue curling during the dehydration. My first inclination is argarose embed (before epon), but I would take any expert advise. Thank you,
DEPARTMENT OF PHYSICS, UNIVERSITY OF ILLINOIS AT CHICAGO
A postdoctoral position is available in the Interface Physics Group at the University of Illinois at Chicago (UIC). Research in the Interface Physics Group focuses on the use atomic resolution imaging and analytical techniques in electron microscopy, coupled with theoretical simulations, to determine the structure-property relationships at internal interfaces on the fundamental atomic scale. Current research programs involve ceramics, high-Tc superconductors and optoelectronic/high-power semiconducting materials and devices. The experimental facilities to perform this research are comprehensive: a JEOL 2010 Field-Emission STEM/TEM featuring a 1.4=C5 probe size, "drift free" stage, high-angle annular dark-field detector (Z-contrast), Gatan Imaging Filter, and Noran EDS; a VG HB501A Field-Emission dedicated STEM with EDS, EELS and Auger spectrometers; a JEOL 3010 conventional TEM with digital imaging capabilities and EDS; a JEOL 6320 =46ield-Emission SEM with EDS and Cathodoluminescence; a JEOL JXA733 microprobe; and a Topometrix AFM/STM. In addition to the electron microscopes, specimen preparation facilities include a Gatan Duo-mill, =46ischione precision ion-mill, SouthBay plasma cleaner and Leica Ultramicrotome. The Interface Physics Group has a Silicon Graphics R10000 Power Indigo workstation with a Molecular Simulations' Cerius 2 package incorporating the CASTEP pseudopotential code. The physics department has additional workstations and access to the UIC Convex Exemplar Supercomputer and the National Center for Supercomputing Applications at UIUC. =20
This position is a joint postdoctoral appointment with Professor Susanne Stemmer in the Department of Physics at UIC. Research performed by the successful candidate for this position will involve the investigation of grain boundaries and defect structures in ionic and mixed ionic/electronic conducting oxide ceramics. The aim of the program is to incorporate experimental results into comprehensive atomic scale models for ionic/electronic transport in these materials.=20 It is anticipated that this position will involve a significant amount of industrial collaboration.
Candidates should be recent Ph.D. graduates in physics, metallurgy, or materials science with a background in the relevent materials issues and an ambition to be part of a developing program pushing at the frontiers of interface physics. Please send a resume, names, addresses of three references and a publication list to Professor Nigel D. Browning at the address below. Prior experience in STEM or TEM is essential. However, consideration will be based on the candidates overall potential for success in the field and applicants with prior experience in related fields are encouraged to apply. Positions are for one year initially, normally renewed for a second year with possibilities existing for further years. Salary is commensurate with experience. UIC is an equal opportunity employer.
} Date: Thu, 7 Jan 1999 15:06:29 -0500 (EST) } From: MICHAEL DELANNOY {delannoy-at-welchlink.welch.jhu.edu} } To: microscopy-at-sparc5.microscopy.com } Subject: flat embedding of vibratome sections } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings, } I need assistance with flat embedding of rat cerebellum } (IEM-DAB), that is 100 um thick and 1.5 x 0.5 cm dimensions. Basically } we are worried about tissue curling during the dehydration. My first } inclination is argarose embed (before epon), but I would take any expert } advise. } Thank you, } } Mike D } } We do this all the time. Sections don't curl. We originally tried to put them into flat molds standing up on their edges, but they would fall over sometimes. We now cut the pointed end off a BEEM capsule; snap the cap on the other end and wrap the junction with a sliver of Parafilm to prevent leaking; put a drop of resin in the lid (now upside down, with the Beem capsule sticking upwards); and lift the thick section into the lid with a wooden applicator stick broken into a flat wedged end with the tip then bent up into the shape of a hoe then fill the capsule with resin. Do this on a light box and under a dissecting scope.
If you need to keep straight which side is which, you can trim the tiny thick section with a razor blade into a funny shape that you will recognize. We use a trapezoid-like shape/state of Nevada shape: __ | \ |___\
That way you can keep the side of interest outward, e.g., confocal scope-selected areas: Miller SE, Levenson RM, Aldridge C, Hester S, Kenan DJ, Howell DN. 1997. Identification of focal viral infections by confocal microscopy for subsequent ultrastructural analysis. Ultrastruc. Pathol. 21:183-193.
Your sections are wider; thus, you will have to embed in a larger mouthed container if you want sections parallel with the face. But the principle is the same. If the sections have enough room to "swim" in the resin, you can gently stretch them out with the applicator stick, i.e., they won't premanently curl even though they will be flimsy as they float around in the resin.
Good luck.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Mike D- we routinely embed flat sections after the immuno proceedures, do any post fixation (OsO4), followed by dehydration, infiltration with resin, then to embed we place the 100 micron section between two glass slides * one of which has been subbed with a gelatin material, the other whioch has been subbed with Trenmittel (we purchase ours from EMS) the trenmittel is something like teflon, it is a release agent. remove all exess resin from around the section, then clamp the two slides together (clothes pins or large paper clips) polymerize as usual, then using a knife blade pry the two glass slides apart, this takes a little practice. finally reembed by filling a gelatin capsule with resin, let it thicken a little, then invert it over thetissue section which remained on the subbed slide, polymerize overnight. to free the section/block; hold slide over an alcohol burner flame (use pliers) then after 6-8 seconds snap off the block with the aid of haemostats or pliers. it really can work quite well. but practice first it is a little tricky -Mike
On Thu, 7 Jan 1999, MICHAEL DELANNOY wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings, } I need assistance with flat embedding of rat cerebellum } (IEM-DAB), that is 100 um thick and 1.5 x 0.5 cm dimensions. Basically } we are worried about tissue curling during the dehydration. My first } inclination is argarose embed (before epon), but I would take any expert } advise. } Thank you, } } Mike D } } }
In a message dated 1/7/99 2:07:07 PM, wesaia-at-iastate.edu wrote:
} Sounds like you may want to check with John Russ about many of these } issues. I thought he was right there in your backyard. But I will offer a } couple thoughts. } } I think you would be better use the same field size for all of your } measurements. Besides, Vf should have the same mean value no matter how } large the field.
Guess I'll throw in my $.02 worth since my name has been taken in vain.... ;-)
Field size does matter, alas. He is trying to measure particle size which means he needs to ignore those which touch the edge of the image field, and this creates several problems: a) large particles are more likely to touch the edge than small ones; b) larger fields will have proportionately fewer edge- touching particles. It can get complicated but the idea is to determine the number of edge-touching features and from that and the size distribution of those measured, adjust the effective area of the image so that the number per unit area is corrected for edge effects. Contact me directly via email if I can be of help
I am new to microscopy - and am trying to figure out what is the best general purpose mountant. I read that Euparal is good and stable for many years, but I can't find any except in Australia. I just want to make some basic slides for now, nothing exotic. I would appreciate any advice from anyone. Thanks.
David W. Bass Appalachian State University Boone, North Carolina dwbass-at-appstate.campuscw.net
Hello All, I have had 21 replies to my original request for information. Many thanks to everybody who responded, and apologies to anybody I misrepresent in this summary!
The state of the art in measurement accuracy was claimed to be close to that of X-ray diffraction by several people. [Of course, it depends what kind of X-ray diffraction you're talking about - I don't personally believe it will ever be possible to get accuracies as good as that of double (triple, etc.) crystal X-ray diffraction from spot patterns, although CBED/HOLZ line analysis comes close.] Jouk Jansen mentioned his paper in Acta Cryst of Jan 1998 on analysis of diffraction patterns. 2% was mentioned as the best day-to-day reproducibility one could hope for without taking special precautions such as evaporating gold onto your sample to include a standard pattern on the same negative as the pattern you want to measure. Lens hysteresis, astigmatism and pincushion/barrel distortion due to poor focusing may make it even worse. Eric Lachowski mentioned the huge effect that being away from eucentric height can have [I came across this myself a little while ago - I was horrified to find that a 50% change of diffraction pattern spacing was possible, even though the image size only changed by 5%, when tilting a sample.] An accuracy of 0.1% in measurement was about the limit, using computer-aided measurements of digital images. The distortion of diffraction patterns due to reciprocal lattice spiking was mentioned by quite a few people as potentially being the limiting factor in measurement. Most people seem to use evaporated gold as a standard for measuring camera length. There seems to be quite a wide variety of measurement methods out there, ranging from the good old fashioned way (i.e. lupe and graticule) to quite sophisticated analysis of digitized images. The software people use seems to fall into two categories; packages used to measure the position of spots and/or rings, and packages which simulate diffraction patterns which can be used for comparison with the real thing. Measurement packages included Gatan's Digital micrograph and NIH-Image. A few people have put a lot of work into producing packages which automatically make measurements: EDP, by Jaap Brink (http://ncmi.bioch.bcm.tmc.edu/~brink), which is free. MacLispix by Dave Bright (http://www-sims.nist.gov/MLx/doc/home.nclk), also free but runs only on a power Mac (sob). Bill Tivol has written plug-ins for the SPIDER image processing package that subtracts circular backgrounds, measure the centre and radius of a ring, and obtain background-subtracted intensities for a spot pattern. Corneliu Sarbu mentioned the free package PATTERN, running on PCs, which can be used as an aid to interpretation of spot patterns (see Microscopy Today 98-9 (Nov 1998)). Analysis packages: Carine, Crystal Designer, CRISP, Desktop microscopist and of course EMS were mentioned as packages which are used to simulate diffraction patterns for a particular crystal and zone axis. Wharton Sinkler has written a FORTRAN program to aid indexing of patterns to a known structure (http://www.lmcp.jussieu.fr/sincris/logiciel/).
Many thanks again to all those who replied.
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK e-mail richard.beanland-at-gecm.com
As far as sectioning, we tend to look for the Tg of the various components. As long as all of the components in your system have Tg well above room temperature, then sectioning at room temperature will result in relatively small deformations. In your case, PVC has a Tg of about 85 C and room temperature sectioning should be OK. In the case of PVB ( I assume B = Butadiene ?) the Tg is bellow RT and therefore it is better to use cryo sectioning for this material. Typically we would section this at about -100C .
As far as staining, osmium should stain the butadiene , but so will RuO4. I am not familiar with a stain for PVC, but you could try etching techniques (e.g. plasma etching). Check Linda Sawyers book on Polymer microscopy .
I hope this helps.
Jordi Marti ---------------------------------- You wrote: What is eveyones favorite stain for enhancing contrast during polymer TEM? OS4 or otherwise? The polymer materials are PVC and PVB.
Also, does anyone out there have some advice on a recommended temperature for ultramicrotomy of the above polymers? We DO have a cryo-ultramicrotome.
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Mike, You shouldn't have much trouble with curling during fixation and dehydration. The problem comes with getting absolutely flat sections for polymerization. Your sections are a bit thicker than I used and we were usually able to cut down the area of interest further but try this procedure. I think it should work fine. I have used it to embed vibratomed x-sections of rat brain which had been subjected to pre-embedding ICC reactions. In this case, since the amount of reaction diminished as you cut further into the section, it was critical to have absolutely flat material for serial sectioning.
Try embedding in droplets of resin on a 22x22mm plastic coverslip (available through most of the EM supply companies). Cover with another coverslip and weight with metal washers or nuts. The weighting is important as it really pressed the sections down insuring that they are very flat and squeezes out extra resin. After 24 hr. polymerization, cut edges of cover slips and they can then easily be separated. Cut the end off of gelatin or beam capsules to give a tube. Place the capsules over areas of interest on the sections, add one small drop of resin and polymerize a number of hours to secure capsules to the sections. Then fill up the capsules and finish polymerization.
When ready to section, blocks are easily broken or cut off of the coverslips. There will be very little extra resin to cut through before getting into the tissue making sectioning fairly easy once you are properly lined up.
An alternative method is to embed between teflon-coated glass slides which are then also weighted. I have found this a bit more cumbersome than using the plastic coverslips but it may work better for thicker specimen material.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
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Greetings, I need assistance with flat embedding of rat cerebellum (IEM-DAB), that is 100 um thick and 1.5 x 0.5 cm dimensions. Basically we are worried about tissue curling during the dehydration. My first inclination is argarose embed (before epon), but I would take any expert advise. Thank you,
Mike D
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Hi, everybody, I am looking for where I can buy an interference microscopy, which can be used to evaluate the thickness of thin film on a glass substrate by comparing the fringe at the edge of the thin film. I like to know also that if there is any kind of glass binder (adhesive material) which can be used inside a vacuum chamber, can last several hours in vacuum before working and can be used as air-tight sealing. Thanks a lot. Zgliu
Liu Zugang Departamento de Fisica Universidade de Aveiro 3810 Aveiro Portugal Fax:+351-34-24965 Email:zugang-at-fis.ua.pt
I would just like to correct one item in Richard's summary: CRISP and (more relevantly) ELD are programs which extract data from experimental images and diffraction patterns (rings and spots), respectively. They are not used to simulate diffraction patterns.
Disclaimer: Calidris sells CRISP and ELD, and we have a vested interest in making sure that microscopists receive accurate information about the programs.
I will be happy to send details to anyone who is interested.
Best wishes for the New Year,
George Farrants.
Richard wrote: } Analysis packages: Carine, Crystal Designer, CRISP, Desktop microscopist and of } course EMS were mentioned as packages which are used to simulate diffraction } patterns for a particular crystal and zone axis.
Does anyone have an answer to those questions? Working with a Variable Pressure SEM; 1- What can we see with it on polymers? 2- Where can I find pictures of polymers on a VP-SEM? 3- Is it possible to see spherolites in polymers without any pre-treatment (etching...) 4- Is it possible to see the "Skin effect" on injected parts in polymers and measure its thickness ?
We have tried to cleave rock salt (1cm cube, purchased from a microscopy supplier) for use as a substrate. We cleaved it with a razor blade, and had hoped to get a near-atomically smooth surface so we could deposit an aluminum film on it. However, the cleaved surfaces appear to be far rougher this, on the order of tens of microns.
Are there any special procedures that we should follow to get a near-atomically smooth surface?
Thank you all for your suggestions.
Mick Thomas
Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
} From: "Ursel Bangert" {USCHI-at-fs2.phy.umist.ac.uk} Organization: Umist To: MTLRMDB-at-LUCS-MAC.NOVELL.LEEDS.AC.UK
Good Morning All,
I am looking for suggestions on dispersing agglomerated crystals. Does anyone have a recommendation on what "dispersing agents" are available for dispersing sub-micron ( {100 nm) crystals?
I have only used ultrasonication of particles in acetone/water so far and that works well for micron sized crystals.
Thank you,
Mohan Kalyanaraman
Sr. Staff Material Scientist Mobil Technology Company PO Box 480 Paulsboro, NJ 08066 mohan_kalyanaraman-at-email.mobil.com
Does anyone have an answer to those questions? Working with a Variable Pressure SEM; 1- What can we see with it on polymers? 2- Where can I find pictures of polymers on a VP-SEM? 3- Is it possible to see spherolites in polymers without any pre-treatment (etching...) 4- Is it possible to see the "Skin effect" on injected parts in polymers and measure its thickness ? 5- Does anyone work on polymers for quality issues?
Does anyone have any tips/protocol/adivce for the preparation of human chromosomes for SEM. I'm just starting the project and I could use anything to get started... Thanks!
Rob Reff } From the Lab of: Professor William J. Perreault Lawrence University
} What is everyones favorite stain for enhancing contrast during polymer } TEM? OS04 or otherwise? The polymer materials are PVC and PVB.
} From the literature, ruthenium trioxide seems to be the most popular. We have used chlorosulphonic acid, but this mainly works for polyolefins, and I think it would chew up PVB (is that poly vinyl butyral?).
} Also, does anyone out there have some advice on a recommended temperature } for ultramicrotomy of the above polymers? We DO have a cryo-ultramicrotome.
The following article is superb. However, the author is out of reprints, so it might be better to use some form of library loan, if you don't take the journal.
*} TI: Reflections on the use of microtomy for materials science specimen preparation AU: Plummer_HK NA: FORD MOTOR CO,RES LAB,MAIL DROP 3028,SRL,DEARBORN,MI,48121 JN: MICROSCOPY AND MICROANALYSIS, 1997, Vol.3, No.3, pp.239-260 IS: 1431-9276 DT: Review
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
------=_NextPart_000_012E_01BE3BBB.A789C660 Content-Type: text/x-vcard; name="Vratislav Richard Eugene Maria John Baptist Bejsak-Collorado-Mansfeld.vcf" Content-Transfer-Encoding: quoted-printable Content-Disposition: attachment; filename="Vratislav Richard Eugene Maria John Baptist Bejsak-Collorado-Mansfeld.vcf"
BEGIN:VCARD VERSION:2.1 N:Bejsak-Collorado-Mansfeld;Vratislav Richard;Eugene Maria John Baptist FN:Vratislav Richard Eugene Maria John Baptist Bejsak-Collorado-Mansfeld ORG:Bayshark Research Laboratory TITLE:director NOTE;ENCODING=3DQUOTED-PRINTABLE:Marketing and = Coaching=3D0D=3D0A=3D0D=3D0ATenebrionidae Orbis and higher taxonomy TEL;WORK;VOICE:(+61 2) 9319 6380 TEL;CELL;VOICE:(+61 414) 540 465 ADR;WORK:;;29 Edward Street;Darlington, SYDNEY;;NSW 2008;Australie LABEL;WORK;ENCODING=3DQUOTED-PRINTABLE:29 Edward = Street=3D0D=3D0ADarlington, SYDNEY NSW 2008=3D0D=3D0AAustralie ADR;HOME;ENCODING=3DQUOTED-PRINTABLE:;;(temporaly address):=3D0D=3D0A32 = Girrawheen Ave;KIAMA;;NSW 2533;AUSTRALIA LABEL;HOME;ENCODING=3DQUOTED-PRINTABLE:(temporaly address):=3D0D=3D0A32 = Girrawheen Ave=3D0D=3D0AKIAMA NSW 2533=3D0D=3D0AAUSTRAL=3D IA URL: URL:http://www.coleoptera.org EMAIL;INTERNET:ricardo-at-login.cz EMAIL;PREF;INTERNET:vratislav-at-bigfoot.com EMAIL;INTERNET:ricardo-at-ans.com.au REV:19990108T233435Z END:VCARD
I am trying to gather information regarding Scanning Transmission = Electron Microscopy. I currently use an Amray 1645 SEM which has the = capacity for STEM work, but I have never used it in this mode. Any = comments or literature citings on this subject would be greatly = appreciated.
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3509.100"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} I am trying to gather information = regarding=20 Scanning Transmission Electron Microscopy. I currently use an Amray 1645 = SEM=20 which has the capacity for STEM work, but I have never used it in this = mode. Any=20 comments or literature citings on this subject would be greatly=20 appreciated. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Thank you for your help, = {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Chris = Holp {/FONT} {/DIV} {/BODY} {/HTML}
A technical difficulty in my lab is coming on the table: How to increase resolution of digitized SEM images, especially for low magnification ( {50X). The pixel size of SEM image (50X)in my machine (LEO-435VP) is 4.5 um. In other word, no matter how good image software performs, the measurement error is at least 4.5 um. We are going to use SEM as a routine measurement tool under 100X, what is the disadvantage? Does any one have excellent idea to solve this problem, in terms of : Increase number of pixels and save as compressed .jpg to reduce file size? Stage mapping? Software solution? What else?
Mick: Maybe the salt could work, but why not use freshly cleaved mica, its not sensitive to humidity, dead easy to cleave and cheaper. I must declare that ProSciTech (and several other EM suppliers) stock mica. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, January 09, 1999 5:13 AM, Mick Thomas [SMTP:mgt3-at-msc.cornell.edu] wrote: } Fellow microscopists, } } We have tried to cleave rock salt (1cm cube, purchased } from a microscopy } supplier) for use as a substrate. We cleaved it with a } razor blade, and } had hoped to get a near-atomically smooth surface so we } could deposit an } aluminum film on it. However, the cleaved surfaces appear } to be far } rougher this, on the order of tens of microns. } } Are there any special procedures that we should follow to } get a } near-atomically smooth surface? } } Thank you all for your suggestions. } } Mick Thomas } } } } } Mick Thomas } UHV-STEM Laboratory } E-1 Clark Hall } Cornell University } Ithaca, NY 14853 } } Phone: 607-255-0650 } Fax: 607-255-7658 } e-mail: mgt3-at-msc.cornell.edu
Chris: Just a couple of the more obvious differences between SEM and STEM (as an attachment to a TEM), using secondary mode - 1. STEM has generally higher kV - greater soft specimen penetration, charging is worse, but on "hard specimen" better resolution. 2. Working distance is low in STEM, greater resolution, poor depths of field. 3. STEM has small sample access and often limited tilt/ rotate facilities. Suitable specimens can give great images, but with more difficulties.
In STEM mode, which is also possible with some SEMs (including yours). The important differences are: The specimen is a section and this is penetrated by the beam. The detector (photo multiplier) is below the specimen. The penetration envelope is not formed because a relative thin section is used and the beam diameter is the most important determinant of image resolution. So when in a conventional SEM resolution in soft (biological) specimen is limited to say 6nm, you could resolve, say 2nm in STEM because the resolution is largely determined by the beam diameter. Better still, in low contrast specimens contrast can be increased at will - until electronic noise takes over. Maybe the best use of STEM is in image analysis of soft specimens. X-ray scattering from the penetration envelope in an SEM at around 15kV will result in spatial X-ray resolution of about 20 micrometer, which is often pretty near useless. In STEM the spatial X-ray resolution is only a fraction thereof. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, January 09, 1999 11:56 AM, Chris [SMTP:cholp-at-ncweb.com] wrote: } I am trying to gather information regarding Scanning } Transmission Electron Microscopy. I currently use an } Amray 1645 SEM which has the capacity for STEM work, but } I have never used it in this mode. Any comments or } literature citings on this subject would be greatly } appreciated. } } Thank you for your help, } } Chris Holp } { { File: ATT00001.html } }
Jena (name also of city), was pre WW2 the Zeiss centre. Zeiss Jena and Zeiss Oberkochen run as two separate companies in East and West Germany. Zeiss Oberkochen later took over the Jena works. Its now known simply as Zeiss. Zeiss should know about these optics, but I doubt that they can or would supply these. Your chances are in the secondhand market. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, January 09, 1999 9:35 AM, ricardo [SMTP:ricardo-at-ans.com.au] wrote: } Dear colleagues } } I am looking for objectives and oculars or other } accessories for microscope } called Technival 2 from old East Germany company JENA.. } } Any help? } } Keep care and be of good cheer. } } Regards } } Vratislav Richard Eugene Maria John Baptiste } of Bejsak (Bayshark)-Collorado-Mansfeld } } Coleoptera - Australia, Tenebrionidae of World } (incl. Lagriinae, Alleculinae) } } Temporally home address: } 32 Girrawheen Ave. } Kiama NSW 2533 } Australia } e-mail: vratislav-at-bigfoot.com } ricardo-at-ans.com.au } (before Ricardo-at-compuserve.com } and ricardo-at-login.cz ) } } http://www.coleoptera.org } phone : 0414 540 465 (Australia) } +61 414 540 465 (International) } } Only after the last tree has been cut down, } only after the last river has been poisoned, } only after the last fish has been caught, } only then will you find that money can not be eaten.' } CREE INDIAN PROPHECY. } } } { { File: Vratislav Richard Eugene Maria John Baptist } Bejsak-Collorado-Mansfeld.vcf } }
Liu: I expect that several of the major microscope manufacturers still make double beam interference microscopes (Certainly Leitz used to). All of these would have scary price tags. Perhaps you can find one second-hand or an alternative method for measuring film thickness.
The properties you seek for sealing air/ glass under vacuum are those of Apiezon T. This item is in our online (page M2) and I must declare an obvious interest. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Friday, January 08, 1999 8:17 PM, Liu Zugang [SMTP:zugang-at-ideiafix.fis.ua.pt] wrote: Hi, everybody, } I am looking for where I can buy an interference } microscopy, which } can be used to evaluate the thickness of thin film on a } glass } substrate by comparing the fringe at the edge of the thin } film. } I like to know also that if there is any kind of glass } binder } (adhesive material) which can be used inside a vacuum } chamber, can } last several hours in vacuum before working and can be } used as } air-tight sealing. } Thanks a lot. } Zgliu } } Liu Zugang } Departamento de Fisica } Universidade de Aveiro } 3810 Aveiro } Portugal } Fax:+351-34-24965 } Email:zugang-at-fis.ua.pt
In a message dated 1/9/99 1:47:00 AM, zhiyuw-at-worldnet.att.net wrote:
} A technical difficulty in my lab is coming on the table: How to increase } resolution of digitized SEM images, especially for low magnification } ( {50X). The pixel size of SEM image (50X)in my machine (LEO-435VP) is 4.5 } um. In other word, no matter how good image software performs, the } measurement error is at least 4.5 um. } ... } Does any one have excellent idea to solve this problem, in terms of : } Increase number of pixels and save as compressed .jpg to reduce file size? } Stage mapping? } Software solution? } What else? } If your beam size and actual imaging resolution is sub-micron, then the 4.5 micron limit is arising from the timing of your ADC, in other words how many samples it takes along each scan line, and from the spacing of the lines. If you can alter than then you can acquire images with more pixels. But DON'T compress them with jpeg or any lossy compression scheme or you will lose the benefits - these methods cause brightness and location shifts for features and will not get the accuracy you want.
If you can't fiddle the acquisition, your other choice is to acquire a series of higher magnification images and stitch them together as a mosaic. This isn't always easy to do, since stage mechanisms aren't very precise and if the sample isn't flat and horizontal you will have magnification that varies from side to side and makes fitting impossible.
On the other hand, what is it that you need to measure that can't be sampled at higher magnification - do you really need One Big Picture?
a Happy and prosperous new year to all! I am a new subscriber and I work at a local branch of the Min. of Health as a veterinary hygiene inspector in Italy. I am working on a substitution of malachite green in aquaculture, but I would like to know more about its nature, use and toxity, how dose it act? I've recently learned that it's used as a dye in staining certain cell tissues; surely somebody has posed himself the problem and has even found some explanation. Can somebody please give me some information or wher I can get it (ex. Web Sites)? Excusing me for having drifted you away from your prevalent work I anticipate my thanks to those how will answer and a good luck to evrybody! regards
emidio fazzini
(...up here from downunder!)
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Jim Darley wrote: ============================================== Maybe the salt could work, but why not use freshly cleaved mica, its not sensitive to humidity, dead easy to cleave and cheaper. I must declare that ProSciTech (and several other EM suppliers) stock mica. =============================================== Jim is of course correct in that salt is sensitive to moisture, and it is that very characteristic that causes freshly cleaved NaCl to be the substrate of choice for some researchers. When studies of epitaxial effects are being done, especially at elevated temperatures, it can be problematic to remove the thin film coating from the substrate, but in the case of NaCl, it can be readily dissolved in water. And even when mica could be used, in many instances, NaCl is also used in parallel since the unit cell dimensions both in size and symmetry are quite a bit different. And they give effects that can be quite different as well.
The impact of the differences in unit cell geometry and unit cell dimensions can also result in a significant difference in annealing effects of small crystals on the surfaces of these substrates. However, from the standpoint of smoothness, for example, if one was making carbon support films, mica would be better (if not also easier and cheaper) than NaCl.
An "old" reference from the literature that shows some of this can be found at Die Makro. Chemie, 113, 246 (1968), "Polyoxymethylene Single Crystals. II. The Effect of Substrate of Annealing Behavior".
Chuck
Disclaimer: SPI is a supplier of both mica and fine single crystal NaCl as are also some of the other main suppliers of consumables to the microscopy and microanalysis market.
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Hello: Can anyone comment on the difference in the volume or resolution of a point EDS analysis at 1kv or even 200V vs. the more standard 10 or 20kV. If we were to use a FE-SEM at 200V could we expect a significant increase in resolution of our EDS for a bulk sample compared our convention SEMs. Any comments are greatly appreciated. Steve
------------------------------ Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
Sure Stephen, lower kV equals better spatial X-ray resolution. But. . . . But what X-rays would you be able to excite with these low voltages? Look at a table of X-ray energies and remember that the very definition of the X-ray energy lines is the minimum voltage required to produce those X-rays. Generally 1.8 x that energy is required to obtain maximum fluorescence. I expect that you would like to analyse light elements (biological samples), and these are in most cases unsatisfactory in SEM because of the poor spatial resolution. For these TEM or STEM with EDS are the answer. (Made a posting on STEM yesterday with more info) Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
} } } Hello: Can anyone comment on the difference in the volume } or resolution of } a point EDS analysis at 1kv or even 200V vs. the more } standard 10 or 20kV. } If we were to use a FE-SEM at 200V could we expect a } significant increase in } resolution of our EDS for a bulk sample compared our } convention SEMs. Any } comments are greatly appreciated. Steve } } } ------------------------------ } Stephen McCartney } Research Associate } Virginia Tech } Materials Institute } 2108 Hahn Hall } Blacksburg, VA 24061-0344 } USA } } TEL: 540-231-9765 } FAX: 540-231-8517 } ------------------------------ }
} Hi Mark, I must be getting real old as I remember my gransfather used to } make chairs out of plant material. Large plants I believe. They wood cut the } larger stems into structural members and glue the parts together into many } different shapes. I sure these were not as functional or beautiful as the } bent steel and plastic used today. Maybe you could find one of these } antiques for your purpose. } Russ
Hi Russ,
your suggestion has stirred some racial memories in my mind. I have a vague recollection of seeing such ancient furniture in scratchy black & white movies (assuming they were not plastic/steel replicas of the cellulose originals).
I will enquire as to the availability of chairs fashioned from our forefather's favourite material, and also purhaps from adobe brick, bamboo, or rock. Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
Hi everybody who interested in the problem of 3-D biological tissue architectures reconstruction. This problem (especially 3-D epithelia structure) is interesting for me too. So I would greatly appreciate a copy of your papers. Also I am inviting you to visit our homepage on same subject. URL is followed: http://members.tripod.com/~Gensav/index.htm Sincerely yours Gennady A. Savostyanov
Dr. Gennady A. Savostyanov E-mail: savost-at-ief.spb.su savost-at-hotmail.com Sechenov Institute for Evolutional Physiology and Biochemistru Russian Academy of Science 44 M. Thorez, 194223 St. Petersburg, Russia
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} } } Hank Adams wrote ........... Dear listservers, We are looking for a dedicated software package for scheduling, in this case, instrument use incorporated into our webpage. We have several microscopes, workstations, microtomes, etc. and would like to have calenders or something similar for each, so users could sign up in advance using the web. It should be possible to assign different levels of privilege to each user. Once an entry is made it could only be changed by an administrator other than the original user. We like would like to then to automatically transfer this information into a billing database, either Access or FileMaker Pro based. Is something like what I have described commercially available
We have just such a system written in house. It has run very smoothly now for 5 years with constant upgrades. You can buy it if you like it.
Just go to the website in my signature.
The introductory material explains how we work.
Click on Booking System, Access to Images and use login {guest} password {guest} to try out the system.
If you are interested get back to me.
Mel Dickson ***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Hi: I am beginning a project on glass and I will be using a contract microscope. Can you explain with diagrams how the phase contrast scope works. Thank you in advance, dsm
High Steve! As far as I remember, the optimum (in sense of intensity) ratio between energy of electrons and excitation energy of X-ray line is from 2 to 3. At that the spatial X-ray resolution is not very high. It can be improved by diminishing this ratio. But the intensity of the line drops very strongly during decreasing of electrons energy to excitation energy of the line. But I think there are not so many X-ray lines interesting for you on EDS spectrum in the range up to 200 eV :-)). Regards. Victor Sidorenko, ANTRON Co. Ltd., Moscow, Russia.
} Hello: Can anyone comment on the difference in the volume or resolution of } a point EDS analysis at 1kv or even 200V vs. the more standard 10 or 20kV. } If we were to use a FE-SEM at 200V could we expect a significant increase in } resolution of our EDS for a bulk sample compared our convention SEMs. Any } comments are greatly appreciated. Steve } } } ------------------------------ } Stephen McCartney } Research Associate } Virginia Tech } Materials Institute } 2108 Hahn Hall } Blacksburg, VA 24061-0344 } USA } } TEL: 540-231-9765 } FAX: 540-231-8517 } ------------------------------ } } }
Hi Zhiyu Wang - Lets hope its me who is confused: I don't care. If a monitor is 100mm across and is represented by 4um pixel (100 divided by 0.004=) 2500 would be required for a single line. If one pixel was missing I would just forget about that, although the percentage error would be constant, regardless of magnification. What I would worry about is the large variation in magnification readings, which is possible because of the SEM's great depths of field and tilt angles. Calibrated latex spheres have been used for decades in TEM. Now larger calibrated spheres are available and these can be routinely and economically applied to SEM and light microscopy specimen to provide a reliable size comparison. Disclaimer: ProSciTech supplies latex particles (page "S2" online) and thus has a vested interest. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, January 09, 1999 4:21 PM, Zhiyu Wang [SMTP:zhiyuw-at-worldnet.att.net] wrote:
} Hi, All: } } A technical difficulty in my lab is coming on the table: } How to increase } resolution of digitized SEM images, especially for low } magnification } ( {50X). The pixel size of SEM image (50X)in my machine } (LEO-435VP) is 4.5 } um. In other word, no matter how good image software } performs, the } measurement error is at least 4.5 um. } We are going to use SEM as a routine measurement tool } under 100X, what is } the disadvantage? } Does any one have excellent idea to solve this problem, in } terms of : } Increase number of pixels and save as compressed .jpg to } reduce file size? } Stage mapping? } Software solution? } What else? } } Thank you for help } } Zhiyu Wang } } } }
Conn's Biological Stains, 9th edition edited by r.d. Lillie, page 248. under Diaminotriphenylmethanes. It is used as a vital dye so I woul 'assume' that it is not very toxic but there was no mention of toxicity.
-- Begin original message --
} From: Emidio FAZZINI {emifax-at-hotmail.com} } Date: Sat, 09 Jan 1999 09:56:24 -0600 } Subject: malachite green in aquaculture } To: Microscopy-at-Sparc5.Microscopy.Com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi everybody, } } a Happy and prosperous new year to all! } I am a new subscriber and I work at a local branch of the Min. of Health } as a veterinary hygiene inspector in Italy. I am working on a } substitution of malachite green in aquaculture, but I would like to know } more about its nature, use and toxity, how dose it act? } I've recently learned that it's used as a dye in staining certain cell } tissues; surely somebody has posed himself the problem and has even } found some explanation. } Can somebody please give me some information or wher I can get it (ex. } Web Sites)? } Excusing me for having drifted you away from your prevalent work I } anticipate my thanks to those how will answer and a good luck to } evrybody! } regards } } } emidio fazzini } } (...up here from downunder!) } } } ______________________________________________________ } Get Your Private, Free Email at http://www.hotmail.com } } } }
-- End original message -- best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
Depending on the size of the spherulites, you can see them readily with polarized light microscopy, with no sample preparation. Also, by using a first order red plate, you can determine the sign of the spherulite.
If you can cross-section the part, you can readily see the skin effect and measure its thickness, again using light microscopy. While EM certainly has a place in the polymer lab, there is a tremendous amount to be learned from light microscopy, with much less sample prep and investment in equipment. I'd really suggest that you start here first.
There are a number of different alternatives, but first you need to make some choices regarding the level of measurement. For very delicate measurements (1/10th - 1/400th of a wave, or on the order of 10-50 nm), you will need to use a multiple beam interferometer with a monochromatic light source. If you just need more standard thicknesses (70 nm or higher), you can go with a regular dual beam interferometer, using a white light source or even just a good narrow band filter.
For general purposes, there are several add-on interferometers which can be used to replace a typical objective (most commonly, the 10x objective). The one I have used most is a small Michelson which used to be available through Hacker in this country. (I will fax you an information sheet).
Regarding multiple-beam/add ons: talk to Nikon. As I remember, in addition to a Michelson, they also make a Tolanksy. The challenge with the Tolansky is getting a reference mirror which has a reflectivity matched to the reflectivity of your sample. I haven't worked with the Nikon system for some years, but I seem to remember that they had a small turret, providing you with a choice of several different R values.
Re: larger systems:
Your choices in Europe tend to be much greater than ours in the US. If you can get your hands on a Reichert Polyvar Met on the used equipment market, they had a very respectable and easily used interferometer accessory. Reichert is now part of the Leica family, but I don't know if they have picked up this neat little attachment. Other, older microscopes which I have used include the Interphako from Jena (now Zeiss) and the Pluta, from PZO. The Interphako was amazingly inexpensive in comparison with its capabilities: in half shade mode it could readily measure to 10 nm. Finally, Leitz used to make the Linnik, the flagship of interferometers. I understand that these are no longer available, but, again, you can probably find one on the used-equipment market.
I am sure that I have missed someone (my apologies), but these comments should give you a starting point.
One more issue: Thin films are transparent and most of these (exception: Linnik and Interphako) work in reflected light. By mounting your sample on a front surfaced mirror, you can overcome this problem.
Phase contrast microscopes have two components: a ring which is placed in the condenser and a special phase-changing plate which is mounted in the objective. They can be viewed in the back focal plane of the objective either by removing the eyepiece and looking down the tube into the back of the objective or replacing the eyepiece with the centering telescope provided with the phase kit. Make sure that the microscope is set up for phase first. That is, make sure that the right ring is rotated or moved into position in the condenser and that you are using the matching phase objective.
In the back focal plane of the objective (BFPo), you will see the smoky phase ring from the objective's plate overlaying the bright ring in the condenser.
First, the principles:
1. a. The basic underlying concept behind phase contrast comes from Nature's kind provision for a fairly predictable delay in the light passing through biological and similar structures. These structures cause the light passing through them to lag by a quarter of a wavelength behind the light which passes through the surrounding material.
b. The microscope image is formed by the interference between light passing through the specimen and light passing through the background. For improved contrast (brighter brights and darker darks), the ideal situation would be for the light from the specimen to either be completely in step or to lag by half a wavelength. The intensity of the light in the image directly proportional to the SQUARE of the amplitude of the wave which results when the two waves are "added together". This is a bit difficult to explain without a drawing, but here's the gist:
If the waves are perfectly in step, the resulting,additive wave has an amplitude twice that of the original components and the light in the image is 2 squared or 4 times brighter.
If the waves are half a wavelength out of step by half a wavelength, then they meet peak to trough and cancel each other. That part of the image will have "zero intensity" and be dark.
All we need to do is design a microscope which controls the light from specimen and background to meet these conditions.
Now, for the purpose of each:
2. The function of the ring in the condenser is to carefully define what will be known as the "background light" and place it very specifically in the smokey area of the phase plate mounted in the objective. The smokey has a channel cut in it so that the background light has to go through less glass
3. When a portion of that light interacts with the sample it (a) scatters, to fill the whole back focal plane of the objective and (b) undergoes approximately a quarter of a wave shift.
4. When that "specimen light" reaches the phase plate, most of it goes through the thicker, non smokey part of the phase plate. The phase plate is designed so that the thicker section adds another quarter of a wave lag to the specimen light. (quarter wave lag from specimen) + (quarter wave lag from phase plate) = half wave lag required for destructive interference and improved contrast.
5. But why is the phase plate smokey? Because the light from the background is MUCH brighter than the light passing through the specimen. That is, its amplitude is much greater. For the two waves to destructively interfer, their amplitudes must match. The manufacturers coat the cut in the phase plate with neutral density material so that the background intensity is brought into range with the light coming from the specimen. You can see the effect in the phase image: the background is cut to about 15% its original intensity, resulting in a soft, pearly gray.
Finally: how can you fine-tune a phase contrast image?
Since the underlying principle is based on a difference in refractive index between the sample and its mounting, one possibility is to change the mountant. For your glass, for instance, you might find that there are terrible haloes around the glass particles when they are mounted in air. Try this test: mount some test samples (all the same refractive index) in (a) water, ri 1.33 (b) glycerin, ri 1.47 and (c) immersion oil, ri 1.55. You will see the image get crisper and cleaner as you move toward the immersion oil, which is a much better match for glass (typically on the order of ri 1.5152).
Secondly, this whole process is wavelength dependent, yet we never specified WHICH wavelength. Look in your phase kit for a green filter (if it is an interference filter, it will appear yellow and mirrored). Place this filter over the light port. It defines the wavelength for which your phase kit was built, usually 548 nm.
For more background (plus important diagrams), may we suggest "Optimizing Light Microscopy for Biological and Clinical Laboratories"? Even though it is biologically oriented, you will find a great deal of sound, basic information which will help you in your microscopy. Details are available on our website:
{ {http://www.MME-Microscopy.com/education} . Also, a reminder that the ACS course on Applied Microscopy for Chemists is just around the corner. Details are also available on the website.
I have been requested to forward this announcement to this group.
Hank Adams Cell Biology Integrated Microscopy Core Baylor College of Medicine One Baylor Plaza Houston, Tx 77030
-----Original Message----- } From: Mancini [SMTP:mancini-at-bcm.tmc.edu] Sent: Sunday, January 10, 1999 6:24 PM To: CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU
Hello !
The female threads , which hold the cone screw that secures the specimen holder in our Ultracut E are partially stripped. One option is for our machine shop to rethread them. This would involve removing the heater and thermocouple wires that are at the top of the bridge. I was wandering if any of you had the same problem and how you went about solving it.
I also checked about replacing the whole bridge, but this option is quite expensive and there is a question about part availability.
I read an interview with a photojournalist who does all digital photography who saves her work on a Jaz drive "in the field"(PhotoMetro interview between ADColeman&Maggie Hallahan). She commented that "...the Jaz only lasts a few months and after that I have to archive everything to CD." Is this true? It's rather alarming as Iomega claims that the storage life of the cartridges(under *ideal* conditions) is 10 years. Someone is planning to supply our microscopy lab with a Jaz drive for image storage, but if it's not archival, I don't think it's worth the investment. Does anyone have experience with long term(at least a year) image storage on Jaz disks?
I am presently looking at the potential purchase of a basic SEM, probably variable pressure for a local museum. Are there any one of you using SEM for public shows? If yes may I have more information on what is exactly done.
The other alternative (more likely) is presentation of a "microzoo" similar to Oceanographic institute of Monaco. Small organisms (plancton, benthos, etc.) are presented via a fully motorized stereoscopic microscope. Any personnal experience to share on the subject? Ciao! -- L.Paul Bedard, ing. Ph.D. DocuScience inc.
Dear Stephen, I have successfully used lower kV electrons to reduce the volume of material excited for x-ray spectroscopy and a good Monte Carlo x-ray program, such as David Joy's, will help you to determine the volume excited. At 5 kV I was able to analyse 0.5 micron layers in a cross-sectioned semi-conductor. However, you must limit your analyses to those elements whose x-rays are excited by this energy of electrons, generally the energy of the x-rays is half or less than the energy of the electrons going in. I don't believe there is much, if any, resolution advantage to lower electron energy. At 200V, there will be very few x-rays excited that an EDX system can detect. You wrote: } Hello: Can anyone comment on the difference in the volume or resolution of } a point EDS analysis at 1kv or even 200V vs. the more standard 10 or 20kV. } If we were to use a FE-SEM at 200V could we expect a significant increase in } resolution of our EDS for a bulk sample compared our convention SEMs. Any } comments are greatly appreciated. Steve } } } ------------------------------ } Stephen McCartney Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
We embed vibratome sections of neural tissue that has been DAB treated between two pieces of Aclar. If a minimum of resin is placed between the sheets, the sections won't move much as a weight is applied to flatten the sections. There are different grades of Aclar, and I had to go directly to Allied Signal to get the one I prefer. The Aclar peels easily away from the polymerized resin, leaving one with a "section" that can be viewed with a LM for subsequent trimming. I super glue it to a blank beem capsule for thin sectioning. Best of luck, Randy -- Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own. http:\\www.uiowa.edu\~cemrf
Are there any references to descriptors for various morphology of grown crystals? We see several morphologies here, but I feel that I am only coming up with vegetable analogies - cauliflower, cabbage, etc..
While they sometimes capture the spirit of the picture, they seem technically inadequate. Are there technical terms that would be more appropriate to describe crystal clusters? Ideally, is there any reference (website etc.) where the terms are defined along with pictures?
If there is interest, I will post a summary. And thanks to those that offered leads on dispersing methods
Mohan Kalyanaraman
Sr. Staff Material Scientist Mobil Technology Company PO Box 480 Paulsboro, NJ 08066 609-224-3989 609-224-3608 (fax) mohan_kalyanaraman-at-email.mobil.com
I'm looking to see if someone knows anyone that prints cell images on slides--I need some black and white reference images of cellular material on a standard slide that can be used to calibrate some equipment. There are the usual problems with bio material that I really don't want to have to address since this is going to serve as a calibration standard. They will be viewed at 4x magnification, so they don't even need to be especially excellent reproductions. The biggest key (besides looking like cells) is that the printed image is *thin* (microns?), as I am trying to accurately determine the thickness of the coverslip/permount conglomeration that gets mounted on top of it.
Can anyone spit a quick Sudan IV protocol for lipid staining (for en bloc or embedded tissues-plant) to my email (or the net if anyone else is curious)?? Much obliged!! Tracey
Tracey Pepper Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
{html} Can anyone spit a quick Sudan IV protocol for lipid staining (for {i} en bloc {/i} or embedded tissues-plant) to my email (or the net if anyone else is curious)?? {br} {x-tab} {/x-tab} Much obliged!! {br} {x-tab} {/x-tab} Tracey {br} {br} {br} {br} {div} Tracey Pepper {/div} {div} Bessey Microscopy Facility {/div} {div} Iowa State University {/div} {div} ph: 515-294-3872 {/div} {div} fax: 515.294.1337 {/div} {/html}
I basically agree with what John says below, only stitching may be your only choice, if you really need a large field of view AND a high resolution.
I suppose, the resolution of 4.5 um comes from the fact, that you digitize a large field of view into a given number of pixels (for example, a field of view of about 9 mm digitized into 2Kx2K would give you such a resolution). The problem is, that going to 4K x 4K resolution may give you a better resolution but more likely is not. The reason is this:
For a digital image acquisition (and I believe, the 435 is digital), you divide your normal x and y sweep into the required number of voltage levels. The total sweep is normally of the order of a few Volts. If you divide that by a few thousand pixels, you end up with a few millivolts per pixel. Any noise of that level will essentially prohibit to keep the beam stationary to the level you would require. This is normally true independent of actual magnification as the digitization is done before the scan amplifier, which translates the x and y sweep into the actual voltages required to activate the scan coils.
So, to stay with the example above (9 mm field of view): if you need to digitize that to, let's say 1 um, you would need a resolution of 9000 x 9000 pixels. As I explained above, you probably would not be able to achive that without image stitching. Also, it would give you an 81 MB image (at 8 bits per pixel) or 162 MB (at 16 bits per pixel). I'm not sure you want to deal with that! John is absolutely right regarding lossy compression: Why spend a lot of money and effort to achieve high resolution to throw it all away through compression? Lossless compression may reduce the files by a factor of 2 or so.
We do have software and hardware for image acquisition and processing, especially but not limited to LEO microscopes, and we migh be able to help you with some of your challenges. If you need further info, please contact me through email.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. } ******************************************************* } ---------- } From: } "DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com[SMTP:"DrJohnRuss-at-aol.com"-at-sparc5. } microscopy.com] } Sent: Saturday, January 9, 1999 6:23 AM } To: zhiyuw-at-worldnet.att.net; Microscopy-at-sparc5.microscopy.com } Subject: Re: Resolution of digital SEM image } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I did not read the article, but perhaps the author was saying that when the jaz disk fills up, she archives to CD to clear out the jaz disk. This may be true, because the relative cost of a few CD's is nothing when compared to a jaz disk (a few dollars vs. over $100US). In this manner, she would only need to purchase one or two jaz disks. I have been archiving images on jaz disks for well over a year now and have had no problems with the file longevity.
Hope this helps,
David A. Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (800) 221-1975x2108
"Pauline C. Yu" {splene-at-pw.usda.gov} on 01/11/99 11:49:06 AM
Please respond to "Pauline C. Yu" {splene-at-pw.usda.gov}
To: Microscopy list {Microscopy-at-Sparc5.Microscopy.Com} cc: (bcc: David Bell/NA/Millipore)
I read an interview with a photojournalist who does all digital photography who saves her work on a Jaz drive "in the field"(PhotoMetro interview between ADColeman&Maggie Hallahan). She commented that "...the Jaz only lasts a few months and after that I have to archive everything to CD." Is this true? It's rather alarming as Iomega claims that the storage life of the cartridges(under *ideal* conditions) is 10 years. Someone is planning to supply our microscopy lab with a Jaz drive for image storage, but if it's not archival, I don't think it's worth the investment. Does anyone have experience with long term(at least a year) image storage on Jaz disks?
I have not used a Jaz drive, but if it is like the Zip (basically a floppy disk inside), I would not consider it an "archive" media. On the other hand, unless the enviroment was really poor, I would expect a much greater life than you mentioned. Also, the equipment capable of retreiving the data is limited. Will you have a working Jaz drive in 10 years? Another down side to the Jaz is the cost of storage compared to a CD-R (650 MB for {$1.50). I would suggest a CD-R archive for cost, longevity, and an increased probability that it can be read a few years down the road. It remains to be seen if the manufacturers are correct, but most rate the CD-R for 100 years or more storage.
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I read an interview with a photojournalist who does all digital photography who saves her work on a Jaz drive "in the field"(PhotoMetro interview between ADColeman&Maggie Hallahan). She commented that "...the Jaz only lasts a few months and after that I have to archive everything to CD." Is this true? It's rather alarming as Iomega claims that the storage life of the cartridges(under *ideal* conditions) is 10 years. Someone is planning to supply our microscopy lab with a Jaz drive for image storage, but if it's not archival, I don't think it's worth the investment. Does anyone have experience with long term(at least a year) image storage on Jaz disks?
by post.its.mcw.edu (8.8.8/8.8.8) with ESMTP id OAA25356 for {microscopy-at-sparc5.microscopy.com} ; Mon, 11 Jan 1999 14:48:35 -0600 (CST) Message-ID: {369A633B.6E10F13A-at-mcw.edu}
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Hi, dear all -
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max size, 2 Afterglow type, 150x135mm CRTs, 1 non-afterglow type120x90mm
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EM Facility Department of Microbiology Medical College of Wisconsin
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I forget the source, but a few months ago I recall a discussion on the web of CD longevity. It had been the conventional wisdom that CDs had a shelf life on the order of 30 years. But people were discovering that CDs 5 years old or so were becoming defective through delamination or other processes, just with age and not necessarily with handling, so there was some skepticism about the 30yr shelf life figure.
Like all media types, there is no question that after 10 years or so you might expect that the ability to read specific disks or cards or whatever would be difficult due to dramatic changes in technology (who has 8-track tapes any longer?) and the natural migration to new technologies that causes older technology to disappear. So any discussion of shelf life for electronic storage is probably a moot point (unlike film, which could be good almost indefinitely with proper handling). I fully expect to have to, sooner or later, transfer all of my archived data over to some new type of storage. Just the way life is...
Larry
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Jim J Darley wrote: } } In STEM mode, which is also possible with some SEMs } (including yours). The important differences are: } The specimen is a section and this is penetrated by the } beam. The detector (photo multiplier) is below the } specimen.
One can also put a parallel EELS detector under the specimen and collect position-tagged spectra. This gives a very high-resolu- tion compositional image.
} Better still, in low contrast specimens contrast can be } increased at will - until electronic noise takes over. } Maybe the best use of STEM is in image analysis of soft } specimens.
Looking at the low-loss region of an EELS spectrum can dif- ferentiate among lipid, protein and nucleic acid, as reported a few years ago by Rich Leapman at MSA. This could be ideal for cryo- specimens or other unstained biological specimens. Yours, Bill Tivol
I am trying to locate Ms. Lisa Hartnell. We were colleagues a few years ago at RMC. After departing from RMC, Lisa worked for a while in Paul Webster's lab at Yale.
If anyone knows of Lisa's whereabouts or has an e-mail address for her, please contact me off-list. Or feel free to forward this message to her.
Thanks for your help!
Bob **************************************** Robert (Bob) Chiovetti, Ph.D. President Microimaging Technologies, Inc. Tucson, Arizona USA Tel. / Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems Research Microscopy Products *****************************************
It might be useful to contact the Royal Microscopical Society. I know that in the past they have displayed material on open access on a SEM with restricted access to controls. I haven't personally seen it, but read about it in the "RMS Proceedings". I think a manual SEM was used with covers over certain controls. A similar effect could be achieved using a simple PC control interface.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: "pbedard-at-saglac.qc.ca"-at-sparc5.microscopy.com {"pbedard-at-saglac.qc.ca"-at-sparc5.microscopy.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Hello all,
I am presently looking at the potential purchase of a basic SEM, probably variable pressure for a local museum. Are there any one of you using SEM for public shows? If yes may I have more information on what is exactly done.
The other alternative (more likely) is presentation of a "microzoo" similar to Oceanographic institute of Monaco. Small organisms (plancton, benthos, etc.) are presented via a fully motorized stereoscopic microscope. Any personnal experience to share on the subject? Ciao! -- L.Paul Bedard, ing. Ph.D. DocuScience inc.
I agree with Larry. If you want to "Archive" images there is only one stable medium - Conventional Film.
Jaz & Zip disks are fine for temporary storage of images. I have been using Jaz disks for a few years without any failures, but due to cost transfer data to CD-R's. The Jaz is only used for rapid access to the images. As pointed out the Jaz/Zip technology will probably be superseded soon ( 1GB drives are gone already ! ) and in a few years it will be difficult to read data if your drive fails. This was a major factor along with cost ) 5 years ago when we decided on a CD-R storage system, over Magneto-Optical which was the current favourite then.
CD-R's are not in themselves a good long-term archive medium. We have experienced a number of failures of CD-R's given to customers over the five years. These failures are usually attributed to environmental factors since the CD-R's are extremely sensitive to bad handling. Our archive has not experienced any failures in this time and the CD-R's are carefully stored in drawers. We always recommend that customers take their own copy so that a second copy exists. Any images lost over the five years were due to hard disk failures prior to storage. CD-R's are so cheap now that it is probably worth writing two disks at a time and storing them separately. At least five years on all the archive can still be read on any computer.
I suppose DVD ( 5.2 GB ) will be the next way to go as soon as industry settle on a universal standard ?
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Larry Allard {l2a-at-ornl.gov} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
I confirm that my address is right. Please add it to the mailing list.
Thank you. --------------------------------------------------- Dr. J. Ricote Laboratoire du Physique et l'Etat Condens=E9 (LPEC) Facult=E9 des Sciences Universit=E9 du Maine-Le Mans Avenue Olivier Messiaen BP 535 72085 Le Mans cedex FRANCE
The 5-year life of CDs is for older technology CDs. Current CDs are nominally rated at 70-200 years, depending on the manufacturer. Magnetic media, regardless of how it is packaged (floppy, winchester, Zip, Jaz, SyQuest, whatever), all fall prey to the same problems that all magnetic media fall to -- archival for a decade or two, less than that under "real world" use.
For a good discussion of the CD longevity debate, see http://www.cd-info.com/CDIC/Industry/news/media-chronology.html.
For a good discussion of how these "100 year" lifetime determinations are made, see
I think that one has to remember that the environmental conditions are exquisitely important when discussing longevity. There *is no* single number that one can look to unless one knows how the media is stored and used.
For an example of how this plays for photographic media, look at http://www.kodak.com/cluster/global/en/consumer/education/imageStability.shtml
While the above is for photographic prints, not CDs or magnetic media, the importance of environmental issues are analogous.
billo
On Mon, 11 Jan 1999, Larry Allard wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Woody and all: } } Just a couple of additional cents... } } I forget the source, but a few months ago I recall a discussion on the web } of CD longevity. It had been the conventional wisdom that CDs had a shelf } life on the order of 30 years. But people were discovering that CDs 5 } years old or so were becoming defective through delamination or other } processes, just with age and not necessarily with handling, so there was } some skepticism about the 30yr shelf life figure. } } Like all media types, there is no question that after 10 years or so you } might expect that the ability to read specific disks or cards or whatever } would be difficult due to dramatic changes in technology (who has 8-track } tapes any longer?) and the natural migration to new technologies that } causes older technology to disappear. So any discussion of shelf life for } electronic storage is probably a moot point (unlike film, which could be } good almost indefinitely with proper handling). I fully expect to have to, } sooner or later, transfer all of my archived data over to some new type of } storage. Just the way life is... } } Larry } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } Hello Pauline, } } } } I have not used a Jaz drive, but if it is like the Zip (basically a floppy } } disk } } inside), I would not consider it an "archive" media. On the other hand, } } unless } } the enviroment was really poor, I would expect a much greater life than you } } mentioned. Also, the equipment capable of retreiving the data is limited. } } Will } } you have a working Jaz drive in 10 years? Another down side to the Jaz is } } the } } cost of storage compared to a CD-R (650 MB for {$1.50). I would suggest a } } CD-R } } archive for cost, longevity, and an increased probability that it can be } } read a } } few years down the road. It remains to be seen if the manufacturers are } } correct, but most rate the CD-R for 100 years or more storage. } } } } Woody } } } } ______________________________ Reply Separator } } _________________________________ } } Subject: Jaz disk archivalness? } } Author: "Pauline C. Yu" [SMTP:splene-at-pw.usda.gov] at CORP } } Date: 1/11/99 10:49 AM } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I read an interview with a photojournalist who does all digital } } photography who saves her work on a Jaz drive "in the field"(PhotoMetro } } interview between ADColeman&Maggie Hallahan). She commented that "...the } } Jaz only lasts a few months and after that I have to archive everything to } } CD." } } Is this true? It's rather alarming as Iomega claims that the storage } } life of the cartridges(under *ideal* conditions) is 10 years. Someone is } } planning to supply our microscopy lab with a Jaz drive for image storage, } } but if it's not archival, I don't think it's worth the investment. } } Does anyone have experience with long term(at least a year) image storage } } on Jaz disks? } } } } thanx } } Pauline Yu } } Microscopist Technician } } USDA-ARS-WRRC } } - - -- --- ----- -------- ------------- --------------------- } } } Dr. Lawrence F. Allard } Senior Research Staff Member } High Temperature Materials Laboratory } Oak Ridge National Laboratory } 1 Bethel Valley Road } Bldg. 4515, MS 6064 } PO Box 2008 } Oak Ridge, TN 37831-6064 } } 423-574-4981 } 423-574-4913 Fax } l2a-at-ornl.gov } } }
They do all sorts of materials printed on glass. We routinely use their image analysis calibration slide and stage micrometers as part of our on-site workshops.
Caveat: MME has no commercial interest in this product.
Suggest you see the Particle Atlas, available from McCrone, now in a very convenient CD Rom format which is quick and easy to search. Try contacting
Dina Mattes at McCrone Associates, Westmont, IL 800-622-8122.
There are also good descriptions of all sorts of crystal families and habits the old reference, Chamot (pronounced Cha-mo) and Mason's Handbook of Chemical Microscopy. I think McCrone Assoc. has also brought this back into print. It's a great reference because it shows you how to control crystal growth of various types under the microscope. Drs. Chamot and Mason reigned over the chemical microscopy facility at Cornell U. for nearly 100 years.... Their book contains lots of good "benchtop wisdom".
Many thanks to all who responded to my question. Below is a summary of the responses to cleaving rock salt:
1) Irradiate the salt (with a Co60 source, for example) to set up point defects in the crystal, making it easier to cleave.
2) Ensure you are using a high quality salt crystal.
3) Use a sharp, brand new razor blade for each cleave.
3) After cleaving, take a piece of lens cleaning paper, place it on a smooth surface, and put a little water on it. Using tweezers, pick up the salt and rub it in a circular motion in the puddle of water.
4) Consider using alternate substrates, depending on their suitability to the particular experiment on hand. Suggestions included mica, Barium sulfate, and Highly Ordered Pyrolytic Graphite (HOPG).
5) Be realistic in your expectations; even a very well cleaved salt crystal will have some cleavage steps.
Thanks again, these suggestions have been very helpful to me.
There was a wonderful microscopy exhibits at both the American Museum of Natural History (NY) and at the Smithsonian in the early-mid 80's. Cecil Fox, then with Armed Forces Institute of Pathology, helped organize both. The Smithsonian exhibit not only had a major display of old and new equipment but a working SEM. I have lost track of Dr. Fox ... last I heard he was still in the Silver Spring area. I have left a message for him and will send you info when available.
The other alternative is to talk to our illustrious listmaster, Nestor. He has a tremendous amount of experience with telemicroscopy which should be easily transferable to this type of situation.
Quoted from "Staining Procedures", 3rd ed. Biological Stain Commission pg 212: Sudan IV to show: suberized walls, cuticle, fat or oil globules
fix: either none or any botanical fixative embedding: paraffin or freehand
Preparation of staining and mounting solutions: Make a saturated solution of Sudan IV in 95% EtOH Add an equal volume of glycerin and filter if stain precipitates out on the sections, dilute further as necessary with a mixture of equal parts glycerine and EtOH Keep in a dropper bottle
Staining schedule: 1) Bring sections into 30% EtOH 2) Mount sections on a slide in a few drops of the staining and mounting solution 3) Seal edges of coverslip with "vas-par"*
Results: Cuticle, cutinized and suberized walls, and fat globules--orange.
*equal parts melted paraffin and vasaline; apply with a metal rod about 1/8th inch in diameter, bent into an L shape with a 6" handle and a 1" leg: warm applicator over a burner/hot plate, etc., then melt and pick up a few drops of vas-par and apply to coverslip edges. pg 239
Reference: Rawlins, T.E. 1933. "Phytopathological and Botanical Research Methods." John Wiley & Sons, NY.
(Pardon me while I go wipe off my keyboard.)
Phil
} Can anyone spit a quick Sudan IV protocol for lipid staining (for en bloc or } embedded tissues-plant) to my email (or the net if anyone else is curious)?? } Much obliged!! } Tracey } } } } Tracey Pepper } Bessey Microscopy Facility } Iowa State University } ph: 515-294-3872 } fax: 515.294.1337
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net
Thanks for the informative links. I was not aware of the extensive "debate" that has gone on regarding CD lifetime expectancy.
It is interesting that models suggest a lifetime of CD-R of 217 years. I think it is safe to assert that no one presently alive will be around to test this hypothesis. And if they were, it is probably safer to assert that there would be no device still in existance that could read such an ancient recording...
Larry
The 5-year life of CDs is for older technology CDs. Current } CDs are nominally rated at 70-200 years, depending on the } manufacturer. Magnetic media, regardless of how it is } packaged (floppy, winchester, Zip, Jaz, SyQuest, whatever), } all fall prey to the same problems that all magnetic media } fall to -- archival for a decade or two, less than that } under "real world" use. } } For a good discussion of the CD longevity debate, see } http://www.cd-info.com/CDIC/Industry/news/media-chronology.html. } } For a good discussion of how these "100 year" lifetime } determinations are made, see } } http://www.cd-info.com/CDIC/Technology/CD-R/Media/Kodak.html. } } I think that one has to remember that the environmental } conditions are exquisitely important when discussing } longevity. There *is no* single number that one can } look to unless one knows how the media is stored and used. } } For an example of how this plays for photographic media, } look at } http://www.kodak.com/cluster/global/en/consumer/education/imageStability.shtml } } While the above is for photographic prints, not CDs or magnetic } media, the importance of environmental issues are analogous. } } billo } } } On Mon, 11 Jan 1999, Larry Allard wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Woody and all: } } } } Just a couple of additional cents... } } } } I forget the source, but a few months ago I recall a discussion on the web } } of CD longevity. It had been the conventional wisdom that CDs had a shelf } } life on the order of 30 years. But people were discovering that CDs 5 } } years old or so were becoming defective through delamination or other } } processes, just with age and not necessarily with handling, so there was } } some skepticism about the 30yr shelf life figure. } } } } Like all media types, there is no question that after 10 years or so you } } might expect that the ability to read specific disks or cards or whatever } } would be difficult due to dramatic changes in technology (who has 8-track } } tapes any longer?) and the natural migration to new technologies that } } causes older technology to disappear. So any discussion of shelf life for } } electronic storage is probably a moot point (unlike film, which could be } } good almost indefinitely with proper handling). I fully expect to have to, } } sooner or later, transfer all of my archived data over to some new type of } } storage. Just the way life is... } } } } Larry } } } } } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Hello Pauline, } } } } } } I have not used a Jaz drive, but if it is like the Zip (basically a floppy } } } disk } } } inside), I would not consider it an "archive" media. On the other hand, } } } unless } } } the enviroment was really poor, I would expect a much greater life than you } } } mentioned. Also, the equipment capable of retreiving the data is limited. } } } Will } } } you have a working Jaz drive in 10 years? Another down side to the Jaz is } } } the } } } cost of storage compared to a CD-R (650 MB for {$1.50). I would suggest a } } } CD-R } } } archive for cost, longevity, and an increased probability that it can be } } } read a } } } few years down the road. It remains to be seen if the manufacturers are } } } correct, but most rate the CD-R for 100 years or more storage. } } } } } } Woody } } } } } } ______________________________ Reply Separator } } } _________________________________ } } } Subject: Jaz disk archivalness? } } } Author: "Pauline C. Yu" [SMTP:splene-at-pw.usda.gov] at CORP } } } Date: 1/11/99 10:49 AM } } } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } I read an interview with a photojournalist who does all digital } } } photography who saves her work on a Jaz drive "in the field"(PhotoMetro } } } interview between ADColeman&Maggie Hallahan). She commented that "...the } } } Jaz only lasts a few months and after that I have to archive everything to } } } CD." } } } Is this true? It's rather alarming as Iomega claims that the storage } } } life of the cartridges(under *ideal* conditions) is 10 years. Someone is } } } planning to supply our microscopy lab with a Jaz drive for image storage, } } } but if it's not archival, I don't think it's worth the investment. } } } Does anyone have experience with long term(at least a year) image storage } } } on Jaz disks? } } } } } } thanx } } } Pauline Yu } } } Microscopist Technician } } } USDA-ARS-WRRC } } } - - -- --- ----- -------- ------------- --------------------- } } } } } } Dr. Lawrence F. Allard } } Senior Research Staff Member } } High Temperature Materials Laboratory } } Oak Ridge National Laboratory } } 1 Bethel Valley Road } } Bldg. 4515, MS 6064 } } PO Box 2008 } } Oak Ridge, TN 37831-6064 } } } } 423-574-4981 } } 423-574-4913 Fax } } l2a-at-ornl.gov } } } } } }
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Sorry Zhiyu Wang, I still have trouble with that concept. A single pixel in a line of 2500 pixel represents an error any microscopist could ignore. The pixel size may be 4.5um - on the charge coupled device; on the specimen, depending on how small the area examined (higher power less field) that pixel may represent rather more. What matters is the observed (enlarged) image and here, if the image width is 100mm, that single pixel represents 100 over 2500= 0.04mm or an 0.04% error. If only part of the image is used, like with a photographic negative the error does not change dramatically. One would not calculate a magnification by ignoring that only half of the enlarged image is used.
If you worry about magnification use a build in calibration, like latex spheres as earlier suggested. With care an SEM may be accurate to 5%, though some people have claimed 1%. In either case those pixel will not be your problem - the way I see it. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Tuesday, January 12, 1999 5:37 PM, Zhiyu Wang [SMTP:zhiyuw-at-worldnet.att.net] wrote: } Hi, Jim: } } Thank you for responding my message. Your calculation is } right, but the } question is that we do not want to ignore the smallest } unit on large scale } of measurement. Say 1/2500 is a small number in } persentage, but its } absolute value is 4 um, my sample is not covered by total } 2500 pixels, } only part of them instead. If I measure multipal samples, } the error should } be +/- 4 um, ie. 8 um, close to 0.01 mm. This is too } rough for quality } control of machinary (in semoconductor industry) } } Basiclly I do not believe SEM is a good machine for } dimension measurement } as many factors involve on imaging system. Therefore, its } high electron } optical resolution and high depth of view attract me and } my boss to do } something different. I am collect information around the } world. I } appreciate your help and we can share some interest idea } later. } } Thanks, } } Zhiyu Wang } ---------- } } From: Jim J Darley {jim-at-proscitech.com.au} } } To: 'Zhiyu Wang' {zhiyuw-at-worldnet.att.net} ; } 'Microscopy-at-sparc5.microscopy.com' } {Microscopy-at-Sparc5.Microscopy.Com} } } Subject: RE: Resolution of digital SEM image } } Date: Monday, January 11, 1999 4:28 AM } } } } } } --------------------------------------------------------- } } --------------- } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } } ml } } ---------------------} } } } } Hi Zhiyu Wang - } } Lets hope its me who is confused: I don't care. } } If a monitor is 100mm across and is represented by 4um } } pixel } } (100 divided by 0.004=) 2500 would be required for a } } single } } line. If one pixel was missing I would just forget about } } } } that, although the percentage error would be constant, } } regardless of magnification. } } What I would worry about is the large variation in } } magnification readings, which is possible because of the } } } } SEM's great depths of field and tilt angles. } } Calibrated latex spheres have been used for decades in } } TEM. } } Now larger calibrated spheres are available and these } } can } } be routinely and economically applied to SEM and light } } microscopy specimen to provide a reliable size } } comparison. } } Disclaimer: ProSciTech supplies latex particles (page } } "S2" } } online) and thus has a vested interest. } } Cheers } } Jim Darley } } ProSciTech } } Microscopy } } PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 7 4774 0370 Fax: +61 7 4789 2313 } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } ********************** www.proscitech.com.au } } ***** } } } } } } } } On Saturday, January 09, 1999 4:21 PM, Zhiyu Wang } } [SMTP:zhiyuw-at-worldnet.att.net] wrote: } } } } } Hi, All: } } } } } } A technical difficulty in my lab is coming on the } } } table: } } } How to increase } } } resolution of digitized SEM images, especially for } } } low } } } magnification } } } ( {50X). The pixel size of SEM image (50X)in my } } } machine } } } (LEO-435VP) is 4.5 } } } um. In other word, no matter how good image } } } software } } } performs, the } } } measurement error is at least 4.5 um. } } } We are going to use SEM as a routine measurement tool } } } under 100X, what is } } } the disadvantage? } } } Does any one have excellent idea to solve this } } } problem, } } in } } } terms of : } } } Increase number of pixels and save as compressed .jpg } } } to } } } reduce file size? } } } Stage mapping? } } } Software solution? } } } What else? } } } } } } Thank you for help } } } } } } Zhiyu Wang } } } } } } } } } } } } } }
} Bill: } } Thanks for the informative links. I was not aware of the extensive } "debate" that has gone on regarding CD lifetime expectancy. } } It is interesting that models suggest a lifetime of CD-R of 217 years. I } think it is safe to assert that no one presently alive will be around to } test this hypothesis. And if they were, it is probably safer to assert } that there would be no device still in existance that could read such an } ancient recording... } } Larry }
I'm not sure how good these models are, though. It's a little like mutagenesis tests on bacteria. Yeah, it's a measure of carcinogenicity, but it's a loose one. I'm much happier using these tests as a relative scale rather than an absolute one.
Hello: I am currently working on mechanical behavior / microstructure correlations in single colony near apha Ti Alloys. I am currently preparing the foils using a dual ion mill (6kV, 12deg, 1mA), but have run into the problem of a very uneven alpha phase morphology, the alpha phase has island formation - we are using a cold stage to minimize the hydride formation at the interface. I am using 3 micron/6micron diamond paste during the dimpling process - the foil itself otherwise is fairly clean in terms of the dislocation content. Could anyone in the microscopy land have some suggestions in terms of what the problem may be?
thanks - if you want you can respond directly to suri.3-at-osu.edu
Pauline Yu wrote: ============================================ I read an interview with a photojournalist who does all digital photography who saves her work on a Jaz drive "in the field"(PhotoMetro interview between ADColeman&Maggie Hallahan). She commented that "...the Jaz only lasts a few months and after that I have to archive everything to CD." Is this true? It's rather alarming as Iomega claims that the storage life of the cartridges(under *ideal* conditions) is 10 years. Someone is planning to supply our microscopy lab with a Jaz drive for image storage, but if it's not archival, I don't think it's worth the investment. Does anyone have experience with long term(at least a year) image storage on Jaz disks? ============================================== When the 1 GIG Iomega drives came out some months ago, I thought then they were the greatest thing since sliced bread. I have used one on my home desktop repeatedly without problems. And ditto for some number of the office desktops (except for one).
But another one that I dedicated for use with my laptop for when I travel has been another story. During the warranty period, when I could get through on the Iomega lines, they did keep replacing the entire drive, which apparently did not survive the bouncing around during travel. The drive itself was packed in my checked luggage but well packed. But the survival rate was about three months. After the warranty period was up, they stopped replacing them.
In any case, my experience seemed to be consistent with that of the photojournalist: If you carry them around, you have problems. But so far as I can tell, after very heavy use of the 1 GIG JAZ drives, if they are permanently installed, we have not had any particular problems. In talking with their customer service people, apparently, something goes wrong with the drive if carried around and when that happens, it does do damage to the replaceable disc when you next try to use it.
Chuck
Disclaimer: In this instance, I have no financial interest in this product, just a satisfied user who wished Iomega had more customer service lines.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
"In Situ Hydride Formation in Zirconium and Titanium during Ion Milling" Graham J.C. Carpenter et al, JMSA Vol.1 No. 4 pp 175-184 1995.
=46rom the paper you will see that a key factor in eliminating hydride formation is having a clean sample free from hydrocarbon contamination. =
This contamination could be remnants from the dimpling process or other pre-thinning steps or it could be caused by the back streaming of diffusi= on pump oil in your ion mill. As titanium has a high chemical affinity for hydrogen, you may want to look over your preparation steps and try to eliminate any areas of possible contamination. If you are still having a=
problem, a quick cleaning of both the specimen and the specimen holder in= a Plasma Cleaner should take care of it.
If you have an interest, I can send you a copy of the above referenced paper. I also have papers on plasma cleaning which may be of interest.
NOTE: South Bay Technology does manufacture the PC150 Plasma Cleaner and therefore I have a vested interest in promoting its use.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Satyarth Suri } Hello: I am currently working on mechanical behavior / microstructure correlations in single colony near apha Ti Alloys. I am currently preparing the foils using a dual ion mill (6kV, 12deg, 1mA), but have run into the problem of a very uneven alpha phase morphology, the alpha phase has island formation - we are using a cold stage to minimize the hydride formation at the interface. I am using 3 micron/6micron diamond paste during the dimpling process - the foil itself otherwise is fairly clean in terms of the dislocation content. Could anyone in the microscopy land have some suggestions in terms of what the problem may be?
thanks - if you want you can respond directly to suri.3-at-osu.edu
Pauline, We have had a horrible time with the Jaz drives (hooked to an SGI archiving confocal images). Apparently when they get near full capacity they screw up. I've sent three unreadable disks to an image retrieval company, with no luck. The told me that when the disk is near full and you try to add another file it will put data back at the begining of the disk, erasing file allocation tables, basically making the disk unreadable. We have several defective disks and I have been forced to the zip drive.
Hello, My lab is currently looking for a used carbon coater for our TEM lab. Our Emitech has passed away and is in England trying to be repaired, but it looks grim. I also have a Denton coater that will not pump down. So I am looking into other options.
If your are selling such a machine or might know someone who is, please write back or drop a line to our website
I need some advice as to the best protocol for fixing fresh plant leaves for routine TEM. My only experience has been with animal tissue. Thanks, MG Engle
We just got a G3 Mac and an Epson digital projector-I have seen some really nice digital slide shows and am excited by the increase in quality and decrease in effort making slides. I expect, however, to spend some effort finding out the best way to put together a slide show, and am wondering what advice or experience might be out there. I would like to show ~10MB images without any computer stuff showing at the same time, but could consider compression (eg., jpeg) if there is no loss of quality. Wondering whether to get into a slide presentation program or just make a stack and open them sequentially (I have 190+ MB of memory). There is also a neat IR pointer that seems to bounce off the screen and feed into software in the projector that allows you to move the cursor around and click/double click with the IR pointer. Too bad they didn't include a laser pointer in this device!....Many thanks for any help or advice!...Tom Reese
Reply to: RE: Ti Alloys - TEM foil preparation Pure titanium has been jet electropolished here at Argonne National = Laboratory for over 10 years. A South Bay 550-B single jet instrument is = used because it has in-situ viewing of the specimen during the process to = speed up the determination of proper electropolishing conditions.
Conditions: -20 degrees C. 70 volts, 35 mA, (one side of 3 mm disc)
Note: After polishing about half way thru the specimen,=
it is cleaned, dried, and a dot of = Microshield stop- off lacquer is placed on the polished "side = one" dimple. The inverted specimen is then remounted on = the polish- ing instrument and thinned to perforation = using the = sensitive optical termination system. It = should be = possible to make several foils per day in = this manner.
Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il, 60439
E-mail: bkestel-at-anl.gov South Bay Technology wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
{HTML} {HEAD} {/HEAD} {BODY} {PRE WIDTH=3D"132"} Reply to: RE: Ti Alloys - TEM foil preparation
{/PRE} {FONT = FACE=3D"Geneva" SIZE=3D3 COLOR=3D"#000000"} Pure titanium has = been jet electropolished here at Argonne = National Laboratory for over 10 years. = A South Bay 550-B single jet instrument is = used because it has in-situ viewing of the = specimen during the process to speed up = the determination of proper electropolishing = conditions. {BR} {BR} Electrolyte: = 30 ml. perchloric acid {BR} = 295 ml. methanol {BR} = 175 ml. butyl cellosolve {BR} {BR} = Conditions: -20 degrees C. {BR} = 70 volts, = 35 mA, (one side of 3 mm disc) {BR} {BR} = Note: After polishing about = half way thru the specimen, {BR} = it is cleaned, dried, = and a dot of Microshield stop- {BR} = off lacquer is placed = on the polished "side one" dimple. {BR} = The inverted = specimen is then remounted on the polish- {BR} = ing instrument = and thinned to perforation using the {BR} = sensitive optical = termination system. It should be {BR} = possible to make = several foils per day in this manner. {BR} {BR} = Bernard Kestel {BR} Materials = Science Division {BR} Argonne National = Laboratory {BR} Argonne, Il, 60439 {BR} {BR} = E-mail: bkestel-at-anl.gov {BR} South = Bay Technology wrote: {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#= 000000"} {BR} >-----------------------------------------------------------------------= - {BR} >The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America {BR} I also have papers on plasma cleaning which = may be of interest. {BR} > {BR} >NOTE: South = Bay Technology does manufacture the PC150 = Plasma Cleaner and {BR} >therefore I have = a vested interest in promoting its use. {BR} > {BR} >Best = regards- {BR} > {BR} >David = {BR} >Writing at 9:38:23 = AM on 1/12/99 {BR} > = {BR} >***************************************************************** {BR} >********** {BR} >************************ {BR} > {BR} >David = Henriks = TEL: {BR} >800-728-2233 (toll = free in the USA) {BR} >South Bay Technology, = Inc. +1-949-492-2600 {BR} >1120 = Via Callejon = FAX: +1-949-492-1499 {BR} >San Clemente, = CA 92673 USA e-mail: {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} henriks-at-southbaytech.com {/U} {= /FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} > {BR} >***************************************************************** {BR} >********** {BR} >************************ {BR} > {BR} > = >>>>> Please visit us = at {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} http://www.= southbaytech.com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} <<<<< {BR} > {BR} >Manufacturers = of precision sample preparation equipment = and supplies for {BR} >metallography, crystallography = and electron microscopy. {BR} > {BR} >Message = text written by Satyarth Suri {BR} >> {BR} >Hello: {BR} >I = am currently working on mechanical behavior = / microstructure {BR} >correlations in single = colony near apha Ti Alloys. I am currently {BR} >preparing = the foils using a dual ion mill (6kV, 12deg, = 1mA), but {BR} >have run into the problem = of a very uneven alpha phase morphology, {BR} >the = alpha phase has island formation - we are = using a cold stage {BR} >to minimize the = hydride formation at the interface. I am = using {BR} >3 micron/6micron diamond paste = during the dimpling process - the {BR} >foil = itself otherwise is fairly clean in terms = of the dislocation {BR} >content. Could = anyone in the microscopy land have some = suggestions {BR} >in terms of what the problem = may be? {BR} > {BR} >thanks - if you want = you can respond directly to {/FONT} {FONT FACE=3D"Geneva" = SIZE=3D1 COLOR=3D"#0000FF"} {U} suri.3-at-osu.edu {/U} {/FONT} {FONT FACE=3D"= Geneva" = SIZE=3D1 COLOR=3D"#000000"} {BR} > {BR} >-satyarth {BR} > {BR} >< {BR} > {BR} > {BR} > {BR} >RFC822 = header {BR} >----------------------------------- {BR} > {BR} > = Received: from dns2.anl.gov (dns2.anl.gov = [146.139.254.3]) by {BR} >horus.et.anl.gov = (8.6.11/8.6.11) with ESMTP id MAA28629; = Tue, 12 Jan 1999 12:21:30 -0600 {BR} > Received: = from Sparc5.Microscopy.Com (sparc5.microscopy.com = {BR} >[206.69.208.10]) by dns2.anl.gov = (8.9.1a/8.6.11) with SMTP id MAA02315; Tue, = 12 Jan 1999 {BR} >12:22:10 -0600 (CST) {BR} > = Received: (from {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = daemon-at-localhost {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} ) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) {BR} >id LAA19027 for dist-Microscopy; = Tue, 12 Jan 1999 11:48:51 -0600 {BR} > Received: = from no_more_spam.com (Sparc5 [206.69.208.10]) = by {BR} >Sparc5.Microscopy.Com (8.6.11/8.6.11) =
Hi Guys, I'm looking for a shareware program for the mac that will allow me to batch process 12 bit images to 8 bit images. Thanks,
--Ciprian ________________________________________________________________ Ciprian A. Almonte Phone: (412) 648-9796 Center for Biologic Imaging FAX: (412) 648-8330 University of Pittsburgh URL:http://sbic6.sbic.pitt.edu Pittsburgh, PA 15261 USA mailto:calmonte-at-pitt.edu ________________________________________________________________
American Chemican Society "Applied Optical Microscopy for Chemists" March 4-5-7 in Beautiful Orlando, Florida Three days of total immersion, hands-on experience with all phases of optical microscopy, including one whole day of Polarized Light (both qualitative and quantitative) Strongly supported with the latest in equipment from the major manufacturers for HANDS-ON labs Special Saturday evening session on video imaging and image analysis
For further details, including registration information, visit the MME website: {http://www.MME-Microscopy.com/education}
Looking forward to seeing you there!
Barbara Foster, Course Coordinator
Microscopy/Microscopy Education ...Educating microscopists for greater productivity. 125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
This is a multi-part message in MIME format. --------------6C36AB32CC464C8C85112D45 Content-Type: text/plain; charset=iso-8859-9 Content-Transfer-Encoding: 7bit
Dear all microscopists;
I am a young electron microscopist who is trying to organise an EM lab in Turkey.
I had a chance to study with a wonderful electron microscopist and also a great teacher, Dr. Robert J.Kayton(the president of PNEMS) in Portland, OR for 1 year. I learned the technical aspects of electron microscopy from him. I have never had much more enjoyable experience than that. Now, I am back and have been asked to organise an EM lab.
All I have in this lab is an old TEM(Carl Zeiss EM 9) and an old ultramicrotome. As you see, I need so many things. I really want to rebuilt a nice EM lab and to apply my experience I have learned in U.S. It will be very hard for me since it is not easy to find financial support for that in my country.
Would you like to help me? I would like to ask you to send me any equipments, appliances, dark room stuffs, books etc. which you don't use anymore in your EM lab. I will prepare a chart for being able to thank all of you on the front door of my lab and write the names of persons, companies, facilities etc. which help me. I also would like to bring small presents from Turkey for you, dear helpfull electron microscopists, when I come to the Microscopy&Microanalysis'1999,in Portland,OR.
I will greatly appreciate any kind of help.
Thanks in advance,
Ranan Gulhan AKTAS, M. D. Trakya University, Faculty of Medicine Pathology Department 22030 Edirne, TURKEY Tel: +90 284 235 44 68 Fax: +90 284 235 76 52 e-mail: ranaoz-at-turk.net
Unless you have a compelling reason to use ion milling for this preparation, I recommend using jet electropolishing, possibly with just a final touch-up in the ion mill. Although both electropolishing and ion milling commonly produce sample preparation artifacts, the artifacts from ion-milling reactive metals such as Ti and Zr are not well understood and can be harder to eliminate. Despite Carpenter et. als work (see D. Henriks posting) implicating hydrocarbon contamination as a source of hydriding during ion milling, my personal experience ion milling Zr samples in 'oil-free' systems indicates that water vapor is the main culprit in the hydridng. Surface irregularities on ion milled Zr/Ti samples (surface terracing, for example) usually have other causes, possibly related to air leaks in the ion mill or oxygen in the feed gas. Another source of irregular surfaces on ion milled samples is embedded polishing compound. Cubic boron nitride (CBN) sometimes gives better results in dimpling metals than diamond, and even the diamond polishing compounds from different suppliers can give different results. Conversely, brief ion milling of electropolished samples--especially at low incidence angles and very low kV--can improve the surface finish and mill away irregularities caused by second-phase particles. Often the 'trick' to eliminating milling artifacts is minimizing time spent in the ion mill.
Larry Thomas Mechanical and Materials Engineering Washington State University Pullman, WA
---------- From: South Bay Technology Sent: Tuesday, January 12, 1999 6:00 PM To: Satyarth Suri; Microscopy ListServer Subject: Ti Alloys - TEM foil preparation
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Dear Satyarth:
An excellent paper on the subject is:
"In Situ Hydride Formation in Zirconium and Titanium during Ion Milling" Graham J.C. Carpenter et al, JMSA Vol.1 No. 4 pp 175-184 1995.
From the paper you will see that a key factor in eliminating hydride formation is having a clean sample free from hydrocarbon contamination. This contamination could be remnants from the dimpling process or other pre-thinning steps or it could be caused by the back streaming of diffusion pump oil in your ion mill. As titanium has a high chemical affinity for hydrogen, you may want to look over your preparation steps and try to eliminate any areas of possible contamination. If you are still having a problem, a quick cleaning of both the specimen and the specimen holder in a Plasma Cleaner should take care of it.
If you have an interest, I can send you a copy of the above referenced paper. I also have papers on plasma cleaning which may be of interest.
NOTE: South Bay Technology does manufacture the PC150 Plasma Cleaner and therefore I have a vested interest in promoting its use.
David Henriks TEL: 800-728-2233 (toll free in the USA) South Bay Technology, Inc. +1-949-492-2600 1120 Via Callejon FAX: +1-949-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Satyarth Suri } Hello: I am currently working on mechanical behavior / microstructure correlations in single colony near apha Ti Alloys. I am currently preparing the foils using a dual ion mill (6kV, 12deg, 1mA), but have run into the problem of a very uneven alpha phase morphology, the alpha phase has island formation - we are using a cold stage to minimize the hydride formation at the interface. I am using 3 micron/6micron diamond paste during the dimpling process - the foil itself otherwise is fairly clean in terms of the dislocation content. Could anyone in the microscopy land have some suggestions in terms of what the problem may be?
thanks - if you want you can respond directly to suri.3-at-osu.edu
I am compiling for the MSA Education Committee a listing of laboratories involved in outreach and/or remote access programs with local schools or just across campus. Please contact me with information about your respective programs. This list will be mounted on the MSA webpage.
I will be compiling lists for scanning electron microscopes, light microscopes(standard and confocal), scanning probes, confocal, and transmission microscopes. I'ld like to know what school levels your program is tartgeted to, what instruments you are using, a web site if you have one, and your email so I can follow up on the information.
Please contact me directly at sbarlow-at-sunstroke.sdsu.edu (NOT THE LISTSERVER LIST) and I will see the information is compiled and posted.
thanks in advance
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
I agree with Chuck. When I took Dr. McKenzie's Digital Microscopy course, he informed us that these things couldn't be banged around. We've had a 1 GB (permanently installed) for over a year and a 2 GB "portable" drive (that never gets moved) for almost a year, and have never had any trouble with either.
I don't know about over filling disks. My fattest one still has 170 MB left and is perfectly fine. For really important files/pictures, I keep 2 backups on separate disks. We have about 20 of these things. We also use Zips as a more portable medium. Go iomega.
No commercial interest; just a satisfied customer.
Sara
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} Dear All, } } Is anyone aware of an electropolishing solution for Mg alloys that (i) } thins very slowly, and (ii) works at room temperature? } } thanks. } Sincerely yours
D. H. Ping
------------------------------------------------------------------------- D. H. Ping, Dr Materials Physics Division National Research Institute for Metals (NRIM) 1-2-1 Sengen, Tsukuba 305-0047, Japan Phone: +81-298-59-2717 FAX:+81-298-59-2701 E-mail: Ping-at-tamamori.nrim.go.jp WWW: http://inaba.nrim.go.jp/apfim/ Home phone: +81-298-59-0918
Can anyone out there tell me a bit about endpoint detection systems for Reactive Ion Etchers? What different types there are, how they work, what situations they are optimal for, and {shudder} approximate price ranges? I was told to write up a summary within the week regarding the possibility of either attaching one to our existing RIE (PlasmaTherm Batchtop) or getting another. (Wow! They actually came to me and offered to buy stuff, instead of me having to beg!!! Either I'm doing stuff really right, or really wrong!)
Much thanks in advance,
Marisa Ahmad R&D Specialist mahmad-at-semiconductor.com
"It's not how hard you fall, it's how high you bounce."
Much as I hate to say it, Microsoft PowerPoint does a pretty good job with digital slide presentation. You definitely do *not* want to simply make a stack of slides and open them independently...it will seem clumsy and will be distracting. PowerPoint opens slides instantly, and you can do a lot of tricks to presumably enhance the production (although sometimes the tricks are somewhat distracting themselves). The best thing is to have a PowerBook with XGA (1024 x 768) resolution, and a projector with the same XGA resolution, to get the best projection results. We have been using digital projection almost exclusively for over a year now with this setup, and it works extremely well. The best thing is that you can be making slides just about up to the minute of your talk...a great capability for those of us who tend to procrastinate just a teeny bit... ;-).
Larry
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Hi Tom, Firstable congratulation on you new G3, awesome machine. I recommend Graphic converter for slide show. It's pretty easy to set up and it allow you to fade down the images. If you need help seeting it up let me know. } } We just got a G3 Mac and an Epson digital projector-I have seen some really } nice digital slide shows and am excited by the increase in quality and } decrease in effort making slides. I expect, however, to spend some effort } finding out the best way to put together a slide show, and am wondering } what advice or experience might be out there. I would like to show ~10MB } images without any computer stuff showing at the same time, but could } consider compression (eg., jpeg) if there is no loss of quality. Wondering } whether to get into a slide presentation program or just make a stack and } open them sequentially (I have 190+ MB of memory). There is also a neat IR } pointer that seems to bounce off the screen and feed into software in the } projector that allows you to move the cursor around and click/double click } with the IR pointer. Too bad they didn't include a laser pointer in this } device!....Many thanks for any help or advice!...Tom Reese
--Ciprian
_____________________________________________________________ Ciprian A. Almonte Phone: (412) 648-9796 University of Pittsburgh FAX: (412) 648-8330 Center for Biologic Imaging http://sbic6.sbic.pitt.edu Pittsburgh, PA 15261 USA mailto:calmonte-at-pitt.edu _____________________________________________________________
The problem with overfilling Jaz disks sounds like more of a software problem than a problem with the Jaz. I have filled a number of Jaz disks to capacity and simply get a message telling me it's too full. This has worked with both PC's and Unix systems.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Sara Miller {saram-at-duke.edu} To: Garber, Charles A. {cgarber-at-2spi.com} Cc: MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com}
At 10.46 -0500 99-01-12, Satyarth Suri wrote: } } Hello: } I am currently working on mechanical behavior / microstructure } correlations in single colony near apha Ti Alloys. I am currently } preparing the foils using a dual ion mill (6kV, 12deg, 1mA), but } have run into the problem of a very uneven alpha phase morphology, } the alpha phase has island formation - we are using a cold stage } to minimize the hydride formation at the interface. I am using } 3 micron/6micron diamond paste during the dimpling process - the } foil itself otherwise is fairly clean in terms of the dislocation } content. Could anyone in the microscopy land have some suggestions } in terms of what the problem may be?
Why can't you electropolish the specimens? For my PhD work we prepared a lot of (commercial purity alpha) Ti specimens using electropolishing with minimal hydride formation. This has been continued by another PhD student who also did work on Ti at the University of Birmingham (England).
We used a non-acid electrolyte developed by B.J. Kestel of Argonne National Lab. He recommends using it in a South Bay Technology single jet electropolisher but we made it work with a Struers Twin jet Tenupol, I guess you could probably make it work with a variety of other jet electropolishing devices. I can give further details of our procedure if anyone wants.
Hope this helps
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren PLEASE NOTE NEW CONTACT DETAILS: Department of Experimental Physics Chalmers University of Technology SE-412 96 G=F6teborg Sweden Tel: +46 31 772 36 33 =46AX: +46 31 772 32 24 email: maclaren-at-fy.chalmers.se or: ianmaclaren-at-hotmail.com ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
I am polishing a low carbon steel that is coated with zinc. The situation i'm experiencing is when i try to etch the base material with 2%nital for grain size determination i seam to get some effect from the zinc in that the area near the surface does not etch. Is it because of polishing practice dragging that zinc onto the surface?
even if you do use something like 'Powerpoint' for the final slide presentation, it might be worth seeing if you can get hold a program such as 'Thumbsplus'. It's shareware for the PC but I don't know if it's available for the Mac. I have found 'Thumbsplus' is much quicker for simple image management and filing images because of it's use of thumbnail images and it can even produce a basic slide show.
Malcolm
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------------------------------------------------------------------------- ----------------- Tom:
Much as I hate to say it, Microsoft PowerPoint does a pretty good job with digital slide presentation. You definitely do *not* want to simply make a stack of slides and open them independently...it will seem clumsy and will be distracting. PowerPoint opens slides instantly, and you can do a lot of tricks to presumably enhance the production (although sometimes the tricks are somewhat distracting themselves). The best thing is to have a PowerBook with XGA (1024 x 768) resolution, and a projector with the same XGA resolution, to get the best projection results. We have been using digital projection almost exclusively for over a year now with this setup, and it works extremely well. The best thing is that you can be making slides just about up to the minute of your talk...a great capability for those of us who tend to procrastinate just a teeny bit... ;-).
Larry } } We just got a G3 Mac and an Epson digital projector-I have seen some really nice digital slide shows and am excited by the } increase in quality and decrease in effort making slides. I expect, however, to spend some effort finding out the best way to } put together a slide show, and am wondering what advice or experience might be out there. I would like to show ~10MB } images without any computer stuff showing at the same time, but could consider compression (eg., jpeg) if there is no loss of } quality. Wondering whether to get into a slide presentation program or just make a stack and open them sequentially (I have 190+ } MB of memory). There is also a neat IR pointer that seems to bounce off the screen and feed into software in the projector that } allows you to move the cursor around and click/double click with the IR pointer. Too bad they didn't include a laser pointer in this } device!....Many thanks for any help or advice!...Tom Reese
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
In message {3306033D.951200C9-at-mauromedia.com} , Michael Draper {quex-at-mauromedia.com} writes } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE: Ti Alloys - TEM foil preparation Re: Electropolishing of Ti alloys = Due to interest in this subject, I will list some additional = electrolytes I have used on Ti alloys. (3mm, jet polishing).
Ti-8 w/% Al 13% HCL -60 degrees C Ti-811 87% methanol 70 to 90 volts Ti-6Al-4V 25-35 mA.
Ti-13 Sn 130 ml HCL -50 degrees C 670 ml methanol 150 volts 100 ml butyl 60 mA.
Ti-3V 60 ml perchloric acid -50 degrees C Ti-20 Zr 590 ml methanol 50 volts Ti-14 Al 350 ml butyl cellosolve 10-15 mA
Ti-8 Al 60 ml perchloric acid -15 t0 -20 C (rolled) 460 ml ethyl alcohol, 95% 180 volts 280 ml butyl alcohol 40 mA. 100 ml butyl cellosolve = Ti 5.3 g. lithium chloride -40 degrees C. nanocrystals, 11.16 g. magnesium 150 volts compacted chloride 30 mA 500 ml. methanol 100 ml. butyl cellosolve (Note: the two salts above must be added to the combined solvents one = "powder" at a time, while stirring. The two dry salts react violently if = mixed together. This electrolyte has nearly as wide an application as = perchloric acid mixtures and is known as B K-2). I suspect that B K-2 will cause the least hydride problem. It was = developed to eliminate hydrides in vanadium TEM foils. The voltages given = would need to be reduced about 20% for a twin jet system due to lower = electrical resistance of that configuration. My work was done on a South = Bay vertical single jet unit. I have no vested interest in the above equipment, but have enjoyed = the ease with which good polishing conditions can be obtained and = reproduced since 1975 or so. Good luck! = Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439
E-mail: {bkestel-at-anl.gov} Thomas, Larry wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
by mail.unixg.ubc.ca with esmtp (Exim 1.92 #2) for Microscopy-at-Sparc5.Microscopy.Com id 100TFD-0006xh-00; Wed, 13 Jan 1999 08:34:12 -0800 X-Sender: ech-at-pop.unixg.ubc.ca Message-Id: {v0300780000e8124c7823-at-[137.82.136.14]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" To: Microscopy-at-Sparc5.Microscopy.Com
Following the discussion about filling Jaz discs:
We collect images from a Bio-Rad MRC 600 confocal to Jaz disc on an IBM computer and transfer to a Jaz drive on a Mac G3 for making projections and plates with NIH Image and Photoshop. We used to transfer by ethernet but it was too slow and taking a disc out of one drive and putting into another drive is by far the quickest approach. Using the Mac G3 to work on the images allows another researcher to collect their images on the confocal and doesn't hold them up while data is being transfered.
We once lost 1GB of data because the user filled up the Jaz disc completely. Now we wouldn't lose that data because as I understand it: When you transfer to a Mac, there is a Mac header put onto the disc. If the disc is full, this header overwrites the PC directory tree. The IBM doesn't recognise the disc and the Mac thinks it is empty. The solution (thanks to Eric Jervis) is to put the Jaz disc back into the IBM. Look for hidden files, delete them, and use Norton Utilities to rebuild the tree directory. We have a rule now which is "Try to leave 10% free on a disc," then this problem never arises.
We use Jaz and Zips regularly as temporary storage and for quick transfers between computers but we always archive to CD. Elaine
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
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Hey Ciprian,
I'm not aware of a shareware tool, however, commercially IPLab (http://www.iplab.com/) can do it, and I believe so can ImportAccess (http://www.desacc.com/). Good luck.
Brian C. Tryon MCP-Hahnemann School of Medicine Ann Preston Hall, Box 551 3300 Henry Avenue Philadelphia, PA 19129 USA
----------------------------------------- "Quantifying is a committing task." - Cruz-Orive, 1994.
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." - Richard Feynman --------------------------------------------------------------
I travel quite a bit with my Jaz, and have not had the same experience mentioned earlier with drive problems. For what it's worth, when traveling I carry 2 carry-ons, one for my laptop, the other (a cool bag from Kensington) carries any drives, media, and accessories. I rarely pack my drives in checked luggage (my paranoia). One reason I carry on all drives (besides brutish bag-manglers, boy, do I have stories) is because I believe this avoids some of the temperature fluctuations that you might experience with checked bags. I've been to some cold regions and noticed luggage items quite cold when upacked. Typically, I'm working on something just prior to catching a flight so going from warm to cold couldn't help drive performance. Likewise, I typically take some dessicant and moisture-proof bags with me, quickly wrap the drive in, throw in a bag of dessicant and I believe this helps avoid condensation on the drive and when I get to my destination, I can get right to work without worry of letting the drive reach room temp or needing a papertowel to drive off the casing.
So, I've had Jaz drives (I currently have 3) since they came out, never a problem with the drive. I do routinely check the disk with disk utility programs, and have occasionally (once every 2 months with routine weekend checks of mission critical data) found some errors but nothing that could not be repaired.
I do store image data (triplicates) on Jaz and CD-ROM, but my main archival tool is CD-ROM, I've had excellent performance from a Pinnacle drive and Toast software and would strongly recommend to others to try CD-ROM storage. Again, I "burn" duplicates of all data stored on Jaz, one copy goes in the safe, the other is kept within easy reach, and the Jaz is used for day to day data working. When not being used, all Jaz are stored in their containers and filed in a clean unit. Most of the computers on my network get a weekend quick cleaning to keep dust and garbage from interrupting the work flow. Hope it helps,
Brian C. Tryon MCP-Hahnemann School of Medicine Ann Preston Hall, Box 551 3300 Henry Avenue Philadelphia, PA 19129 USA
----------------------------------------- "Quantifying is a committing task." - Cruz-Orive, 1994.
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." - Richard Feynman --------------------------------------------------------------
My colleague here at Polaroid has recently purchased an old microscope. He is interested in finding out as much information about it as possible. The markings on the microscope are the following: Waldemar Straufs Berlin, S.W. 68 N0. 3692 Note that the S and F is written in a strange script and may not be translated correctly. It is a black stand microscope with "brass accents" and a brass optical tube. It has a condenser which slides out, a polarizing stage, a mirror for capturing the light, a pivot arm in the stand. There are three objectives on the nose piece with the following markings: J. Thamm A.G. Berlin N.W. 6, one is an oil immersion. It came with three eye pieces as well. Pls. let us know if you have any information on this microscope. You can contact my colleague directly, Russ Gaudiana Gaudiar-at-Polaroid.com
We have a graduate student who is looking at concrete & the weathering of the steel rebar inside of it. He needs to section the steel rebar. We have never worked with steel, we are kinda biological oriented. So I'm asking for help. What's the best way for him to get thin sections of steel? Any & all suggestions gratefully accepted, I will pass them on to him.
Steeling myself for your replies,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
I am one of those people who moved from the bench to management (in USDA), and just can't seem to give up my roots (mycology/plant pathology) so keep plugging along with my self-funded research. I have been using glass knives for LM sections, and as part of a recent trade picked up a new EdgeCraft Standard Ultrathin 2.7mm diamond knife. Turns out that I am pretty happy with my glass knife sections, and would be better off with something else or some money instead of the diamond knife. The knife sells for about $2000, but I would be open to offers of about half that, or trades (or trade + cash) involving a better sterio microscope(s) - I am pretty happy with my B&L SZ5, but keep dreaming of a Wild - other areas of interest are older Leitz microscopes (170 mm Ortholux, Dialux, etc.), as well as objectives and components. If you want more information on the diamond knife (model, angle, etc.) - I can provide that. My objective is to find a home for something that I don't need, and in the process keep up the support for my own work.
RE: Ti Alloys - TEM foil preparation 1/13/99 3:47 PM
Bernard Kestel wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America = = } electrolytes I have used on Ti alloys. (3mm, jet polishing). } } Ti-8 w/% Al 13% HCL -60 degrees C } Ti-811 87% methanol 70 to 90 volts } Ti-6Al-4V 25-35 mA. } } Ti-13 Sn 130 ml HCL -50 degrees C } 670 ml methanol 150 volts } 100 ml butyl 60 mA. } } Ti-3V 60 ml perchloric acid -50 degrees C } Ti-20 Zr 590 ml methanol 50 volts } Ti-14 Al 350 ml butyl cellosolve 10-15 mA } } Ti-8 Al 60 ml perchloric acid -15 t0 -20 C } (rolled) 460 ml ethyl alcohol, 95% 180 volts } 280 ml butyl alcohol 40 mA. } 100 ml butyl cellosolve } Ti 5.3 g. = lithium = } chloride -40 degrees C. } nanocrystals, 11.16 g. magnesium 150 volts } compacted chloride 30 mA } 500 ml. methanol } 100 ml. butyl cellosolve } (Note: the two salts above must be added to the combined solvents = one = } "powder" at a time, while stirring. The two dry salts react violently if = mixed = } together. This electrolyte has nearly as wide an application as = perchloric acid = } mixtures and is known as B K-2). } I suspect that B K-2 will cause the least hydride problem. It was = } developed to eliminate hydrides in vanadium TEM foils. The voltages = given would = } need to be reduced about 20% for a twin jet system due to lower = electrical = } resistance of that configuration. My work was done on a South Bay = vertical single jet unit. } I have no vested interest in the above equipment, but have enjoyed = the = } ease with which good polishing conditions can be obtained and reproduced = since = } 1975 or so. Good luck! } Bernard Kestel } Materials Science Division } Argonne National Laboratory } Argonne, Il., 60439 } } E-mail: {bkestel-at-anl.gov} } Thomas, Larry wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } = To = } Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Satyrith- } } } } Unless you have a compelling reason to use ion milling for this = preparation, I } } recommend using jet electropolishing, possibly with just a final touch-= up = } in the } } ion mill. Although both electropolishing and ion milling commonly = produce } } sample preparation artifacts, the artifacts from ion-milling reactive = metals } } such as Ti and Zr are not well understood and can be harder to eliminate.=
} } Despite Carpenter et. als work (see D. Henriks posting) implicating = hydrocarbon } } contamination as a source of hydriding during ion milling, my personal } } experience ion milling Zr samples in 'oil-free' systems indicates that = water } } vapor is the main culprit in the hydridng. Surface irregularities on = ion = } milled } } Zr/Ti samples (surface terracing, for example) usually have other causes,=
} } possibly related to air leaks in the ion mill or oxygen in the feed gas. } } Another source of irregular surfaces on ion milled samples is embedded = } polishing } } compound. Cubic boron nitride (CBN) sometimes gives better results in = dimpling } } metals than diamond, and even the diamond polishing compounds from = different } } suppliers can give different results. Conversely, brief ion milling of } } electropolished samples--especially at low incidence angles and very low = } kV--can } } improve the surface finish and mill away irregularities caused by second-= phase } } particles. Often the 'trick' to eliminating milling artifacts is = minimizing } } time spent in the ion mill. } } } } Larry Thomas } } Mechanical and Materials Engineering } } Washington State University } } Pullman, WA } } } } } } ---------- } } From: South Bay Technology } } Sent: Tuesday, January 12, 1999 6:00 PM } } To: Satyarth Suri; Microscopy ListServer } } Subject: Ti Alloys - TEM foil preparation } } } } ------------------------------------------------------------------------=
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America = } To = } Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.= html } } -----------------------------------------------------------------------.=
} } } } } } Dear Satyarth: } } } } An excellent paper on the subject is: } } } } "In Situ Hydride Formation in Zirconium and Titanium during Ion Milling"=
} } Graham J.C. Carpenter et al, JMSA Vol.1 No. 4 pp 175-184 1995. } } } } From the paper you will see that a key factor in eliminating hydride } } formation is having a clean sample free from hydrocarbon contamination. = } } This contamination could be remnants from the dimpling process or other } } pre-thinning steps or it could be caused by the back streaming of } } diffusion } } pump oil in your ion mill. As titanium has a high chemical affinity = for } } hydrogen, you may want to look over your preparation steps and try to } } eliminate any areas of possible contamination. If you are still having } } a } } problem, a quick cleaning of both the specimen and the specimen holder } } in a } } Plasma Cleaner should take care of it. } } } } If you have an interest, I can send you a copy of the above referenced } } paper. I also have papers on plasma cleaning which may be of interest. } } } } NOTE: South Bay Technology does manufacture the PC150 Plasma Cleaner = and } } therefore I have a vested interest in promoting its use. } } } } Best regards- } } } } David } Writing at 9:38:23 AM on 1/12/99 } } } = } } *************************************************************** } ** } } ********** } } ************************ } } } } David Henriks TEL: = } } 800-728-2233 (toll free in the USA) } } South Bay Technology, Inc. +1-949-492-2600 } } 1120 Via Callejon FAX: } } +1-949-492-1499 } } San Clemente, CA 92673 USA e-mail: } } henriks-at-southbaytech.com } } } } = } } *************************************************************** } ** } } ********** } } ************************ } } } } } } } } } Please visit us at http://www.southbaytech.com { { { { { } } } } Manufacturers of precision sample preparation equipment and supplies = for } } metallography, crystallography and electron microscopy. } } } } Message text written by Satyarth Suri } } } } } Hello: } } I am currently working on mechanical behavior / microstructure } } correlations in single colony near apha Ti Alloys. I am currently } } preparing the foils using a dual ion mill (6kV, 12deg, 1mA), but } } have run into the problem of a very uneven alpha phase morphology, } } the alpha phase has island formation - we are using a cold stage } } to minimize the hydride formation at the interface. I am using } } 3 micron/6micron diamond paste during the dimpling process - the } } foil itself otherwise is fairly clean in terms of the dislocation } } content. Could anyone in the microscopy land have some suggestions } } in terms of what the problem may be? } } } } thanks - if you want you can respond directly to suri.3-at-osu.edu } } } } -satyarth } } } } { } } } } } } } } } } RFC822 header } } ----------------------------------- } } } } Received: from dns2.anl.gov (dns2.anl.gov [146.139.254.3]) by = } } horus.et.anl.gov (8.6.11/8.6.11) with ESMTP id SAA02218; Tue, 12 Jan = 1999 18:17:43 -0600 } } Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com = } } [206.69.208.10]) by dns2.anl.gov (8.9.1a/8.6.11) with SMTP id SAA19261; = Tue, 12 Jan 1999 = } } 18:18:22 -0600 (CST) } } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = } (8.6.11/8.6.11) } id RAA22399 for dist-Microscopy; Tue, 12 Jan 1999 17:41:= 01 -0600 } } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by = } } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id RAA22384 for = } } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 12 Jan 1999 17:40:30 -= 0600 } } Received: from Sparc5AAEM (aaem.amc.anl.gov [146.139.72.3]) by = } } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA22377 for = } } {microscopy-at-sparc5.microscopy.com} ; Tue, 12 Jan 1999 17:40:17 -0600 } } Received: from pnl.gov (relay.pnl.gov [130.20.128.34]) by Sparc5AAEM = } } (8.6.11/8.6.11) with ESMTP id RAA14848 for {MICROSCOPY-at-aaem.amc.anl.gov} ;= Tue, 12 Jan = } 1999 } 17:52:22 -0600 } } Received: from pnlmse1.pnl.gov by pnl.gov (PMDF V5.1-12 #28154) } } with ESMTP id {01J6GJ2FP8R4970ZC9-at-pnl.gov} for MICROSCOPY-at-aaem.amc.anl.= gov; } } Tue, 12 Jan 1999 15:54:41 PST } } Received: by PNLMSE1.pnl.gov with Internet Mail Service (5.5.2232.9) } } id {CR51M2B5} ; Tue, 12 Jan 1999 15:54:42 -0800 } } Content-return: allowed } } Date: Tue, 12 Jan 1999 15:54:36 -0800 } } From: "Thomas, Larry" {Larry.Thomas-at-pnl.gov} } } Subject: RE: Ti Alloys - TEM foil preparation } } To: Satyarth Suri {suri-at-mse.eng.ohio-state.edu} , } } Microscopy ListServer {MICROSCOPY-at-aaem.amc.anl.gov} , } } "'South Bay Technology'" {Henriks-at-CompuServe.COM} } } Message-id: {7A04D1887189D2118EEB00A024BF29DA41CB17-at-pnlmse2.pnl.gov} } } X-Mailer: Internet Mail Service (5.5.2232.9) } } Errors-to: Microscopy-request-at-sparc5.microscopy.com } } Content-Type: text } } Content-Length: 5172 } } Status: } } } } } RFC822 header } ----------------------------------- } } Received: from dns2.anl.gov (dns2.anl.gov [146.139.254.3]) by = } horus.et.anl.gov (8.6.11/8.6.11) with ESMTP id KAA07881; Wed, 13 Jan 1999 = 10:30:15 -0600 } Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com = } [206.69.208.10]) by dns2.anl.gov (8.9.1a/8.6.11) with SMTP id KAA11490; = Wed, 13 Jan 1999 = } 10:30:55 -0600 (CST) } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.= 11) = } id JAA28860 for dist-Microscopy; Wed, 13 Jan 1999 09:57:58 -0600 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id JAA28855 for = } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 13 Jan 1999 09:57:27 -= 0600 } Received: from Sparc5AAEM (aaem.amc.anl.gov [146.139.72.3]) by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id JAA28848 for = } {microscopy-at-sparc5.microscopy.com} ; Wed, 13 Jan 1999 09:57:14 -0600 } Received: from horus.et.anl.gov (horus.et.anl.gov [146.139.240.27]) by = } Sparc5AAEM (8.6.11/8.6.11) with ESMTP id KAA15405 for {MICROSCOPY-at-aaem.= amc.anl.gov} ; = } Wed, 13 Jan 1999 10:09:20 -0600 } Received: from 146.139.240.184 (et212mac184.et.anl.gov [146.139.240.184])= by = } horus.et.anl.gov (8.6.11/8.6.11) with SMTP id KAA07653; Wed, 13 Jan 1999 = } 10:11:07 -0600 } Message-Id: {199901131611.KAA07653-at-horus.et.anl.gov} } Date: 13 Jan 99 10:11:49 -0500 } From: Bernard Kestel {kestel-at-anl.gov} } Subject: RE: Ti Alloys - TEM foil preparation } To: "'South Bay Technology'" {Henriks-at-CompuServe.COM} , } "Thomas, Larry" {Larry.Thomas-at-pnl.gov} , } Microscopy ListServer {MICROSCOPY-at-aaem.amc.anl.gov} , } Satyarth Suri {suri-at-mse.eng.ohio-state.edu} } X-Mailer: QuickMail Pro 1.5.4 (Mac) } X-Priority: 3 } MIME-Version: 1.0 } Reply-To: Bernard Kestel {kestel-at-anl.gov} } Content-Transfer-Encoding: quoted-printable } Errors-to: Microscopy-request-at-sparc5.microscopy.com } Content-Type: text/plain; charset=3D"US-Ascii" } Content-Length: 10127 } Status: = } --====56524855544856555650===1 Content-Type: text/html; charset="US-Ascii" Content-Transfer-Encoding: quoted-printable
{HTML} {HEAD} {/HEAD} {BODY} {PRE WIDTH=3D"132"} RE: Ti Alloys - TEM foil preparation 1/13/99 3:47 PM
{/PRE} {FONT = FACE=3D"Geneva" SIZE=3D2 COLOR=3D"#000000"} {BR} Bernard Kestel wrote: {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >-----------------------------------------------------------------------= - {BR} >The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America {BR} Reply to: RE: Ti Alloys - TEM = foil preparation {BR} > Re: Electropolishing = of Ti alloys {BR} > Due to interest = in this subject, I will list some additional = {BR} >electrolytes I have used on = Ti alloys. (3mm, jet polishing). {BR} > {BR} > = Ti-8 w/% Al 13% HCL = -60 degrees C {BR} > Ti-811 = 87% methanol = 70 to 90 volts {BR} > Ti-6Al-4V = 25-35 = mA. {BR} > {BR} > Ti-13 Sn = 130 ml HCL -50 degrees = C {BR} > = 670 ml methanol 150 volts {BR} > = 100 ml butyl = 60 mA. {BR} > {BR} > = Ti-3V 60 ml perchloric = acid -50 degrees C {BR} > Ti-20 = Zr 590 ml methanol = 50 volts {BR} > Ti-14 Al = 350 ml butyl cellosolve 10-15 mA {BR} > {BR} > = Ti-8 Al 60 ml perchloric = acid -15 t0 -20 C {BR} > (rolled) = 460 ml ethyl alcohol, 95% = 180 volts {BR} > = 280 ml butyl alcohol = 40 mA. {BR} > = 100 ml butyl cellosolve {BR} > = Ti = 5.3 g. lithium {BR} >chloride = -40 degrees C. {BR} > nanocrystals, = 11.16 g. magnesium 150 = volts {BR} > compacted = chloride 30 mA {BR} > = 500 ml. methanol {BR} > = 100 ml. butyl = cellosolve {BR} > (Note: the two salts = above must be added to the combined solvents = one {BR} >"powder" at a time, = while stirring. The two dry salts react = violently if mixed {BR} >together. This = electrolyte has nearly as wide an application = as perchloric acid {BR} >mixtures and is = known as B K-2). {BR} > I suspect that = B K-2 will cause the least hydride problem. = It was {BR} >developed to eliminate hydrides = in vanadium TEM foils. The voltages given = would {BR} >need to be reduced about 20% = for a twin jet system due to lower electrical = {BR} >resistance of that configuration. = My work was done on a South Bay vertical = single jet unit. {BR} > I have no vested = interest in the above equipment, but have = enjoyed the {BR} >ease with which good = polishing conditions can be obtained and = reproduced since {BR} >1975 or so. Good = luck! {BR} > Bernard Kestel {BR} > = Materials Science Division {BR} > = Argonne National Laboratory {BR} > = Argonne, Il., 60439 {BR} > {BR} > = E-mail: < {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = bkestel-at-anl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} > {BR} >Thomas, Larry = wrote: {BR} >>-------------------------------------------------------------------= ----- {BR} >>The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America >To {BR} >Subscribe/Unsubscribe = -- Send Email to {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {= U} ListServer-at-MSA.Microscopy.Com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >>On-Line Help = {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} http://www.msa.= microscopy.com/MicroscopyListserver/FAQ.html {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >>-------------------------------------------------------------------= ----. {BR} >> {BR} >> {BR} >>Satyrith- {BR} >> {BR} >>Unless = you have a compelling reason to use ion = milling for this preparation, I {BR} >>recommend = using jet electropolishing, possibly with = just a final touch-up {BR} >in the {BR} >>ion = mill. Although both electropolishing and = ion milling commonly produce {BR} >>sample = preparation artifacts, the artifacts from = ion-milling reactive metals {BR} >>such = as Ti and Zr are not well understood and = can be harder to eliminate. {BR} >>Despite = Carpenter et. als work (see D. Henriks posting) = implicating hydrocarbon {BR} >>contamination = as a source of hydriding during ion milling, = my personal {BR} >>experience ion milling = Zr samples in 'oil-free' systems indicates = that water {BR} >>vapor is the main culprit = in the hydridng. Surface irregularities = on ion {BR} >milled {BR} >>Zr/Ti samples = (surface terracing, for example) usually = have other causes, {BR} >>possibly related = to air leaks in the ion mill or oxygen in = the feed gas. {BR} >>Another source of = irregular surfaces on ion milled samples = is embedded {BR} >polishing {BR} >>compound. = Cubic boron nitride (CBN) sometimes gives = better results in dimpling {BR} >>metals = than diamond, and even the diamond polishing = compounds from different {BR} >>suppliers = can give different results. Conversely, = brief ion milling of {BR} >>electropolished = samples--especially at low incidence angles = and very low {BR} >kV--can {BR} >>improve = the surface finish and mill away irregularities = caused by second-phase {BR} >>particles. = Often the 'trick' to eliminating milling = artifacts is minimizing {BR} >>time spent = in the ion mill. {BR} >> {BR} >>Larry = Thomas {BR} >>Mechanical and Materials = Engineering {BR} >>Washington State University {BR} >>Pullman, = WA {BR} >> {BR} >> {BR} >> ---------- {BR} >> From: = South Bay Technology {BR} >> Sent: Tuesday, = January 12, 1999 6:00 PM {BR} >> To: = Satyarth Suri; Microscopy ListServer {BR} >> Subject: = Ti Alloys - TEM foil preparation {BR} >> {BR} >> ------------------------------------------------------------------= ------ {BR} >> The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America > To {BR} >Subscribe/Unsubscribe = -- Send Email to {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {= U} ListServer-at-MSA.Microscopy.Com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >> On-Line Help = {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} http://www.msa.= microscopy.com/MicroscopyListserver/FAQ.html {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >> ------------------------------------------------------------------= -----. {BR} >> {BR} >> {BR} >> Dear = Satyarth: {BR} >> {BR} >> An excellent = paper on the subject is: {BR} >> {BR} >> "In = Situ Hydride Formation in Zirconium and = Titanium during Ion Milling" {BR} >> Graham = J.C. Carpenter et al, JMSA Vol.1 No. 4 pp = 175-184 1995. {BR} >> {BR} >> From = the paper you will see that a key factor = in eliminating hydride {BR} >> formation = is having a clean sample free from hydrocarbon = contamination. {BR} >> This contamination = could be remnants from the dimpling process = or other {BR} >> pre-thinning steps or = it could be caused by the back streaming = of {BR} >>diffusion {BR} >> pump oil = in your ion mill. As titanium has a high = chemical affinity for {BR} >> hydrogen, = you may want to look over your preparation = steps and try to {BR} >> eliminate any = areas of possible contamination. If you = are still having {BR} >>a {BR} >> problem, = a quick cleaning of both the specimen and = the specimen holder {BR} >>in a {BR} >> Plasma = Cleaner should take care of it. {BR} >> {BR} >> If = you have an interest, I can send you a copy = of the above referenced {BR} >> paper. = I also have papers on plasma cleaning which = may be of interest. {BR} >> {BR} >> NOTE: = South Bay Technology does manufacture the = PC150 Plasma Cleaner and {BR} >> therefore = I have a vested interest in promoting its = use. {BR} >> {BR} >> Best regards- {BR} >> {BR} >> David = > Writing = at 9:38:23 AM on 1/12/99 {BR} >> = > {BR} >>*************************************************************** {BR} =
>** {BR} >>********** {BR} >> ************************ {BR} >> {BR} >> David = Henriks = TEL: {BR} >> 800-728-2233 = (toll free in the USA) {BR} >> South = Bay Technology, Inc. = +1-949-492-2600 {BR} >> 1120 Via = Callejon = FAX: {BR} >>+1-949-492-1499 {BR} >> San = Clemente, CA 92673 USA e-mail: {BR} >> {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} henriks-at-southbaytech.com {/U} {= /FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >> {BR} >> {BR} >>*************************************************************** {BR} =
>** {BR} >>********** {BR} >> ************************ {BR} >> {BR} >> = >>>>> Please visit us = at {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} http://www.= southbaytech.com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} <<<<< {BR} >> {BR} >> Manufacturers = of precision sample preparation equipment = and supplies for {BR} >> metallography, = crystallography and electron microscopy. {BR} >> {BR} >> Message = text written by Satyarth Suri {BR} >> > {BR} >> Hello: {BR} >> I = am currently working on mechanical behavior = / microstructure {BR} >> correlations = in single colony near apha Ti Alloys. I = am currently {BR} >> preparing the foils = using a dual ion mill (6kV, 12deg, 1mA), = but {BR} >> have run into the problem = of a very uneven alpha phase morphology, {BR} >> the = alpha phase has island formation - we are = using a cold stage {BR} >> to minimize = the hydride formation at the interface. = I am using {BR} >> 3 micron/6micron diamond = paste during the dimpling process - the {BR} >> foil = itself otherwise is fairly clean in terms = of the dislocation {BR} >> content. = Could anyone in the microscopy land have = some suggestions {BR} >> in terms of = what the problem may be? {BR} >> {BR} >> thanks = - if you want you can respond directly to = {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} suri.3-at-osu.edu {/= U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >> {BR} >> -satyarth {BR} >> {BR} >> < {BR} >> {BR} >> {BR} >> {BR} >> {BR} >>RFC822 = header {BR} >>----------------------------------- {BR} >> {BR} >> = Received: from dns2.anl.gov (dns2.anl.gov = [146.139.254.3]) by {BR} >>horus.et.anl.gov = (8.6.11/8.6.11) with ESMTP id SAA02218; = Tue, 12 Jan 1999 18:17:43 -0600 {BR} >> = Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com = {BR} >>[206.69.208.10]) by dns2.anl.gov = (8.9.1a/8.6.11) with SMTP id SAA19261; Tue, = 12 Jan 1999 {BR} >>18:18:22 -0600 (CST) {BR} >> = Received: (from {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = daemon-at-localhost {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} ) by Sparc5.Microscopy.Com = {BR} >(8.6.11/8.6.11) >id RAA22399 for = dist-Microscopy; Tue, 12 Jan 1999 17:41:01 = -0600 {BR} >> Received: from no_more_spam.com = (Sparc5 [206.69.208.10]) by {BR} >>Sparc5.Microscopy.Com = (8.6.11/8.6.11) with SMTP id RAA22384 for = {BR} >>" {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = MicroscopyFilteredEmail-at-msa.microscopy.com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} "; Tue, 12 Jan 1999 = 17:40:30 -0600 {BR} >> Received: from = Sparc5AAEM (aaem.amc.anl.gov [146.139.72.3]) = by {BR} >>Sparc5.Microscopy.Com (8.6.11/8.6.11) = with ESMTP id RAA22377 for {BR} >>< {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} microscopy-at-sparc5.microscopy.= com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} >; Tue, 12 Jan 1999 = 17:40:17 -0600 {BR} >> Received: from = pnl.gov (relay.pnl.gov [130.20.128.34]) = by Sparc5AAEM {BR} >>(8.6.11/8.6.11) = with ESMTP id RAA14848 for < {/FONT} {FONT FACE=3D"Geneva" = SIZE=3D1 COLOR=3D"#0000FF"} {U} MICROSCOPY-at-aaem.amc.anl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} >; Tue, 12 Jan {BR} >1999 = >17:52:22 -0600 {BR} >> Received: = from pnlmse1.pnl.gov by pnl.gov (PMDF V5.1-12 = #28154) {BR} >> with ESMTP id < {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} 01J6GJ2FP8R4970ZC9-at-pnl.gov {/= U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} > for {/FONT} {FONT FACE=3D"= Geneva" = SIZE=3D1 COLOR=3D"#0000FF"} {U} MICROSCOPY-at-aaem.amc.anl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} ; {BR} >> Tue, 12 = Jan 1999 15:54:41 PST {BR} >> Received: = by PNLMSE1.pnl.gov with Internet Mail Service = (5.5.2232.9) {BR} >> id <CR51M2B5>; = Tue, 12 Jan 1999 15:54:42 -0800 {BR} >> = Content-return: allowed {BR} >> Date: = Tue, 12 Jan 1999 15:54:36 -0800 {BR} >> = } From: "Thomas, Larry" < {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} Larry.Thomas-at-pnl.gov {/U} {/= FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} > {BR} >> Subject: = RE: Ti Alloys - TEM foil preparation {BR} >> = To: Satyarth Suri < {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#= 0000FF"} {U} suri-at-mse.eng.ohio-state.edu {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} >, {BR} >> = Microscopy ListServer < {/FONT} {FONT FACE=3D"Geneva" = SIZE=3D1 COLOR=3D"#0000FF"} {U} MICROSCOPY-at-aaem.amc.anl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} >, {BR} >> = "'South Bay Technology'" < {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} Henriks-at-CompuServe.COM {/U} {/= FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} > {BR} >> Message-id: = < {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = 7A04D1887189D2118EEB00A024BF29DA41CB17-at-pnlmse2.pnl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} > {BR} >> X-Mailer: = Internet Mail Service (5.5.2232.9) {BR} >> = Errors-to: {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = Microscopy-request-at-sparc5.microscopy.com {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >> Content-Type: = text {BR} >> Content-Length: 5172 {BR} >> = Status: > {BR} > {BR} > {BR} > {BR} >RFC822 = header {BR} >----------------------------------- {BR} > {BR} > = Received: from dns2.anl.gov (dns2.anl.gov = [146.139.254.3]) by {BR} >horus.et.anl.gov = (8.6.11/8.6.11) with ESMTP id KAA07881; 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by mtiwmhc02.worldnet.att.net (InterMail v03.02.07 118 124) with ESMTP id {19990114001451.EPZL10402-at-worldnet.att.net} ; Thu, 14 Jan 1999 00:14:51 +0000 Message-ID: {369D37CA.56F575DD-at-worldnet.att.net}
Dear Paula,
The success of thin sections will depend on the blade type and size which will then be dictated on the type of saw you have. Slow speed diamond saws have a blade capacity of 5" Diameter which can limit the sample size to be cut and take a long time to cut.. High speed saws can accomodate larger blades and perform cuts much faster (5 minutes or less) with the same accuracy as a slow speed saw. Feed pressure can affect the blade during the cut on either saw. If the blade is too thin, too much feed pressure will cause the blade to wander form the original cut line affecting the thickness of the section. This can be remedied by using a large flange or more rigid, thicker blade.
CBN is the only choice for cutting steels on a low speed saw without destroying the blade or sample, but can also be used on a high speed saw. Aluminum Oxide blades cut the most effieciently on a high speed saw and cost much less. DO NOT attempt cutting with diamond blades, they will only become loaded with steel and this will prohibit the sample from being cut. Even frequent dressing will not enhance the performance. Aluminum Oxide blades are better suited for steels and Allied HighTech has a wide selection of blades for any saw you may have.
I know of two labs at LBL that have a high speed saw and can get you this information should you require it. In additon, if you have any application questions, please feel free to contact me at 800-675-1118 to further discuss your interest.
Good Luck,
Gary Liechty
Paula Sicurello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hey there boarders, } } We have a graduate student who is looking at concrete & the } weathering of the steel rebar inside of it. He needs to section the steel } rebar. We have never worked with steel, we are kinda biological oriented. } So I'm asking for help. } What's the best way for him to get thin sections of steel? Any & } all suggestions gratefully accepted, I will pass them on to him. } } Steeling myself for your replies, } } Paula :-) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } phone: 510-642-2085 } fax: 510-643-6207 } http://biology.berkeley.edu/EML
-- Gary Liechty Product Application Specialist
Allied High Tech Products, Inc. 2376 E. Pacifica Place Rancho Dominguez, CA 90220 800-675-1118 310-635-2466 310-762-6808 Fax
Equipment and Consumable Products for Materialographic, SEM and TEM Sample Prepration
Try Graphic converter, it's a shareware that can read and translate almost every format. It can be able to solve your problem. You can find it on every info-mac mirror.
Eric LEROY Dr. Laboratoire de Chimie Metallurgique des Terres Rares UPR 209 - CNRS Groupe des Laboratoires de Thiais 2-8, rue Henri Dunant 94320 THIAIS cedex
A few years ago I got hold of a beta version of a SEM Macintosh simulation program, SuperSEM. It was a Macintosh program that was developed with Supercard software. It was misplaced during the process of upgrading my computers. Does anyone know if it exists?
Thanks in advance, Lou Ross Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html
Colin raises a good point of media being orphaned that we feel the pinch of already. Our HP 1300T MO drive (~650 MB each side) failed on us a while back. It will no longer accept the cartridges - it keeps kicking them out. We had already offloaded much of the data to CD but we have a few cartridges that I would still like to retrieve. However, we appear to be the only ones on campus that ever purchased the HP drive.
Is there anyone out there with a working HP MO drive (gathering dust or not) that could help us retrieve the last few cartidges?
At 07:10 AM 1/12/99 +0000, you wrote: {snip} } As pointed out the Jaz/Zip technology will probably be superseded } soon ( 1GB drives are gone already ! ) and in a few years it will be } difficult to read data if your drive fails. This was a major factor } along with cost ) 5 years ago when we decided on a CD-R storage system, } over Magneto-Optical which was the current favourite then. {snip} } Colin Reid, } Electron Microscope Unit, } Trinity College Dublin,
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
Please remember that 3/4ths of your 10 MB image won't be doing you any good during the slide show. If your projector supports true 24-bit color (as opposed to 16-bit or 8-bit) and 1024x768 pixels, you will only be displaying 2.25 MB of image. You might as well match your image size to the projector as you make up your presentation. It would help file size and the speed of transition between slides, but these fast computers are handling that pretty well.
I concur with the others. Most imaging programs have a slide show capability built in. They may not have all the fancy transitional effects, but they should do the show. Even my shareware LViewPro on the PC had that.
At 05:29 PM 1/12/99 -0400, you wrote: } } We just got a G3 Mac and an Epson digital projector-I have seen some really } nice digital slide shows and am excited by the increase in quality and } decrease in effort making slides. I expect, however, to spend some effort } finding out the best way to put together a slide show, and am wondering } what advice or experience might be out there. I would like to show ~10MB } images without any computer stuff showing at the same time, but could } consider compression (eg., jpeg) if there is no loss of quality. Wondering } whether to get into a slide presentation program or just make a stack and } open them sequentially (I have 190+ MB of memory). There is also a neat IR } pointer that seems to bounce off the screen and feed into software in the } projector that allows you to move the cursor around and click/double click } with the IR pointer. Too bad they didn't include a laser pointer in this } device!....Many thanks for any help or advice!...Tom Reese } ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
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Reply to: RE: Ti Alloys - TEM foil preparation Re: Electropolishing of Ti alloys = Due to interest in this subject, I will list some additional = electrolytes I have used on Ti alloys. (3mm, jet polishing).
Ti-8 w/% Al 13% HCL -60 degrees C Ti-811 87% methanol 70 to 90 volts Ti-6Al-4V 25-35 mA.
Ti-13 Sn 130 ml HCL -50 degrees C 670 ml methanol 150 volts 100 ml butyl 60 mA.
Ti-3V 60 ml perchloric acid -50 degrees C Ti-20 Zr 590 ml methanol 50 volts Ti-14 Al 350 ml butyl cellosolve 10-15 mA
Ti-8 Al 60 ml perchloric acid -15 t0 -20 C (rolled) 460 ml ethyl alcohol, 95% 180 volts 280 ml butyl alcohol 40 mA. 100 ml butyl cellosolve = Ti 5.3 g. lithium chloride -40 degrees C. nanocrystals, 11.16 g. magnesium 150 volts compacted chloride 30 mA 500 ml. methanol 100 ml. butyl cellosolve (Note: the two salts above must be added to the combined solvents one = "powder" at a time, while stirring. The two dry salts react violently if = mixed together. This electrolyte has nearly as wide an application as = perchloric acid mixtures and is known as B K-2). I suspect that B K-2 will cause the least hydride problem. It was = developed to eliminate hydrides in vanadium TEM foils. The voltages given = would need to be reduced about 20% for a twin jet system due to lower = electrical resistance of that configuration. My work was done on a South = Bay vertical single jet unit. I have no vested interest in the above equipment, but have enjoyed = the ease with which good polishing conditions can be obtained and = reproduced since 1975 or so. Good luck! = Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439
E-mail: {bkestel-at-anl.gov} Thomas, Larry wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE: Ti Alloys - TEM foil preparation Re: Electropolishing of Ti alloys = Due to interest in this subject, I will list some additional = electrolytes I have used on Ti alloys. (3mm, jet polishing).
Ti-8 w/% Al 13% HCL -60 degrees C Ti-811 87% methanol 70 to 90 volts Ti-6Al-4V 25-35 mA.
Ti-13 Sn 130 ml HCL -50 degrees C 670 ml methanol 150 volts 100 ml butyl 60 mA.
Ti-3V 60 ml perchloric acid -50 degrees C Ti-20 Zr 590 ml methanol 50 volts Ti-14 Al 350 ml butyl cellosolve 10-15 mA
Ti-8 Al 60 ml perchloric acid -15 t0 -20 C (rolled) 460 ml ethyl alcohol, 95% 180 volts 280 ml butyl alcohol 40 mA. 100 ml butyl cellosolve = Ti 5.3 g. lithium chloride -40 degrees C. nanocrystals, 11.16 g. magnesium 150 volts compacted chloride 30 mA 500 ml. methanol 100 ml. butyl cellosolve (Note: the two salts above must be added to the combined solvents one = "powder" at a time, while stirring. The two dry salts react violently if = mixed together. This electrolyte has nearly as wide an application as = perchloric acid mixtures and is known as B K-2). I suspect that B K-2 will cause the least hydride problem. It was = developed to eliminate hydrides in vanadium TEM foils. The voltages given = would need to be reduced about 20% for a twin jet system due to lower = electrical resistance of that configuration. My work was done on a South = Bay vertical single jet unit. I have no vested interest in the above equipment, but have enjoyed = the ease with which good polishing conditions can be obtained and = reproduced since 1975 or so. Good luck! = Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439
E-mail: {bkestel-at-anl.gov} Thomas, Larry wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
by hawk.csrv.uidaho.edu (8.8.6 (PHNE_14041)/) with ESMTP id IAA24952 for {Microscopy-at-Sparc5.Microscopy.Com} ; Thu, 14 Jan 1999 08:45:37 -0800 (PST) Received: from OAK/SpoolDir by oak.csrv.uidaho.edu (Mercury 1.43); 14 Jan 99 08:47:23 -0800 (PST) Received: from SpoolDir by OAK (Mercury 1.43); 14 Jan 99 08:46:51 -0800 (PST)
I am looking for a used, but functional, EDX specimen holder for a JEOL 1200 EX TEM. If anyone has one for sell, please contact me at: jfb-at-uidaho.edu (208)885-6656
Thank you.
Franklin Bailey Holm Research Center University of Idaho Moscow, ID 83844-2204
A pair of postdocs in Plant Pathology came in to the facility to ask about a cold stage for a light microscope, which no one at this university seems to have. They want to look at cryostat sections of wood, and need digital images for image analysis, and the sections must remain frozen. I have here a light microscope fitted with a digital camera hooked to the Mac with NIH Image. I have liquid nitrogen, styrofoam, and a machine shop. I also have high humidity. Can anyone tell me how to kludge together a cold enough stage that we can get images of these frozen sections while preventing frost? I have some general ideas, but I'd appreciate any specific plans or ideas.
The idea is to look at the number of vacuoles filled with gas vs the number filled with water. Up to now they have had images taken with an SEM with a cold stage in Canada, and would prefer to do them locally, at lower mag, and cheaper.
Thanks in advance for any advice!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
A colleage from an industrial SEM lab contacted me about a problem with contamination on the final aperture on an variable pressure SEM. This SEM was recently purchased by one of their branch plants to examine semiconductor products. Although it is a variable pressure SEM, they have only operated it in the high vac mode like a standard SEM.
The problem they are experiencing is that the final aperture presumably gets so dirty they have to change the aperture every two weeks. Unfortunately the manufacturer has not been able to shed any light on this situation.
If the vacuum system is alright, I suggested that the problem might be with the samples. Supposedly the branch lab operators are following the same sample prep protocol established in the local SEM lab.
Any other ideas out there?
Thanks in advance, Lou Ross Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html
Products like Dazzle can digitize the video over to MPG files which should be transportable. We bought a unit for about $200 and are still coming up to speed with it. If you have a CD-R, you should be able to cut the movies onto a CD which they could read over there.
Disclaimer: We have no interest in Dazzle other than its the unit we bought. There are many other products on the market which can also accomplish the task.
Warren S.
At 12:15 PM 1/14/99 -0500, you wrote: } I have in-situ results currently stored on VHS tape that I need to present } in Europe. } } Any recipes available? } } Thanks. } David E. Luzzi } Professor } Department of Materials Science } University of Pennsylvania
To all We have a client trying to convert from glass plates to sheet film. Can anyone help us to obtain film adapters for the Siemens 1A camera? Thank you. Peter Stolzenberg
I'm just posting to ask if anyone has developed a fixation protocol for Drosophila embryos suitable for use in TEM studies. Is it necessary to remove the chorion and vitelline membranes as in light microscopy procedures. Also, I'm wondering if the use of methanol is strictly taboo, or can it be tolerated if the exposure time is short?
Does anyone have a good protocol or reference for virus isolation = from food other than shellfish? We are trying to isolate and concentrate = Norwalk virus from some foods implicated in a poisoning outbreak in = Detroit. The foods in question are salami, garbanzo beans and some kind of = cheese (I think provolone). We need to concentrate to at least 1 million = particles/ml in order to use our prep for negative staining and TEM = observation. Any suggestions would be greatly appreciated!
Sincerely, Peggy Casey Michigan Dept. of Community Health Lansing, Mich. phone: 517-335-8102 e-mail: caseym-at-state.mi.us
I have been told to search for "M6", an adhesive which was used in the somewhat distant past for bonding samples for cross-sectioning. ..I cannnot find a company which has even heard of this. Any suggestions as to where I can find it?
__ _-==-=_,-. /--`' \_-at---at-.-- { Tim (TJ) LaFave Jr. `--'\ \ {___/. Department of Physics \ \\ " / University of North Carolina, Charlotte } =\\_/` { Charlotte, NC 28223 ____ /= | \_|/ _' `\ _/=== \___/ (704)547-3392 [x4] `___/ //\./=/~\====\ (704)509-6622 [Hm] \ // / | ===: http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ \/ \\ \\`--| / \\ ---------- +*+ ---------- | _ \\: /==:-\ `.__' `-____/ |--|==: Such that the future be theirs \ \ ===\ :==:`-' to shape and direct. _} \ ===\ /==/ -----------------------------------------------------------------
} Hello, } } A colleague is interested in examining rat kidney tubules at the TEM level. } The tubules are approximately 40 um in length. Unfortunately, once they } are placed in fixative in a 1 ml centrifuge tube, the tubules are no longer } visible which creates processing problems. We are going to try placing } the tubules in agar as a way of transporting the tubules through the } processing procedure so as not to lose them in transit. } } Also, the few cells (not tubules) we did see appeared to be washed out. We } used the following solutions: } } 2% glutaraldehyde in sodium cacodylate buffer } 2% osmium tetroxide } alcohol } propylene oxide } polybed } } Any suggestions would be greatly appreciated. } } Thank you, } } Ginger Baker } EM Lab Manager } Oklahoma State University College of Osteopathic Medicine } lizard-at-osu-com.okstate.edu } 918-561-8232
Through the years I have approached, evaluated and re-evaluated "hole sizing" from SEM micrographs for various technologies: biology, electron optics, polymers, chemistry and microelectronics. The paramount objectives are always the same: 1) spatial resolution(of digitizing hardware and software) 2) repeatability 3) SEM background contrast (gray scales) differentiation 4) Maximizing Automation 5) Minimizing user judgement (auto-thresholding, etc.) 6) Manual extraction of artifacts 7) Stereological considerations to mention a few. Also such techniques were attempted to improve the above such as: Image projection, edge detection, pre- and post- image or frame(or pixel) averaging and processing. Well once again this re-evaluation in light of current advances in hard and soft digital technologies is required. I thought I would see if commercial and non-commercial members of this forum would share any current technological information with me. Hopefully after the first 5 or 10 responses, to avoid driving subscribers crazy , you could respond directly to my email. I will gather and redistribute this data in any fashion requested at a later date to avoid congesting the mailservor. Thank you for your interest. Jeff Day/ 'JD' DBA Texas Industrial Mesquite, Texas Email: WA5EKH-at-juno.com
___________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com/getjuno.html or call Juno at (800) 654-JUNO [654-5866]
} } } I have been told to search for "M6", an adhesive which was used in the } somewhat distant past for bonding samples for cross-sectioning. ..I } cannnot find a company which has even heard of this. Any suggestions as } to where I can find it?
Great stuff! Does work well. M6 is a short abbreviation for M-Bond 610 which was originally desighned for strain gages. Can be purchased from "MM" Micro Measurement Division at Raleigh North Carolina USA. Phone: (919) 365 3800 Fax: (919) 365 3945
Standard disclaimer.
Mr. S H Coetzee Electron Microscope Unit Private bag X3 Wits Johannesburg 2050 Tell: +27 11 716 2419 Fax : +27 11 339 3407 E-mail stephan-at-gecko.biol.wits.ac.za
Tim (TJ) LaFave Jr. wrote: ================================================= I have been told to search for "M6", an adhesive which was used in the somewhat distant past for bonding samples for cross-sectioning. ..I cannnot find a company which has even heard of this. Any suggestions as to where I can find it? ================================================== Could it be that you are looking for M-Bond(TM) 610? It is all described on our website below, if that is what you had in mind. Maybe there was a predecessor called M6 but in any case, M-Bond 610 is perhaps what you had in mind? The product is manufactured by a division of Vishay Technology.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
} I have in-situ results currently stored on VHS tape that I need to present } in Europe. } } Any recipes available? } } Thanks. } David E. Luzzi } Professor } Department of Materials Science } University of Pennsylvania } 3231 Walnut Street } Philadelphia, PA 19104-6272 } } 215-898-8366 } 215-573-2128 - fax } luzzi-at-lrsm.upenn.edu {mailto:luzzi-at-lrsm.upenn.edu} } } Hi David,
Here in Europe we use different standards in different countries, however, we also have multi-standard machines that will play the 3 commonest standards (NTSC, PAL and SECAM). I suggest that before you go to too much trouble check if your hosts can play NTSC. VHS is the commonest format here anyway. Regards, Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
I did my first bifringant crystals with Polaroid light to night. My experiment in marginal at best. The swift is not too swift and my polarizing setup was a Grey photo polarize and the resolver was a broken pair of brown sun glasses. Probably the worst choice I could have come up with.
But it was a hoot.
I am trying a little slower drying solution and hope for larger salt crystals.
I have gone through my stock of Siemens spares, and have found 23 cut film inserts to fit the Elmiskop 1a plate holders. They are second-hand, but appear to be in very good condition. If you are interested please Email me directly.
Regards,
Bob ********************************************** Bob Phillips, MicroServiS Electron Microscopy Services, 11 Grafton Close, St. Ives, Huntingdon, Cambs. PE17 6DL United Kingdom. Email: microservis-at-dial.pipex.com ****************************************** -----Original Message----- } From: "PESTOEM-at-aol.com"-at-Sparc5.Microscopy.Com {"PESTOEM-at-aol.com"-at-Sparc5.Microscopy.Com} To: Microscopy-at-Sparc5.Microscopy.Com {Microscopy-at-Sparc5.Microscopy.Com} Cc: uelzen-at-erols.com {uelzen-at-erols.com}
O.k., all you good microscopists, I'm looking for a source for either a Uranium Glass Block or a suitable replacement for visualizing/teaching the light path on a light microscope. The block is placed in the microscope stage and the illumination path way can be viewed inside the block - particularly as the condensor is focused and as either the stage aperature or field aperature are varied.
I know this can be visualize with a dilute milk solution but I really would prefer something a little more stable (and non-perishable).
Thanks!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meeting point for developers and users working in these rapidly evolving fields and play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic developments in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
If you pulse-spiin the tubules in an Eppendorf centrifuge and then osmicate (I would use 1% OSO4 in 0.1M sodium cacodylate, 30 min to 1 hr, room temp), then spin again, you should be able to see them. At that point you can embed them in a small drop of 2% agarose and continue the processing. Good luck.
On Thu, 14 Jan 1999, Ginger R Baker wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hello, } } } } A colleague is interested in examining rat kidney tubules at the TEM } level. } } The tubules are approximately 40 um in length. Unfortunately, once they } } are placed in fixative in a 1 ml centrifuge tube, the tubules are no } longer } } visible which creates processing problems. We are going to try placing } } the tubules in agar as a way of transporting the tubules through the } } processing procedure so as not to lose them in transit. } } } } Also, the few cells (not tubules) we did see appeared to be washed out. } We } } used the following solutions: } } } } 2% glutaraldehyde in sodium cacodylate buffer } } 2% osmium tetroxide } } alcohol } } propylene oxide } } polybed } } } } Any suggestions would be greatly appreciated. } } } } Thank you, } } } } Ginger Baker } } EM Lab Manager } } Oklahoma State University College of Osteopathic Medicine } } lizard-at-osu-com.okstate.edu } } 918-561-8232 } } } }
I am a new faculty member in a small department of veterinary pathology. I am interested in obtaining the equipment necessary to produce good quality ultrathin sections for TEM examination at our University's central EM facility. I am particularly interested in ultratomes (1 or 2), vacuum oven, embedding molds, and other necessities. Equipment from laboratories that are down-sizing would be considered ideal. Please respond directly to my e-mail address: sfrasca-at-canr1.cag.uconn.edu
Richard: Here is a summary I had saved of a previous uranyl glass thread. Hope it helps. Tom
--------------------------------------
Newport Industrial Glass, Inc. 1631 Monrovia Ave. Costa Mesa, CA 92627 Tel: 714-642-9980 Fax: 714-645-6800 Contact person: Bill Larsen (you can tell him I sent you). Sold as a 6.5x6.5" sheet (you can specify 1 mm height), so you may want to form a "consortium" to have Newport pre-cut a sheet to slide size (nominal extra cost, but your lab only needs one or two slides). If there is a lot of interest, my company may start selling single slides.
As for references and the Shading Correction equation: please see my article in the 11/94 issue of Journal of NIH Research 6(11): 80 (usual Internet disclaimer: yes, that is an ad from my company on the facing page). Also look at Jericevic et al (1989) Methods in Cell Biology 30:47-83.
MutliSpeck beads: The Molecular Probes pre-mounted slide kits should be ideal for DAPI and Fluorescein. I believe they were optimized for Rhodamine, but should still work ok for Texas Red. If your problem is with mounting, Mol. Probes now sells the beads in solution, so you can 'sprinkle' some on your specimens. If you have a different problem with the current MultiSpeck's, Mol. Probes may be able to work something out for you.
Sorry, but I usually buy my reference material from Mol. Probes and don't keep close track of other slide manufacturers.
Sincerely,
Dr. George McNamara Universal Imaging Corporation George_M-at-Image1.com
} Dear confocalers,
} I read through the archives for information on uranyl glass test slides, } but couldn't figure out if there is a definitive answer to the question, } Does anyone still manufacture uranyl glass, and where could I get some?
You can obtain micro slide sized pieces of uranyl glass from:
Newport Industrial Glass, Inc. 1631 Monrovia Avenue Costa Mesa, CA 92627 (714) 645 - 1500 (714) 645 - 6800 [FAX]
They have this glass (glass #3750) in large stock pieces, so it must be cut down to the size of a micro slide. The will grind it down to whatever thickness you desire, as well. We ordered two micro slide sized peices in July of 1993. The cost was $86.
Good luck!
Cheers, Bill Bug
************************* * Bill Bug * * Dept. of Biology * * Swarthmore College * *************************} O.k., all you good microscopists, I'm looking for a source for either a Uranium Glass } Block or a suitable replacement for visualizing/teaching the light path on } a light } microscope. The block is placed in the microscope stage and the } illumination path way } can be viewed inside the block - particularly as the condensor is focused } and as either } the stage aperature or field aperature are varied. } } I know this can be visualize with a dilute milk solution but I } really would prefer } something a little more stable (and non-perishable). } } Thanks! } } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax) --============_-1295708639==_ma============ Content-Type: text/enriched; charset="us-ascii"
Richard: Here is a summary I had saved of a previous uranyl glass thread. Hope it helps. Tom
--------------------------------------
} From: Not Specified { {Kris_Kavanau-at-dmcmail.ucsf.edu} Attn: George M
Return-Path: Kris_Kavanau-at-dmcmail.ucsf.edu To: Microscopy-at-aaem.amc.anl.gov (M)
Organization: University of California, SF Division of Molecular Cytometry Date: Fri, 03 Feb 1995 10:03:40 PST
OFFICE MEMO Uranyl Glass/FM Stds. Date: 2/3/95
Dear Microscopists,
Does anyone have any uranyl glass, or know where it might be obtained? I have been told that it is no longer manufactured commercially. It might be an excellent "generic" fluorescence microscopy control. Are there any commercially available, pre-mounted fluorescence standards besides "MultiSpeck" from Molecular Probes? They are very convenient, but they are not ideal for our applications as DAPI, fluorescein, and Texas Red specific controls. Unfortunately, Flow Cytometry Standards Co. no longer makes pre-mounted standards.
I have been managing the UCSF core flow and image cytometry facility ("Lab for Cell Analysis") for 2 years, but I had no real QC for our 2 occasionally used fluorescence microscopes. Now I need to establish QC protocols for 6 additional multi-user, computerized fluorescence (one scanning confocal) microscopes in the "National Molecular Cytogenetics Resource." I was surprised that so few standards (and journal references) seem to be available.
Any suggestions or comments would be greatly appreciated. Thank you very much. Kris Kavanau; kavanau-at-dmc.ucsf.edu
To: Microscopy-at-AAEM.amc.anl.gov
} From: George M { {George_M-at-image1.com}
Reply to: RE} Uranyl Glass/FM Stds.
Hi Kris,
Uranium glass slides can be purchased from:
Newport Industrial Glass, Inc.
1631 Monrovia Ave.
Costa Mesa, CA 92627
Tel: 714-642-9980
Fax: 714-645-6800
Contact person: Bill Larsen (you can tell him I sent you). Sold as a 6.5x6.5" sheet (you can specify 1 mm height), so you may want to form a "consortium" to have Newport pre-cut a sheet to slide size (nominal extra cost, but your lab only needs one or two slides). If there is a lot of interest, my company may start selling single slides.
As for references and the Shading Correction equation: please see my article in the 11/94 issue of Journal of NIH Research 6(11): 80 (usual Internet disclaimer: yes, that is an ad from my company on the facing page). Also look at Jericevic et al (1989) Methods in Cell Biology 30:47-83.
MutliSpeck beads: The Molecular Probes pre-mounted slide kits should be ideal for DAPI and Fluorescein. I believe they were optimized for Rhodamine, but should still work ok for Texas Red. If your problem is with mounting, Mol. Probes now sells the beads in solution, so you can 'sprinkle' some on your specimens. If you have a different problem with the current MultiSpeck's, Mol. Probes may be able to work something out for you.
Sorry, but I usually buy my reference material from Mol. Probes and don't keep close track of other slide manufacturers.
} I read through the archives for information on uranyl glass test slides, but couldn't figure out if there is a definitive answer to the question, Does anyone still manufacture uranyl glass, and where could I get some?
} Thanks,
} Chi-Bin Chien
} chien-at-jeeves.ucsd.edu
Chi-Bin Chien,
You can obtain micro slide sized pieces of uranyl glass from:
Newport Industrial Glass, Inc.
1631 Monrovia Avenue
Costa Mesa, CA 92627
(714) 645 - 1500
(714) 645 - 6800 [FAX]
They have this glass (glass #3750) in large stock pieces, so it must be cut down to the size of a micro slide. The will grind it down to whatever thickness you
desire, as well. We ordered two micro slide sized peices in July of 1993. The cost was $86.
Good luck!
Cheers,
Bill Bug
*************************
* Bill Bug *
* Dept. of Biology *
* Swarthmore College *
*************************} O.k., all you good microscopists, I'm looking for a source for either a Uranium Glass
} Block or a suitable replacement for visualizing/teaching the light path on a light
} microscope. The block is placed in the microscope stage and the illumination path way
} can be viewed inside the block - particularly as the condensor is focused and as either
} the stage aperature or field aperature are varied.
}
} I know this can be visualize with a dilute milk solution but I really would prefer
} something a little more stable (and non-perishable).
by zahn.acpub.duke.edu (8.8.5/Duke-4.7.0) with ESMTP id KAA20344; Fri, 15 Jan 1999 10:36:52 -0500 (EST) Received: (from saram-at-localhost) by godzilla4.acpub.duke.edu (8.8.5/Duke-4.7.0) id KAA15860; Fri, 15 Jan 1999 10:36:49 -0500 (EST)
On Thu, 14 Jan 1999, Margaret Casey wrote:
} Date: Thu, 14 Jan 1999 16:02:26 -0500 } From: Margaret Casey {CaseyM-at-state.mi.us} } To: microscopy-at-sparc5.microscopy.com } Subject: TEM:virus isolation from foods for negative staining } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Fellow Listers, } } Does anyone have a good protocol or reference for virus isolation from food other than shellfish? We are trying to isolate and concentrate Norwalk virus from some foods implicated in a poisoning outbreak in Detroit. The foods in question are salami, garbanzo beans and some kind of cheese (I think provolone). We need to concentrate to at least 1 million particles/ml in order to use our prep for negative staining and TEM observation. Any suggestions would be greatly appreciated! } } Sincerely, } Peggy Casey } Michigan Dept. of Community Health } Lansing, Mich. } phone: 517-335-8102 } e-mail: caseym-at-state.mi.us } The best way to detect these kinds of viruses in dilute concentrations is PCR. In this case, viral numbers are likely to be very low, and getting enough to see by negative staining, unless you aggregate them with antiserum, may be difficult. If you want to proceed anyway, see concentration methods in Hayat and Miller, Negative Staining, McGraw-Hill.
Under separate cover, I will send you the name of an expert who does PCR on Norwalk. She is on the faculty at UNC and the CDC.
SM
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
How fast can you work and how long will the sections stay frozen? We l= ooked at frozen samples on a light microscope by covering the scope in a glov= e bag (~$15 from scientific supply houses) and doing all our work inside the = bag. We started by flooding the bag with dry nitrogen for a couple of hours = to drive out all humidity. We were able to freeze our samples in the bag,=
transfer them to the stage and back to the LN2 when we thought they wer= e starting to melt. You might be able to transfer the frozen sections in= to the bag and keep them frozen with some LN2 and quickly transfer them to the= stage of the microscope. This is a very kludgy setup, but it worked for us a= nd was dirt cheap.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-3537 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com =
since a couple of weeks I have to face a problem with our LEO EM912 which I could not solve up to now: I can't correct for the astigmatism in diffraction mode any more. If I use the "Image Stig xy" buttons they influence the appearence but the possible range of this potentiometers is not large enough for the correction. In addition the zero-beam moves very slowly across the screen (approx. 0.5mm/min on the final screen). I did a thorough alignment of all the settings of the microscope and the problem persists. We had service here and were told that it is probably some dirt particle in a part of the column which we can't clean. The only possibility would be to try to "burn" the particle in low-mag mode with high illumination aperture and "playing" with the illumination tilt in diffraction mode or the illumination shift in spot mode. And in fact, this helps for a short time, but the problem reappears latest after half an our again.
We have no problems with astigmatism in the image mode.
Does anybody out there has an idea what we could do, check, exchange ... to help solving the problem?
Greetings,
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu Visit our WWW site! http://www.crpcu.lu/~wahlbrin
Lou - } } A few years ago I got hold of a beta version of a SEM Macintosh simulation } program, SuperSEM. It was a Macintosh program that was developed with } Supercard software. It was misplaced during the process of upgrading my } computers. Does anyone know if it exists?
Might you be thinking of Brian Griffen's "Virtual SEM"? It's a great simulation, and you'll find it listed in the CD-ROM section of the bibliography on Project MICRO's web page (URL below).
Caroline }
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Reply to: RE: Ti Alloys - TEM foil preparation Re: Electropolishing of Ti alloys Due to interest in this subject, I will list some additional = electrolytes I have used on Ti alloys. (3mm, jet polishing).
Ti-8 w/% Al 13% HCL -60 degrees C Ti-811 87% methanol 70 to 90 volts Ti-6Al-4V 25-35 mA.
Ti-13 Sn 130 ml HCL -50 degrees C 670 ml methanol 150 volts 100 ml butyl 60 mA.
Ti-3V 60 ml perchloric acid -50 degrees C Ti-20 Zr 590 ml methanol 50 volts Ti-14 Al 350 ml butyl cellosolve 10-15 mA
Ti-8 Al 60 ml perchloric acid -15 t0 -20 C (rolled) 460 ml ethyl alcohol, 95% 180 volts 280 ml butyl alcohol 40 mA. 100 ml butyl cellosolve Ti 5.3 g. = lithium chloride -40 degrees C. nanocrystals, 11.16 g. magnesium 150 volts compacted chloride 30 mA 500 ml. methanol 100 ml. butyl cellosolve (Note: the two salts above must be added to the combined solvents one = "powder" at a time, while stirring. The two dry salts react violently if = mixed together. This electrolyte has nearly as wide an application as = perchloric acid mixtures and is known as B K-2). I suspect that B K-2 will cause the least hydride problem. It was = developed to eliminate hydrides in vanadium TEM foils. The voltages given = would need to be reduced about 20% for a twin jet system due to lower = electrical resistance of that configuration. My work was done on a South = Bay vertical single jet unit. I have no vested interest in the above equipment, but have enjoyed = the ease with which good polishing conditions can be obtained and = reproduced since 1975 or so. Good luck! Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439
E-mail: {bkestel-at-anl.gov} Thomas, Larry wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } = To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Satyarth: } } An excellent paper on the subject is: } } "In Situ Hydride Formation in Zirconium and Titanium during Ion Milling" } Graham J.C. Carpenter et al, JMSA Vol.1 No. 4 pp 175-184 1995. } } From the paper you will see that a key factor in eliminating hydride } formation is having a clean sample free from hydrocarbon contamination. } = This contamination could be remnants from the dimpling process or other } pre-thinning steps or it could be caused by the back streaming of } diffusion } pump oil in your ion mill. As titanium has a high chemical affinity for } hydrogen, you may want to look over your preparation steps and try to } eliminate any areas of possible contamination. If you are still having } a } problem, a quick cleaning of both the specimen and the specimen holder } in a } Plasma Cleaner should take care of it. } } If you have an interest, I can send you a copy of the above referenced } paper. I also have papers on plasma cleaning which may be of interest. } } NOTE: South Bay Technology does manufacture the PC150 Plasma Cleaner and } therefore I have a vested interest in promoting its use. } } Best regards- } } David } Writing at 9:38:23 AM on 1/12/99 } } } ******************************= *********************************** } ********** } ************************ } } David Henriks TEL: } 800-= 728-2233 (toll free in the USA) } South Bay Technology, Inc. +1-949-492-2600 } 1120 Via Callejon FAX: } +1-949-492-1499 } San Clemente, CA 92673 USA e-mail: } henriks-at-southbaytech.com } } } ***************************************************************** } ********** } ************************ } } } } } } } Please visit us at http://www.southbaytech.com { { { { { } } Manufacturers of precision sample preparation equipment and supplies for } metallography, crystallography and electron microscopy. } } Message text written by Satyarth Suri } } } Hello: } I am currently working on mechanical behavior / microstructure } correlations in single colony near apha Ti Alloys. I am currently } preparing the foils using a dual ion mill (6kV, 12deg, 1mA), but } have run into the problem of a very uneven alpha phase morphology, } the alpha phase has island formation - we are using a cold stage } to minimize the hydride formation at the interface. I am using } 3 micron/6micron diamond paste during the dimpling process - the } foil itself otherwise is fairly clean in terms of the dislocation } content. Could anyone in the microscopy land have some suggestions } in terms of what the problem may be? } } thanks - if you want you can respond directly to suri.3-at-osu.edu } } -satyarth } } { } } } } } RFC822 header } ----------------------------------- } } Received: from dns2.anl.gov (dns2.anl.gov [146.139.254.3]) by } horus.et.= anl.gov (8.6.11/8.6.11) with ESMTP id SAA02218; Tue, 12 Jan 1999 18:17:43 -= 0600 } Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com } [206.69.208.= 10]) by dns2.anl.gov (8.9.1a/8.6.11) with SMTP id SAA19261; Tue, 12 Jan = 1999 } 18:18:22 -0600 (CST) } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.= 11) } id RAA22399 for dist-Microscopy; Tue, 12 Jan 1999 17:41:01 -0600 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by } Sparc5.= Microscopy.Com (8.6.11/8.6.11) with SMTP id RAA22384 for } "= MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 12 Jan 1999 17:40:30 -= 0600 } Received: from Sparc5AAEM (aaem.amc.anl.gov [146.139.72.3]) by } Sparc5.= Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA22377 for } {microscopy-at-= sparc5.microscopy.com} ; Tue, 12 Jan 1999 17:40:17 -0600 } Received: from pnl.gov (relay.pnl.gov [130.20.128.34]) by Sparc5AAEM } (8.= 6.11/8.6.11) with ESMTP id RAA14848 for {MICROSCOPY-at-aaem.amc.anl.gov} ; Tue,= 12 Jan 1999 } 17:52:22 -0600 } Received: from pnlmse1.pnl.gov by pnl.gov (PMDF V5.1-12 #28154) } with ESMTP id {01J6GJ2FP8R4970ZC9-at-pnl.gov} for MICROSCOPY-at-aaem.amc.anl.= gov; } Tue, 12 Jan 1999 15:54:41 PST } Received: by PNLMSE1.pnl.gov with Internet Mail Service (5.5.2232.9) } id {CR51M2B5} ; Tue, 12 Jan 1999 15:54:42 -0800 } Content-return: allowed } Date: Tue, 12 Jan 1999 15:54:36 -0800 } From: "Thomas, Larry" {Larry.Thomas-at-pnl.gov} } Subject: RE: Ti Alloys - TEM foil preparation } To: Satyarth Suri {suri-at-mse.eng.ohio-state.edu} , } Microscopy ListServer {MICROSCOPY-at-aaem.amc.anl.gov} , } "'South Bay Technology'" {Henriks-at-CompuServe.COM} } Message-id: {7A04D1887189D2118EEB00A024BF29DA41CB17-at-pnlmse2.pnl.gov} } X-Mailer: Internet Mail Service (5.5.2232.9) } Errors-to: Microscopy-request-at-sparc5.microscopy.com } Content-Type: text } Content-Length: 5172 } Status: }
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by pilsener.ucs.ualberta.ca (8.9.1a/8.9.1) with ESMTP id MAA10247; Fri, 15 Jan 1999 12:46:33 -0700 (MST) Received: from localhost (mingchen-at-localhost) by gpu2.srv.ualberta.ca (8.8.5/8.8.5) with SMTP id MAA67494; Fri, 15 Jan 1999 12:46:33 -0700
Hi Mary,
I have done a lot of fixation for plant material for more than 25 yrs. I think 2% paraformaldehyde-2.5% glutaraldehyde fixative in 0.2 M cacodylate or phosphate buffer (pH 7.2) is the best.
Good luck.
Ming
On Tue, 12 Jan 1999, Mary Gail Engle wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I need some advice as to the best protocol for fixing fresh plant leaves } for routine TEM. My only experience has been with animal tissue. } Thanks, } MG Engle } } }
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * #1074B Dentistry Pharmacy Building * * University Of Alberta. * * Edmonton, Alberta, Canada T6G 2N8 * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Lou Ross wrote: } } } } A colleage from an industrial SEM lab contacted me about a problem with } contamination on the final aperture on an variable pressure SEM. This SEM } was recently purchased by one of their branch plants to examine } semiconductor products. Although it is a variable pressure SEM, they have } only operated it in the high vac mode like a standard SEM. } } The problem they are experiencing is that the final aperture presumably } gets so dirty they have to change the aperture every two weeks. } Unfortunately the manufacturer has not been able to shed any light on this } situation. } } If the vacuum system is alright, I suggested that the problem might be with } the samples. Supposedly the branch lab operators are following the same } sample prep protocol established in the local SEM lab. } } Any other ideas out there? } } Thanks in advance, } Lou Ross } Senior Electron Microscope Specialist } Room 101 } Department of Geological Sciences } University of Missouri } Columbia, MO 65211 } (573) 882-4777 } (573) 882=5458 fax } www.missouri.edu/~geosclmr/ebaf.html
Dear Lou Ross,
Based on your message concerning the contaminated aperture, my feeling is that the problem probably rests somewhere other than the aperture itself. But since there is the slightest possibility that there might be a problem with the aperture perhaps we can assist you. Ladd produces the vast majority of apertures/microholes used in the United States and the aperture you have may very will be a Ladd aperture. To make sure there is no residual contamination on an aperture we have a post production protocol that is designed to eliminate any contamination that results during production. In fact we have some apertures/microholes used in fluid and gas control that are required to be absolutely contamination free. We have a protocol to ensure that. It is also important that an aperture be shipped or stored in a glass or non-contaminating vial and that the aperture surface should not touch tissue or even lint free cloth. We would be glad to examine the aperture to see if the contamination was a result of packaging, handling or the production process. Please contact us if you if you wish us to do that.
John Arnott Chairman
-- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
My institution needs to purchase a new cryostat in the next several months. I would appreciate any comments from cryostat users about their satisfation/ dissatisfaction with particular makes and models of relatively new cryostats.
We do not anticipate particularly heavy usage of this equipment, but the tissues that will need to be sectioned range from soft tissues such as brain and heart all the way to mouse and rat bone. Some investigators would like to collect sections in excess of 30 microns for confocal microscopy projects, and others would like to serial section whole organs from transgenic mice. Are disposable knives sufficient for this range of usage?
Also, cryostat vendors are welcome to email me directly. Surprisingly, it has been like pulling teeth to get some companies to talk to a ready and willing customer.
Thanks for your help,
Dr. Steve Fields Oklahoma Medical Research Foundation 825 N.E. 13th Street Oklahoma City, OK 73104
Good Morning, Lou! There are several reasons why the final aperture may be getting dirty on a variable pressure SEM, and why this may be more noticeable on semiconductor samples. Several causes contribute to contamination within SEM chambers. If this system has been used for any other analysis other than semiconductor, at low vac modes, residues from prior analysis may be lining the chamber walls - these are difficult to remove even by baking the chamber out. Additionally, this system probably has oil-cooled pumps - the deleterious effect of backstreaming oils into chamber which "gums" onto the final aperture is well established, but can be diminished with a strategically located dry N2 purge port in the sample exchange chamber. Mounting media is also of concern - since most conductive paints outgas, these vapors can also contaminate the F.A. quickly, especially if the material is not exceedingly well dried. Simple cleanliness may also be the culprit - by handling a metal sample fixture with bare hands, oils and sweats are deposited on the fixture. These deposits love to attach to the F.A. (and cold tip of EDS detectors too!) As most semiconductor analysis requires fairly high magnification, and often lower KeV, these problems are exacerbated. I've delt with these and many other problems concerning the optimal performance of SEM's with semiconductor materials, and would be pleased to look over his system for performance improvements. Let me know if I may be of assistance!
In a message dated 1/14/99 8:41:46 PM Mid-Atlantic Standard Time, RossLM-at-missouri.edu writes:
{ { A colleage from an industrial SEM lab contacted me about a problem with contamination on the final aperture on an variable pressure SEM. This SEM was recently purchased by one of their branch plants to examine semiconductor products. Although it is a variable pressure SEM, they have only operated it in the high vac mode like a standard SEM.
The problem they are experiencing is that the final aperture presumably gets so dirty they have to change the aperture every two weeks. Unfortunately the manufacturer has not been able to shed any light on this situation.
If the vacuum system is alright, I suggested that the problem might be with the samples. Supposedly the branch lab operators are following the same sample prep protocol established in the local SEM lab.
Any other ideas out there?
Thanks in advance, Lou Ross Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html } }
I am asking for help. We bought a JSM 5410 SEM (3.5 nm at 30kv 8mm working distance) and JEOL 2010 microscopes (without FEG) here. Both microscopes has almost finished installation. For JEOL 2010 I checked our quotation written by JEOL that they should provide working pressure of 6x10-6 Pa or better at specimen as read by the ion pump. But now we got best vacuum is 1.5x10-5 Pa. Is there anyone who has JEOL 2010 TEM and can tell me your best working value ASAP? Thank you very much in advance.
As for JSM 5410, is there anybody who has JSM 5000 series SEM and can send me your resoultion shooting photos by JEOL engineer (resolution 3.5nm) to let me have a look. I will pay the Fedex fee and send you back by Fedex express. Thank you very much.
Sincerely yours,
Weilie Zhou (Ph.D)
************************************************ Advanced Materials Research Institute University of New Orleans Science Building 2021 New Orleans, LA 70148 Tel:(504) 280-5570 Fax:(504) 280-3185 ************************************************
I have given the following advice to several users of variable pressure SEMs who have dirty microscopes and contamination, and they all said it cured the problem.
Don't leave it in high Vacuum mode all the time. Some variable pressure SEMs are very dirty due to backstreaming at high vacuum. They are designed for low vacuum not high vacuum. By placing the system into low vacuum mode, the chamber is placed into viscous flow vacuum dynamics which stops backstreaming and purges out contaminants. Try leaving it in low vacuum mode overnight for a month and check the results.
Let me know if this helps.
Ronald Vane XEI Scientific
Disclaimer: XEI is in the anti-contamination business. } } -----Original Message----- } From: Lou Ross {RossLM-at-missouri.edu} } To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com} } Date: Thursday, January 14, 1999 1:48 PM } ----------------------------------------------------------------------. } } } } } } A colleage from an industrial SEM lab contacted me about a problem with } } contamination on the final aperture on an variable pressure SEM. This SEM } } was recently purchased by one of their branch plants to examine } } semiconductor products. Although it is a variable pressure SEM, they have } } only operated it in the high vac mode like a standard SEM. } } } } The problem they are experiencing is that the final aperture presumably } } gets so dirty they have to change the aperture every two weeks. } } Unfortunately the manufacturer has not been able to shed any light on this } } situation. } } } } If the vacuum system is alright, I suggested that the problem might be with } } the samples. Supposedly the branch lab operators are following the same } } sample prep protocol established in the local SEM lab. } } } } Any other ideas out there? } } } } Thanks in advance, } } Lou Ross } } Senior Electron Microscope Specialist } } Room 101 } } Department of Geological Sciences } } University of Missouri } } Columbia, MO 65211 } } (573) 882-4777 } } (573) 882=5458 fax } } www.missouri.edu/~geosclmr/ebaf.html } } } } }
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Happy New Year all,
A question for the New Year: what are EM laboratories using for pure water = these days? Is it possible to use deionized water, or even the distilled = water from Rite Aid or Arrowhead? In the past I had a still which was fed = from a deionized water supply. We had no problems at all with this.
If the preferred purification process turns out to be deionized water, = which quality is best - Type 1 (HPLC etc.) or will lower quality suffice? = Also has anyone had problems with resin in the water?
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/apw.htm
microscopy-at-Sparc5.Microscopy.Com, tina-at-pbrc.hawaii.edu MMDF-Warning: Parse error in original version of preceding line at rfdata.net
Another quick cheap fix is dress warm and do the work in a walk in freezer. The glove bag would work well to keep your breath from freezing on the scope.
When I was working at Ag Engineering we had and etymology and vet student that spent the summer in a walk in cooler monitor tick behavior from freezing to 55 degrees F.
Gordon
Gordon Couger gcouger-at-couger.com Owner PRAG-L PRactical AGriculture List www.couger.com/prag-l Stillwater, OK 405 624-2855 GMT -6:00
How fast can you work and how long will the sections stay frozen? We looked at frozen samples on a light microscope by covering the scope in a glove bag (~$15 from scientific supply houses) and doing all our work inside the bag. We started by flooding the bag with dry nitrogen for a couple of hours to drive out all humidity. We were able to freeze our samples in the bag, transfer them to the stage and back to the LN2 when we thought they were starting to melt. You might be able to transfer the frozen sections into the bag and keep them frozen with some LN2 and quickly transfer them to the stage of the microscope. This is a very kludgy setup, but it worked for us and was dirt cheap.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-3537 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com
} From Microscopy-request-at-sparc5.microscopy.com Thu Jan 14 14:44:05 1999 } } Date: Thu, 14 Jan 1999 09:11:29 -1000 (HST) } From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } To: Microscopy Listserver {Microscopy-at-Sparc5.Microscopy.Com} } Subject: Cold stage for LM } } Happy New Year to you all } } .......... I have liquid nitrogen, styrofoam, and a machine } shop. I also have high humidity. Can anyone tell me how to kludge } together a cold enough stage that we can get images of these } frozen sections while preventing frost? I have some general ideas, but } I'd appreciate any specific plans or ideas.............. } } } Aloha, } Tina } }
Tina, We have done some work with a cold stage for cathodoluminescence (CL) studies of minerals, ceramics, etc. A simple vacuum chamber (mechanical pump - pressures of 30 to 100 millitorr) is used for the CL work and the chamber sits on the stage of the light microscope. For cold stage work we use a simple system consisting of a chillable plate that is cooled in liquid nitrogen outside the chamber and then inserted and pumped down. Temperatures low enough to dramatically change the CL behaviour are achieved for about 10 - 12 minutes and there is minimum condensation. The vacuum helps to provide thermal insulation for the chillable plate and also minimizes the amount of water vapor that can condense. The plate itself is designed to minimize thermal transfer to the other parts of the vacuum chamber.
Don Marshall
(Claimer: RELION is in the business of CL instrumentation for light microscopes and chillable sample plates are one of the items we offer.)
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
A three month (June, July, and August) position is open for a microscopy oriented technician at the Marine Biological Laboratory, Woods Hole, MA. We would like to attract someone with some knowledge of biological preparative techniques and experience in laser scanning confocal microscopy, TEM, SEM, and/or LM.
The technician will assist in the Central Microscopy Facility. The technician's duties will be to check out incoming investigators in the usage of our equipment and then to supervise its continuing usage and to perform contract work for investigators. This may include fixation, embedding, sectioning, scope use, darkroom work, etc. The technician will also provide routine maintenance.
This is a short term and scientifically rewarding position. Salary will be in the $7 to $10/hour range. Housing may be available to rent through MBL.
For more information, including a more detailed position description, please contact Louis Kerr at: MBL, 7 MBL Street, Woods Hole, MA 02543. Telephone, 508-289-7273; or at Email: lkerr-at-mbl.edu. Please apply to: Human Resources, MBL, 7 MBL Street, Woods Hole, MA 02543. or resume-at-mbl.edu.
An Equal Opportunity/Affirmative Action Employer/ Non-smoking workplace.
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX)
} SUMMER MICROSCOPY TECHNICIAN POSITION AVAILABLE } } A three month (June, July, and August) position is open for a microscopy } oriented technician at the Marine Biological Laboratory, Woods Hole, MA....
} ...This is a short term and scientifically rewarding position. Salary will be } in the $7 to $10/hour range. Housing may be available to rent through MBL... } } ...For more information, including a more detailed position description, } please contact Louis Kerr at: MBL, 7 MBL Street, Woods Hole, MA 02543. } Telephone, 508-289-7273; or at Email: lkerr-at-mbl.edu. } Please apply to: Human Resources, MBL, 7 MBL Street, } Woods Hole, MA 02543. or resume-at-mbl.edu.
I once had a senior student (at UC Berkeley) who took this job. He was recruited into a top MD/PhD program and met his wife, all in a summer of hard work at MBL. Apply, folks!
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Perhaps a little off-topic, but for those of you who archive microscopy images to recordable CD-ROM, does anyone know of a strategy for placing a "software lock" on the CD-ROM which would require a "key" to access, view, or copy the CD-ROM contents? I'd rather not compress or encrypt images and use a password to access but to have a security option initiated upon mounting of the CD-ROM.
Thanks very much for any info,
Brian C. Tryon MCP-Hahnemann School of Medicine Ann Preston Hall, Box 551 3300 Henry Avenue Philadelphia, PA 19129 USA
----------------------------------------- "Quantifying is a committing task." - Cruz-Orive, 1994.
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." - Richard Feynman --------------------------------------------------------------
I am a school counselor at the elementary level. Our leadership team is spending some of our grant money on microscopes for a Science lab which we are developing. I would like to know basic information on purchasing microscopes for the elementary student. We hope to purchase five-six scopes depending on the price. Can you help with this information? Please e-mail me at diorado-at-stc.net Thanks! Diane
Tina: I am afraid I will not provide detailed plans, just a couple of hints. I expect that the easiest way would be to built a suitable, insulated enclosure with a minimal opening for the objective lens. On top of that small opening a resistance heater loop may be required. Somewhere you would have an inlet for dry industrial nitrogen gas and with a small sealed chamber and the only opening near the lens, no air and therefore no moisture can enter. Gas flow could be quite low - if the outlet hole is not too large. Gasflow would need to be activated well before freezing. Temperature control and nitrogen flow are other problems, but avoiding frost tends to be the greatest challenge. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
Hi all
Here is a 'WISH' list of bits that we want for our labs, if anyone has these bits that they want to giveaway, trade, sell, we look forward to hearing from you.
Spares for a Gatan model 600 ion beam thinner
Spares for JEOL 35-CF Spares for JEOL 100-CX
EDXS for a JEOL 35-CF with with electronics. It doesn't have to have excellent energy resolution. EELS for a JEOL 2010 or JEOL 100-CX Four quadrant Back Scatter detector (UHV Compatible)
An oven Any specimen preparation gear for physical and biological specimens Microtome and accessories
Enlargers working or broken Timers for enlargers Print dryer
Printers - B&W and color
Dessicators (vacuum)
Chemical storage cupboard
I know some of these are extravagant, but I have to ask. The Gods just might smile upon me today.
Thank you George G. Theodossiou Dept Applied Physics RMIT Email: George-at-bunyip.ph.rmit.edu.au ph:+61 3 9925 3394 fax:+61 3 9925 5290
Hi all Best wishes for the new year to all. We have a small problem that needs fixing and would like to get a few = other ideas on the matter before the user spends money on fixing it. The user has an ISI SEM which has the typical back streaming problems. = This oil is contaminating the ED detector window and therefore causing = absorption of the light x-rays. Other than cleaning the window regularly, which carries the danger of = blowing the window and detector, we were looking at methods to stop this = contamination. We are fitting a foreline trap to the Rotary pump, but = experience shows that you still get oil back streaming. We could try a = Ln2 cold finger but this is a hassle to keep toped up and can only be = fitted above the diff pump which makes engineering a problem. A SEM clean gas leak system would also help but they are very short on = cash. ( surprise surprise) They also need to work at high mag and so = whilst the SEM is in use the SEM clean system could not be effective.=20 The only other idea we have is to fit a peltier cold finger to the SEM = chamber in the hope of attracting most of the oil to the cold finger = and thus not contaminating the detector. This we though could be = controlled to have a fairly low temp whilst the SEM is in use and = frequent sample changes are required, then turn up the power for when = the SEM is not in use to be more efficient.
Has any one fitted a peltier to their EM before ? What experiences have = you had with them and what size and power would work.=20 All ideas welcome.=20 Thanks =20 Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
Hello fellow listservers, I've recently discovered a problem with a batch of a particular type of cold setting epoxy resin. I have been using this batch of resin on an infrequent basis for about 2 years (and is probably about 4 years old). When I analysed some of the cured resin by microanalysis last week appreciable bromine (Br) was detected as well as the usual chlorine content (Br} Cl). The 'introduction' of the Br is a recent thing because I also analysed some resin (from the same batch) prepared approximately one year ago. The analysis of this showed no Br with Cl the only detectable element present (with the Be window of the EDS detector in place that is). Is it possible therefore for the resin to in some way deteriorate with time and for Br compounds to form? If so, what are the chemical reactions going on here. I'm very certain that the Br is not a contaminant. Also, I should point out that if this is a chemical degradation of the resin then the physical characteristics of the cured resin are normal. I would appreciate any thoughts on the matter. Regards
Martin Roe Macaulay Land Use Research Institute Aberdeen AB15 8QH Scotland United Kingdom
Greetings All, } } I have been reading this list for well over a year and am confident that this is the place to come if one has a question regarding microscopy...so here goes. } } The Chemical Hygiene Officer of the company I work for has decided that all laboratory personnel must wear full eye protection when in a lab. } This presents a problem to me, because of my near sightedness, I prefer to look through the optical/IR microscope without glasses and this results in a side splash safety hazard. Does anyone have any suggestions on how to fully protect myself from side impact and satisfy the safety requirements and view my microscopy images? Thank you all in advance and if anyone wants a summary of responses, I will be more than happy to oblige. } } } } } Deborah Hills-Haney } Research Analytical Services/NMR Lab } International Flavors and Fragrances R&D } 1515 US Highway 36 } Union Beach, NJ 07735 } } Phone: (732) 335-2663 } Fax: (732) 335-2591
______________________________________________________ Get Your Private, Free Email at http://www.hotmail.com
} Hello fellow listservers, } I've recently discovered a problem with a batch of a particular } type of cold setting epoxy resin. I have been using this batch of } resin on an infrequent basis for about 2 years (and is probably } about 4 years old). When I analysed some of the cured resin by } microanalysis last week appreciable bromine (Br) was detected as well } as the usual chlorine content (Br} Cl). The 'introduction' of the Br } is a recent thing because I also analysed some resin (from the same } batch) prepared approximately one year ago. The analysis of this } showed no Br with Cl the only detectable element present (with the Be } window of the EDS detector in place that is). Is it possible } therefore for the resin to in some way deteriorate with time and for } Br compounds to form? If so, what are the chemical reactions going on } here. I'm very certain that the Br is not a contaminant. Also, I } should point out that if this is a chemical degradation of the resin } then the physical characteristics of the cured resin appear to be normal. I } would appreciate any thoughts on the matter. Regards } } Martin Roe } Macaulay Land Use Research Institute } Aberdeen } AB15 8QH } Scotland } United Kingdom } } }
} I am a school counselor at the elementary level. Our leadership team is } spending some of our grant money on microscopes for a Science lab which } we are developing. I would like to know basic information on purchasing } microscopes for the elementary student. We hope to purchase five-six } scopes depending on the price. Can you help with this information? } Please e-mail me at diorado-at-stc.net Thanks! Diane
I'm happy to help; that's what Project MICRO is all about. You'll find detailed general purchase and evaluation advice on the MICRO web page (URL below), plus a list of possible sources. For elementary science, I strongly favor monocular "dissecting" scopes; you can get good ones for under $80. Plus perhaps one conventional 3-objective (4, 10, 40x) compound scope, for $120. Where are you located? I may be able to get you some advice from a local MSA member.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Martin J. Roe wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello fellow listservers, } I've recently discovered a problem with a batch of a particular } type of cold setting epoxy resin. I have been using this batch of } resin on an infrequent basis for about 2 years (and is probably } about 4 years old). When I analysed some of the cured resin by } microanalysis last week appreciable bromine (Br) was detected as well } as the usual chlorine content (Br} Cl). The 'introduction' of the Br } is a recent thing because I also analysed some resin (from the same } batch) prepared approximately one year ago. The analysis of this } showed no Br with Cl the only detectable element present (with the Be } window of the EDS detector in place that is). Is it possible } therefore for the resin to in some way deteriorate with time and for } Br compounds to form? If so, what are the chemical reactions going on } here. I'm very certain that the Br is not a contaminant. Also, I } should point out that if this is a chemical degradation of the resin } then the physical characteristics of the cured resin are normal. I } would appreciate any thoughts on the matter. Regards } } Martin Roe } Macaulay Land Use Research Institute } Aberdeen } AB15 8QH } Scotland } United Kingdom
Martin, One possibility that comes to mind is the storage cabinet of the resin: if you store bromine in the same cabinet and the resin is packed in a polymer flask, I can think of some diffusion out of the bromine into the resin bottle.
An other possibiliy is that there is a solvent used with a part of the resin, which contains Br. If the resin is growing older, there may be decomposition/degradation of the solvent, leaving a non volatile Br conaining component behind. Check each single component of the resin.
Hope this helps -- Gerhard Frank UNIVERSITAET ERLANGEN-NUERNBERG Phone: +49 9131 85 28606 Institut fuer Werkstoffwissenschaften VII Fax: +49 9131 85 28602 Cauerstr. 6, D-91058 Erlangen, Germany frank-at-ww.uni.erlangen.de
I ordinarily wear bifocals with an astigmatism correction. My company has bought me distance-only safety glasses, which I use irregularly but satisfactorily in place of my bifocals at the LM. The main advantage is the lack of the bifocal line.
If you can't bear the glasses, you might propose writing a "job hazard analysis" in which you analyze the hazards involved in your specific operation and propose specific solutions other than safety glasses, e.g., a splash shield between you and other workers, use of safety glasses for certain operations but not for viewing, after you have assured that risk of splash etc. during viewing is minimal.
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
Our laboratory is in the process of 'remodelling' and have found several items which we would be able to part with at this time. These items include: 1. A Polaron Critical Point Dryer 2. An Arkay CD-80 cabinet dryer 3. A Gatan model 673 mark 2 wide angle TV system with a Data Translation 3851 A/D converter board 4. A Gatan model 622 fiber optically coupled TV system
This equipment was 100% functional the last time it was used. If you are interested in any of this equipment please contact me off line.
Jon Charlesworth, Coordinator Electron Microscopy Core Facility Mayo Clinic 1426 Guggenheim Building Rochester, MN 55905 ph: (507) 284-3148 fax: (507) 284-9349 email: charlesworth.jon-at-mayo.edu
Is anyone familliar with specific fixation- and/or staining techniques for rat and/or chicken eyes. Someone on our laboratory wants to to do some research on eyes. He wants to stain specific membranes like Bowmann and Desmett membrane.
Hi, I am looking for a program to convert weight % to Atomic % and vise versa preferably on a Power Mac.. An old program we have works on pre '90 Mac's but not on the newer ones. Anyone have any ideas? Thanks- Mike
I am looking for a 8cm by 10cm (TEM film format) film holder for a Leafscan 45 negative scanner.
I've been told that the company which makes the Leafscan was bought out. Does anyone know by whom, or who might now deal in parts?
Thanks, Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
Hope that there is room on this list for a bit of pre-millenial philosophy.
Before I saw John White's prototype laser scanner in 1986, I had spent my time trying to improve instrumentation for high-resolution SEM and TEM. Though capable of "seeing molecules" and even labelling them, these instruments and have one tremendous disadvantage for the biologist: because they form their images using quanta that carry (much) more than 5 electron volts of energy, the act of imaging destroys molecules. Consequently, these instruments can only be used to image dead cells.
So, even when the confocal microscope was seen primarily as a method of making 3D images of biological structures that had been fixed and stained, my own focus was on using it to image living specimens.
Early confocal microscopes were so inefficient in their use of the light from the specimen that one could only image a "living" specimen at high magnification for a very sort time (one frame?) before it was well on the way to death. However, the wide-field/deconvolution contingent under Agard and Sedat showed that 3D imaging of living cells was possible, so we tried harder.
Even though instrumental improvements have now reduced the amount of damage produced per image by about 100x, and even though "the hidden agenda" of the Second edition of the Confocal Handbook was to give researchers the tools needed to view living specimens optimally, it seemed (seems?) to me that the large preponderance of confocal images is still made on dead cells.
There were several important reasons:
* Not clear which biological questions could be best approached in this way. * Hard to keep cells alive on microscope stage. * Results tedious to obtain and outcome uncertain. * Unclear which dyes were most suitable for live-cell operation in terms of cytotoxicity and bleaching * The choice of operational parameters was complicated by interactions between them. If pixel and raster-size, NA, sampling time and, laser power all have to be optimized to produce the best signal-to-noise and signal-to-damage ratios. * Single=photon or multi-photon. * Finally, there seemed to be a geography problem. While individual groups were making great strides, their isolation kept the practical knowledge that they gained by trial and error from reaching many of those who had good biological problems but lacked "a place to start from".
This Email Listserve was a response to the isolation and, I believe, it has been a major factor in the development of the field. Thanks Bob and now Steve!!
However, sometimes one needs a more personal type of interaction than is yet possible over email. In an attempt to fill this perceived gap, 5 years ago I decided to organize a 10-day course devoted to the 3D microscopy of living cells. Although the course would also cover advanced 2D techniques, the bulk of the time would be devoted to hands-on, laboratory sessions with no more than 3-4 students to a 3D Workstation. To focus attention, students were encouraged to design and carry out a 3D Live-cell Microscopy project during 5 evening sessions in the 3D part of the course and them present the results on the last morning.
Since that time, the course has served over 80 students from 18 countries, and will be offered again this year June 16 -27 (with a 3D Image Processing Workshop from June 29 - July 1. More info at http://www.cs.ubc.ca/spider/ladic/course/bulletin.html).
My question is this:
Now that the field has progressed and so many groups are actively publishing in the field (and helping out on the Email Net!), is there still need for such a course?
Although organizing 15 faculty members to come to beautiful Vancouver isn't so difficult, it costs the manufacturers an estimated $200,000 to send the equipment and personnel for the 11, 3D Workstations and you know who pays for that in the end!!
In addition, the controversy about who can, and cannot, do 2-photon excitation and under what conditions becomes ever more strident. Besides making organizing more onerous, this is particularly sad because it is clear that the main advantage that 2-photon imaging has over other methods is in imaging live cells, where conventional anti-bleach agents cannot be used, and where objects thicker than 20 micrometers must sometimes be viewed.
Last year, this controversy was almost certainly a factor in the last minute non-appearance of a multi-photon instrument (we did have 2 others) and this year it may lead to the total absence of one our major sponsors from earlier years (we will have at least 9 others).
I ask whether we have outlived our usefulness because I want to know. And I don't any more knowledgeable group to ask.
Although the student evaluations have been predominantly very positive, and the course itself has definitely improved as we found out what worked best, the trend in student numbers has not been encouraging: the first year we had over 50 applications for 28 places, the second about 35 applications and last year we had only 24 students after 4 dropped out at the last moment. Applications this year are about the same as last year but the March 1 deadline is still far away so this doesn't mean much.
I ask for your thoughts in planning beyond this year, because, given the "informal" nature of the founding of the course, there is no "institutional support" and tuition and expenses are handled through my personal account. So far, we are solvent but I personally can't afford to make good losses, should they occur.
And just to make sure there is no misunderstanding: I am not talking about this year's course (1999). I am asking for the future.
The Emory University Neurology Microscopy Laboratory, the University of Georgia Botany Department EM Laboratory, and Leica Microsystems, Inc.
Present a Cryo Techniques and Immunogold Workshop.
April 18-23, 1999 at the Campus of Emory University in Atlanta, GA.
Objectives
1. To provide researchers the opportunity to learn the theory and practice of cryo techniques for biological sample preparation and immunogold labeling.
2. To permit participants to process their own samples using these techniques under expert guidance.
Scitex America owned Leaf and its products. The address for Scitex was: One O'Hare Center 6250 River Rd. Rosemont, IL 60018 (847) 692-6000
The film holders were negative carriers made by Beseler for the Beseler 45M series and CB7 enlargers. Why not contact Beseler either directly or through a dealer to get a carrier for, say, 35 mm film. Then you can have your shop cut the appropriate opening in the carrier for your film.
Below is the text of a message I received from Joe Peng at Scitex: --------------------------------------------------------
Dear Joost,
Are you doing these eyes for TEM or LM? We do mice eyes for TEM (in plastic) and have gotten some really beautiful shots of Desmett's membrane. We have a whole fixation and embedding protocol worked out. The membranes are perfect. If this is for TEM, let me know and I'll send you our fixation and embedding protocol.
Regards!!
Lesley Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
I think Tracy and Jean would be a good start. There are some other TEM labs on campus and in the Vet College and Labs. They should be able to tell you about them, too. IThat's about as much as I know since I am over on the Engineering/Materials side of campus.
Name: PEPPER TRACEY M Phone(w): 515-294-3872 Phone(w): 515-769-2471 Fax: 515-294-3932 Email(w): tpepper-at-iastate.edu Office: Address: 001 BESSEY City/State: AMES, IA 50011-1020
Name: OLSEN JEAN ANN Phone(w): 515-294-1009 Phone(w): 515-233-2696 Fax: 515-294-1401 Email(w): jaolsen-at-iastate.edu Department: VETERINARY MEDICAL RESEARCH INSTITUTE Office: Address: VMRI BLDG 2 City/State: AMES, IA 50011-1240
At 03:36 PM 1/19/99 -0600, you wrote: } } Who would be a contact person at Iowa State Univ. for biological TEM ? } Possibly involving Immuno EM. Thanks. } } Rick L. Vaughn }
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Hi,
Does anyone out there have a vacuum evaporator that is taking up space? I = have lots of space but no evaporator. =
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/apw.htm
with Novell_GroupWise; Wed, 20 Jan 1999 11:28:14 +1100 Message-Id: {s6a5bdce.092-at-rsbs.anu.edu.au} X-Mailer: Novell GroupWise 5.2
We are just setting up TEM EDXA in a multidisciplinary unit, and need to = acquire standards covering a wide range of applications. Any recommendatio= ns, pitfalls etc? (Philips 430 TEM up to 300kV, Oxford ISIS ultra-thin = window detector).=20 thanks Sally
Dr Sally Stowe Facility Coordinator Australian National University EM Unit Research School of Biological Sciences Box 475 ACT 2601, Canberra, Australia FAX 06 (0)2 6279 8525
This message is in MIME format. The first part should be readable text, while the remaining parts are likely unreadable without MIME-aware tools. Send mail to mime-at-docserver.cac.washington.edu for more info.
I have a series of se images of a serial sectioned material which I would like to display as a volume rendered image. I have aligned the images, using an nih-image macro, however the brightness and contrast differs between them. As I understand it I need to perform histgram equalisation to correct for these differences.
I've noticed photoshop allows the user to load a transfer function using the "curves" dialog box, but it doesn't give the desired result.
Is this because the function which you specify is not the cumulative transfer function based on the reference histogram area??
Is there anyone who has done this before? I am using nih-image and photoshop for these tasks. As for the reconstruction, well any suggestions there would be apreciated also.
Thanks
---------------------------- Travis Baroni (PhD Student) | tbaroni-at-cyllene.uwa.edu.au | The University of Western | Australia, Nedlands, WA. | 6907. | ----------------------------
As Curriculum & Technology Specialist for the Alliance for Education, I take a Personal Scanning Electron Microscope (manufactured by RJ Lee, Pittsburgh, PA) to schools and museums. It is easily operated by anyone who can use a joystick and a mouse. I generally have students work in teams. Since our major funder is the Air Force, I refer to the responsibilities in flying terms, such as pilot and copilot.
We coordinate with the teacher to select specimens appropriate to their curriculum. Since we are using 17 inch monitors, it's not a particularly flashy activity for a large group. With large screens and appropriate sound systems, it could be. The main thing, though, is to choose a system that can be operated hands-on by young people.
For more information, you can call me at (937) 222-2934 between 9 and 5 EST.
I have a series of se images of a serial sectioned material which I would like to display as a volume rendered image. I have aligned the images, using an nih-image macro, however the brightness and contrast differs between them. As I understand it I need to perform histgram equalisation to correct for these differences.
I've noticed photoshop allows the user to load a transfer function using the "curves" dialog box, but it doesn't give the desired result.
Is this because the function which you specify is not the cumulative transfer function based on the reference histogram area??
Is there anyone who has done this before? I am using nih-image and photoshop for these tasks. As for the reconstruction, well any suggestions there would be apreciated also.
Thanks
Travis Baroni
---------------------------- Travis Baroni (PhD Student) | tbaroni-at-cyllene.uwa.edu.au | The University of Western | Australia, Nedlands, WA. | 6907. | ----------------------------
Is anyone famillair with good fixation and staining techniques for rat and chicken eyes. Is Davidson's fluid THE best fixative? Someone in our lab wants to stain slides in order to examin Desmett's and Bowman's membrane. Is it possible, to stain these kind of membranes?
Try using the excellent piece of Software "Electron Flight Simulator" version 3.1-E. It is marketed by D, Chernoff of Small Woprld somewhere in the US. I am running it on a PC and I am not sure its works on a Mac. Contact David Joy at {djoy-at-utk.edu} as he has written much of the software and will give you an adress for Chernoff.
Patrick Echlin Director, Multi-Imaging Centre University of Cambridge UK
On Tue, 19 Jan 1999, by way of Nestor J. Zaluzec wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Hi, I am looking for a program to convert weight % to Atomic % and vise } versa preferably on a Power Mac.. An old program we have works on pre '90 } Mac's but not on the newer ones. Anyone have any ideas? Thanks- Mike } } } }
Is anyone familiar with the re-embedding methods of paraffin embedded tissues in resin? I was asked to re-embed human lung tumor specimens into resin and reclassify them on electron microscope level. I will appreciate if you could send me your reembedding protocol.
Thanks in advance,
Regards,
Ranan
Ranan Gulhan AKTAS, M. D. Trakya University, Faculty of Medicine Pathology Department Edirne 22030 TURKEY
Paul:=20 I have two rather old evaporators which still get 10-6 torr. I will give=20 then to you but you must pay the shipping costs !
Patrick E=A3chlin Multi-Imaging Centre University of Cambridge UK
On 19 Jan 1999, Paul Webster wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Hi, } =20 } Does anyone out there have a vacuum evaporator that is taking up space? = I have lots of space but no evaporator. =20 } Paul Webster, Ph.D } House Ear Institute } 2100 West Third Street } Los Angeles, CA 90057 } phone:213 273 8026 } fax: 213 413 6739 } e-mail: pwebster-at-hei.org } http://www.hei.org/htm/apw.htm } =20 } =20 } =20
Martin Roe discovered some Br in epoxies and asked for the source. I gave him an answer off line. However I see some other answers and I think that I should add my two cents for everybody:
I have a good friend who worked a long time as microscopist for CIBA before it came Novartis=8A
He told me about 5 years ago that bromine is voluntarily added (for some reasons=8A) in certain types of epoxies. So it is not a contamination, nor a new thing! So he (or his supplier) has probably just changed his epoxy source.
We used some compound sample made of A alloysl bits in epoxy to show to our students how dangerous it can be to work only at low kV in EDS. You have a nice superposition of Al Ka and Br L lines. If the uncertainty on sulfur/molybdenum is quite well known, that on Al/Br confuses quite a lot of people because Br is often unexpected!
Yours
Philippe Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, I am looking for a program to convert weight % to Atomic % and vise } versa preferably on a Power Mac.. An old program we have works on pre '90 } Mac's but not on the newer ones. Anyone have any ideas? Thanks- Mike
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
In a message dated 1/19/99 10:19:06 PM, tbaroni-at-cyllene.uwa.edu.au writes:
} As I understand it I need to perform histgram equalisation } to correct for these differences.
In your case what you should really do is apply a transfer function that is the cumulative histogram of ALL of the images in the stack to each image, not equalize them individually.
UNIVERSITY OF LEEDS School of Process, Environmental and Materials Engineering
POSTDOCTORAL RESEARCH FELLOW
Applications are invited for this post funded by the EPSRC (Waste Minimisation Managed Programme) to join an active research group working with Dr. Mark A. Keane (Department of Chemical Engineering) and Dr. Rik Brydson (Department of Materials) on the development of supported metal catalysts for the "environmentally friendly" treatment of chlorinated aromatics (see, for example Appl. Catal. B: Environmental 18 (1998) 241-250). The project is a fundamental study of catalytic hydrodechlorination and will involve the preparation, testing and characterisation of a range of catalyst systems with an emphasis on developing an understanding of reaction kinetics and mechanism. The successful applicant should have a PhD in Chemistry, Chemical Engineering or Materials with a strong background= in Heterogeneous Catalysis: some experience in catalyst characterisation is a decided advantage. The post is available from 1 April 1999, or as soon as possible thereafter, for a period of three years
Salary Range: RA1A3 =A3 15,462-=A317,226
Informal Enquiries may be made to Dr. Mark A. Keane , Tel: +44 113 233 2428, E-mail: chemaak-at-leeds.ac.uk
Interested candidates should apply in writing with a detailed CV and the names of two referees to Dr. Mark A. Keane, Department of Chemical Engineering, University of Leeds, Leeds LS2 9JT, UK
Closing Date: 1 March 1999 __________________________________________________________ Dr. Rik Brydson, University Research Fellow, Electron Optical Unit, Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
There is a good shareware product named GraphicWorkshop Pro from Alchemy Mindworks in Ontario, Canada. It is a very good batch converter for Windows XXX.XXX based operating systems. They have a friendly web site to down load a copy:www.midworkshop.com. We of course have no financial interest in this product. Consider the Canadian dollar and our American friends can get it for about 65% of cost which I think is $40.00 if you register it. And we all register shareware, don't we? Jon McGovern J. P. McGovern and Associates jon-at-microscopy.net
available immediately in Department of Neuroscience and Neurology, University of Kuopio.
We are seeking suitably qualified persons to join a progressive multidisciplinary group investigating Alzheimer=92s disease. The experimental projects will investigate the efficacy of estrogen replacement therapy in preventing structural and functional impairment in different animal models of Alzheimer=92s disease.
We are looking for applicants who have experience in at least one of the following specialities:
Histopathology/histochemistry, electron microscopy, autoradiography.
Salary and date of commencement are negotiable.
For further information please contact:
Research Director Paavo Riekkinen, Jr. M.D., PhD. Dept. Neuroscience and Neurology Univ. Kuopio FIN-70211 Kuopio Finland Tel. 358-17-162016 Fax. 358-17-162048 E-mail: paavojr.riekkinen-at-uku.fi
Riitta Miettinen, Ph.D. Dept. Neuroscience and Neurology Univ. Kuopio P.O.Box 1627 FIN-70211 Kuopio FINLAND E-mail: Riitta.Miettinen-at-uku.fi Tel. 358-17-162761 Fax: 358-17-162048
I join John Russ in saying that you do NOT want to apply histogram equalization to each image individually, although I am not quite sure that what he suggested is quite what you want.
I suppose that there may be some differences in exposure between the images for whatever reason. The challenge will be to get the same features to the same level of gray. That is, if you have some light features on a dark background, you always want those light features to be at the same light gray level and the background to be at the same darker gray level. Histogram equalization is definitely NOT the tool you want to use since it will move gray levels depending on their population in the image. It normally increases contrast between some pixels and decreases it between others and hardly gives you a uniform transfer function for all images.
I am not very familiar with Photoshop. I thought the tool you would look for was called something like "levels". I think you would want to primarily use the gamma adjustment to bring two reference features (e.g., light features and dark background) to the same gray levels in all images, then maybe use some tweaking of brightness and contrast to finish the match (per John Mackenzie's recipe). I will leave it to you to figure out which tool to use (histogram or a pixel cursor or ...) to determine when the images are matched. In my own work I do not need a quantitative match, but only a qualitative, visually apparent match. Therefore, I just eyeball a match between one reference image and all the others as I adjust them.
WES
At 10:37 AM 1/20/99 +0800, you wrote: } } Hi all, } } I have a series of se images of a serial sectioned material which I would } like to display as a volume rendered image. I have aligned the images, } using an nih-image macro, however the brightness and contrast differs } between them. As I understand it I need to perform histgram equalisation } to correct for these differences. } } I've noticed photoshop allows the user to load a transfer function using } the "curves" dialog box, but it doesn't give the desired result. } } Is this because the function which you specify is not the cumulative } transfer function based on the reference histogram area?? } } Is there anyone who has done this before? I am using nih-image and } photoshop for these tasks. As for the reconstruction, well any suggestions } there would be apreciated also. } } Thanks } ---------------------------- } Travis Baroni (PhD Student) | } tbaroni-at-cyllene.uwa.edu.au | } The University of Western | } Australia, Nedlands, WA. | } 6907. | } ----------------------------
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
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Hi, Dear all -
I have an AO Reichert Ultracut and a LKB knifemaker. They have been in a storage for a while and need to be cleaned and lubricated. Does anyone has such experience in lubricating them and can give me some suggestions about what kind of lubricator(s) that I should use and how to do that?
Thank you in advance.
Gang Ning EM Facility Medical College of Wisconsin
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We have two old Reichert-Jung OMu2 microtomes which are in good working order with one exception - we are in need of a "female" allen key part which is used to tighten the block into the chuck. Does anyone have any they no longer need? Or know where to get one? Or has found a substitute which is easily available?
Thanks in advance,
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-med.mcgill.ca
Hi Joost- you could refer to an article published in Cornea 12(3): 255-260, 1993 (Rock et al.,) we used a modified Karnovsky's 2.5% glut, 2.0% paraform, in a 300-momol, in cacodylate bfr., post fixed in OsO4, dehydrated, embedded in epon.
then using a Mallory's Azure II, methylene-blue followed by a basic fuchsin counter stain protocol we developed a fat and effective method for identifying and differentiating various regions of the cornea. Descemet's membrane stains blue, Bowman's layer stains pink, the collagen of the stroma striated pink & blue, scar tissue blue, and keratocytes blue. staining differences in the various regions are presumably attributed to the proteoglycan content and/or the various collagen types
hope this helps -Mike Rock
On Wed, 20 Jan 1999 J.Bruyntjes-at-voeding.tno.nl-at-Sparc5.Microscopy.Com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi there } } Is anyone famillair with good fixation and staining techniques for rat and } chicken eyes. Is Davidson's fluid THE best fixative? } Someone in our lab wants to stain slides in order to examin Desmett's and } Bowman's membrane. Is it possible, to stain these kind of membranes? } } Thanks in advance } } Joost Bruyntjes } TNO Zeist } The Netherlands } }
Dear All, I need to obtain some TEM analytical standards for stainless steel (for example, a standard containing just Ni, Cr, and Fe, in a well characterized composition). I was hoping for something along the lines of Cr ~ 28%, Ni ~22% and Fe ~ 50%. It would be great if I could find a sample that was TEM "ready", so that I don't have to make a new sample. If anyone knows of a source for such a sample, I would be grateful for a reference. Thanks in advance, Dorrance
Does anyone have epifluorescence attachments for a Nikon Optiphot that they wish to sell? I'm looking for a complete epi set-up.
Thanks, Glen
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
The box said "Requires Windows98 or better.", so I bought a Macintosh.
We have a Microm HM 500 0 sold by Zeiss. It has been very reliable for three years and gives excellent sections with minimal adjustment. The only caveat is the refrigeration system. A compressor was replaced under warranty and recently we experienced a leak of refrigerant. Local refrigeration people can do the work (Zeiss is not licensed to do it). I might add that refrigeration problems are common in this building due to various factors. Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
I will be receiving a sample consisting of a culture grown on a polycarbonate membrane (Transwell permeable). I need to know any pitfalls working with the membrane! Is this membrane compatible with acetone dehydration? Is it compatible with a Spurrs resin? Does anyone have experience in this area?
Thanks - Sally Burns
Sally Burns Center for Electron Optics B5 Pesticide Research Center (517) 355-5004
I have processed many filter membranes for TEM and SEM. I follow standard procedures with the following changes;
-dehydrate in EtOH, not acetone (eats the polycarbonate) -embed in Epon 812 or equivalent (I use Ted Pella's Eponate 12) -times can be shortened conciderably (Fix 20 min, wash and dehydrate steps 5 min, resin changes around 30 min.) -do a couple more resin changes than normal
I carry out the entire process (including embedment) in a 24 well plate and cut the filters out with a jeweler's saw after polymerization.
Thomas Moninger moninger-at-emiris.iaf.uiowa.edu University of Iowa Central Microscopy Research Facility http://www.uiowa.edu/~cemrf Views expressed are mine.
On Wed, 20 Jan 1999, Sally Burns wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I will be receiving a sample consisting of a culture grown on a } polycarbonate membrane (Transwell permeable). I need to know any pitfalls } working with the membrane! } Is this membrane compatible with acetone dehydration? Is it compatible with } a Spurrs resin? Does anyone have experience in this area? } } Thanks - } Sally Burns } } Sally Burns } Center for Electron Optics } B5 Pesticide Research Center } (517) 355-5004 } } burnssal-at-pilot.msu.edu } }
We have recently acquired a LEO 438 and have need, for safety reasons, to provide small particle (1 micron) filtration at the inlet side of both the roughing and turbo pumps. I believe we can use conventional filters in-line on the roughing pump, but probably cannot afford to restrict flow at the turbo pump inlet. Does anyone have advice or ideas on how we might provide small particle filtration, perhaps via a commercial or custom built free-flow filtering device placed just under the perforated screen at the floor of the chamber. Servicing is a consideration. Also, any advice on filtering the roughing pump would also be much appreciated.
Thank you very much for your consideration.
Bruce Cunningham High Explosives Application Facility (HEAF) Lawrence Livermore Laboratory
-------------------------------------- Bruce Cunningham Lawrence Livermore National Laboratory P.O. Box 808, L-282 Livermore, CA 94550 cunningham1-at-llnl.gov TEL: 510-423-0135 FAX: 510-424-3281
I am looking for the company that makes ACLAR. I need to contact them. I don't need vendors I need the manufacturer of this wonder substance. So if anybody out there knows who it is, please let me know.
Tired of changing my fingerprints with borken slides,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
} With a good camera for initial camera, the results can be better than any } mid range color camera!! Definitely worth it.
What filter sets would people recommend? And for light microscopy I assume the filters are usually placed out of the focal plane, maybe somewhere near the field diaphram?
Gary Radice 804-289-8107 Department of Biology 804-289-8233 (FAX) University of Richmond gradice-at-richmond.edu Richmond VA 23173
HI I am attempting to examine embrylogical samples of a marine bivalve using SEM. But any containers I have used during the processing proceedure have proved indequate. I need containers which will hold samples down to a size of 30um. I have used filters placed in processing capsules but they are very awkward, and the sample has more often than not been lost during the proceedure. Any ideas would be greatly appreciated. Jean Raleigh, Marine Zoology Dept., Natioal University Galway Ireland
COMPUTER VIRUS NOW INFECTING HUMANS! Trojan Horse Supervirus Could Be Worse Than AIDS, Warn Docs -- And TWICE As Deadly! by Kevin Creed/Weekly World News
CHICAGO, ILL. -- Concerned scientists say the dreaded "Trojan Horse" computer virus has made the jump to humans -- and the brain-eating bug may soon be sweeping through America, claiming even more human lives than the AIDS epidemic! An unidentified 38-year-old man known only as Patient Zero has been diagnosed with the virus that had been heretofore found only in PCs.
"We've been dreading this day," said noted virologist Dr. Frederick Attingale who made the terrifying diagnosis. "We knew it was only a matter of time. That's how these things work. "At some point, Acquired Immune Deficiency Syndrome -- AIDS -- made the jump from monkeys to man. Now, in a similar way, the Trojan Horse virus has worked its way into the human population. Both viral transmissions were bound to happen sooner or later."
Dr. Attingale will not name the Chicago-area hospital where Patient Zero is being held. But the prognosis is not good. "People whose PCs have been infected know the virus eats away the hard disks and 'nervous systems' before anyone is aware that their computers have been infected.
"I'm afraid the same thing is happening in this man's body."
The patient, a junior executive with a large investment firm, is suffering from nerve spasms, hearing and vision loss and severe deterioration of the parts of the brain that govern memory, reasoning, math and language. His brain and nerves are being literally eaten away.
"There's no known cure and the illness continues to worsen," Dr. Attingale said. "He can barely communicate with us now."
The alarming situation came to light early last month when the patient went to his family physician complaining of headaches and memory lapses.
The doctor, suspecting a virus, referred him to Dr. Attingale.
For more information on this deadly virus, including how it enters the body and interviews with other doctors, pick up the current edition of Weekly World News (1/26/99), at your local supermarket or newsstand! Or better yet, why not subscribe and have it mailed to your home or office!
I have no experience with the Ultracut but I am aware that the surface between the cutting head and the main structure of the LKB Knifemaker (LKB 7801A) must NOT be lubricated. The metals are supposed to provide the correct level of friction to allow glass to be clamped at the correct pressure. So perhaps my message is don't lubricate anything until you are certain.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: Gang Ning To: microscopy-at-sparc5.microscopy.co
Aclar is made by and can be purchased in bulk from :
Pro Plastics Inc 1190 Sylvan Street P.O. Box 1489 Linden NJ 07036
201-925-5555
Area code may have changed At 04:48 PM 01/20/1999 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} With a good camera for initial camera, the results can be better than any } mid range color camera!! Definitely worth it.
What filter sets would people recommend? And for light microscopy I assume the filters are usually placed out of the focal plane, maybe somewhere near the field diaphram?
Gary Radice 804-289-8107 Department of Biology 804-289-8233 (FAX) University of Richmond gradice-at-richmond.edu Richmond VA 23173
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would appreciate hearing from anyone with an ESEM-FEG. Please respond to me personally as I would like to talk to you re: this instrument.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
by imo29.mx.aol.com (IMOv18.1) id DMMGa01223; Thu, 21 Jan 1999 09:38:42 -0500 (EST) Message-ID: {cb9380fd.36a73bf2-at-aol.com}
Not only should the knifemaker not be lubricated but all sliding surfaces between the clamping head and pedestal should be periodically cleaned with acetone or alcohol. Any lubrication will attract dirt which will affect the gravity drop of the pins onto the glass,it will also cause the clamping head to move up when pressure is applied to make the break. Many times poor knives can be eliminated by this simple cleaning procedure. It will necessitate removing the clamping head from the pedestal Good luck, Norm Woodside former LKB product specialist
Out of curiosity I mentioned this to a chemist colleague. He indicated that bromine is sometimes added to resins as a flame retardant. In some cases antimony is also added to boost the effect of the bromine. Everett Ramer Federal Energy Technology Center
} } } "Martin J. Roe" {mi596-at-mluri.sari.ac.uk} 01/18/99 07:11am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello fellow listservers, I've recently discovered a problem with a batch of a particular type of cold setting epoxy resin. I have been using this batch of resin on an infrequent basis for about 2 years (and is probably about 4 years old). When I analysed some of the cured resin by microanalysis last week appreciable bromine (Br) was detected as well as the usual chlorine content (Br} Cl). The 'introduction' of the Br is a recent thing because I also analysed some resin (from the same batch) prepared approximately one year ago. The analysis of this showed no Br with Cl the only detectable element present (with the Be window of the EDS detector in place that is). Is it possible therefore for the resin to in some way deteriorate with time and for Br compounds to form? If so, what are the chemical reactions going on here. I'm very certain that the Br is not a contaminant. Also, I should point out that if this is a chemical degradation of the resin then the physical characteristics of the cured resin are normal. I would appreciate any thoughts on the matter. Regards
Martin Roe Macaulay Land Use Research Institute Aberdeen AB15 8QH Scotland United Kingdom
I have a question about the KEVEX 7700 XRF system. We have been trying = for some time to get the output of the unit into a PC through the = printer port. Apparently it can be done and we would like to know if = anyone out there has tried this and what sort of parameters and software = have they used to connect to the 7700 unit. Any help would be greatly = appreciated. Currently all we get from the printer output to the PC via = a hyperterminal link is just garbeld. Thanks in advance.
______________________ Roberto Garcia Senior Analyst, Metallography North Carolina State University Analytical Instrumentation Facility Box 7531, Room 303 EGRC Raleigh, NC 27695-7531 rgarcia-at-unity.ncsu.com
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD} {META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {STYLE} {/STYLE}
{META content=3D'"MSHTML 5.00.0910.1309"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT size=3D2} I have a question about the KEVEX 7700 XRF system. = We have=20 been trying for some time to get the output of the unit into a PC = through the=20 printer port. Apparently it can be done and we would like to know if = anyone out=20 there has tried this and what sort of parameters and software have they = used to=20 connect to the 7700 unit. Any help would be greatly appreciated. = Currently all=20 we get from the printer output to the PC via a hyperterminal link is = just=20 garbeld. Thanks in advance. {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D2} ______________________ {BR} Roberto Garcia {BR} Senior = Analyst,=20 Metallography {BR} North Carolina State University {BR} Analytical = Instrumentation=20 Facility {BR} Box 7531, Room 303 EGRC {BR} Raleigh, NC 27695-7531 {BR} {A=20 href=3D"mailto:rgarcia-at-unity.ncsu.com"} rgarcia-at-unity.ncsu.com {/A} {/FONT} {= /DIV} {/BODY} {/HTML}
You can get ACLAR from Allied Signal, Inc. Westwood Road. Pottsville, PA. 17901. The phone number that we have appears to no longer be the correct one but I am sure you can find the number using information.
Good Luck.
--------------------------------------- David McKemy Dept. of Pharmacology/318 University of Nevada School of Medicine Howard Bldg. Rm.#214 Reno, Nevada 89557 Phone: (775) 784-4537 Fax: (775) 784-1620 Email: ddmckemy-at-med.unr.edu
On Wed, 20 Jan 1999, Paula Sicurello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hey Boarders, } } I am looking for the company that makes ACLAR. I need to contact } them. I don't need vendors I need the manufacturer of this wonder } substance. So if anybody out there knows who it is, please let me know. } } } Tired of changing my fingerprints with borken slides, } } } Paula :-) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } phone: 510-642-2085 } fax: 510-643-6207 } http://biology.berkeley.edu/EML } } }
I routinely refer to two articles regarding this type of SEM prep.
Preparation of Microbiological Specimens for Scanning Electron Microscopy, L.P. Watson, A.E. Mckee, and B.R. Merrell. Scanning Electron Microscopy/1980/II, pages 45-56.
Specimen Preparation Techniques for Aquatic Organisms, T.K. Maugel, D.B. Bonar, W.J. Creegan and E.B. Small. Scanning Electron Microscopy/1980/II, pages 57-77.
Hope this helps, Louie
At 11:16 AM +0000 1/21/99, Jean Raleigh wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX)
Soon I may need to study an optical fiber with a periodic refractive index (doped). Since I am just beginning TEM studies, I am hoping that someone may be able to suggest the typical method of observing optical fibers with a TEM (these optical fibers are typically 100-125 microns in diameter). I suspect that thinning the fibers lengthwise may be a possible means, however, I would like to see a reference where optical fibers have been studied...or perhaps similar geometries like biological hair samples (though optical fibers are generally much thinner).
cross-sectional thinning (circular samples) is useless since we need to study period structures along the length of the fiber.
Thanks
__ _-==-=_,-. /--`' \_-at---at-.-- { Tim (TJ) LaFave Jr. `--'\ \ {___/. Department of Physics \ \\ " / University of North Carolina, Charlotte } =\\_/` { Charlotte, NC 28223 ____ /= | \_|/ _' `\ _/=== \___/ (704)547-3392 [x4] `___/ //\./=/~\====\ (704)509-6622 [Hm] \ // / | ===: http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ \/ \\ \\`--| / \\ ---------- +*+ ---------- | _ \\: /==:-\ `.__' `-____/ |--|==: Such that the future be theirs \ \ ===\ :==:`-' to shape and direct. _} \ ===\ /==/ -----------------------------------------------------------------
OK, so we should not lube the knifemaker at the clamping head, but what should we do with ours that squeeks so bad it makes your teeth hurt? Our LKB 7800 has some serious squeeking when you increase the upward pressure for the break (and release it). Can we lube the arm that causes the pressing up motion (the one on the front of the machine)? It's driving us all crazier than we already are. So any suggestions will be greatly appreciated.
I was thinking of taking the cover off the knifebreaker and see if I could find any lubrication points, is that a bad thing to do?
Squeeking loudly in Berkeley,
Paula :-)
} Not only should the knifemaker not be lubricated but all sliding surfaces } between the clamping head and pedestal should be periodically cleaned with } acetone or alcohol. Any lubrication will attract dirt which will affect the } gravity drop of the pins onto the glass,it will also cause the clamping head } to move up when pressure is applied to make the break. Many times poor knives } can be eliminated by this simple cleaning procedure. It will necessitate } removing the clamping head from the pedestal } Good luck, } Norm Woodside } former LKB product specialist
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Does anyone knows where can I get high temperature conductive glue for TEM sample preparation? I need to glue a ceramic sample to Cu grid and Ion-mill it then heat treat to 1000 C in air before TEM observation. Thank you in advance.
Maoxu Qian
**************************** * Maoxu Qian, Ph.D. * * Dept of MSE, box 352120 * * University of Washington * * Seattle, WA 98195 * * mxq-at-u.washington.edu * * (206)616-3973(phone) * * (206)543-3100(fax) * ****************************
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I would think that it would be a simple enough issue. I have done similar things in the past. In fact you are probably pretty close to the solution.
It sounds like you probably have a mismatch in the baud rate between the two computers - it would give you garbage. The PC side is fairly easy to change for experimentation while the Kevex side is harder to change. I don't think the old DEC serial cards supported more than 19,200 baud. We had some old printers that required the rate set down to 2400. You could check your printer or Kevex manuals to see if there is any info in there. Otherwise, you could just keep setting the baud rate down on the PC side and reprinting until something sensible comes through.
The specs on the port are usually 8-bit, no parity, 1 stop bit. That should be the same as the default PC setting. Most DEC stuff was setup to use XON/XOFF software handshaking as oppsed to DTR or other hardware handshaking.
Good luck, and please let me know how it works.
WS
At 10:48 AM 1/21/99 -0500, you wrote: } } I have a question about the KEVEX 7700 XRF system. We have been trying for } some time to get the output of the unit into a PC through the printer port. } Apparently it can be done and we would like to know if anyone out there has } tried this and what sort of parameters and software have they used to connect } to the 7700 unit. Any help would be greatly appreciated. Currently all we get } from the printer output to the PC via a hyperterminal link is just garbeld. } Thanks in advance. } } ______________________ } Roberto Garcia } Senior Analyst, Metallography } North Carolina State University } Analytical Instrumentation Facility } Box 7531, Room 303 EGRC } Raleigh, NC 27695-7531 } {mailto:rgarcia-at-unity.ncsu.com} rgarcia-at-unity.ncsu.com
Hello, everyone Is there anyone that has "PC-PDF" JCPDS-ICDD PDF-2 DATABASE files of RuO4 and RuO2 or knows from where we can get these files? We have a very old PC-PDF file that has not been upgraded for several years. I would appreciate any information from anyone.
With best regards, Pipat Prayoonthong Graduate student Materials Science & Engineering Department. Stevens Institute of Technology Hoboken, NJ 07030
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Greetings Everyone, At one time or another, i have recieved an email from you and am just letting you know my email addy has changed from pclypool-at-sgi.net to claypool-at-serve.com
This email might concern the SX-50 Users group, contacts dealing with a Task8verC Microprobe, or you might just be a personal friend (or all the above hehe).
The following procedure may sound a little obvious, but I imagine you may be facing a rather limited access to all those books and journals, and I would feel really happy if this could help. I do wish you all the best in your EM lab-raising mission!
First, you will have to select smaller, "EM-size", portions of your paraffin-embedded specimens, cut them out and thoroughly deparaffinate. I would recommend at least 2 hours in xylene with frequent changes of xylene and some agitation; you should be able to see when it's all gone. Then several changes of 100% (anhydrous) ethanol, at least 1-1.5 h total time, and then gradually down the ethanol concentrations, like 95-85-70-50, 30 min or more at each step. Rinse 3x5 min in your choice of phosphate or cacodylate buffer, and then osmicate for 1-2 h, 1% OsO4 in the same buffer. Then dehydrate and embed for EM as you normally would.
Needless to say, even with the best original fixation the material will look pityful, but most of those diagnostically significant cell-to-cell junctions, filaments, etc. must still be there.
Best of luck! Sincerely, Vlad.
Vladislav V. Speransky Postdoctoral Research Associate School of Marine Sciences University of Maine 5722 Deering Hall Orono ME 04469-5722 Ph: 207 581 2998 FAX: 207 581 2969 Email: vladis-at-maine.maine.edu
I have a slip that was attached to a roll of ACLAR giving specifications and the following company name and address: Engineered Plastics Specialty Plastic Films Pottsville, PA 17901
I have no knowledge of whether this company still exists. I would guess the roll of ACLAR was 5 years old or more.
Missy Josephson
Eleanor Josephson, DVM, PhD University of Connecticut Health Center Department of Anatomy MC-3405 263 Farmington Ave. Farmington, CT 06030-3405 Ph.(860)679-2463 Fax (860)679-1274 ejosephs-at-neuron.uchc.edu
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I have not seen a reply to your posting so try this.
Astigmatism in an image of a modern TEM is always down to cleanlyness. I= f you have a gross change in astigmatism, if everything is working correctl= y, it has to be dirt. Lets look at typical symptoms for dirt in a system:-
1. An increase in astigmatism levels due to the charge on the dirt affecting the beam.
2. Beam movement, gradual in one direction as the charge grows then rapid in the return direction as the problem is discharged. When the charge reaches a level sufficient to touch earthed components within the system it discharges, the problem starts all over again.
3. The problem will vary depending upon the number of electrons hitting the area that is charging and their energy. The greater the numb= er of electrons you place in the charging area the faster it will charge. T= he higher the kV the more chance you have of the beam penetrating the contamination and finding an earth; less or no charge. The mode of operation in which you are working will also change the charge rate. In the normal imaging mode the setting of the diffraction lens does not plac= e the beam near the problem area of your coulmn - no noticable astigmatism change in this mode. In the diffraction mode the diffraction lens focal length is dramatically changed hence electrons in larger numbers strike t= he problem area and we see what happens; trouble.
There is only one solution as the problem will get worse - strip the column. I am sorry for the engineer because this is a rare move on a modern machine BUT IT MUST BE DONE!
How to make friends with service engineers this is not, but it is THE onl= y solution!
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Here is the method used by the pathology people around here.
- Remove tissue of interest from parafin embedded specimen, cut in 1-2 mm cubes. - Leave the specimens in fresh 1% OsO4 in XYLENE for 1,5 - 2 hrs. - Wash in xylene 2 times 10 min - One part resin(we use Epon-Araldite) + two parts xylene for 1 hr. - 1 part resin + 1 part xylene for 1 hr. - Resin for 30 min to 1 hour without a lid on the glass
All this steps on a carousel.
Embedd as usual, but keep the specimens in the resin over night before polymerization.
Good luck!
Best regards Randi Olsen
Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso MH-Breivika N-9037 TROMSO NORWAY
HI I am attempting to examine embrylogical samples of a marine bivalve using SEM. But any containers I have used during the processing proceedure have proved indequate. I need containers which will hold samples down to a size of 30um. I have used filters placed in processing capsules but they are very awkward, and the sample has more often than not been lost during the proceedure. Any ideas would be greatly appreciated. Jean Raleigh, Marine Zoology Dept., Natioal University Galway Ireland
For the SEM preparations of spores and other small samples we put nets made of metal or nylon, meshs 20 =B5m, in our holder for CPD. For very small things, e.g. bacteria, zoospores, we introduce the samples with syringe and needle into LUMitainers. Lumitainers are little balls D=3D2,5-3,5 mm made of a kind of cellulose. They are useful for both SEM and TEM. In Germany you can buy them by Plano W. Planet GmbH, Marburgerstr. 90, 35043 Marburg, Fax 06425-51173. They are not cheap! I do not know where to get it abroad? Good luck! Anne Heller _____________________ Dr. Anne Heller Institut fuer Botanik (210) Universitaet Hohenheim Garbenstr.30 D-70593 Stuttgart Tel.0049-711-459-2180 Fax 0049-711-459-3355
Try folding small (10 x 10 mm) envelopes from lens tissue. Close with a small staple. These envelopes are permeable enough for processing. They work very well for pollen and erythrocyte samples.
Some types of lens tissue have rather large pores - I have seen holes up to 30 microns in some makes of tissue - check yours before using.
} I am attempting to examine embrylogical samples of a marine bivalve } using SEM. But any containers I have used during the processing } proceedure have proved indequate. I need containers which will hold } samples down to a size of 30um. I have used filters placed in processing } capsules but they are very awkward, and the sample has more often than not } been lost during the proceedure. Any ideas would be greatly appreciated. } Jean Raleigh, Marine Zoology Dept., Natioal University Galway Ireland
Prof Jan Coetzee Head: Lab for Microscopy and Microanalysis Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-362-5150 Pretoria 0002, South Africa http://www.up.ac.za/science/electron/emunit1.htm
I was interested in the use of osmium in xylene that you described. Does the tissue blacken as in a water-solution or does it simply become yellowish/brown? I am curious because the latter is what happens in the absence of water during freeze-substitution using e.g. 1% in acetone at -80 *C, although the colour change probably happens when warming up from low temperature.
Hello. I am looking for a company that is willing to build optical u-scope objectives with wavelength specific AR coatings. All leads will be greatly appreciated.
The Advanced Materials Processing and Analysis Center (AMPAC) at the University of Central Florida will be sponsoring two TEM specimen preparation short courses in conjunction with the FL AVS/FSM conference and vendor participation.
AMPAC's UCF/Cirent Materials Characterization Facility is offering two short courses in March.
"Tripod Polisher" will be offered March 12 and 13 instructor: Ron Anderson, IBM co-sponsored by UCF and South Bay Technology
"Focused Ion Beam Specimen Preparation" will be offered March 18 and 19. Instructors: Fred Stevie, Lucent Technologies and Lucille Giannuzzi, UCF co-sponsored by FEI and Micro Optics
Both courses will be held at the MCF located at 12443 Research Parkway, Suite 305, Orlando in Research Park {1 mile from the UCF campus. The registration fee is $750 per course or $1200 for both courses. Fees include course materials and meals (breakfast/lunch/breaks) all days. Registration deadline is March 1 and space is limited.
Additional information and registration forms for all of these events can be found at the AMPAC website: http://pegasus.cc.ucf.edu/~ampac
Please contact Dr. Giannuzzi below for any additional questions.
We look forward to seeing you in March!
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-mail.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
The 2nd Annual AMPAC Golf Scholarship Invitational will be held on Sunday, March 14, at the Ekana Golf Club in Oviedo, Florida, in conjunction with the FL AVS/FSM conference. The tournament starts at 8:00 a.m. Entry fee is $135 per player or $500 per foursome. There will be lots of prizes and food! Hole sponsorships (both corporate and individual) are also available. A corporate donation of $1000 will also include 4 golfers. A corporate donation of $500 will include 2 golfers. Individual hole sponsors may donate $100. Deadline for entries is February 26. The tournament is limited to the first 80 golfers so register today! Proceeds will benefit graduate student fellowships in Materials Science and Engineering at UCF.
Additional information and registration forms for all of this event can be found at the AMPAC website: http://pegasus.cc.ucf.edu/~ampac
We look forward to seeing you in March!
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-mail.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
This is easy and cheap to make: _ /_____\ _ | | | | | | | | {-- BEEM capsule |_|_____ |_| \ / ^ ^ |____|__ plankton netting
(I hope the ascii "art" survives the 'net...)
Cut out the center of the lid of a BEEM capsule, leaving a ring that still snaps onto the body of the capsule as usual. Close the lid, trapping a taut bit of plankton netting under it.
Cut off the lid from a second BEEM capsule and cut out its center as above. Cut of the pyramidal end from the first BEEM capsule, and snap the cut-off end over the new end of this capsule, again trapping a bit of plankton netting.
The netting comes in various sizes, so you can get one small enough for your needs, but use the largest size mesh you can to ease fluid flow.
Problem: when changing solutions before drying, the mesh will trap air. The best way to deal with this is to leave the 2nd end of the capsule open, and place the capsule mesh-end down in a 4mL Wheaton vial or similar. Change solutions by sucking out the old solution from the vial outside the capsule, and pipetting in the new solution into the open end of the capsule. Don't fill the capsule/vial completely, or the specimens might float out into the vial.
When ready to dry, snap on the 2nd lid with netting, suck some of the final fluid change into the capsule so there is no air bubble, then CPD.
I've also dried marine gastropod larvae from HMDS, no CPD with good results. These capsules work well for that also, and don't need the 2nd lid then.
Phil
} I am attempting to examine embrylogical samples of a marine bivalve } using SEM. But any containers I have used during the processing } proceedure have proved indequate. I need containers which will hold } samples down to a size of 30um. I have used filters placed in processing } capsules but they are very awkward, and the sample has more often than } not been lost during the proceedure. } Any ideas would be greatly appreciated. } Jean Raleigh, } Marine Zoology Dept., } Natioal University Galway } Ireland
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net
Jean Raleigh wrote: ============================================= I am attempting to examine embrylogical samples of a marine bivalve using SEM. But any containers I have used during the processing proceedure have proved indequate. I need containers which will hold samples down to a size of 30um. I have used filters placed in processing capsules but they are very awkward, and the sample has more often than not been lost during the proceedure. Any ideas would be greatly appreciated. ============================================= Have you tried what are called "Microporous Specimen Capsules"? They are available from SPI as well as several other of the main suppliers to the microscopy and histology market. They come in three pore sizes, the smallest being 30 um. They were designed and engineered just for your kind of application.
As the pores get smaller and smaller, the exchange times of fluids during the processing becomes longer. However, in this case patience can be a virtue since at 30 um, we believe these capsules to be far superior to the alternatives. And of course, 30 um entities are contained within the volume of the capsule.
Disclaimer: SPI Supplies is a supplier of these microporous capsules and would like to see their use increased! More information about these particular capsules are available on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
I am looking for a cpu-based aid for teaching crystallography at the undergraduate level.
Is anyone in ListLand familiar with CaRIne Crystallography software from France? This is popular in our chemistry dept., but I'm wondering whether other packages exist with comparable (or superior) libraries and algorithms to demonstrate 3D structures, rotate them, make substitutions, correlate structural data with XRD or CBED data, etc, etc.
You can reply to me off-list. If others are interested, I can summarize responses.
Thx, all--
Ann Hein Lehman, EM Facility Mgr Trinity College, Hartford, CT v 860-297-4289 f 860-297-2538 email: ann.lehman-at-exchange.cc.trincoll.edu
Hello to you all! Thanks to everyone for kindly responding to my recent enquiry about the Br in epoxy resins; I apologise for not responding sooner. Your answers were informative and on the basis of most of the suggestions, I favour the idea that the Br is from the resin itself and is not from an external i.e. contaminant source. It appears that Br is added by manufacturers to some types of epoxies. With regard to my particular resin, the company I got it from has still to get back to me. Regards
Hi everybody! I am looking for a used EDS, in good conditions, for installation on Jeol SEM. Any offer? Please inform price and I will estimate the transport & handling costs. Thanks in advance, Sincerely, Nelson Fava/Geosciences Institute/U. Brasilia, Brazil. MIcroprobe CAMECA/SX50#359 and SEM Lab.
3 D imaging of in vitro bio samples is a young technology with many new and exciting facets.
For example: The university of Linz Austria has made great strides in both 1- single molecule fluorescence and in 2- images characterizing the antibody antigen binding forces
Workshop beginning next Week end. See Web: http//:www.molec.com/linz
George
-----Original Message----- } From: James Pawley {jbpawley-at-facstaff.wisc.edu}
Hi Folks,
I was quite surprised that I had the only listing in answer to the call o= f Petra Wahlbring (with an EM912) who had problems of dirt in the TEM imagi= ng system!
Techniques for the identification and positioning of contamination within=
an EM column should be part of the general knowledge of every EM Unit. A= re there people who would like to take on a little more in terms of EM maintenance? Eight years ago we took the Royal Microscopical Society course "Monitoring and Maintaining the Electron Microscope" to Australia and it now runs each October at the University of Sydney, it has also tak= en place in South Africa and Hong Kong.
If there is a demand in the United States, and an organisation who feel they could host such a course, we would be pleased to help.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Thought I'd send out the responses to my post regarding aperture contamination in a VPSEM operated in the high vac mode.
I want to thank everyone who responded. Suggestions covered both improper sample preparation and microscope design problems due to using it in the high vac mode. I have passed the suggestions on and encouraged the SEM lab to contact me if they arrive at a solution so I could post it to the listserver.
Also if any one wants to contact an individual listed below, drop me an email and I can provide their address.
Lou
----- Harry Ekstrom wrote:
So, presumably, the signal is attenuated so much that simply by replacing the final apertures...the problem goes away? Is the inside of the chamber clean? ie Any contamination possibly from the diffusion pump? Other possibilities are outgassing samples causing contamination or a vacuum leak. How is the filament life? ---- Charles Garber wrote:
If these are semiconductor products, unless they are looking at photo resist , there should not be anything to really crud up their column or their apertures.
The only time I have seen this happen is when people were not cleaning the aperture holder and insertion rod properly and contamination was migrating down to the apertures and contaminating them.
The discover of this phenomenon was the basis of the Zaluzec patent and the plasma cleaning interest.
If the vacuum system is acceptable, then you might want to suggest an extra special cleaning of the aperture holder and insertion mechanism. I don't want you to think that I am suggesting this in order to sell them a plasma cleaner. Perhaps this might be of sufficient interest to Nestor Zaluzec at Argonne that your friend could pay him some fee to actually test out if doing a plasma cleaning of his insertion mechanism and aperture holder assembly actually would lead to a reduced contamination rate.
I could of course be wrong about this so you might want to ask Nestor what he thinks of my suggestion. But I think that at Argonne they do have some mechanism for helping industrial problems in that kind of a situation.
------ Steve Chapman wrote:
Dirty final apertures must be down to problems with the gas from the specimen or specimen chamber.
1. Could they use a bigger aperture without any problems? Most people do not push the instrument to its limit so the "normal apertures" could be too small for the application. A cure rather than a prevention!
2. If the specimen chamber has become highly contaminated try this. Pump down the microscope and use an industrial hot air (hair) dryer on ALL the metal areas. The vacuum will almost certainly fall back but as long as you are not trying to heat up an area right by an "O" ring this is what you should expect from the outgasing. Get the chamber hot is the secret.
3. Use an inflow of dry nitrogen gas to cut down the filth that may enter from the environment.
4. Check the rotary pump fluid and its efficiency?
5. What is the vacuum level, have all the LV inlet systems closed correctly?
---- Robert Foglia wtote:
I used to work for a company called High Yield Technology and they were selling an InSitu particle monitoring probe sensor, which could be used in high vacuum. I know they had developed an application for a Hitachi SEM which enabled the user to real time monitor the cleanliness of their system. They initially requested this sensor because they were seeing large amounts of contamination within their system, and the probe sensor was used to monitor while they cycled different valves and moved the robotics around (the robotic arm was scraping). The company is located in Silicon Valley and they might be an option to explore. By the way, they usually place the sensor on the exhaust port in order to get as close to the wafer as possible.
------
John Arnott wrote:
Based on your message concerning the contaminated aperture, my feeling is that the problem probably rests somewhere other than the aperture itself. But since there is the slightest possibility that there might be a problem with the aperture perhaps we can assist you. Ladd produces the vast majority of apertures/microholes used in the United States and the aperture you have may very will be a Ladd aperture. To make sure there is no residual contamination on an aperture we have a post production protocol that is designed to eliminate any contamination that results during production. In fact we have some apertures/microholes used in fluid and gas control that are required to be absolutely contamination free. We have a protocol to ensure that. It is also important that an aperture be shipped or stored in a glass or non-contaminating vial and that the aperture surface should not touch tissue or even lint free cloth. We would be glad to examine the aperture to see if the contamination was a result of packaging, handling or the production process. Please contact us if you if you wish us to do that.
----- Thomas C. Isabell wrote
Regarding the cleaning of SEM apertures, have you considered plasma cleaning?? We have had great success in removing the aperture strip from the microscope, plasma cleaning it in our Model 1020 Plasma Cleaner, and replacing it in the scope. Some of our results were published in a recent MRS proceedings - MRS Symp. Proc. Volume 523, pp 31-38. If you are interested in a copy, I could send one your way. Plasma cleaning returns the aperture strips to their "as new" state and eliminates the need to constantly replace them.
To take this one step further, if the apertures are indeed getting dirty because of dirty specimens, our plasma cleaner has also been shown to effectively clean specimens prior to insertion into the microscope. Some of these results are also shown in the aforementioned MRS Proceedings. Our instrument allows not only the cleaning of the specimen, but also the specimen stub, and if needed, the specimen stage.
If you have any further questions about the technique or our instruments, please do not hesitate to contact me. Additional information can be found on our website, at www.fischione.com .
------ Lisa Montanaro wrote:
There are several reasons why the final aperture may be getting dirty on a variable pressure SEM, and why this may be more noticeable on semiconductor samples. Several causes contribute to contamination within SEM chambers. If this system has been used for any other analysis other than semiconductor, at low vac modes, residues from prior analysis may be lining the chamber walls - these are difficult to remove even by baking the chamber out. Additionally, this system probably has oil-cooled pumps - the deleterious effect of backstreaming oils into chamber which "gums" onto the final aperture is well established, but can be diminished with a strategically located dry N2 purge port in the sample exchange chamber. Mounting media is also of concern - since most conductive paints outgas, these vapors can also contaminate the F.A. quickly, especially if the material is not exceedingly well dried. Simple cleanliness may also be the culprit - by handling a metal sample fixture with bare hands, oils and sweats are deposited on the fixture. These deposits love to attach to the F.A. (and cold tip of EDS detectors too!) As most semiconductor analysis requires fairly high magnification, and often lower KeV, these problems are exacerbated. I've delt with these and many other problems concerning the optimal performance of SEM's with semiconductor materials, and would be pleased to look over his system for performance improvements. Let me know if I may be of assistance!
----- Randy Nestor wrote:
I'll take a stab at the problem. Most of these types of SEM's have vacuum gradients from the chamber to the gun, even when run in the "normal" mode. The worst vacuum is in the chamber, the best in the gun. If your friend has any outgassing of the sample, the contaminants are proalby finding their way to the final aperature. It seems these apts are one way of controlling vacuum levels in different parts of the scope. We have seen similar problems with our Hitachi 2460N. What scope does your friend have?
-----
Ronald Vane wrote:
I have given the following advice to several users of variable pressure SEM who dirty microscopes and contamination, and they all said it cured the problem.
Don't leave it in high Vacuum mode all the time. Some variable pressure SEM are very dirty due to backstreaming at high vacuum. They are designed for low vacuum not high vacuum. By placing the system into low vacuum mode, the chamber is placed into viscous flow vacuum dynamics which stops backstreaming and purges out contaminants. Try leaving it in low vacuum mode overnight for a month and check the results. --- Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html
To : scientist in MSA I'm a graduate student at Electronic Ceramics Lab. Dept. of Ceramic engineering Yonsei University. I've investigated electron diffraction of gas clusters in Korea Institute of Science and Technology. Please let me know electron-atomic scattering factor of In, Zn, and, Ar. Thank you for your effort.
Sung Han p.s. my e-mail address is pibhan-at-kistmail.kist.re.kr
At least for the specimens I happened to deal with so far (vertebrate tissues, cell culture, bacteria, microalgae), the material normally will not really blacken until you wash and start dehydrating it AFTER osmication in traditional aqueous OsO4 solutions. While in aqueous OsO4, it will remain brownish. (Unless, of course, you are using that simultaneous, single solution glutaraldehyde/OsO4 fixation, or a reducing buffer like PIPES, and have passed the time when it all turns black...)
The explanation used to be that, at the stage of osmication, while some of the OsO4, indeed, oxidizes nonsaturated lipids, etc., a lot of it simply dissolves in the lipids of the specimen as nonreduced OsO4. Then, say, the ethanol reduces that specifically accumulated OsO4, to form the so called "osmium black".
As for making FRESH solutions of OsO4 in xylene (or CCl4, etc.), I would just never have a spare OsO4 ampule (and enough paraffine blocks to EM-reevaluate) to try that! :-)
Sincerely, Vlad.
Vladislav V. Speransky Postdoctoral Research Associate School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Ph: 207 581 2998 FAX: 207 581 2969 Email: vladis-at-maine.maine.edu
I'm trying to adapt the TUNEL procedure of labeling cleaved DNA in apoptotic cells for TEM level observation. Basically I just want to rework it to utilize colloidal gold and localize the tagged 3'-OH ends of the degraded DNA. TUNEL has been done for several years at the LM level, but I've only come up with a handful of studies that successfuly nailed it with plastic embedded tissues and gold conjugates... and of those few if any seem to be A+ work (sometimes the specificity of the labelling looks a bit sketchy.) Anyone out there have any experience with this? I'm using a line of chronic myelogenous leukemia cells that has a well characterized apoptotic response to etoposide... previous ultrastructural studies show classic apoptotic changes. Any input will be appreciated.
I'm interested in finding out if there are any variable pressure SEMs available for paid use in the NY/NJ area. Specifically, I'm interested in doing some long-term imaging of large, uncoated specimens at very low mags (10X and below).
Many thanks in advance.
Best regards,
Angela
--------------------------------------------- Angela V. Klaus
Manager, Core Microscopy Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
www.coleoptera.org is working now, please be so kind and send me your comments. There is also large index of entomologist who worked or working on Tenebrionidae...
I apologise for crossposting.
Keep care and be of good cheer.
Regards
Vratislav Richard Eugene Maria John Baptiste of Bejsak (Bayshark)-Collorado-Mansfeld
Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
Temporally home address: 32 Girrawheen Ave. Kiama NSW 2533 Australia e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
I have received a letter asking if I knew of anyone who would donate two used light microscopes for use by clinics in northern Burma. The doctors there are currently using instruments with cracked lenses and no light source and have a desperate need for a better instrument.
If you have a microsacope you could donate please contact me privately and I can give you the details.
Probably the microscopes should have an oil objective and a built-in light source though the latter is not absolutely essential.
We are working with stereo microscope. We are more concern about the ope= rator health of fatigue after peeping into the microscope. The color temp= erature is about 3200K to 3950K. Anyone out there can suggest how long ca= n one (normal person) looks into the microscope without causing fatigue ?= We are now proposing for every 2 hours of looking into a stereo microsco= pe, an operator will rest for 10 minutes. Anyone make any research to fin= d out these ? We do not want our operator to experience epilepsy. Please help. Lim Hian Ho Senior Marketing And Application Engineer Wisma QES, No.6 Jalan USJ 9/5P, UEP Subang Jaya, 47620 Selangor, West Malaysia Tel : 603-7241188 ext 207 Fax: 603-7244488 e-mail : hhlim-at-qes.po.my
Hi everybody in microworld! I am looking for a second hand EDS, simple or equiped with an image soft, to attach to a JEOL SEM. Any offer? Thanks in advance, Sincerely, Nelson Fava. EPMA CAMECA SX50 Lab. Geosciences Institute/U. Brasilia/Brazil.
I am interested in using backscatter elelctron imaging to evaluate the distribution of iron in 1 um plastic sections of biological material. My intent is to provide an independent measure of the distribution of iron in my samples and compare this distribution with the distribution of iron as evidenced through use of the ferrocyanide reaction on adjacent sections. Any comments on the likelihood of success of such an endeavor and the pitfalls of using BEI for sectioned material would be appreciated.
Thank you very much,
WB Jaeckle
============================== Will Jaeckle Department of Biology Illinois Wesleyan University P.O. Box 2900 Bloomington, IL 61702-2900 TELE: 309-556-3779 FAX: 309-556-3864 EMAIL: wjaeckle-at-titan.iwu.edu =============================
Hi everybody in microworld! I am looking for a second hand EDS, simple or equiped with an image soft, to attach to a JEOL SEM. Any offer? Thanks in advance, Sincerely, Nelson Fava. EPMA CAMECA SX50 Lab. Geosciences Institute/U. Brasilia/Brazil.
Having been a subscriber to this list for half a year now, I'm convinced that someone can help me with the following problem. I want to use heparin conjugated with gold, without any "bridge" such as albumin in between, but I cannot find any commercial source. Since I'm a beginner in the gold business, is there any good recepy for the preparation of heparin-gold?
In a message dated 99-01-26 05:11:00 EST, hhlim-at-qes.po.my-at-sparc5.microscopy.com writes:
{ { Anyone out there can suggest how long can one (normal person) looks into the microscope without causing fatigue ? We are now proposing for every 2 hours of looking into a stereo microscope, an operator will rest for 10 minutes. } }
Lim,
I do not have any research on the subject, but the two most common causes of fatigue are: (1) eyestrain and (2) the physical strain on the neck, upper back and arms from staying in one position for such a long time. As far as eyestrain, I think your proposal for two hour blocks of time is realistic.
You don't mention the type of microscope, but if it is a fairly modern one you can probably obtain a tilting "ergotube" for viewing, so the operator can find the most comfortable position and adjust the viewing angle as needed. If this is not possible, you can also get an ergonomic table that can be raised and lowered either mechanically or via a motor. You can place the scope on the table and adjust the height to suit the operators.
You might also consider putting a video system on the microscope so the operator can choose to either look through the eyepieces or at a high resolution monitor. There are also video inspection stations that can take the place of the microscope entirely.
Good luck to you.
Best regards,
Bob ****************************** Robert (Bob) Chiovetti, Ph.D. President Microimaging Technologies, Inc. Tucson, Arizona USA Tel./Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems Clinical, Research & Industrial Microscopy Cytology/Histology/Pathology/EM *******************************
I thought the "gold standard" for apoptosis is the ultrastructural changes that can be seen by TEM and that the DNA labeling was developed in order to view apoptosis/vs necrosis at the light level-over a larger area of tissue? At least that is the idea I've gotten from articles that I have read. Could someone please clarify this? Thanks in advance.
Karen Pawlowski Sr. Research. Assoc. UT Southwestern Med. Ctr. PhD candidate, UT Dallas
On Mon, 25 Jan 1999, Doug Matthews wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi everyone, } } I'm trying to adapt the TUNEL procedure of labeling cleaved DNA in } apoptotic cells for TEM level observation. Basically I just want to rework } it to utilize colloidal gold and localize the tagged 3'-OH ends of the } degraded DNA. TUNEL has been done for several years at the LM level, but } I've only come up with a handful of studies that successfuly nailed it with } plastic embedded tissues and gold conjugates... and of those few if any seem } to be A+ work (sometimes the specificity of the labelling looks a bit } sketchy.) } Anyone out there have any experience with this? I'm using a line of } chronic myelogenous leukemia cells that has a well characterized apoptotic } response to etoposide... previous ultrastructural studies show classic } apoptotic changes. Any input will be appreciated. } } Doug Matthews } } }
We offer the 1.4 nm Nanogold=AE gold cluster label with several reactivities for covalently labeling biomolecules. If you can selectively oxidise the terminal reducing sugar of heparin to give an aldehyde residue, it should react with Mono-amino-Nanogold=AE (you then reduce the resulting Schiff base and purify the conjugate by gel filtration). Alternatively, if you can selectively expose an amino- group, you can conjugate this with Mono-Sulfo-NHS-Nanogold=AE.
Protocols for using these reagents are posted on our web site (http://www.nanoprobes.com/Inf2021.html and http://www.nanoprobes.com/Inf2025.html). I hope this helps,
Rick Powell Nanoprobes, Incorporated
} } Dear all, } } Having been a subscriber to this list for half a year now, I'm convinced } that someone can help me with the following problem. } I want to use heparin conjugated with gold, without any "bridge" such as } albumin in between, but I cannot find any commercial source. Since I'm a } beginner in the gold business, is there any good recepy for the preparation } of heparin-gold? } } Thanks in advance! } } Anne
****************************************************************** * NANOPROBES, Incorporated | US Toll-free: (877) 808-2101 * * 25 East Loop Road, Suite 113 | Tel: (516) 444-8815 * * Stony Brook, NY 11790-3350, | Fax: (516) 444-8816 * * USA | rpowell-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
Hi everybody in microworld! I am looking for a second hand EDS, simple or equiped with an image soft, to attach to a JEOL SEM. Any offer? Thanks in advance, Sincerely, Nelson Fava. EPMA CAMECA SX50 Lab. Geosciences Institute/U. Brasilia/Brazil.
Hi everybody in microworld! I am looking for a second hand EDS, simple or equiped with an image soft, to attach to a JEOL SEM. Any offer? Thanks in advance, Sincerely, Nelson Fava. EPMA CAMECA SX50 Lab. Geosciences Institute/U. Brasilia/Brazil.
The Science of Biological Specimen Preparation for Microscopy
Edited by Marek Malecki and Godfried M. Roomans
Proceedings of the 14th Pfefferkorn Conference, Belleville, IL
Hardbound book with 31 papers; 466 + xii pages. Table of Contents available on request.
Published by and available from: Scanning Microscopy Intl., Box 66507, AMF O'Hare (Chicago), IL 60666-0507= , USA FAX: (847) 985-6698 / E.mail: 73211.647-at-compuserve.com
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BOOK REVIEW (published in January 1999 issue of the Journal of Biomedical=
Optics; vol 4 pages 191-192):
Advance in our knowledge of the molecular organization of living material=
rests on two kinds of technological development: (1) Microscope instrumentation (2) Specimen preparation. This includes new techniques=
appropriate for a specific mode of microscopy, and information on any structural alterations produced during such specimen preparation.
The present volume contains the proceedings of the 14th Pfefferkorn Conference organized by Scanning Microscopy International and dedicated t= o the late Prof. G.E. Pfefferkorn.
The organizors and editors of this conference brought together 31 leading=
scientists in this field from the USA, Europe and Japan, to present and discuss new techniques to prepare biological specimens for microscopy. =
Discussions with a panel of reviewers are added to each paper. There is not space for a detailed discussion of each paper. I shall try to indica= te the general substance of the contributions.
In general, three unique features of this book are worth stressing.
The first, it reflects current trends in science towards interdisciplinar= y approaches in solving important biological questions. An essential part = of incorporating microscopy into these interdisciplinary approaches is development of reporter molecules which will have features precisely determined in biochemistry and molecular biology labs and which will rema= in stable in changing environment of living cells being labeled for various modes of microscopy. Chapters by Heinfeld - nanogold, Kessels - boronate= d antibodies, Malecki - organo-metallic ligands, and Swartz - fluorescent derivatives of proteins are concerned with the development of such new probes which are based upon covalent bonds, therefore they can serve as reliable markers for structures of interest. Moreover, chemically define= d features can be translated into their functional architecture e.g., as shown for atomic force microscopy in the chapter by Woodward and Zasadzinski or for life time imaging by Lakowicz. The detailed protocols=
will allow an investigator to have them easily modified for a particular new application. This book also demonstrates how fruitful this interdisciplinary approach can be e.g., as demonstrated in chapters by DeBault and Gu, Malecki, and Nuovo, polymerase chain reaction developed f= or DNA amplification in molecular biology labs can be modified to determine morphological localization of selected sequences. The book clearly demonstrates, that in order to solve a biological question, it is nearly impossible to categorize approaches and restrain an investigator to traditional research techniques. To the contrary, the interdisciplinary approaches create backgrounds and precedents for breaking the boundaries = of traditional and specialized areas of science and opening new possibilitie= s.
The second, this book is also an attempt toward integrated microscopy. =
While many already published books were dedicated to very specialized are= as of one kind of microscopy, this one stands out as an eclectic (in a positive sense), but comprehensive review of the most recent developments=
in various areas of microscopy. This is particularly close to my scientific preferences, since I promoted this approach for many years. =
Integrated microscopy, allows us to overcome technical limitations of one=
kind of microscopy alone e.g., light microscopy of a living cell is limit= ed by resolution ~250 nm, while electron microscopy having atomic level of resolution can be pursued only on frozen or fixed cells, therefore studyi= ng the same cells with both types of microscopy creates an opportunity to study living cell phenomena at the molecular level. This approach is elegantly exemplified in the chapters by Malecki, Peachey et al., Ralston=
and Ploug. It also demonstrates how beneficial and cross-fertilizing thi= s integrating approach can be e.g., in the chapters by Lyubchenko where a technique of functional modifications of substrate surface used in TEM an= d SEM, now finds new applications in AFM. These chapters create not only a= n excellent starting point for a reader, but also collecting many other techniques in one book, opens for an investigator a compendium of choices= . =
The newest developments in specimen preparation for two-photon excitation=
fluorescence microscopy, atomic force microscopy, life-time imaging, ener= gy filtering transmission electron microscopy, etc. are all covered in this one volume. Therefore, scientists attempting to solve a life sciences problem with tools of modern microscopy can make a choice from the vast variety of techniques and examples presented in this book. The detailed hands-on specimen preparation protocols will guide them through. The discussions with the reviewers will provide them with the critical evaluation of choices.
The third, the book paves the road for future developments in microscopy.= =
The chapters and discussions with the reviewers, fairly define current technical limitations of various modes of microscopy and suggest possible=
ways towards overcoming these difficulties. In many cases, authors clear= ly state the directions of their future research. Therefore, not only it is=
updated information concerned with the status-quo, but also a proposal fo= r future research.
Specifically, the first group of papers deals with methods to study chromatin and nucleic acids. In the first paper M. Malecki describes new=
methods of gene transfer, which were developed based upon incorporation o= f reporter molecules allowing us imaging of cellular pathways in living cells, and in cryo-immobilized cells by energy filtered TEM from the cell=
surface to the chromatin. G.V. Childs describes methods to identify mRNA=
and proteins in the same cell, in tissue sections. L.E. De Bault and J. = Gu present detailed protocols for in situ hybridization, in situ transcripti= on and in situ polymerase chain reaction. M. Thiry developed the in situ terminal transferase - immunogold technique to pinpoint specific nucleic acid regions in thin sections. Scanning probe microscopy is represented = by five contributions. Hydration Scanning Tunneling Microscopy (Heim et al.= ) is based on conductivity of surface adsorbed water molecules and can imag= e hydrophilic insulators and biological specimens such as collagen IV molecules, TMV, and cryo-sectioned bovine tendon.
For atomic force microscopy imaging of macromolecules, a strong attachmen= t to the substrate is essential. Lyubchenko et al. show that treatment of mica with aminopropyltriethoxy- silane will hold DNA in place for imaging=
even in water. They also introduced other chemically reactive mica surfaces, hydrophobic or charged. Mueller-Reichert and Gross discuss DNA=
and DNA-protein assembly analyzed by TEM, Scanning Tunneling Microscopy (STM) and Atomic Force Microscopy (AFM). An interesting new application = of STM is imaging of freeze-fracture replicas (Woodward and Zasadzinski). I= t also can examine interior interfaces and provides quantitative informatio= n about the vertical dimension of interior structures.
Four contributions discuss light microscope techniques for the imaging of=
living cells. The first of these by N.S. Allen and M.N. Bennet use Alfal= fa root hairs before and after treatment with Nod factors (produced by Rhizobia) to study in live and fixed cells the role of actin and endoplasmic reticulum in growth form change. Imaging was with a confocal=
laser scanning microscope. The paper by Peachey et al. describes techniques to image cultured cells by phase, epifluorescence and confocal=
microscopy, and after fixation and critical point drying image the same cell as whole mount with the Jeol 400kV-EX intermediate voltage TEM, correlating structures seen in the living cell with the EM image. The paper by D.R. Swartz on covalent labeling of proteins with fluorescent compounds for imaging applications beautifully illustrates with alpha-actinin how a protein can be covalently linked to a fluorophor without interfering with its normal chemical interactions in the living cell. This paper is important for all who need fluorescently labeled cel= l proteins for imaging applications. L. Edelman and A. Ruf describe a simp= le treatment to stabilize freeze-dried cells during or after low temperature=
embedding in Lowicryl, to prevent loss of material from thin sections during wet cutting.
J.F. Hainfeld describes the techniques for labeling with Nanogold, Undecagold and FluoroNanogold. This paper is essential for anybody who requires gold labeling. The increasing availability of energy filtered TEMs will facilitate immuno-cytochemistry by providing electron spectroscopic imaging. Kessels et al. describe the development of organo-boron compounds which can be incorporated into organic molecules such as Fab'-boronated peptide conjugates for immunochemistry.
The last paper by H. Sitte contains a masterful critical review of cryofixation and the blueprints on how to build a cryomicrotome that can provide useful cryosections. I think that these seventy pages contain th= e most valuable part of this volume.
Finally, this volume obviously contains a wealth of stimulating and usefu= l information. It bears testimony that microscopy is alive and well.
Hans Ris, Zoology and Integrated Microscopy Resource, University of Wisconsin, Madison, WI
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Copies of several other reviews are available on request.
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Price: $95 (US delivery by uninsured mail) or $105 (elsewhere by uninsure= d mail); insured delivery, air mail, etc., are available for additional cos= t, please inquire.
REPRINTS or photo-copies of papers are available for (rates for delivery = by cheapest mail method); EACH: $5 (up to 8 pages); $10 (9-16 pages); $15 (17-24 pages) and $20 (25+ pages).
Recently we have been having problems with a resin mixture that we have been using for two years, until recently with good results. I am wondering if anyone else has any ideas about what might be the problem.
The resin is a mixture of Araldite 6005 (about 23% by wt.) SPIpon 812(about 23%) and DDSA (about 54%). We store the unopened resin bottles in the refrigerator, but warm them fully before opening. The opened, capped bottles are stored in a vented cabinet at room temperature. We typically make about 55 ml of resin at a time and store it without the accelerator (DMP-30) in the freezer. Again, we always make sure the bottle is completely warmed to room temp before opening it to use. To embed, we dehydrate through propylene oxide and then 50:50 PO:resin overnight. The next day the blocks are rolled for 4 hours or so in 100% resin + 1.5% DMP-30, then polymerized in fresh resin with accelerator for 24-48 hours at 60=B0.
Several months ago we began getting blocks which are difficult to trim smoothly; they seem to chip and crack. Although the resin sections well, when sections are picked up they tended to disintegrate, or at best, showed rifts and cracks throughout the resin. Increasing the curing time helps a bit, but does not eliminate the problem. We have tried new bottles of all the resins and the accelerator, without improvement. However, in some cases the new bottles may be from the same lots as the old (we haven't kept records of resin lot numbers).
Does this sound familiar? Specifically, we are wondering about storage and shelf life of the resins. Are we inviting problems by storing unopened bottles in the fridge (we have done so for years, but with the old Epon 812)? What about shelf life? The unopened bottles are not more than a year old. Do you store your opened resin bottles in desiccators? At what temperatures? How do you store resin mixtures? Finally, should DMP-30 be added at the 50:50 stage? We had not done this in the past, did for a year or so, then stopped doing it again and thought this wasn't a problem, but perhaps it is. Any ideas would be welcome. Thanks
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 =46ax: 860-4861936=20
Colleagues, can anyone help this person? Reply directly to him ...
Nestor Your Friendly Neighborhood SysOp -------------------------
Email: micnaut-at-aol.com Name: julius simon School: retired chemist school volunteer
State: florida
Question: i would like a description of a procedure for smear preparations of onion root tips to demonstrate the varios stages of mitosis-that could be quickly mastered by ahigh school bio student. I have a good scope with an automatic ca mera
Hello Doug, I have tried the TUNEL technique for EM. I got a very clean and specific labelling of heterochromatin in ALL nuclei, normal and apoptotic. My explanation is that colloidal gold labels only on the surface of the section. And there will be a lot DNA breaks at the surface due to sectioning and that is where the reaction will occur. As far as I can remeber, I could find only two references for EM. One of them (Thiery?) used this technique as a staining for all DNA. If you want more info, please contact me and I will dig deeper to find the references and the protocol I used (it worked beautifully on paraffin sections). Yours sincerely,
Sarka Lhotak
Hamilton Regional Cancer Centre Hamilton, Ontario, Canada lhotaks-at-mcmaster.ca
One of the most common causes of brittle tissue is exposure to propylene oxide (or a mixture containing it) in the presence of air, i.e. allowing the tissue to become exposed when in a propylene oxide (or PO:resin mixture) stage.
One or more of your ingredients going "off" could also be the cause of your problem, as you suggest. However, in over 30 years of TEM embedding with a whole variety of raw materials, many of which have stood on the shelf for years, we have very seldom experienced this sort of problem.
For a reliable protocol and resin mixture how about trying the one described in a paper I published several years ago? The reference is: Cross, R.H.M. (1989) A reliable epoxy resin mixture and its application in routine biological transmission electron microscopy. Micron & Microscopica Acta, 20(1), 1 - 7.
Good luck!
Robin
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za (or eurc-at-giraffe.ru.ac.za) tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
This is similar to a situation I was in many years ago. In my case it was self-induced because I was experimenting to find a mixture which would cut very large ultrathin sections. In that instance (as far as I can remember - it was late 1970's) I used a high anhydride:epoxy ratio combined with a partially hydrated anhydride. The latter was obtained by exposure to air over a long period, like the bottle was 10 years old before I got to it and it was pretty viscous.
Maybe your mixture already has the high A:E ratio and the anhydride has aged. Although I would not have thought that brittlenss would be the problem, DDSA is the long chain component whereas I was using both DDSA and MNA in my recipe and the MNA was the component which really gave hardness.
Brittleness was a sign of good cutability but it could be overdone! Then the sections would break up on cutting. One block would cut 3 mm square sections, and if you went down to grey in colour, they would simply dissolve on the water! They left bits of fixed tissue which were sort of de-embedded (I never loooked at these).
Brittleness was also induced by a long infiltration time - 48 hours at 35 Centigrade, then 24 hours at 45, followed by 24 hours at 60 C. This procedure induces sterical hindrance which catches the big molecules in certain configurations whereby they cannot form complete cross-linkage. A sign of this is that the blocks are thermoplastic - if you heat them in an oven, they get soft, you can impress the surface with tweezer tips etc. IS THIS TRUE OF YOURS?
Possibly you have aged components, or they have taken up water.
Alternatively, maybe your dehydration isn't complete. Water in the alcohol or acetone, or maybe propylene oxide (? don't know about this possibility, we haven't used it for 20 years or more because of the health aspect). Try adding molcular sieve to a sample of your dehydrating agent?
First of all it would be useful to clarify whether we are talking about a true stereo microscope or a binocular microscope. I personally find stereo microscopes a little more comfortable to work with - perhaps because you are looking at something with a more natural perspective and light than when examining a slide in transmitted light.
I would certainly think that a two hour working period may be possible but I would suspect that there might be problems with general fatigue, attention span, and even RSI (repetitive strain injury) if stage controls are being rotated very regularly. It would probably be necessary during that time to encourage 'micro-breaks' (meaning breaks of a few seconds every 10 or 15 minutes - rather than 'microscope breaks') as you should for display screen equipment/computers. The general rule of thumb with computer rest periods is little and often rather than saving time up over a couple of hours and I suspect that the problems are similar with microscopes.
Other considerations should include ergonomics (Microscope, work area, seating and the people using it - which of course is part of the ergonomics equation).
I hope this helps.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: "RCHIOVETTI-at-aol.com" To: hhlim; microscopy
Dear Marie,
Firstly, I 'm not quite sure how to store SPIPON-812, you should ask the manufacturer, but with my knowlege, all epoxy resins are stored at room temperature in a cool and away from direct sun light. Low temperature storage is not recommended for epoxy resins, especially araldites and anhydrides, such as Araldite 502, 6005, DDSA, NMA, NSA. DER... Meantime, DMP-30, DMAE, BDMA, they are contained amino group (-NH-), storing them in the refrigerator will increase its shelf life. Storing epoxy resins in the desiccator is the best. Secondly, you can store the resin mixture (without the addition of DMP-30) in the refrigerator for later use, but only 3 to 4 weeks is maximum, with well protected from contamination with moisture. After all, I think your sections problem is associated with the storage condition of your resins. For more general tips for embedding media, please refer to Electron Microscopy Sciences catalog XIII, page 48.
Sally Stowe wrote: } } We are just setting up TEM EDXA in a multidisciplinary unit, and need to acquire standards covering a wide range of applications. Any recommendations, pitfalls etc? (Philips 430 TEM up to 300kV, Oxford ISIS ultra-thin window detector). } thanks
Dear Sally, Chuck Fiori published an article about using lithium borate glass standards for biological work--the matrix is about the same as that for resin/organic material. There may be someone who sells these or similar standards. For frozen-hydrated work, one can dissolve a known amount of an appropriate compound in a sucrose solution which approximates the composition of tissue and cut cryo-sections. For materials work, there are some commercially available standards. It is important that the matrix of the standard match that of the unknown, so one type does not fit all. Good luck. Yours, Bill Tivol
Dear Anyone: I'm interested in finding out programs to simulate and to index electron diffraction patterns. If it is possible, for windows. Thaks in advance
Patricia Bozzano Comision Nacional de Energia Atomica Buenos Aires, Argentina.
Cytotechnologists who screen Pap smears, among other things, sit at a microscope up to 8 hours daily. Comfort is obviously important. Among the usual recommendations: sit upright, eyes forward (no bending of the neck), use elbow or forearm rest pads (e.g., "Wedgies"), feet flat on the floor (though some use a foot rest), break for 10 minutes hourly to stimulate circulation and minimize the vigilance decrement, maintain room light at a level comparable to the illumination intensity to minimize contrast differences and eyestrain, and look "through the eyepieces" (i.e., at infinity, relaxed accommodation), rather than "in the microscope".
I've coined a term to describe the collective response to the various ills (e.g., varicose veins, hemorrhoids, panorama butt, elbow calluses, repetitive strain injury, self-inflicted stab wounds from the dotting pens, eyepiece grooves under the eyes, and bruised orbits) that can beset a long sitting cytotechnologist: hyperchronokathistophobia, which translates to "fear of sitting for extended periods of time." Think we can claim workman's compensation and take early retirement?
Gary Gill
-----Original Message----- } From: "RCHIOVETTI-at-aol.com"-at-sparc5.microscopy.com [mailto:"RCHIOVETTI-at-aol.com"-at-sparc5.microscopy.com] Sent: Tuesday, January 26, 1999 11:32 AM To: hhlim-at-qes.po.mysparc5.microscopy.com; microscopy-at-sparc5.microscopy.com
In a message dated 99-01-26 05:11:00 EST, hhlim-at-qes.po.my-at-sparc5.microscopy.com writes:
{ { Anyone out there can suggest how long can one (normal person) looks into the microscope without causing fatigue ? We are now proposing for every 2 hours of looking into a stereo microscope, an operator will rest for 10 minutes. } }
Lim,
I do not have any research on the subject, but the two most common causes of fatigue are: (1) eyestrain and (2) the physical strain on the neck, upper back and arms from staying in one position for such a long time. As far as eyestrain, I think your proposal for two hour blocks of time is realistic.
You don't mention the type of microscope, but if it is a fairly modern one you can probably obtain a tilting "ergotube" for viewing, so the operator can find the most comfortable position and adjust the viewing angle as needed. If this is not possible, you can also get an ergonomic table that can be raised and lowered either mechanically or via a motor. You can place the scope on the table and adjust the height to suit the operators.
You might also consider putting a video system on the microscope so the operator can choose to either look through the eyepieces or at a high resolution monitor. There are also video inspection stations that can take the place of the microscope entirely.
Good luck to you.
Best regards,
Bob ****************************** Robert (Bob) Chiovetti, Ph.D. President Microimaging Technologies, Inc. Tucson, Arizona USA Tel./Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems Clinical, Research & Industrial Microscopy Cytology/Histology/Pathology/EM *******************************
Marie, Our lab uses a variety of resins, ie, Spurrs, EMbed 812, Eponate 12, araldite, some very old Epon 812, and we store all of it at room temp. away from any water. We've had no problems with any of it. We also mix all ingredients including accelerator, for at least 15-20 min. on a stir plate. The more viscous resins need to be mixed first with a tongue depressor or applicator stick first for a minute or so. We do add the accelerator (BDMA, DMP-30, or DMAE)to the resin for the overnight 50:50 PO:resin step. The next morning we add 2 changes of fresh resin (again, with accelerator) for an hour each to make sure all the PO is gone and then embed at 60 degrees for 48 hrs. The only problem I've ever encountered similar to yours was from using PO that had been opened alot and was nearly empty. It evidently had gotten moisture in it. You might also check your absolute alcohol for moisture.
Mary Gail Engle Electron Microscopy & Imaging Facility] University of Kentucky
This may have nothing to do with your problem but we used to have seasonal changes in our block/resin quality before we got a proper air conditioning/heating system. Every summer when it was humid the resultant blocks would be soft and sticky, every winter hard and brittle. To help alleviate this problem our last step before actually embedding was to place the tissues in pure resin and let them sit in a vacuum for an hour or so. This might help if for some reason your humidity level in the lab has changed lately.
Pat Hales Dept. of Anatomy & Cell Biology McGill University hales-at-med.mcgill.ca
Institute of Physics EMAG '99 Conference - 25-27th August 1999, Sheffield= U.K. (Co-sponsored by RMS and IOM)
Electron and scanning probe microscopy and related techniques.
Scientific Content This biannual, three-day conference aims to bring together international S= cientists and Engineers both in Industry and Academia who employ electron and scanning p= robe microscopies, together with associated techniques such as surface science,= in both imaging and analytical applications. All aspects of Electron and Scanning Probe Micros= copy/ Spectroscopy will be discussed including:
o New Instrumentation (particularly Field Emission systems), Imaging and A= nalytical Techniques o High resolution Electron Microscopy and Electron Crystallography o Advanced SEM, Scanning Probe and Surface Science o Advances in Microanalysis and Elemental/Chemical Imaging o Microscopy of Catalysts, Sensors and Environmental Materials o Ferrous/ Non-ferrous metals and Intermetallics o Carbons, Ceramics, Electroceramics and Composites. o Semiconductors, Superconductors and Magnetic Materials. o Polymers o Microscopy of Interfaces and Surfaces
Sessions will be arranged by topic and through international keynote plena= ry lectures, invited and contributed papers, both oral and poster, current state-of-the-art iss= ues in the field will be highlighted and discussed. Papers from postgraduate students are particula= rly encouraged; correspondingly registration fees will be kept low and student bursaries w= ill be available through the Institute of Physics EMAG group.
o On Tuesday 24 August, prior to the conference, there will also be an Adv= anced School on HIGH RESOLUTION ELECTRON MICROSCOPY. Contact: c.j.hetherington-at-sheffield.ac.uk for further details
o A major trade exhibiton will be mounted in the purpose built Octagon Cen= tre which will run in parallel with the conference. Contact: i.m.reaney-at-sheffield.ac.uk for further details
SUBMISSION OF ABSTRACTS - DEADLINE MARCH 19 1999 Both oral and poster contributions are invited. Abstracts should be approx= . 300 words in length and may include figures and diagrams.
Please submit abstracts either by:
o WWW see http://www.iop.org/Confs/EMAG
o Email send blank email to confs-at-ioppublishing.com with "EMAG instructions" as th= e subject
o FTP download the files "readme.txt" and "EMAG.tem" from ftp.ioppublishing.com/outgoing/conferences/submisssions/EMAG/
In all cases please indicate whether you would prefer either ORAL or POSTE= R presentation and also which sessions you would prefer your contribution to be included = within.
Nominal Sessions o Interfaces and Surfaces (INT) o Electron Crystallography (ECR) o Catalysts/Sensors/Env. Materials (CAT) o High Resolution Electron Microscopy (HRM) o Semiconductors and Superconductors (SSC) o SEM/EBIC/CL (SEM) o Analytical Electron Microscopy (AEM) o Advanced Scanning Probe Techniques (SPT) o Ceramics/Carbons/Composites (CCC) o Metals/Intermetallics (MET) o Plenary (PLE)
STUDENT BURSARIES Students and young scholars are encouraged to apply for an EMAG bursary wh= ich will help meet costs incurred in attending the conference. Applications should = be sent to Dr R. Brydson, Department of Materials, School of Process, Environmental a= nd Materials Engineering, University of Leeds, Leeds LS2 9JT. email: mtlrmdb-at-leeds.ac.uk
GENERAL ENQUIRIES Contact: conferences-at-iop.org
_____________________________ Dr. Rik Brydson, University Research Fellow, Electron Optical Unit, Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
The methods that I mention have been used for several years with undergraduate students and can produce quite good results. Success seems to depend, in part, on choosing the right sort of onion tips at the right time.
GROWING ROOT TIPS We have tried both seeds and bulbs and have only really had good results using root tips from onion bulbs which have been left to sprout over suitable beakers of water (the bulb should rest on the neck of the beaker and almost touch the water and after 1 to 2 weeks you should have a lot of tips - CAUTION supermarket onions may be incapable of sprouting roots you may have to try fresh ones). It is also possible that mitosis may vary at different times of day so you may have to experiment with 'harvesting times'.
STAINING METHODS These are not my methods but have been handed down since the dawn of time - the results are temporary mounts which should be examined soon after preparation. Caution the stains are harmful and acids are used to hydrolyse the cell walls/tissue. Also both methods use acetic acid solutions so the smell can be overpowering in an open lab. DO YOUR RISK ASSESSMENT FIRST. FEULGEN STAIN TECHNIQUE 1 Cut 1cm of root tip off onion, 2. fix in acetic alcohol (3:1) for at least 30 min 3. Hydrolyse in 1 molar/normal HCl at 60 deg C 10 - 15 min 4 Stain in Feulgen reagent for minimum of 60 min (IN THE DARK) - root tip will become purple in meristem 5. Without letting specimen dry out = place the end 2-3mm of root tip on a clean slide with a drop of 45% acetic acid 6. Cut root tip into 4 longitudinally then gently and carefully squash under a coverslip - protecting your thumb several layers of paper towel. 7. Examine for mitosis ACETIC ORCEIN TECHNIQUE 1, Cut 1cm of root tip off onion, 2. put into watch glass with concentrated HCl and absolute ethanol (1:1) for 10 min - NB take care when mixing this reagent 3. transfer root tips to watch glass with 45% acetic acid for 5 min 4. Without letting specimen dry out = place the end 2-3mm of root tip on a clean slide with a drop of ACETIC ORCEIN 5. Cut root tip into 4 longitudinally then gently and carefully squash under a coverslip - protecting your thumb several layers of paper towel. 6. Examine for mitosis
Good luck and take care.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
---------- } From: micnaut To: Microscopy
Colleagues, can anyone help this person? Reply directly to him ...
Nestor Your Friendly Neighborhood SysOp -------------------------
Email: micnaut-at-aol.com Name: julius simon School: retired chemist school volunteer
State: florida
Question: i would like a description of a procedure for smear preparations of onion root tips to demonstrate the varios stages of mitosis-that could be quickly mastered by ahigh school bio student. I have a good scope with an automatic ca mera
by smtp.uky.edu (8.8.8/8.8.8) with ESMTP id OAA12519 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 27 Jan 1999 14:52:35 -0500 (EST) Received: from mdc.novel.uky.edu ([128.163.67.41]) by pop.uky.edu (8.8.8/8.8.8) with SMTP id OAA13421 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 27 Jan 1999 14:52:35 -0500 (EST) Message-Id: {3.0.5.32.19990127145132.007f1700-at-pop.uky.edu} X-Sender: mgengle-at-pop.uky.edu X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.5 (32)
Marie, Our lab uses a variety of resins, ie, Spurrs, EMbed 812, Eponate 12, araldite, some very old Epon 812, and we store all of it at room temp. away from any water. We've had no problems with any of it. We also mix all ingredients including accelerator, for at least 15-20 min. on a stir plate. The more viscous resins need to be mixed first with a tongue depressor or applicator stick first for a minute or so. We do add the accelerator (BDMA, DMP-30, or DMAE)to the resin for the overnight 50:50 PO:resin step. The next morning we add 2 changes of fresh resin (again, with accelerator) for an hour each to make sure all the PO is gone and then embed at 60 degrees for 48 hrs. The only problem I've ever encountered similar to yours was from using PO that had been opened alot and was nearly empty. It evidently had gotten moisture in it. You might also check your absolute alcohol for moisture. Good luck!
Mary Gail Engle Electron Microscopy & Imaging Facility] University of Kentucky mgengle-at-pop.uky.edu
Marie, Our lab uses a variety of resins, ie, Spurrs, EMbed 812, Eponate 12, araldite, some very old Epon 812, and we store all of it at room temp. away from any water. We've had no problems with any of it. We also mix all ingredients including accelerator, for at least 15-20 min. on a stir plate. The more viscous resins need to be mixed first with a tongue depressor or applicator stick first for a minute or so. We do add the accelerator (BDMA, DMP-30, or DMAE)to the resin for the overnight 50:50 PO:resin step. The next morning we add 2 changes of fresh resin (again, with accelerator) for an hour each to make sure all the PO is gone and then embed at 60 degrees for 48 hrs. The only problem I've ever encountered similar to yours was from using PO that had been opened alot and was nearly empty. It evidently had gotten moisture in it. You might also check your absolute alcohol for moisture. Good luck!
Mary Gail Engle Electron Microscopy & Imaging Facility] University of Kentucky mgengle-at-pop.uky.edu
1. I suggest that you not store your resins in the refrigerator. Room temp is best, tightly closed
2. Make sure that your propylene oxide is dry. Keep it strictly closed. Do not use small amounts which have been sitting in bottles.
3. Look up the dehydration schedule for your tissue in a textbook, or a paper. For instance, liver and tongue (same size blocks) need vastly differing dehydration and infiltration schedules.
4. Crucially important: Where did you get your formulation for your resin mixture? Does it meet the requirements of WPE weights of the components for a good TEM block? Have you contacted the manufacturer about any changes they might have made recently? Are you dealing with companies that are specialists for TEM supplies and carefully redistill all components once they get them from the manufacturer? (Resins bought after manufacture may contain many impurities at totally random times). Why are you using 6005? It is possible that your formulation is only marginally good, and is easily "derailed" by minor differences in environments.
5. Also crucially important: All resin coming into contact with tissue either during infiltration or during the final steps must be accelerated. Accelerating only part of the resin coming into contact with tissue used to be popular in the 60s. It is no longer considered good practice, because it yields uneven infiltration.
6. Is your accelerator dry? Your accelerator should not be older than 6 months. It should be kept strictly closed and at room temp.
We have 5 direct drive vacuum pumps (Hitachi CuteVac) that are in need of repair. They are leaking oil (suspect the oil seal is shot) but otherwise pump fine. Does anyone know where we could have them repaired? - Hitachi does not do this type of work and I hate to throw them away at $3,000 each.
Thanks,
John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I am hoping that somebody knows the answer to this;
I have been asked to look at some chromosomes in the TEM, and am wondering if someone knows of a good reference on stains that can be used for this work.
If you know of a guide or papers on the preparation tasks involved that would be of great help.
Many thanks
David
Dr. David C. Bell Room 13-1818 E-mail: dcb-at-MIT.EDU Center for Mat. Sci. and Eng. PH: (617) 253-3317 Massachusetts Institute of Technology FAX: (617) 258-6478 77 Massachusetts Ave, Cambridge, MA 02139
Postdoctoral positions at Karolinska Institutet, Stockholm
Membrane protein structure/Electron crystallography
Two postdoctoral positions are immediately available for work on structure determinations of membrane-bound enzymes using cryo electron microscopy. Two-dimensional crystals are available of proteins both in their native states and in the form of reaction intermediates. The work will initially be concentrated on data collection and processing. Previous experience in the field of electron crystallography is advantageous.
The Center for Structural Biochemistry (CSB) is a unit at the Department of Biosciences, Karolinska Institutet and comprises research groups with facilities for X-ray crystallography, NMR spectroscopy, electron microscopy and molecular modelling. CSB is located at the South Campus of Karolinska Institutet close to Stockholm, encompassing the Novum Research Center and Huddinge University Hospital. Excellent resources will be provided.
The postdoctoral positions/research associates are initially for two years with possibilities for extensions for up to four years.
Applications should be sent to Dr. Hans Hebert, Karolinska Institutet, Department of Biosciences at Novum, S-141 57 Huddinge, Sweden together with a CV and the names of two referees. Inquiries: Hans.Hebert-at-csb.ki.se or Philip.Koeck-at-csb.ki.se.
Qualifications (education, certification, language, etc.) and Experience required: A candidate with a BS or MS degree in polymer science, material science or chemistry is preferred with some prior experience in electron microscopy. Good written and oral communication skills and the ability to work both independently and in a team environment are extremely important.
Job Overview:
The Surface/Microscopy/X-ray (SMX) Group in Dow's Corporate R&D Analytical Science Laboratory has two professional level full time openings for Polymer Microscopists, one position at each of the Dow's facilities in Midland, MI and Freeport, TX. The primary responsibilities include working with partners to support research projects involving new and existing products in Dow's polymer businesses.
Key responsibilities will include:
1. Extensive problem solving. 2. Microscopy preparation technique experience including ultramicrotomy and cryo-ultramicrotomy. 3. Operation of transmission electron microscope. 4. Interpretation of images. 5. Documentation of work. 6. Compliance with safety and quality programs. 7. Active member of project and SMX work teams.
Interested: Please e-mail or send your resume and cover letter, with reference to this ad to: Email: R&D-at-Dow.com or The Dow Chemical Company, Workforce Planning 98-289, P. O. Box 150, Plaquemine, LA 70765. E-mail respondents must list Job 98-289 and their last name as the first and second items on the Subject line. Only those selected for an interview will be contacted. Only U.S. citizens or aliens who are authorized to work in the United States will be considered for employment.
We are an equal opportunity employer and offer a competitive compensation and benefits package including 401k, stock purchase, tuition reimbursement and performance incentives. The Dow Chemical Company is the fifth largest chemical company in the world with annual sales of US$20billion. Dow manufactures and supplies chemicals, plastics and agricultural products for customers in 164 countries and employs approx. 43,000 people worldwide. For more news and information about Dow, please visit our web site at www.dow.com.
Bob Cieslinski Microscopy & Microanalysis 1897 E Bldg. (517) 636-6875
John J. Bozzola wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Vacuum-folks, } } We have 5 direct drive vacuum pumps (Hitachi CuteVac) that are in need of } repair. They are leaking oil (suspect the oil seal is shot) but otherwise } pump fine. Does anyone know where we could have them repaired? - Hitachi } does not do this type of work and I hate to throw them away at $3,000 each. } } Thanks, } } John } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } #################################################################### Dear John,
I have never had any luck with anyone rebuilding these pumps at an economical price. The companies that rebuild these pumps complain about the parts availability. Moreover, the rebuild doesn't last more than one year. At $600.00 to $1000.00 a rebuild it is cheaper in the long run to buy a replacement. I have used Alcatel Model 2010 with sucess.
There are several companies that offer rebuilding of vacuum pumps. One that I know of that is very reputable is Torr International in New Windsor, NY. You can reach them at ph: 914-565-4027 or visit their web sight at www.torr.com. I am sure they can help bring your pumps back up to spec. The contacts at Torr Int'l are Dr. Masud Naraghi and Mr. Jeff Terranova.
Good luck,
Steve Collins South Bay Technology East 4019 S. 16th St. Arlington, VA 22204
} } } } } Please visit us at http://www.southbaytech.com} } } } }
Celebrating our 35th year of Manufacturing Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy
John J. Bozzola wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Vacuum-folks, } } We have 5 direct drive vacuum pumps (Hitachi CuteVac) that are in need of } repair. They are leaking oil (suspect the oil seal is shot) but otherwise } pump fine. Does anyone know where we could have them repaired? - Hitachi } does not do this type of work and I hate to throw them away at $3,000 each. } } Thanks, } } John } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ####################################################################
Hi !! One of my collagues is interested to found out a reprit of the paper: Identification of a new Zr-Nb- Fe Phase O.T.Woo and G.J.C.Carpenter. Proc. of the 12th Int. Congress for Electron Micoscopy. San Fransisco Press, 132(1990) Thank in advance
} } I was interested in the use of osmium in xylene that you described. Does the tissue blacken as } in a water-solution or does it simply become yellowish/brown? I am curious because the latter is } what happens in the absence of water during freeze-substitution using e.g. 1% in acetone at -80 } *C, although the colour change probably happens when warming up from low temperature.
Hello Keith.
According to the people here that most often works with this (Irene Lund) the blocks are not as dark as with standard methods, but light brown. We haven't done freeze-substitution with osmium, so it's not easy to compare directly. For this purpose we buy ampoulas with 0,1 gram OsO4.
Best regards Randi Olsen Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso MH-Breivika N-9037 TROMSO NORWAY
Just made a trip to our Physics machine shop. They have sent pumps to Kurt J. Lesker in PA for repair. The cost to rebuild a 25 liter pump in 1997 was ~$600.
Kurt J. Lesker 1515 Worthington Ave. Clairton, PA 15025 1-800-245-1656
Hope this helps, Lou Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html
Duniway Stockroom (http://www.duniway.com) lists a rebuild for a Hitachi VP-160 at $480. I assume that would come with shaft seals. Or you could order the shaft seals from Hitachi and rebuild the pumps yourself. The part number(s) ought to be in the CuteVac manual that came with the pump/microscope. I did this several years ago on a pump from a Hitachi S-570.
However, parts availability from Hitachi, in my experience, is not quick or easy (even with part numbers).
Carl
Earl Weltmer wrote: } } John J. Bozzola wrote: } } Hi Vacuum-folks, } } } } We have 5 direct drive vacuum pumps (Hitachi CuteVac) that are in need of } } repair. They are leaking oil (suspect the oil seal is shot) but otherwise } } pump fine. Does anyone know where we could have them repaired? - Hitachi } } does not do this type of work and I hate to throw them away at $3,000 each. } } } } Thanks, } } } } John } Dear John, } } I have never had any luck with anyone rebuilding these pumps at an } economical price. The companies that rebuild these pumps complain about } the parts availability. Moreover, the rebuild doesn't last more than one } year. At $600.00 to $1000.00 a rebuild it is cheaper in the long run to } buy a replacement. I have used Alcatel Model 2010 with sucess. } } Good Luck } } Earl Weltmer
====================================== Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA (734) 936-1550 FAX (734) 763-4690 ======================================
Potential lead: Capital Equipment Co. (or Capital Vacuum) in the northern Virgina area offered repair for many brands of equipment. I cannot locate the address and do not know if they are still in business. A quick search did turn up a company in Herndon, VA (named Capital...), but I don't know if that is the correct one.
If the leak is not too fast, a tray under the pump is a cheap solution.
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Hi Vacuum-folks,
We have 5 direct drive vacuum pumps (Hitachi CuteVac) that are in need of repair. They are leaking oil (suspect the oil seal is shot) but otherwise pump fine. Does anyone know where we could have them repaired? - Hitachi does not do this type of work and I hate to throw them away at $3,000 each.
Thanks,
John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Internet Mail Server 2.1); Thu, 28 Jan 1999 12:59:39 -0500 Mime-Version: 1.0 Content-Type: multipart/alternative; boundary="============_-1294576389==_ma============" Message-Id: {v04011727b2d654a02b0d-at-[128.206.162.35]}
Assistant or Associate Director Molecular Cytology Core Facility
The Molecular Biology Program at the University of Missouri is seeking an individual with experience in some or all of the following areas:
* confocal scanning laser microscopy * bright field microscopy * wide field fluorescence microscopy * low light video microscopy * image processing/analysis * deconvolution * all types of microtomy * immunocytochemistry * in situ hybridization * Adobe Photoshop
The Assistant/Associate Director will be responsible for training users, maintaining instruments and developing protocols for a campus-wide multi-user light micrscopy facility. Electron Microscopy is not a part of this facility. PhD desirable but not required for individuals with extensive experience. Although an ideal candidate would have experience in all of the areas listed above, candidates with extensive experience in selected areas and who have the desire and capacity to learn the additional areas will be considered. Excellent oral and written communication skills are essential. Experience in a multi-user core facility would be viewed positively. Women and minority candidates are especially encouraged to apply. Review of applications will begin immediately and continue until an appropriate candidate is hired.
Address applications (CV and 3 letters of reference) or inquires to:
Thomas E. Phillips, Ph.D. Division of Biological Sciences 3 Tucker Hall, University of Missouri Columbia, MO 65211-7400. 573-882-4712 PhillipsT-at-missouri.edu.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax) --============_-1294576389==_ma============ Content-Type: text/enriched; charset="us-ascii"
{bold} {bigger} {bigger} {bigger} Assistant or Associate Director
Molecular Cytology Core Facility
{/bigger} {/bigger} {/bigger} {/bold} The Molecular Biology Program at the University of Missouri is seeking an individual with experience in some or all of the following areas:
* confocal scanning laser microscopy
* bright field microscopy
* wide field fluorescence microscopy
* low light video microscopy
* image processing/analysis
* deconvolution
* all types of microtomy
* immunocytochemistry
* in situ hybridization
* Adobe Photoshop
The Assistant/Associate Director will be responsible for training users, maintaining instruments and developing protocols for a campus-wide multi-user light micrscopy facility. Electron Microscopy is not a part of this facility. PhD desirable but not required for individuals with extensive experience. Although an ideal candidate would have experience in all of the areas listed above, candidates with extensive experience in selected areas and who have the desire and capacity to learn the additional areas will be considered. Excellent oral and written communication skills are essential. Experience in a multi-user core facility would be viewed positively. Women and minority candidates are especially encouraged to apply. Review of applications will begin immediately and continue until an appropriate candidate is hired.
Address applications (CV and 3 letters of reference) or inquires to:
Folks: I am trying to get hold of a dual replica device for a Cressington freeze fracture machine. An alternate possibility would be a dual replica device from a Balzers 400 If anyone, anywhere, has one tucked in a drawer gathering dust etc. etc. and would like to part with it (for the purchase price, or whatever they feel is appropriate) please let me know as soon as possible Thanks Simon
---------------------------------------------------------------------------- -------- Simon C. Watkins Ph.D. MRC Path Associate Professor Cell Biology and Physiology Director, Center for Biologic Imaging BSTS 225 University of Pittsburgh Pittsburgh PA 15261 Tel: 412-648-3051 Fax: 412-648-8330 Mobile: 412-607-3534 URL: http://sbic6.sbic.pitt.edu ---------------------------------------------------------------------------- -----
---------- } Van: HASWELL Malcolm {malcolm.haswell-at-sunderland.ac.uk} } Aan: Microscopy {microscopy-at-sparc5.microscopy.com} } Onderwerp: RE: retired chemist school volunteer ne
} GROWING ROOT TIPS ..CAUTION supermarket onions may be incapable of sprouting roots you } may have to try fresh ones).
This seems to become more and more a problem as onions are frequently treated with growth inhibitors. Instead of onions, garlic (Alium sativum) can be used! There was an article in MIKROKOSMOS some years ago, stating that garlic isn't treated that way, at least not in Germany (Europe?).
} STAINING METHODS
Some of my modifications:
} FEULGEN STAIN TECHNIQUE } 1 Cut 1cm of root tip off onion, } 2. fix in acetic alcohol (3:1) for at least 30 min
fix in ethylalcohol 95%-acetic acid 3/1 (v/v) for about 30 min
} 3. Hydrolyse in 1 molar/normal HCl at 60 deg C 10 - 15 min
when an incubator or a water bath isn't available: use HCl, 5N, 40 min at RT
} 4 Stain in Feulgen reagent for minimum of 60 min (IN THE DARK) - root tip
} will become purple in meristem } 5. Without letting specimen dry out = place the end 2-3mm of root tip on a } clean slide with a drop of 45% acetic acid } 6. Cut root tip into 4 longitudinally then gently and carefully squash under } a coverslip - protecting your thumb several layers of paper towel. } 7. Examine for mitosis
} ACETIC ORCEIN TECHNIQUE } 1, Cut 1cm of root tip off onion
fix in ethylalcohol 95%-acetic acid 3/1 (v/v) for about 30 min
} 2. put into watch glass with concentrated HCl and absolute ethanol (1:1) for } 10 min - NB take care when mixing this reagent
Wash in running water for at least 10 min
} 3. transfer root tips to watch glass with 45% acetic acid for 5 min
} 4. Without letting specimen dry out = place the end 2-3mm of root tip on a } clean slide with a drop of ACETIC ORCEIN
Slightly heat the slide untill steam apears, let cool, repeat this several times until the meristem comes dark red-brown.apply new staining solution when necesary. Don't let dry!
} 5. Cut root tip into 4 longitudinally then gently and carefully squash under } a coverslip - protecting your thumb several layers of paper towel. } 6. Examine for mitosis
Slides can be made permanent. There are lots of methods, I use these: For ACETIC ORCEIN-stained slides: use slides treated with Haupt's adhesive. Put the slides after examination in a jar of water, the cover slip down. Wait until the coverslip comes of. Wash slides and coverslips careful but toroughly in distilled water, dehydrate in ethylalcohol, 2-propanol. Treat with xylene and mount in DPX (or another resin). Most of the cells remain on the slide (coverslip).
For cells stained with Feulgen: Use slides treated with Haupt's adhesive. Put the slides after examination in a jar of water, the cover slip down. Wait until the coverslip comes of. Wash slides and coverslips careful but toroughly in distilled water, dehydrate in ethylalcohol 70%, stain in Light Green or Fast Green FCF in ethylalcohol 70% (concentration not critical, about .5% will do), dehydrate in ethylalcohol 90% and differentiate the green in it, 2-propanol. Treat with xylene and mount in DPX (or another resin). Most of the cells remain on the slide (coverslip). DNA: violet, RNA (nucleoli): green (Light Green) or blue-green (Fast Green FCF).
} Malcolm Haswell } Electron Microscopy } School of Sciences } Fleming Building } University of Sunderland } SUNDERLAND SR1 3SD } Tyne and Wear } UK } } Tel (0191) 515 2872 } e-mail: malcolm.haswell-at-sunderland.ac.uk
I am surprised that Hitachi will not repair the pumps for you. We had three pumps repaired ( Oil seals ) by Hitachi 1.5 years ago at minimal cost during our TEM move. The pumps are still working perfectly & have no leaks. It is not a major job to replace these seals yourself and in the past we have replaced a couple, with no difficulty getting parts from Hitachi (UK). The problem with replacement is seating the seals in evenly. It requires patience which is probably why Martin ( Hitachi, UK ) did a better job than us.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Carl Henderson {chender-at-umich.edu} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
I am still using Eric Shelden's Presentit {shelden-at-umich.edu} to do my digital presentations. It takes a folder full of PICT's and Quicktime's and place them in name order. No fancy transitions or anything, but I find that overkill. The main advantage is that it plays quicktimes cleanly and at maximum frame rate with easy access to the quicktime controls for frame by frame play. I keep a copy on my desktop so I can drag and drop a movie or image on it anytime I need to look at something. Doing quicktime in Powerpoint is tricky at best and requires a session with tech support to find out how to make it work correctly. For making text slides, and playing simple images Powerpoint is great although I still liked Persuasion better since you could apply multiple formats to a single presentation. The best of that lot may be Astound which is the only quicktime application that implements a speed control for quicktime movie playback rate. Dave
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Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax)
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REGARDING post-doc at EMAT, Belgium
Dear colleagues,
as of April 1, 1999, a 2-year TEM post-doc position will be available at = the EMAT group in Antwerp. The research will consist of using advanced = TEM techniques (HREM, EDX, EELS, low-dose, ...) for the study of new = photographic materials in close colaboration with the Agfa-Gevaert = company, one of the leading manufacturers of photographic material. The = position is open to EC citizens or equalized persons.
I look forward to your applications,
Nick Schryvers e-mail: schryver-at-ruca.ua.ac.be fax: 32-3-2180257
by upei.ca (PMDF V5.1-9 #20757) with ESMTP id {01J741DZDB3WA8CEC4-at-upei.ca} for microscopy-at-msa.microscopy.com; Fri, 29 Jan 1999 11:47:40 AST Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.43); Fri, 29 Jan 1999 11:51:38 -0400 Received: from SpoolDir by AVCN1 (Mercury 1.43); Fri, 29 Jan 1999 11:51:07 -0400
Hi! I have a question regarding the limits of Reichter-Jung Ultracut E ultramicrotome. One of the students wants to cut 1 micrometer sections of the blocks almost 5x5mm in diameter. So the sections would be 1 micrometer thick, 5 mm wide, 5 mm high. My questions is is what is the maximum size of the block face that can be safely (not destroying the machine) cut on that type of the microtome. I need to know numbers to back up my argument. Thanks in advance Dorota
Dr. Bozzola, I am not sure that my reply to your posting went through as I unknowingly did not follow protocol for this list server. I can rebuild these inexpensively, return them quickly, test ultimate vacuum, and provide a guarantee. I have rebuilt these pumps a couple hundred times. You are right the seals harden over time due to heat from the pump. Your also right in that it is silly to replace a perfectly good pump for $3K that can be rebuilt for a couple hundred. Give me a call. J. McClintock (606)257-1242 (606)277-6507 jcmcclin-at-pop.uky.edu
Dear John and all, In the USA the Hitachi service company Hitachi Instruments, Inc. (HII) used to have its Hitachi vacuum pumps rebuilt by Dunniway Stockroom (800) 446-8811 in Mountain View, CA. They (HII) found it uneconomic to do so and stopped. Now for warranty and service repairs of Hitachi pumps they replace them with Edwards pumps. The Hitachi pumps are thrown away! More of the new EMs from Japan are showing up with the Edwards pumps new. Dunniway has told me that the Hitachi pumps are hard to repair, and that there is no secondary market for them. You can buy used/rebuilt pump from Dunniway for a reasonable price. The Edwards replacement model for most Hitachi SEMs is E2M12 with a rebuilt price of $1400.
Disclaimer: I am a only a Customer of Dunniway with no financial interest.
Ron Vane XEI Scientific. -----Original Message----- } From: John J. Bozzola {bozzola-at-siu.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Dear fellow microscopists,
We are considering the purchase of a new semi-in-lens FESEM. Quality of service is of paramount importance to me in terms of the selection process. I would like to hear from others in the Pittsburgh area (say 500 mile radius) about the quality of their SEM service (not necessarily FESEM instruments) from the following vendors: LEO, JEOL, Hitachi, and Philips. I'm interested in the good, the bad, and the ugly kind of stories.
The types of things that I'm interested in is time it took for installation, timeliness (i.e. response time from the initial call), downtime, satisfaction with their work, your confidence in the service engineers, helpfulness in diagnosing problems and solving them over the phone without a service call, etc.
Please respond directly to me. I will keep your responses confidential.
Thanks in advance.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Hello All, Does anyone out there have any suggestions for getting good SEM cross= sectional images from dried wood? I have a student who would like to image= the microstructure of several types of wood. All the references I have= found so far suggest cutting fresh or moist wood with a new razor blade to= achieve good cross-sections. The student is studying the effects of two= different drying techniques, and therefore soaking the wood in water is out= of the question. Cutting the dried wood with razor blades has resulted in= crushed, damaged cross sections.=20
Thanks, Scott F. Scott Miller Electron Microscopy Lab smiller-at-umr.edu University of Missouri-Rolla =20 223 McNutt Hall voice: 573 341 4727 Rolla, MO 65409 USA fax: 573 341 6934
I am looking for a mounting medium with a high optical index (around 1.7). Once I used something called piperin (n=1.68). Could you please let me know where can one obtain such products?
Fellow Microscopists, I am soliciting your comments on what you consider the best approach to damp vibrations when using a light microscope. We are in the process of redesigning our light microscopy/image analysis work area and have a choice between a conventional marble table and a Newport BenchTop vibration isolation system (pneumatic). Which should it be? Are there other possibilities to consider?
Thanks for a moment of your time. Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
I would like to draw your attention to the materials science symposium being held as part of the Scanning 99 meeting in April this year. The abstract deadline is February 15th (the same as MSA) and the abstract format is the same as the MSA format. It is intended that there will be contributed presentations as part of the program, and depending on the number of submissions, the symposium may be extended for another day. Please forward this e-mail to anyone you think may be interested in attending the symposium.
{bold}
"Analyzing Materials Interfaces at Atomic Resolution" {/bold}
There will be a Materials Science symposium at Scanning 99 entitled "Analyzing Materials Interfaces at Atomic Resolution". Scanning 99 is being held at the Hyatt Regency O'Hare in Rosemont, IL from April 11-14 1999. The "Analyzing Materials Interfaces at Atomic Resolution" symposium is tentatively scheduled for Tuesday April 13th and will consist of invited and contributed presentations. The details of the conference and deadlines/forms for abstract submissions can be found at http://www.scanning.org or details can be requested from Mary Sullivan (e-mail: scanning-at-fams.org, tel:201-818-1010). Registration for members of the Midwest Microscopy and Microanalysis Society are at the reduced rates of $150 for the whole conference or $50 for a single day (all attendees of the MMMS symposium held at UIC last May are members of MMMS).
A preliminary list of invited speakers and the titles for their presentations is shown below.
{bold} Prof Ondrej Krivanek-University of Washington {/bold}
Title "Towards sub-Angstrom electron probes by Cs-corrected STEM."
{bold} Dr Max Haider-CEOS GmbH {/bold}
Title "Towards sub-Angstrom resolution by correction of spherical aberration"
{bold} Dr J. Murray Gibson -Argonne National Lab {/bold}
Title "Statistical Measurement of Electron Scattering Fluctuations in Amorphous
Materials - A new Structural Tool"
{bold} Prof Laurie Marks-Northwestern University {/bold}
Title "Picometer structure determination using Electron Diffraction"
{bold} Prof Marija Gajdardziska-Josifovska-University of Wisconsin at Milwaukee {/bold}
Title "Quantitative surface microscopy and diffraction over the length scales:
Morphology and crystallography of polar oxide surfaces. "
{bold} Dr David Muller-Lucent Technologies {/bold}
Title "The end of the Roadmap for Silicon Dioxide: Atomic resolution EELS of
of Hyper-Thin Gate Oxides"
{bold} Prof David Williams-Lehigh University {/bold}
Title "Atomic-Resolution X-ray Microanalysis in the TEM"
{bold} Dr Ed James-University of Illinois at Chicago {/bold}
Title "Atomic resolution scanning transmission electron microscopy on the 200kV
FEGTEM"
{bold} Dr Stephen Pennycook-Oak Ridge National Lab {/bold}
Title "Atomic scale analysis of interfaces by Z-contrast STEM, EELS and
If the wood is very dry, you might try simply fracturing a piece. A surface so prepared will have an extreme amount of relief, but it may provid you with a less deformed microstructure.
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Hello All, Does anyone out there have any suggestions for getting good SEM cross sectional images from dried wood? I have a student who would like to image the microstructure of several types of wood. All the references I have found so far suggest cutting fresh or moist wood with a new razor blade to achieve good cross-sections. The student is studying the effects of two different drying
techniques, and therefore soaking the wood in water is out of the question. Cutting the dried wood with razor blades has resulted in crushed, damaged cross sections.
Thanks, Scott F. Scott Miller Electron Microscopy Lab smiller-at-umr.edu University of Missouri-Rolla 223 McNutt Hall voice: 573 341 4727 Rolla, MO 65409 USA fax: 573 341 6934
It may well depend on the type of wood. Years ago I had some luck with Quercus blocks by cleaning up the top surface with a sliding microtome (e.g. AO 860) with a properly sharpened (and long) microtome knife set to a high shear angle and using a small advance. There were still some crumbs in the vessels but you could find good areas too.
I've never had much but frustration using razor blades on woody plants of any kind and lately it seems most brands of single edge blades we use around here for trimming blocks are softer and duller then they used to be.
cheers, John Heckman
} Hello All, } Does anyone out there have any suggestions for getting good SEM cross } sectional images from dried wood? I have a student who would like to image } the microstructure of several types of wood. All the references I have } found so far suggest cutting fresh or moist wood with a new razor blade to } achieve good cross-sections. The student is studying the effects of two } different drying techniques, and therefore soaking the wood in water is } out of the question. Cutting the dried wood with razor blades has } resulted in crushed, damaged cross sections. } } Thanks, } Scott } F. Scott Miller } Electron Microscopy Lab smiller-at-umr.edu } University of Missouri-Rolla } 223 McNutt Hall voice: 573 341 4727 } Rolla, MO 65409 USA fax: 573 341 6934
I am studying about Hf-Ta-V alloy. So, I want to make a TEM sample for that alloy(Hf and Ta of 1/3, V of 2/3) by jet-electropolishing technique using the apparatus made by Struers. I would like to know the experimental condition including solution, temperature and cathode. Please tell me about it who have experienced in that alloy.
won
Won-yong Kim Department of Materials Science and Engineering University of Pennsylvania, LRSM building 3231 Walnut St., Philadelphia, PA 19104
} redesigning our light microscopy/image analysis work area and have a choice } between a conventional marble table and a Newport BenchTop vibration } isolation system (pneumatic). Which should it be? Are there other } possibilities to consider? You are going to get lots of responses for cheap solutions using tennis balls or spaldeen balls or inner tubes from bicyles or wheelbarrows or layers of felt and sheets of concrete. Having no experience with these, I can't comment on their efficacy. However, the Newport or TMC table floating on air or N2 is going to be far superior to the marble table. Guarranteed. ******************************************************* * Michael Cammer Analytical Imaging Facility * * Albert Einstein College of Medicine * * Bronx, NY 10461 (718) 430-2890 * * work URL: http://www.ca.aecom.yu.edu/aif/ * * personal URL: http://cammer.home.mindspring.com/ * *******************************************************
Years ago, upon recommendation by folks out at Sharon Vacuum Company in Brockton MA, I bought a two component epoxy kit called A-12 Epoxy Adhesisve from the Armstrong Products Company on Argonne Road in Warsaw, Indiana, for repairing leaks in vacuum systems. It also proved useful for sealing cracked plastic parts and metal fittings on cryogenic systems, and for many other repairs. Great stuff, and it cured to a beautiful milk chocolate brown tone.
I'm just about out now, and I've written to Armstrong Products and they are no longer in existence,letter was not forwardable, etc.
Anybody out there know of their whereabouts? Or could you recommend any other suitable replacement product?
Thanks,
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
I'm interested in doing some immuno work with the SEM. Basically tagging colloidal gold onto a surface antigen on cultured cells. I've got some old information and papers but as for up-to-date methodologies, I'm a little sketchy. Does anyone have any good review papers on the use of immuno surface markers w/ SEM (preferably colloidal gold?) Nothing too fancy. In case it helps, I'm specifically looking at phosphotidylserine exposed on the outer membrane leaflet during apoptosis in a cultured line of CML cells. Thanks in advance.
Gib Ahlstrand wrote: =============================================== Years ago, upon recommendation by folks out at Sharon Vacuum Company in Brockton MA, I bought a two component epoxy kit called A-12 Epoxy Adhesisve from the Armstrong Products Company on Argonne Road in Warsaw, Indiana, for repairing leaks in vacuum systems. It also proved useful for sealing cracked plastic parts and metal fittings on cryogenic systems, and for many other repairs. Great stuff, and it cured to a beautiful milk chocolate brown tone.
I'm just about out now, and I've written to Armstrong Products and they are no longer in existence,letter was not forwardable, etc.
Anybody out there know of their whereabouts? Or could you recommend any other suitable replacement product? ================================================= It is definitely not the same thing, but we (and a number of our customers) have had excellent results with VACSEAL® High Vacuum Leak Sealant. The product can withstand repeated temperature cycling from liquid helium temperatures to 450° C over long intervals of time.
It is described on URL http://www.2spi.com/catalog/vac/vacleak.html
Disclaimer: SPI Supplies offers this product for this kind of application so we would have a vested interest in seeing it used more widely.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Hello Paul. Believe it or not 4 small inner tubes, inflated so that they have no real tension before having a table top put on them work very well. I have used this trick for Atomic force microscopy (AFM) & holography (on a wooden table). Check out how pneumatic legs are made, aside from the automatic leveling (option) they are not much different. An improvement is to build a base consisting of layered, carpet, plywood, 8x8x16" bricks (commonly called cinder blocks around here) up to the height needed. There are a lot of optical tables on this planet set up this way. You can get small tubes (6-8" dia) from cart & dolly vendors. Another trick is to to build a sand box. Check books on do it yourself holography Yea, I know these ideas don't look great but they do work & save lots of money.
HI Gib. Varian Vacuum Products has been selling "Torr Seal" for years. Don't know about it's cryo properties.
Bruce Brinson Rice U.
Gib Ahlstrand wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Microscopy Listers, } } Years ago, upon recommendation by folks out at Sharon Vacuum Company in Brockton } MA, I bought a two component epoxy kit called A-12 Epoxy Adhesisve from the } Armstrong Products Company on Argonne Road in Warsaw, Indiana, for repairing } leaks in vacuum systems. It also proved useful for sealing cracked plastic parts } and metal fittings on cryogenic systems, and for many other repairs. Great } stuff, and it cured to a beautiful milk chocolate brown tone. } } I'm just about out now, and I've written to Armstrong Products and they are no } longer in existence,letter was not forwardable, etc. } } Anybody out there know of their whereabouts? Or could you recommend any other } suitable replacement product? } } Thanks, } } Gib Ahlstrand } Electron Optical Facility, University of Minnesota, Dept. Plant Pathology } 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 } 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Scott - I never found razor blades satisfactory on fresh wood and rather suggest using a microtome with a permanent knife (not a disposable). I have not tried myself using a tungsten carbide knife but expect that this would give still better results. Small triangular TC knives which fit ultramicrotome chucks are an obvious and cheaper solution than the big TC knives.
Diamond knife sections would be too thin for SEM purposes but diamond knives can be used for facing a block. However, those knives are expensive and the operation is risky.
Another alternative is the use of a series of wet and dry papers on a lap. Use the paper wetted with kerosene or another slowly evaporating solvent and when finished use absolute alcohol to rinse. These papers are available to about 2000 gird, which is still not fine enough for SEM. For the final finish I suggest diamond lapping film. This material is expensive but the diamond particles are embedded with the plastic. The particles will rarely contaminate the specimen, the particle size is available down to 0.1 micrometre and the film is long lasting.
Using the grinding powder that is normally used for cutting of rock, makes a mess of a specimen like wood. Disclaimer PST is a supplier of some mentioned products.
Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, January 30, 1999 3:18 AM, Scott Miller [SMTP:smiller-at-umr.edu] wrote: } } Hello All, } Does anyone out there have any suggestions for getting } good SEM cross sectional images from dried wood? I have a } student who would like to image the microstructure of } several types of wood. All the references I have found } so far suggest cutting fresh or moist wood with a new } razor blade to achieve good cross-sections. The student } is studying the effects of two different drying } techniques, and therefore soaking the wood in water is } out of the question. Cutting the dried wood with razor } blades has resulted in crushed, damaged cross sections. } } Thanks, } Scott } F. Scott Miller } Electron Microscopy Lab smiller-at-umr.edu } University of Missouri-Rolla } 223 McNutt Hall voice: 573 341 4727 } Rolla, MO 65409 USA fax: 573 341 6934
These were once very popular, especialy for preparations of diatoms, but they seem very hard to find these days! (I have posted a question regarding mounting media with high RI on Histonet a few weeks ago: received only one answer...) The only source I know of is a Dutch company called Euromex Microscopes. They sell Naphrax (RI = 1.710) in 25 ml bottles. Catalog nr = PB0267. You can contact them trough email: info-at-euromex.nl Yvan Lindekens.
---------- } Van: SROUBEK {pasroube-at-mtu.edu} } Aan: Microscopy-at-sparc5.microscopy.com } Onderwerp: help on mounting medium } Datum: vrijdag 29 januari 1999 20:24 I am looking for a mounting medium with a high } optical index (around 1.7). Once I used something } called piperin (n=1.68). Could you please let me know } where can one obtain such products? } } Thanks } } Pavel Sroubek } } pasroube-at-mtu.edu }
Could the sample be planed at low temperature? Such as in a cryostat or cryoultramicrotome? The problem is that it would have to be returned to room temperature without condensation forming, maybe in a "sealed" container with dry nitrogen or air+dessicant.
Maybe it could be impregnated with a non-aqueus medium. I am wondering if 1-hexadecene (from Sigma) could be used. It is used as a mdium for filling gas spaces etc in leaves before high pressure freezing is carried out. It is a non-aquous, non-toxic, non-osmotically active type of paraffin. I am not sure about its removal afterwards.
Something that would sublime would be favoured! Maybe one of the media used instead of critical point drying? Hexamethyldisilazane? Combined with low temperature sectioning?
Another primitive approach would be to put it in liquid nitrogen in a styrofoam contained with a metal "plate" on the bottom and fracture the sample across with a cooled razor blade (held in tweezers) whacked with a hammer. Then it needs rewarming as above, in a container with dessicant (fresh phosphorus pentoxide, in fine powder form, is my favourite - this is a vicious chemical so beware of inhalation of the dust), preferably with dried nitrogen gas leaking through to prevent air plus atmosheric moisture ingressing.
At the recent resin cutting workshop in Seefeld (Austria), Prof. Sitte (who has had a lot to do with the design of Reichert/Leica ultramicrotomes) said that it is possible on modern instruments to cut 5 or 6 mm across by 12 mm in length. For TEM examination, ther grid size is the limiting factor! I have cut 2.5 x 2.5. mm ultrathinns for TEM. Someone else here has cut roughly the same size 0.5-1.0 micrometers thick, without a "boat"/water bath, of freeze-dried tissue, dry mounted, for x-ray microanalysis.
I would say that 5 x 5 mm, 1 micron thick should not be a problem for the instrument providing that the approach and trimming is done carefully and that the specimen is not very hard. The specimen should section easily, if it "sticks" on the knife and doesn't pass to cut a section then I would think again.
Hi Pavel: I don't know about Piperine, maybe somebody else can help with that. However, Meltmounts are available in a range of several refractive indices, including 1.68 and 1.704. The Meltmount quickstick is applied to a slide on a hotplate and after mounting the coverslip, the medium instantly solidifies at room temperature. The coverslip may be removed at anytime by re-heating the slide. Disclaimer: Meltmount-Quickstick are available from PST and other microscopy suppliers. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, January 30, 1999 5:24 AM, SROUBEK [SMTP:pasroube-at-mtu.edu] wrote: } } I am looking for a mounting medium with a high } optical index (around 1.7). Once I used something } called piperin (n=1.68). Could you please let me know } where can one obtain such products? } } Thanks } } Pavel Sroubek } } pasroube-at-mtu.edu
Dorota, I also have a Reichert ultracut and it does fine with very large thick sections. Knives tend to suffer however. Depending on your sample, you may find that you'll need a diamond histo knife. Glass however will also work . MG Engle Electron Microscopy & Imaging Facility University of Kentucky
Have you been experiencing trouble with vibration before now?
I'm on the 4th floor of my building, which has a lot of activity during the day. In my lab there are 4 sectioning stations with ultramicrotomes lined up in the same room, sited against the same wall. Three are on marble tables, one is on a 'floating' table.
Hands down, the N2-damped table beats out the marble tables.
For any critical work, the microtome on the floating table is usable at any hour of the day, while those on marble tables usually must be used after-hours, after traffic in the building has eased. The vibration in the floor is easily visible in the water's surface on any of the 3 marble tables; it is damped out in the N2-table. Well worth the investment.
Good luck! Ann Lehman EM Facility Mgr Trinity College Hartford, CT v 860-297-4289 f 860-297-2538 e ann.lehman-at-exchange.cc.trincoll.edu
-----Original Message----- } From: Gerroir, Paul J [mailto:Paul.Gerroir-at-crt.xerox.com] Sent: Friday, January 29, 1999 2:44 PM To: Microscopy-at-Sparc5.Microscopy.Com
January 29, 1999
Fellow Microscopists, I am soliciting your comments on what you consider the best approach to damp vibrations when using a light microscope. We are in the process of redesigning our light microscopy/image analysis work area and have a choice between a conventional marble table and a Newport BenchTop vibration isolation system (pneumatic). Which should it be? Are there other possibilities to consider?
Thanks for a moment of your time. Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
We do a lot of extremely sensitive micromanipulation work here with the LM and have a few very effective anti-vibration systems.
We placed a heavy slab (approx. 400lb) 2' x 3' of Boiler Plate Steel on top of about 150 tennis balls on a well supported bench. The steel is well-protected and finished to make a clean, safe work area. Total cost was about $250. Cdn.
If you'd like more details, you can contact me off-line.
Karen Rethoret Microscopy Lab York University Toronto, Ont. 416-736-2100 x33289
On Fri, 29 Jan 1999, Gerroir, Paul J wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } January 29, 1999 } } Fellow Microscopists, } I am soliciting your comments on what you consider the best approach to damp } vibrations when using a light microscope. We are in the process of } redesigning our light microscopy/image analysis work area and have a choice } between a conventional marble table and a Newport BenchTop vibration } isolation system (pneumatic). Which should it be? Are there other } possibilities to consider? } } Thanks for a moment of your time. } Paul } } Paul J. Gerroir } Microscopy } Materials Characterization } Xerox Research Centre of Canada } 2660 Speakman Drive } Mississauga, Ontario L5K 2L1 } } Phone: (905) 823-7091, ext. 216 } FAX: (905) 822-7022 } e-mail: paul.gerroir-at-crt.xerox.com } }
Research Opportunities Mount Sinai School of Medicine is a leader in medical research and education. The establishment of our new Microscopy Center has created opportunities for experienced Microscopists with a BS/BA or MS in Biology or Life Sciences. All applicants should have excellent organizational and communication skills, an understanding of basic laboratory procedures, and the ability to manage a large and varied workload.
Electron Microscopist The successful candidate will participate in ultrastructural studies of various biological systems. Qualifications include at least 2 years of experience in routine electron microscopy procedures (TEM, SEM), ultramicrotomy, immunogold labelling, specimen preparation, photographic darkroom work, and routine maintenance of equipment. Individuals with immunofluorescence and confocal microscopy experience are desirable. Code: EM
Light Microscopist The successful candidate will participate in biomedical studies that use a variety of advanced light microscopic techniques. Duties will include maintaining equipment, instructing users in equipment operation, and sample preparation. Qualifications include at least 2 years of experience in fluorescence and confocal microscopy, immunofluorescence labelling, in situ hybridization, digital imaging and analysis, cell culture, and histological techniques. Strong computer skills are essential. Code: LM
We offer a salary commensurate with experience and excellent benefits. For consideration, please mail your resume, which must indicate code for position of interest, to:
Scott Henderson, Ph.D., Director, Microscopy Center, Box 1007, Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, NY 10029-6574
http://www.careermosaic.com/mountsinai
We are an equal opportunity employer fostering diversity in the workplace.
______________________________
Scott Henderson, Ph.D. Director of Microscopy, Mount Sinai School of Medicine, Dept. of Cell Biology & Anatomy, Box 1007, One Gustave L. Levy Place, New York, NY 10029-6574
Paul, we use a combination of marble tables and pneumatic shock absorbers on our microscopes. We have been very happy with the results. One of our microscopes has the bench top vibration table which is also very good.
At 02:44 PM 1/29/99 -0500, Gerroir, Paul J wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Doug: Try this paper. You will have to adapt the protocol to fit your needs. If you have any questions please feel free to call.
Coller, Barry S., Kutok, J.L., Scudder, L.E., Galanakis, D.K., West, S.M., Rudomen, G.S., Springer, K.T. . Studies of Activated GPIIb/IIIa Receptors on the Luminal Surface of Adherent Platelets: Paradoxical Loss of Luminal Receptors When Platelets Adhere to High Density Fibrinogen. J. Clin. Invest. (1993) Vol. 92, pp. 2796-2806.
Doug Matthews wrote:
} Hi everyone, } } I'm interested in doing some immuno work with the SEM. Basically } tagging colloidal gold onto a surface antigen on cultured cells. I've got } some old information and papers but as for up-to-date methodologies, I'm a } little sketchy. Does anyone have any good review papers on the use of immuno } surface markers w/ SEM (preferably colloidal gold?) Nothing too fancy. In } case it helps, I'm specifically looking at phosphotidylserine exposed on the } outer membrane leaflet during apoptosis in a cultured line of CML cells. } Thanks in advance. } } Doug Matthews } Providence College } }
-- Regards, Gregory Rudomen Technical Specialist University Microscopy Imaging Center State University of New York at Stony Brook 516-444-3126 Greg-at-umic.sunysb.edu *************************************** Standard disclaimer: The opinions expressed in this communication are my own and do not necessarily reflect those of the University Microscopy Imaging Center. ***************************************
Hi! Thanks to all of you who responded to my posting. All suggestions are very helpful. I forgot to add that the tissue (lung from rat) is embedded in Epon/Araldite. It is not cryosectioning. Thanks again Dorota
I am submitting a request for a faculty member who is not a member of the list. The researcher wishes to locate, for sale, antibodies and/or conjugated antibodies for Salmonella, Clostridium and Campylobacter.
Any replies may be directed to my e-mail address and not to the list.
With Best Wishes,
Bill Monroe
Bill Monroe EM Center Mississippi State University (601)-325-3019 Lab Fax 325-0246
Greetings, I'm looking for a conductive epoxy (for materials science use) in which I can embed metal samples, sand and polish, and then look at them in a field emission SEM. I have seen this used at other facilities, so I know it exists. I found something at Electron Microscopy Sciences that I thought might work, but the product has been discontinued. Does anyone know of an epoxy that can be used for the above application?
Any replies can be directed to me personally.
Thank you, Kevin Croat tkc-at-howdy.wustl.edu Dept. of Physics Washington University in St. Louis
Thanks to all those who responded to my question about resin problems. I got a number of good suggestions, mostly related to storage (most people thought storing resins in the fridge was unnecessary and probably a bad idea), use of accelerator (several people suggested that I switch to BDMA or add DMP-30 to all infiltrating steps) or water contamination in any of the components (several people suggested replacing all resins). We will definitely be following up. Many thanks for taking the time to reply.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-4861936
The Microscopy Society of America (MSA) and the MSA Technologists' Forum are the sponsors of the Professional Technical Staff Awards (PTSA) to provide assistance on a competitive basis to full-time professional staff who submit papers for presentation at Microscopy and Microanalysis '99 (M&M '99). The deadline (15 February 1999) is fast approaching! Eligible applicants: you are encouraged to submit an abstract and supporting documentation. Managers: you are encouraged to support eligible staff members in this effort. Please read the following, taken from the M&M '99 Registration Bulletin, for more information.
It is the intent of this award to stimulate attendance at M&M '99 for professional technical staff who ordinarily might not participate in the meeting, and to encourage employers to support their staff in professional activities. Awardees will be selected based on the quality of an abstract submitted for presentation at M&M '99. The applicant must be the first author of the submitted abstract. Applicants must be full paid-up members of MSA at the time of application. The awards consist of free full registration for the meeting, a copy of the Proceedings and the Sunday evening social event. MSA will reimburse awardees up to $600 for travel, lodging and other expenses. There will be a maximum of four awards given. Successful applicants must present their abstracts personally at M&M '99 in order to receive the award. They are expected to attend and participate in the entire meeting. Former winners will not be eligible for another award. Applications shall consist of: (1) a supporting letter from the applicant's employer, manager or supervisor, attesting to the applicant's status as a full-time professional staff member; (2) a scientific abstract (original and 4 photocopies) for presentation, as described in the Registration Bulletin, accompanied by a completed Data Form (available on-line at http://resolution.umn.edu:591/MandM/DataEntry.html, or if inaccessible, by calling the Meeting Manager at 708/361-6045; electronic submission of the Data Form is encouraged); (3) a copy of the abstract to be sent by 15 February 1999 to the Chair of the Technologists' Forum: Ms. Beverly E. Maleeff, SmithKline Beecham Pharmaceuticals, Safety Assessment-US, UE0462, 709 Swedeland Road, King of Prussia, PA 19406. Phone: 610/270-7987; Fax: 610/270-7202; e-mail: Beverly_E_Maleeff-at-sbphrd.com. In order to be considered, completed applications must be received by 15 February 1999. Abstracts will be judged by the MSA Technologists' Forum. All applicants will be notified of the outcome in early March. Applicants not receiving the award will have the opportunity to withdraw their abstract if necessary.
Regards, Bev Maleeff Chair, MSA Technologists' Forum
I'd like to thank all those who responsed to my question. It is clear now that we should use double glid (Ni) instead glue to hold the sammple. Attached are messages I've received.
} From zrdai-at-u.washington.edu Mon Feb 1 16:06:29 1999
You might consider the following, (a thick, rather crude adhesive). It is: Sauereisen Insa-Lute adhesive, No. 1 paste. We used it to hold U-Si slices for polishing mechanically. The as mounted slices were then ion irradiated in a UHV vacuum chamber at 350 C approximately. The glue held pretty well, but can't be dissolved-unless the company has a special solvent. It outperformed several "high temp" glues that I tried on hotplate tests. Source: Sauereisen 160 Gamma Drive Pittsburgh Pennsylvania, 15238-2989
Phone: (412) 963-0303 FAX: (412) 963-7620
Bernard Kestel Materials Science Div. 9700 So. Cass Ave. Argonne, Ill., 60439
} From stephan-at-gecko.biol.wits.ac.za Mon Feb 1 16:06:29 1999
I was wondering if anyone in the microscopic community could recommend an outfit that can polish petrographic thin sections for EMP work. I need to can the samples processed within a month timeframe and I am willing to pay some for this. Many thanks
The Emory University Neurology Microscopy Laboratory, the University of Georgia Botany Department EM Laboratory, and Leica Microsystems, Inc.
Present a Cryo Techniques and Immunogold Workshop.
April 18-23, 1999 at the Campus of Emory University in Atlanta, GA.
Objectives
1. To provide researchers the opportunity to learn the theory and practice of cryo techniques for biological sample preparation and immunogold labeling.
2. To permit participants to process their own samples using these techniques under expert guidance.
I was requested to identify a crystal phase of small (5-7 micron) alumina particles embedded into copper. There are only a few particles, and all of them are on the surface. Any ideas how it can be done?
Thanks,
Alexander Titkov Millennium Inorganic Chemicals PO Box 245 Bunbury WA 6231 Australia Ph 61 8 9780 8505 W FAX 61 8 9780 8500 E-mail: atitkov-at-micl.com.au
Do any of you probers out there have a spare TAP crystal for a JEOL 8600/733 probe that they may be willing to sell? ..we've had something of an accident..
Thanks,
Stu -------------------------------------------- Stuart Kearns Electron Microbeam Laboratories Department of Earth Sciences University of Bristol Queens Road Bristol BS8 1RJ UK tel: (0)117 954 5435 fax: (0)117 925 3385 e-mail: Stuart.Kearns-at-bristol.ac.uk http://eugf.gly.bris.ac.uk --------------------------------------------
When I was an undergraduate research assistant at Wash U, I worked in the McDonnell Center for the Space Sciences. They were using an epoxy called E-7 which I have been using ever since. It comes in two parts (A & B) which you mix in a 2 to 1 ratio. Curing time is 2 hours at 150 degrees F. If cured right, there is really no outgassing or beam damage, even at higher micrprobe currents. Contact Pat Swan on the 4th floor, he might still use it. You can buy it from:
Techkits PO Box 105 Demerest, NJ 07627 201-768-7334
The latest price was $17.25/set for {10 sets. Hope this helps, Lou Ross Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html
American Lab will be running an extensive review of new equipment being displayed at the upcoming PITTCON meeting (March 7-11, Orlando). For the first time, microscopy and related imaging will have its own section, as part of the on-going FOCUS ON MICROSCOPY column. If you are: (1) showing products which have not been exhibited or presented at a prior PITTCON and/or (2) introducing new products at this PITTCON please send copies of press kits, press releases, slides/product shots, and any other information to me at the address below. Indicate on the envelope that this is for the PITTCON Microscopy/Imaging review.
This article will need to go to press shortly after the meeting, so if I can get a head start on your materials, I would greatly appreciate it. Also, I will be on the show floor, following through on any materials received, so please enclose your booth number, name of a pertinent contact, and, if possible, a selection of times when they might be in your booth.
This article will appear in the May issue of American Lab.
Please call/email if you have any questions. Thanks in advance for your assistance.
Best regards, Barbara Foster Microscopy/Marketing & Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Contributing editor to American Lab ("Focus on Microscopy" column)
"Jeff" wrote: ============================================== Good Day,
Does anyone know of a dry etch that has a high selectivity to etch SiO2 preferentially to Si? We need to image these samples in a SEM. ================================================= This is usually done in a plasma etcher, using CF4 or some other reactive F gas of the more expensive types. You can get more information about this on our website given below. Typically, in a 100 watt barrel etcher, you can remove 1 um of SiO2 in about 30 minutes. The process is completely dry and is used in failure analysis laboratories.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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We are beginning to use the Historesin plus embedding medium supplied by Leica for immunocytochemistry of the retina by light microscopy. We will be using the images to count photoreceptor and retinal pigment epithelial cells of animals fed special diets to examine the effects of nutrition on the retina. I would like to communicate with others who have used this product to learn the best ways of storing material to preserve immunoreactivity for long periods of time and to share information on other technical issues.
You can respond to me directly at the address below.
Max Snodderly, Ph.D. Schepens Eye Research Institute 20 Staniford Street Boston, MA 02114, USA
In a message dated 2/1/99 9:03:47 PM Eastern Standard Time, kjtobin-at-uic.edu writes:
{ { I was wondering if anyone in the microscopic community could recommend an outfit that can polish petrographic thin sections for EMP work. I need to can the samples processed within a month timeframe and I am willing to pay some for this. } }
We recommend:
Mineral Optics Laboratory 29 "A" Street, P. O. Box 828 Wilder, Vermont 05088 USA 802-295-9373 802-295-7540 (FAX)
Yours truly, Steve Stokowski Stone Products Consultants Concrete Petrographers 10 Clark Street, Suite A Ashland, Massachusetts, 01721 USA 508-881-6364 http://members.aol.com/CrushStone/petro.htm
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Hello to all!
I am looking for a set of negative cassettes (30 plates), a cassette magazine box, and a cassette receiver box for Hitachi-600 TEM. Please let me know if any of you or your friends have a Hitachi TEM being taken down apart and wants to give away or trade these things.
Thanks in advance.
Gang Ning EM Facility Medical College of Wisconsin 414-456-8344
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Hello to all!
I am looking for a set of negative cassettes (30 plates), a cassette magazine box, and a cassette receiver box for Hitachi-600 TEM. Please let me know if any of you or your friends has a Hitachi TEM being taken down apart and wants to give away or trade these things.
Thanks in advance.
Gang Ning EM Facility Medical College of Wisconsin 414-456-8344
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Colleagues will be growing Vero cells (Green Monkey Kidney Cells) and intestinal epithelial cells on cover slips for subsequent ultrathin sectioning and TEM examination. I am interested to know which cover slips are best to use for this purpose. Can they be purchased sterile? I have checked the Ted Pella and EMS catalogs, they both sell non sterile cellulose acetate cover slips. Are there compatibility issues with resins and solvents? I am interested in any tips or tricks to smooth the way. Thanks, Sally Burns
Sally Burns Center for Electron Optics B5 Pesticide Research Center Michigan State University East Lansing, MI 48823 (517) 355-5004 Phone (517) 353-5598 FAX
I need some microneedles to dislodge some very fine particles (micron size) on my sample but still flexible enough to bend. Any information is appreciated.
As an electron microscopist who has come from a neurophysiology background, I have used various fine needles for "dusting" off debris. You need to find the one that feels right.
Cat whiskers are long, pointy, strong, and flexible. They are particularly good for chasing tiny bubbles out of microelectrodes or capillary tubes.
Finely drawn glass. Heat a pipet or rod over a Bunsen burner and draw it out until it breaks. Find somewhere along the long string that has the right size and flexibility, but won't break and leave more debris!
Cactus spines. They come in many shapes and sizes. Also useful for pinning down things for dissection that can't come into contact with metal.
Find someone in neurobiology who does microelectrode recording and get them to make some electrodes, which are capillary tubes drawn to a very fine point. You can probably get some in the micron range. Beveled, even!
Insect Minutin pins mounted on a stick are very strong, but may scratch your substrate. They can be ground down for a finer point.
Eyelashes, beard hair, and other body hairs each have different properties. Have fun experimenting.
Good luck!
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
We have used tungsten needles of about that dimension with our high- voltage electron microscope.. what we were looking for was a rather rigid "needle point" on which to mount micron sized objects. It was found that when we attempted to move these small specimens around and to get them to stick to the needles, the needles would bent very easily when they contacted the glass slides our specimens were prepared on. So you might try tungsten. We made them by electrolytically etching 5 mil. tungsten wires in NaNo3 solution, connecting the tungsten wire and a small graphite rod (1/8 inch dia.) to each of the terminals of a 6 volt AC transformer ( yes AC! ) and dipping both into the solution.The graphite rod was dipped well into the solution but the wtungsten would be dipped only an 1/8 inch or less below the surface. After a number minutes the wire would etch to a very small point, which could be examined under a light microscope until it was small enough for the application. Good luck.
For TEM of cultured cells, we grow the cultures on "Thermanox" tissue culture coverslips. From Nalge Nunc INternational, 50 sterile coverslips, 13 mm in diameter is catalog 174950. EMS also sells these thermanox cover slips in a variety of sizes (see page 143 of their cat XIII). The coverslips can be treated with all the same chemistry as tissue including propylene oxide and Spurrs epoxy, which are two components which solubilize polystyrene. These thermanox coverslips can be sunk cell side up in freshly made Spurrs, then following polymerization the coverslips can be removed by first sawing a small area of the epoxy/cell/substrate, then immersing in liquid nitrogen for a few seconds, then prying away the substrate. Now the embedded cells are immediate on the epoxy. Re-embed two fragments of the culture face to face for cross-sections, or cut the block parallel to the face for tangential sections. We particularly like the round 13 mm thermanox coverslips for immunocytochemistry of cultured cells since they can be floated cell side down in a drop of 100 microlitters antibody - gold conjugate, conserving reagents.
If you would like to grow a larger culture, you could also use "permanox" culture dishes, which are equally resistant to chemicals common in TEM processing. These are also available through EMS and other suppliers.
Good luck,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Microscopy Unit 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 503-221-3434 DRK-at-shcc.org
Orlando in March!!!! To Register follow the local meetings in www.vacuum.org or http://www.msa.microscopy.com
Over 30 Invited Talks, 40 vendors, and over 40 student posters!
Golf Tourny on Sunday March 14 - www.pegasus.cc.ucf.edu/~ampac
AVS short courses and UCF/Vendor Sponsored Short Courses in Tripod Polisher and FIB TEM Specimen Preparation see www.pegasus.cc.ucf.edu/~ampac -----------------------------
Monday March 15, 1999
Opening and Keynote Address
8:00-8:15 am Welcome and Introduction, Lucille Giannuzzi, 1999 FL AVS/FSM Program Chair
8:15-9:00 am Keynote Address, Sam Durrance, Astronaut, Professor, University of Central Florida
Monday, March 15, 1999
Session I: Thin Films
Session Moderator: Maggie Puga-Lambers, University of Florida
9:00-9:25 am Tim Anderson, Chemical Eng. Dept., University of Florida
9:25-9:50 am CONCENTRATION DEPTH PROFILING OF IMPURITIES AND DOPANTS IN FLAT PANEL DISPLAYS AND GLASSES BY SIMS, Temel Buyuklimanli, Evans East
9:50-10:15 am MODIFICATION AND CHARACTERIZATION OF DEFECT STATES IN ZnO FILMS, Gregory J. Exarhos and Charles F. Windisch, Jr., Pacific Northwest National Laboratory
10:15-10:45 am Coffee Break
10:45-11:10 am FUNDAMENTALS OF TUNGSTEN CMP DURING CMOS DEVICE FABRICATION, Rajiv Singh, Engineering Research Center for Particle Science and Technology, University of Florida
11:10-11:35 am SURFACE ANALYSIS APPLICATIONS IN SEMICONDUCTOR TECHNOLOGY, Bridget Rogers, Vanderbilt University
11:35-12:00 pm THE INFLUENCE OF AIR ON THE PROPERTIES OF THIN FILMS DEPOSITED FROM BEAMS OF NANOPARTICLES USING A COPPER SOURCE, F. K. Urban, III, A. Khabari, A. Housseini-Tehrani, P. Griffiths, and G. Fernandez
12:00-1:00 pm Lunch Monday, March 15, 1999
Session II: Microscopy of Biological Samples
Session Moderator: Karl Muffly, University of South Florida Jo Ann Moore, University of South Florida
9:00-9:25 am DIGITAL MANIPULATION OF ACQUIRED IMAGES, WHAT IS POSSIBLE VS. WHAT IS ETHICAL, John Kinnamon, University of Denver and The Rocky Mountain Taste and Smell Center
9:25-9:50 am THE USE OF CORRELATIVE MICROSCOPY IN BIOLOGICAL PROBLEM SOLVING, Ralph Albrecht, University of Wisconsin
9:50-10:15 am APPLICATIONS OF A 300 KV, FIELD EMISSION ELECTRON MICROSCOPE IN STRUCTURAL BIOLOGY, Kenneth A. Taylor, Florida State University Institute for Molecular Biophysics
10:15-10:45 am Coffee Break
10:45-11:10 am DECONVOLUTION VS STANDARD FLUORESCENCE MICROSCOPY, Karl Muffly, University of South Florida College of Medicine
11:10-11:35 am SURFACE AND MICROSCOPIC ANALYSIS OF BIOMATERIALS AS THEY CHANGE IN VIVO: HOW FAR ARE WE FROM NEEDED DATA?, Chris Batich and Anthony Brennan, University of Florida
11:35-12:00 pm CARBOHYDRATE DEPOSITION PATTERNS IN ETIOLATED SOYBEAN SEEDLINGS GROWN IN MICROGRAVITY, E.C. Stryjewski, NASA Gravitational Biology Laboratory, Dynamac Corp., K.M. Johnson, National Research Council, NASA/KSC, W.C. Piastuch1, H.G. Levine1, and L.H. Levine, NASA Gravitational Biology Laboratory, Dynamac Corp., O. Martynenko3, and V. Prima,, Institute for Molecular Biology and Genetics, National Academy Of Sciences, Ukraine
12:00-1:00 pm Lunch
1:30-3:30 pm WORKSHOP ON MANIPULATING DIGITAL IMAGES, John Kinnamon, University of Denver and The Rocky Mountain Taste and Smell Center
3:30-4:00 pm Florida Society for Microscopy Annual Business Meeting
3:30-6:00 pm Competition Session and Student Competition (Session IV) Monday, March 15, 1999
Session III: Surface Science and Analysis
Session Moderators: Sudipta Seal, University of Central Florida
1:00-1:25 pm WETTABILITY AND INTERFACES IN METAL/NITRIDE SYSTEMS, Natalia Sobczak, Foundry Research Institute, Cracow, POLAND
1:25-1:50 pm SOFT X-RAY FLUORESCENCE SPECTROSCOPY IN MATERIALS SCIENCE, E. Joseph Nordgren, Uppsala University, Uppsala, Sweden
1:50-2:15 pm MAGNETIC PHASE DIAGRAMS OF ULTRA-THIN FILM BINARY ALLOYS FOR SPIN-VALVE APPLICATIONS, Brian Tonner, University of Central Florida
2:15-2:40 pm PRACTICAL ASPECTS OF HIGH RESOLUTION XPS WITH MONOCHROMATIC AlKA X-RAYS, A.C. Miller, Lehigh University
2:40-3:05 pm STUDIES OF OXIDATION AND CORROSION USING AN ANAEROBIC CELL APPROACH, Peter M.A. Sherwood, Kansas State University
3:05-3:30 pm MICRO-ESCA/NEXAFS AT A THIRD GENERATION SYNCHROTRON LIGHT SOURCE, J. H. Underwood, U. Kleineberg, S. Mrowka, P. J. Batson, S. B. Rekawa, M. S. Jones, R. C. C. Perera, P. N. Ross, University of California, Berkeley
3:30-4:00 pm Coffee Break
3:30-6:00 pm Competition Session and Student Competition (Session IV)
Monday, March 15, 1999
Session IV: Poster Session
Session Moderators: Larry Plew, Cirent Semiconductor
CHARACTERIZATION AND SURFACE ANALYSIS OF HYDROXOCARBONATE COMPOUNDS, B. B. Adhyaru, K. R. Williams, and V.Y. Young, University of Florida
INTERFACIAL REACTIONS BETWEEN METAL/P-GaN FOR FORMATION OF OHMIC CONTACTS, M. Ahonen, B. Liu, P.H. Holloway, University of Florida
SURFACE METASTABLE STRUCTURE OF KTa03 (001) BY HELIUM ATOM SCATTERING, E. A. Akhadov, T. W. Trelenberg, J. A. Li, J. G. Skofronick, S. A. Safron, Florida State University and L. A. Boatner, Oak Ridge National Laboratory
SIMS STUDY OF DIFFUSION PHENOMENA OF METAL ELEMENTS IMPLANTED INTO SILICON, Elvira V. Anoshkina,a,b) Hughes Francois-Saint-Cyr,a,b,c) Ashfaq Hussain,a,b) , Isaiah Oladeji,d) Fred A. Stevie,e) Lee Chow,b,d) Dan Zhou a-dUniversity of Central Florida, eCirent Semiconductor
FAILURE ANALYSIS : AN AES-SEM STUDY, K. R. Beaulieu, (UNDERGRADUATE) A. S. Kale, S. Seal, V. Desai, University of Central Florida
IDENTIFICATION OF SURFACE CHEMICAL FUNCTIONAL GROUPS IN POLYMER MEMBRANES: AN X-RAY PHOTOELECTRON SPECTROSCOPY STUDY, Sharon D. Beverly 1,2, Satyajit V. Shukla, 3,4 Seungkwan Hong, 2 and Sudipta Seal3, 4, 1NASA, 2-4University of Central Florida
STUDY OF CORROSION FAILURES UNDER ATMOSPHERIC CONDITIONS, L.A. Bracho, V. H. Desai, S. Seal, University of Central Florida
ANISOTROPIC PATTERN TRANSFER IN GaN BY PHOTO-ENHANCED WET ETCHING, Hyun Cho(1), S.M. Donovan(1), C.R. Abernathy(1), J. Han(2), R.J. Shul(2), F. Ren(3) and S.J. Pearton(1), (1) & (3) University of Florida (2) Sandia National Laboratories
NOVEL EMITTER BASE SELF-ALIGNED PROCESS FOR AlGaN/GaN HETEROJUCTION BIPOLAR TRANSISTORS, X. A. Cao1, H. Cho1, S. J. Pearton1, C. R. Abernathy1, F. Ren1, J. Han2, R. J. Shul2, and A. G. Baca2, 1 University of Florida, 2 Sandia National Laboratories
HELIUM ISOLATION IMPLANT FOR GALLIUM NITRIDE BASED FIELD EFFECT TRANSISTORS, G. Dang1, X. Cao1, F. Ren1, S.J. Pearton1, J. Hang2, and R. J. Shul2, 1University of Florida, 2Sandia National Labs
CAPILLARIZATION OF SKELETAL MUSCLE IN RATS UNDERGOING HEART FAILURE, Hans Degens, Rebecca K. Anderson, Karl E. Muffly and Stephen E. Always, University of South Florida
UN-ANNEALED AND ANNEALED PD ULTRA-THIN FILM ON SIC CHARACTERIZED BY ATOMIC FORCE MICROSCOPY, SCANNING TUNNELING MICROSCOPY, AND X-RAY PHOTOELECTRON SPECTROSCOPY, K. Elshot, Mechanical Materials and Aerospace Engineering, University of Central Florida, Orlando, FL 32826, W. Lu, D.T. Shi,E. Bryant, K. Lafate, H. Chen, A. Burger, W. E. Collins, Department of Physics, Fisk University, Nashville, TN 37208
FUNDAMENTAL PROPERTIES ON E-BEAM EVAPORATED ZnS:Mn AND Zn1-xMgxS:Mn ELECTROLUMINESCENT THIN FILMS, Tao Feng, Mark Davidson, Paul Holloway, University of Florida
DESIGN, DEVELOPMENT AND TESTING OF A COMPUTERIZED DATA ACQUISITION AND CONTROL SYSTEM FOR A NANOPARTICLE DEPOSITION SYSTEM, Frank K. Urban and G Fernandez, Florida International University, Miami, Florida
CHARACTERIZATION OF THE DIFFUSION PROPERTIES OF Mg, Cl, K, Ge, AND Mo IN SILICON BY SIMS, Hughes Francois-Saint-Cyr,a,b,c) Elvira V. Anoshkina,a,b) Ashfaq Hussain,a,b) ,Fred A. Stevie,e) Lee Chow,c,d) Kathleen Richardson, a,b,c) and Dan Zhou a,b), a-dUniversity of Central Florida, eCirent Semiconductor
A STUDY OF THE SURFACE COMPOSITION OF KTAO3 DOPED WITH CA, BA, SR, AND NB, P.W. Gresser Florida State University
NOZZLE DESIGN IN A DIFFERENTIALLY PUMPED NANO-PARTICLE INSTRUMENT, Peter D. Griffiths and Frank K. Urban, Florida International University
INTERFACIAL CHRACTERISTICS OF AlN TO Si, SiC AND GaN, K. Harris, B. P. Gila, F. Ren, J. Deroaches, K. N. Lee, J. D. MacKenzie, C. M. Zitterling+, M. Ostling+, S. N. G. Chu**, C. R. Abernathy, and S. J. Pearton, University of Florida, Gainesville, FL, +KTH, Royal Institute of Technology, Stockholm, **Bell Laboratories, Lucent Technologies
SELECTIVE DRY ETCHING USING INDUCTIVELY COUPLED PLASMAS: GaAs/AlGaAs AND GaAs/InGaP, D.C. Hays, H. Cho, K.B. Jung, Y.B. Hahn*, C.R. Abernathy, S.J. Pearton, and F. Ren, University of Florida, W.S. Hobson, Bell Laboratories, Lucent Technologies
HOT FILAMENT CVD OF DIAMOND THIN FILMS, Ashfaq Hussain, Lee Chow, and Dan Zhou, University of Central Florida
QUANTITATIVE COMPARISON OF VON WILLBRAND FACTOR (VWF) EXPRESSION IN HUMAN NON-MALIGNANT AND MALIGNANT TISSUE USING CONFOCAL LASER SCANNING MICROSCOPY (Undergraduate), D Janarious, J Biggerstaff, JL Francis, Walt Disney Memorial Cancer Institute
DIETARY MODIFICATION AND THE ALZHEIMER'S-LIKE PHENOTYPE IN mAPP/mPS1 TRANSGENIC MICE, P.T. Jantzen, M.N. Gordon and D.G. Morgan, University of South Florida
COMPARISON OF CHARACTERISTICS DRY ETCHING OF LaCaMnO3, NiMnSb AND NiFe THIN FILMS, K. B .Jung(1), Hyun Cho(1), J. J. Wang(1), J. Caballero(1), Tao Feng(1), J. R. Childress(2), K. H. Dahmen(3) and S. J. Pearton(1), (1)University of Florida, (2)IBM Almaden Research Center (3)Florida State University
FABRICATION OF DC-MAGNETRON SPUTTERED 70Ti-30Al THIN FILMS, A. S. Kale, K. R. Beaulieu, K. B. Sundaram, V. H. Desai, S. Seal, University of Central Florida
THE ATOMIC FORCE MICROSCOPY INVETIGATION OF THIN FILM DEPOSITED FROM NANOPARTICLE SOURSE, F.K. Urban, A. Khabari, Florida International University
COMPARISON OF ZnS:TbOF THIN FILMS DEPOSITED BY R.F MAGNETRON SPUTTERING USING ZnS, TbOF MIXED TARGET AND SEPERATED ZnS,TbOF TARGETS, Jongpyo Kim, Mark Davidson, Barbara Speck, David Moorehead, Qing Zhai, and Paul Holloway, University of Florida
ELECTRON BEAM DEGRADATION OF OXIDE AND SULFIDE THIN FILM PHOSPHORS FOR FIELD EMMISION DISPLAYS, Caroline A. Kondoleon, Billie Abrams, Philip Rack* and Paul Holloway, University of Florida, Rochester Institute of Technology
ELECTRON CYCLOTRON RESONANCE CHEMICAL VAPOR DEPOSITED SILICON NITRIDE FOR T-GATE PASSIVATION, J. LaRoche1, F. Ren1, S.J. Pearton1, J. R. Lothian2, J.W. Lee3, and D. Johnson3, 1University of Florida, 2Multiplex Inc, 3Plasma-Therm, Incorporated
DRY ETCHING OF BaSrTiO3 AND LaNiO3 THIN FILMS IN INDUCTIVELY COUPLED PLASMAS, K. P. Lee, K. B. Jung, A.Srivastava, D. Kumar, R. K. Singh and S. J. Pearton, University of Florida
EFFECTS OF Ni THICKNESS ON Ni/Ti/Ag OHMIC CONTACT TO p-GaN, B. Liu, M. H. Ahonen and P. H. Holloway, University of Florida
TITANIUM MAGNESIUM NICKEL ALLOY AND HYDROGEN STORAGE, Janice K. Lomness, Sudipta Seal, Michael D. Hampton, and Meredith Stowell (Undergraduate), University of Central Florida
(Undergradutate) CHARACTERIZATION OF BEAM PRODUCED BY PULSED ARC CLUSTER ION SOURCE, Samantha A. Moore and Anne J. Cox, Eckerd College
COMPARISON OF THE MICROSTRUCTURE AND ELECTROLUMINESCENT PROPERTIES OF ZnS:Mn DEPOSITED BY SPUTTERING AND ATOMIC LAYER EPITAXY, David J. Moorehead Karen E. Waldrip, M.R. Davidson*, J.H. Lee, B. Pathangey*, M.Puga-Lambers*, K.S. Jones, P.H. Holloway, University of Florida, and S.S. Sun and C.N. King, Planar Systems
TRI-LAYER ELECTRON BEAM RESIST FOR SUBMICRON T-GATE GaN BASED FIELD EFFECT TRANSISTORS, M. Mshewa, H. Hudspeth, F. Sharifi, S. J. Pearton, and F. Ren, University of Florida
A MODIFICATION OF THE VARIABLY DAMPED LEAST SQUARE ALGORITHM ASSISTED BY MEASUERED DATA POINTS AND DERIVATIVE PRESELECTION FOR IMPROVEMENT OF SOLUTIONS, J. Pontillo, F.K. Urban, Florida International University
PULSED CURRENT ELECTROMIGRATION, Cross Reardon and Rolf Hummel, University of Florida
ILLUSTRATION OF CAPILLARY PERFUSION IN HYPERTROPHIED CARDIAC MUSCLE USING THE FLUORESCENT DYE, THIOFLAVIN-S, Corey A. Schoder, Hans Degens, Don R. Hilbelink, Karl E. Muffly University of South Florida
FORMATION OF SILVER SULFIDE NANO-PARTICLES BY NOVEL SOL-GEL METHOD FOR INDUSTRIAL APPLICATIONS, S. Shukla and S. Seal, University of Central Florida
CO-LOCALIZATION OF GLIAL FIBRILLARY ACIDIC PROTEIN (GFAP), OX-42 AND OX-6 IN SPINAL CORD FOLLOWING SCIATIC NERVE DAMAGE, Stacy Sinibaldi, Edward Haller, and Samuel Saporta University of South Florida
CHARACTERIZATION OF THE CONSORTIUM BETWEEN THE RED TIDE CAUSING DINOFLAGELLATE GYMNODINIUM BREVE, AN UNIDENTIFIED BACTERIUM AND A PHAGE, Theresa R. Slifko, Lee Houchin, Anthony Greco, and John H. Paul, University of South Florida
INVESTIGATIONS INTO CHEMICAL MECHANICAL POLISHING OF TUNGSTEN USING VARIOUS ELECTROCHEMICAL AND SURFACE SCIENCE TECHNIQUES, Dnyanesh Tamboli, Sudipta Seal, Vimal Desai University of Central Florida
THE MINERAL CONTENT AND CELLULAR STRUCTURE OF HETEROTOPIC BONE, Gregory S. Taylor University of Florida College of Medicine, and Melinda K. Harman and W. Andrew Hodge, Orthopaedic Research Laboratory
EFFECTS OF FIB OPERATING CONDITIONS ON BEAM DAMAGE OCCURRING IN SILICON TEM SAMPLES, C.A. Urbanik, B.I. Prenitzer, L.A. Giannuzzi, S.R. Brown1, R.B. Irwin1, B. Rossi2 , T.L. Shofner2, and F.A. Stevie1, University of Central Florida, 1Cirent Semiconductor, 2The Bartech Group
IMPROVED PERFORMANCE IN THIN FILM ELECTROLUMINESCENT PHOSPHORS BY DONOR DOPING, K.E. Waldrip, J.S. Lewis, III, Q. Zhai, D. Moorehead, M.R. Davidson*, and P.H. Holloway, University of Florida, and S.S. Sun, Planar Systems, Inc.
EFFECTS OF FLUX AGENTS ON THE MICROSTRUCTURE AND ELECTROLUMINESCENCE OF SPUTTERED ZnS:Mn THIN FILM PHOSPHORS, Qing Zhai, Karen Waldrip, Jay Lewis, Jungpyo Kim, David Moorhead, Jinghong Li, Kevin Jones, and Paul Holloway, Maggie Puga-Lambers, Barbara Speck, and Mark Davidson, Microfebritech, University of Florida
ICP Cl/Ar PLASMA DAMAGE ON GaN SCHOTTKY, A. Zhang1, H. Cho, F. Ren1, S.J. Pearton1, J. M. Van Hove2, P. P. Chow2, R. Hickmand2, J. J. Klaasen2 and R. J. Shul3, 1University of Florida, 2SVT Associates, 3Sandia National Labs
MICROSTRUCTURE EVALUATION OF STRESS CORROSION CRACKING USING FIB TECHNIQUES, Hanlin Zhang, Brian Kempshall, Carrie Urbanik and Lucille A. Giannuzzi, University of Central Florida
Tuesday, March 16, 1999
Session V: Microscopy of Physical Samples
Session Moderator: Lucille A. Giannuzzi, University of Central Florida
8:30-8:55 am ANALYTICAL MICROSCOPY IN THE REAL SEMICONDUCTOR PROCESSING WORLD, Ron Anderson, IBM Analytical Services
8:55-9:20 am FIB APPLICATIONS IN MATERIAL SCIENCE PROBLEMS, M.W. Phaneuf, J. Li, G.A. Botton*, L. Weaver, Fibics Incorporated, *Materials Technology Laboratory
9:20-9:45 am FOCUSED ION BEAM IMAGING OF MICROELECTRONIC STRUCTURES, Ann N. Campbell, Sandia National Laboratories
9:45-10:10 am DETERMINATION OF THE COMPOSITION OF GRAIN BOUNDARIES AND INTERFACES BY X-RAY MICROANALYSIS, David B. Williams, Lehigh University
10:10-10:40 am Coffee Break
10:40-11:05 am MATERIALS APPLICATIONS OF ELECTRON HOLOGRAPHY, Altaf Carim, The Pennsylvania State Univeristy
11:05-11:30 am PHASE MAPPING AND PHASE IDENTIFICATION USING ELECTRON BACKSCATTER DIFFRACTION, David J Dingley and Stuart I Wright, TexSEM Laboratories
11:30-11:55 pm AFM, Phil Russell, North Carolina State University
12:00-1:00 pm Lunch and Vendor Talks (Session VI)
Tuesday, March 16, 1999
Session VI: Vendor Presentations
Session Moderators: Fred Stevie, Cirent Semiconductor
12:10-12:20 pm LOW ENERGY ION MILLING FOR TEM, David Henriks, South Bay Technologies
12:20-12:30 pm INNOVATIONS IN MASS FLOW CONTROLLERS FROM UNIT, Greg Vaughan, Schoonover, Inc.
12:30-12:40 pm RECENT DEVELOPMENTS IN ATOMIC FORCE MICROSCOPY, Matt Thompson, Digital Instruments
12:40-12:50 pm NEW PRODUCTS FROM SPECS: Er-LEED, ULTRA HIGH RESOLUTION EEES: DELTA0.5, AND PLASMA UV SOURCE UV300, Dietrich von Diemar, SPECS U.S.A.
Tuesday, March 16, 1999
Session VII: Electronic Materials
Session Moderator: Drew Hoff, University of South Florida
1:00-1:25 pm LOW FIELD ELECTRON EMISSION FROM UNDOPED NANO-STRUCTURED DIAMOND, W. Zhu, G. P. Kochanski and S. Jin, Bell Laboratories, Lucent Technologies
1:25-1:50 pm SYNTHESIS AND APPLICATIONS OF NANOCRYSTALLINE DIAMOND FILMS, Dieter M. Gruen, Argonne National Laboratory
1:50-2:15 pm CHARACTERIZATION OF SHALLOW JUNCTIONS USING SECONDARY ION MASS SPECTROMETRY, Charles W. Magee, I.M. Abdelrehim, T.H. Buyuklimanli, J.T. Marino and W. Ou, Evans East
2:15-2:40 pm NOVEL PROCESSING OF ELECTRONIC MATERIALS, W. V. Lampert, Air Force Research Laboratory, WPAFB, OH
2:40-3:05 pm GaN BASED ELECTRONICS FOR HIGH TMEPERATURE APPLICATION, F. Ren1, S. J. Pearton1, C. R. Abernathy1, J. M. Van Hove2, P. P. Chow2, R. Hickmand2, J. J. Klaasen2, J. Han3, A. G. Baca3, and R. J. Shul3, 1University of Florida, 2SVT Associates, 3Sandia National Labs
3:05-3:30 pm LOW ENERGY IMPLANTATION AND SHALLOW JUNCTIONS IN Si, Kevin Jones, University of Florida
3:30-4:00 pm Coffee Break, Student Awards and Door Prizes
Tripod Polisher, instructor: Ron Anderson March 12-13 FIB, instructors: Lucille Giannuzzi and Fred Stevie March 18-19
in conjunction with the FL AVS/FSM meeting week of March
for information see www.pegasus.cc.ucf.edu/~ampac or contact lag-at-mail.ucf.edu
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-mail.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Tong - You might find that a glass needle will work for your purpose and you can easily make them yourself to almost any desired thinness. Simply take a thin 8 or 10 inch long glass rod (can be hollow too) and heat near the center with a bunsen burner or other source. After glowing pull the ends apart and you will draw the glass down to a suitable thickness. At the micron level glass is relatively strong for moving particles and as you will find quite flexible.
Dave Joswiak Dept. of Astronomy Univ. of Washington Seattle, WA 98195 (206)543-7702
On Tue, 2 Feb 1999, Tong Wang wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } I need some microneedles to dislodge some very fine particles (micron size) } on my sample but still flexible enough to bend. } Any information is appreciated. } } Tong } } } } }
Postdoctoral Positions in Nuclear Materials Science - PD994571
The Materials Science and Processing Group (NMT-11) at Los Alamos National Laboratory is seeking qualified applicants to fill two postdoctoral positions in the science of nuclear materials. Successful candidates will work with technical staff members in the Materials Science and Technology (MST) and Nuclear Materials and Technology (NMT) Divisions. Both positions require the willingness to work on radioactive materials and the ability to obtain a DOE Q clearance (which usually requires US citizenship).
Position One: Electron Microscopy of Plutonium and Ceramic Actinide Waste Forms. The individual will have access to (a) the Electron Microscope Facility in MST for microstructural studies of unirradiated and surrogate materials and (b) the Materials Characterization Facility in NMT for analysis of radioactive materials. Techniques available for this research include SEM, OIM, TEM, HRTEM and STEM. This work will support research efforts in radiation effects in ceramics and plutonium characterization for weapons programs. Candidates must have prior experience in electron microscopy and microanalysis. Contact Jeremy Mitchell (505-665-3934, jeremy-at-lanl.gov) or Kurt Sickafus (505-665-3457 kurt-at-lanl.gov) for further technical information.
Position Two: Radiation Damage in Metals, Self-radiation Damage in Plutonium Metal, Alloys and Compounds. This work is basic research in support of plutonium metallurgy and chemistry for the weapons programs. Candidates must have prior experience in x-ray and/or neutron diffraction and the associated data analysis with GSAS, knowledge of mechanisms of radiation damage in metals and some knowledge of condensed matter physics. The individual will also have access to calorimeters, SEM, OIM, and TEM. Contact Luis Morales (505-665-7703 lmorales-at-lanl.gov) for additional technical information.
A Ph.D. in Materials Science, Metallurgical Engineering, Geosciences, or a related field completed within the last three years or soon to be completed is required. Candidates may compete for a Director's Fellowship and outstanding candidates may be considered for the prestigious J. Robert Oppenheimer, Richard P. Feynman or Frederick Reines Fellowships. Further details about the Postdoctoral Program may be found at: http://www.hr.lanl.gov/postdoc/ For consideration, submit a resume and publications list along with a cover letter outlining current research interests to postdoc-jobs-at-lanl.gov (no attachments, please!)
OR SUBMIT TWO COPIES to:
Postdoc Program Office, PD994571 MS P290 Los Alamos National Laboratory Los Alamos, NM 87545
NOTE: Advertisement #PD994571 must be referenced in the e-mail Subject line (or the address) and cover letter.
Affirmative Action/Equal Opportunity Employer. Individuals with disabilities needing reasonable accommodation should call (505) 667-8622. A Teletype Device for the Deaf (TDD) is available by calling (505) 665-5357. Los Alamos National Laboratory is operated by the University of California for the US Department of Energy. ============================ Jeremy N. Mitchell MS G730, NMT-11 Los Alamos National Laboratory Los Alamos, NM 87545 Phone: 505-665-3934 Fax: 505-665-4013
Dear Sally, The coverslips that everyone uses is my Thermanox plastic coverslps which are sterile and come in many different sizes depending on your need. They may be found wither in our hard copy catalog or on our website at www.emsdiasum.com. Go to our online catalog and click chapter 7. They are listed under Thermanox. In our hard copy catalog XIII they may be found on page 143. Please let me know if I may be of further assistance to you.
Sincerely, Electron Microscopy Sciences 215-646-1566
The best way I know of preparing pigments for TEM analysis is by blowing an aerosol of the sample dispersed in alcohol/water solution over a hot plate and collecting the sample on a grid placed at the far end of the hot plate. The company I work for is concerned about the safety issues surrounding this so does anybody know of any less hazardous sample preps. or a simple containment facility for the aerosol spray? I thought about building a glass box for containment but are there any commercially available containment/sample prep. units?
Thanks
Brian Manzor e-mail: brian.manzor-at-grmouth.zeneca.com
The best way I know of preparing pigments for TEM analysis is by blowing an aerosol of the sample dispersed in alcohol/water solution over a hot plate and collecting the sample on a grid placed at the far end of the hot plate. The company I work for is concerned about the safety issues surrounding this so does anybody know of any less hazardous sample preps. or a simple containment facility for the aerosol spray? I thought about building a glass box for containment but are there any commercially available containment/sample prep. units?
Thanks
Brian Manzor e-mail: brian.manzor-at-grmouth.zeneca.com
Hi Michael! To avoid tissue curling try 50% or 30% ETOH in the first dehydrating step. You can do dyhradation in cell-culture multiwells. You=B4ll have to remove the specimens if you=B4d like to use Propylene ox= ide because this interacts with the multi-well plastic. If you like to embed the sections flat, do it on silicated glass slides(o= r try a slide, coverd with slightly fatted aluminium foil) , put a drop of Epon/araldite on it, leave it at 45=B0C in the oven (12-24h) and then put= BEEM Capsules filled with Epon/araldit on the top(take care of bubbles!) , polymerize 2 days at 60=B0C. If you can=B4t remove the glass slide easily= , try it with fluid nitrogen. Good luck, Michael
MICHAEL DELANNOY schrieb:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------. } } Greetings, } I need assistance with flat embedding of rat cerebellum } (IEM-DAB), that is 100 um thick and 1.5 x 0.5 cm dimensions. Basically } we are worried about tissue curling during the dehydration. My first } inclination is argarose embed (before epon), but I would take any exper= t } advise. } Thank you, } } Mike D
Sprays can be contained to some extent within a box for spraying thin-layer chromatographic plates, 20 x 20 cm or a little larger. This has to be placed in a hood, since it won't catch and retain the smallest droplets. They come in cardboard and plastic versions. I can't find one in VWR, but any big Web catalog with a search capability should lead you to one.
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
Hi all. We have identified a need for some hands on WDS training here in South = Africa. A number of our clients have either changed operators or = upgraded to new integrated systems. So we would like to know if there is = any one who would be able to help with presenting a WDS course here in = South Africa. No need to panic, South Africans do have money and are = willing to pay, not too much mind you.=20
We are open to the course content, however would like to cover a bit of = customer care operations, calibration and set up and then theory and = hands on operation.=20 We will have a Jeol 733 with an Advanced Microbeam / Edax integrated = system available by about June onwards.=20
If any one is interested please let us know so we can discuss details. Thanks
Luc Harmsen=09 Anaspec, South Africa International technical support on microscopy. Tel: +27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290 anaspec-at-icon.co.za
Hello All, Thanks to all those who offered suggestions for the damping of vibrations sometimes experienced when using a light microscope. Once again you have proven to be an ingenious lot! Users have their microscope sitting on platforms supported by any of the following; tennis balls, "sandwiches" of bubble pack and aluminum plates or inner tubes all of which indicates that the pneumatic tables or benchtop isolation systems are preferable to the marble tables. An alternative approach is to support the microscope on a platform beneath which are sandwiched layers of Sorbothane (elastomer) and aluminum plates. One simple suggestion was to "plant" each leg of a conventional table; supporting your microscope, into its own little box of sand. I'm a little wary of this last suggestion as some of the laboratory wildlife might find the sand an attractive litter box!
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
To Dr. Naseem: TEM results on paraffin processed tissue will only be as good as the processing procedure used by the histology lab. In my experience, most histology labs use a very aggressive protocol, which causes extraction of cellular components to the point where only nuclei and possibly cell membranes might be recognizable. Laboratories that handle large volumes and are concerned with quick turnaround generally dehydrate and clear the heck out of specimens so they don't have to go back and reprocess if something was too big, not well fixed, etc. Our laboratory provides good fixation, gentle dehydration, clearing and infiltration of routine histology specimens so that when I have to do EM on one of these, there is usually enough cellular matrix left to make the effort worthwhile. I have been able to resolve Birbeck bodies, tonofilaments, junctions, microvilli and immune complex deposits (in glomeruli) in paraffin processed specimens. Good luck! Bob Santoianni Emory University Hospital Atlanta, Georgia robert_santoianni-at-emory.org
To process cell cultures for grown on round, 13 mm Thermanox tissue culture cover slips for TEM, we use the following hardware:
For fixation, we use porcelain multi-well dishes. These are what most people refer as "staining dishes". They are white, measure about 3.5 x 4.5" and have 12 depressions. We use these for glutaraldehyde, buffer rinses and OsO4. OsO4 can be completely rinsed from the dishes. Once in the last buffer after OsO4 and prior to ethanols and propylene oxide, we transfer the disks to 50 ml polypropylene culture tubes, such as Fisher 05-539-7 (these are sterile, but certainly it is not necessary to pay for sterile containers). There is plenty of room to enter a pipette for fluid exchange without touching the culture disks, and the culture surface will not touch the walls of the cylindrical tube so there is no worry that the cells will be rubbed off. Given the depth of the tube, we do not worry too much that the culture will dry out as fluids are exchanged, since the atmosphere within the tubes is fairly saturated with solvent vapor. Still, fluid exchange is done quickly. The polypropylene tubes are resistant to P.O. and Spurrs. Do not use tubes made from polystyrene as they will dissolve. Once infiltrated with the last change of epoxy, we fill a resin-resistant container with epoxy to a minimum depth of 5 mm, sink the disk so that the cells face up, then polymerize the epoxy. We steal the polypropylene lids from wheaten snap-cap vials (Fisher #0333520E) which are of the appropriate diameter for embedding 13 mm coverslips. Wearing a dust mask, We use a jewelers saw to free small blocks of embedded culture, loosen the cover slip with liquid nitrogen, then remove the disk which exposes the culture to the surface of the block. We then either re-embedded (in some of the same batch of media which was used to infiltrate the culture) face to face for cross sections, or cut the blocks parallel to the culture surface for tangential sections.
Good luck,
Doug Keene EM Facility Shriners Hospital for Children DRK-at-shcc.org
} } Hello All, } Does anyone out there have any suggestions for getting good SEM } cross sectional images from dried wood? I have a student who would } like to image the microstructure of several types of wood. All the
} } Thanks, } Scott
Hi Scott,
I think the classic paper you want is Exley et al. (1973 ?) J. Microscopy, vol 101, 21-30 "Preparation of wood specimens for the scanning microscope".
I don't have a copy handy to check, but I think this paper emanated from good ol' New Zealand! As I recall, the authors were able to cut fresh lignified wood satisfactorily using a fresh razor blade for each cut, and then cleaned the surfaces with hypochlorite solution before drying and coating. Most specimens were OK with just air drying but delicate features needed CPD. The authors often cut the one specimen in two planes to good effect, e.g. transverse and radial longitudinal.
If you want to look at long-dead wood it might be too hard to cut cleanly. You might need to soak it in something first. Exley et al. might have suggestions.
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department Faculty of Medicine & Health Sciences University of Auckland Private Bag 92019 Auckland New Zealand
-----Original Message----- } From: Gerroir, Paul J {Paul.Gerroir-at-crt.xerox.com}
} One simple suggestion was to "plant" each leg of a } conventional table; supporting your microscope, into its own little box of } sand. I'm a little wary of this last suggestion as some of the laboratory } wildlife might find the sand an attractive litter box!
Mix red pepper with the sand that will keep the kitty out of the sand.
Gordon
Gordon Couger gcouger-at-couger.com Owner PRAG-L PRactical AGriculture List www.couger.com/prag-l Stillwater, OK 405 624-2855 GMT -6:00
I posted a notice several months back about a Siemens 101 Transmission Electron Microscope that was up for grabs. Although we had some interest, no one decided to take it. At this point, we have a major space crunch and need to dismantle and get rid of it. I've been told it contains 25 gms of liquid mercury which should be removed before disposal. Can anyone refer me to a technician familiar with this equiment in the NY area who could assist us in this project. Thanks in advance for your help. Please reply directly to me. I am also posting this notice on the safety list. ---------------------------------------- Gerry Griffin Environmental Services NYU Medical Center Email: Gerry.Griffin-at-med.nyu.edu
I'm studying the transport and localization of some proteins in the cytoplasm of cultured cells. The cells are fixed in paraformaldehyde and stained with a avidin-biotin-fluorescein system. I need to do double staining for this protein and the Golgi, so I intend to use the Bodipy-tr-ceramide probe, as suggested by Molecular Probes .
I would like to know if someone has experience or worked with this specific probe. Particularly I need to know if this probe may be used with fixed cells, that is, after the cells are fixed.
I will be very grateful for any information
Thanks in advance
Francisco Javier Hernandez-Blazquez IARC-WHO Unit of Multistage Carcinogenesis 150 Cours Albert Thomas Lyon - France
We are interested in buying a variety of used AFM and STM equipment. To see our wish list, go to our web page (or contact us directly) http://a1.com/asm/wantused.htm
thanks Don Chernoff
Advanced Surface Microscopy, Inc. E-Mail: asm-at-indy.net 6009 KNYGHTON RD. Voice: 317-251-1364 INDIANAPOLIS IN 46220 Toll free: 800-374-8557 (in USA) web: http://www.a1.com/asm Fax: 317-254-8690 (note: "a1"= letter "a", numeral "1")
I am looking for a postdoc/visiting scientist who will do developmental work on Direct Methods for structure determination using dynamical electron diffraction data. If you know of anyone who has: a) A good diffraction background b) A good crystallography background c) Good computer skills Please ask them to contact me. The position is available now.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
I just got an Amray 1600 with turbo and would like to correspond with others who are using this instrument. If there are any quirks about it, I would like to know what they are. If anyone has brought one up after several years of storage, I'd REALLY like to hear from you.
I'm hoping to be able to take 4x5" sheet film pix and 120 roll film pix.
Any info on the care and feeding of the turbo pump, roughing pump and inevitable column cleaning would be appreciated. I'm told that this SEM can take good pictures. I'm hoping to find out if this is true.
Having serviced Siemens microscopes I can tell you that the source of th= e mercury is a mercury diffusion pump. This pump is situated between the rotary pump and the "oil" diffusion pump in the vacuum circuit.
So that is where the mercury is now to dispose of it is another problem?
Stay safe.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
I am currently in the process of acquiring a TEM with accessory equipment. We are starting a new TEM-lab here at the Swedish Museum of Natural History.
One of the instruments I am considering is the LEO 912 AB. Anyone on the list with experience of that microscope? We will primarily be doing morphology including ESI imaging of thick sections, but not analytical work or cryo.
Also: any recommendations on which ultramicrotome to choose.
TIA
Ulf Jondelius
Ulf Jondelius, Invertebrate Zoology Swedish Museum of Natural History P.O.B. 50007, 104 05 Stockholm, Sweden phone: Int + 46 (0)8 666 5160 fax: Int + 46 (0)8 666 4125 e-mail: ulfj-at-nrm.se
Here's a few comments to follow from Doug Keene's reply to you celss on coverslips question.
You can process and section Thermanox quite easily with cells grown on them.
Orientation problems can be overcome if you cut the coverslip to fit a BEEM capsule before you innoculate with cells. I have seen good results with various cell types, and if you taper the cut coverslip to fit into the BEEM pot, trimming the block is pretty quick too. If you need, you can also bend up the coverslip at the non-tapered end to indicate which side the cells are growing.
Spurrs resin seems pretty good, but there can be some delamination between the coverslip and resin, but not always!
Hope this helps.
Plymouth Electron Microscopy Unit University of Plymouth Drake Circus Plymouth Devon UK PL4 8AA Tel: 01752 233092 Fax: 01752 233092 email: pbond-at-plymouth.ac.uk
I need to colorize some SEM pictures but I do not know which programm to use. If someone can help me, I use PC or Macintosh computer. Thanks a lot for your help Best regards
Didier
-------------------------------------------- Didier Le Thiec I.N.R.A. Centre de Recherches Forestieres Unite d'Ecophysiologie Forestiere Laboratoire de Pollution Atmospherique 54280 Champenoux - France
To save a long search can anyone remember which months archive of the listserver the discussion about preparing cd-roms for sem is in??
Chris
Chris Gilpin Experimental Officer Biological Sciences EM Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 0161 275 5170 Fax +44 0161 275 5171 Chris Gilpin http://www.empgu.man.ac.uk
I am now running a clinical path, diagnostic EM lab. but must reduce my storage. What are the clia and cap regs on how long to keep transmission em blocks, thick section slides, grids and phot mic's? I am being reduced from a 3 room lab to a room and a closet. thanks for you help. S. Drew
I would like to draw your attention to the materials science symposium being held as part of the Scanning 99 meeting in April this year. The abstract deadline is February 15th (the same as MSA) and the abstract format is the same as the MSA format. It is intended that there will be contributed presentations as part of the program, and depending on the number of submissions, the symposium may be extended for another day. Please forward this e-mail to anyone you think may be interested in attending the symposium.
{bold}
"Analyzing Materials Interfaces at Atomic Resolution" {/bold}
There will be a Materials Science symposium at Scanning 99 entitled "Analyzing Materials Interfaces at Atomic Resolution". Scanning 99 is being held at the Hyatt Regency O'Hare in Rosemont, IL from April 11-14 1999. The "Analyzing Materials Interfaces at Atomic Resolution" symposium is tentatively scheduled for Tuesday April 13th and will consist of invited and contributed presentations. The details of the conference and deadlines/forms for abstract submissions can be found at http://www.scanning.org or details can be requested from Mary Sullivan (e-mail: scanning-at-fams.org, tel:201-818-1010). Registration for members of the Midwest Microscopy and Microanalysis Society are at the reduced rates of $150 for the whole conference or $50 for a single day
The discussion was last September, maybe on into October. I will send you a copy of the summary that I have on hand.
Warren S.
At 02:18 PM 2/5/99 +0000, you wrote: } } Hi all } } To save a long search can anyone remember which months archive of the } listserver the discussion about preparing cd-roms for sem is in?? } } Chris Gilpin } Experimental Officer } Biological Sciences EM Unit } G452 Stopford Building } Oxford Road } Manchester } M13 9PT } phone +44 0161 275 5170 } Fax +44 0161 275 5171 Chris Gilpin } http://www.empgu.man.ac.uk }
Just a reminder:ACS's Applied Optical Microscopy will be held in conjunction with PITTCON, March 5-7, in Orlando, Florida.
This is a three-day, hands-on total immersion workshop. Although this workshop is offered under the American Chemical Society Short Course programming, it is not just for chemists. The curriculum focuses on key techniques to help you be more effective in the lab. Topics range from alignment, care and cleaning to contrast techniques and basic measurement. There is also a special Saturday evening program on video and digital imaging and a full day on qualitative and quantitative Polarized Light Microscopy. Participants are encouraged to bring samples.
For details and registration information, visit the MME website {http://www.MME-Microscopy.com/education} or call the course coordinator, Barbara Foster
Barbara Foster Course Coordinator ACS Applied Optical Microscopy Course c/o Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 (413)746-6931 Fax: (413)746-9311 email: mme-at-map.com
According to some of the messages I saved, the dates are mid-August and September '98. Hope this helps.
Chris Gilpin wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi all } } To save a long search can anyone remember which months archive of the } listserver the discussion about preparing cd-roms for sem is in?? } } Chris } } Chris Gilpin } Experimental Officer } Biological Sciences EM Unit } G452 Stopford Building } Oxford Road } Manchester } M13 9PT } phone +44 0161 275 5170 } Fax +44 0161 275 5171 Chris Gilpin } http://www.empgu.man.ac.uk
I was just asked to provide the equation for gamma adjustment to image contrast. I had always assumed that it was just
Output = (Input)^gamma i.e. a simple power law.
Thus for gamma = 0.5 you get a square root curve and for gamma = 2.0 you have a parabola. However, the contrast curves in various books I've checked do not appear to be a power law, but look more like circular arcs. Unfortunately these books only describe gamma but do not provide the equation. Are the curves just artist's license or is my understanding of the gamma correction wrong?
If you include Contrast and Brightness, I would expect the general equation to be
Output = B + C*(Input)^gamma
Where B = Brightness and C = Contrast.
Thanks, Henk Colijn Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu An optimist believes that we live in the best of all possible worlds. A pessimist fears that this is true.
Following a very successful year, NORAN Instruments is pleased to announce employment opportunities within its Confocal Sales Division.
The position of District Sales Manager will be responsible for all confocal sales activities within a predefined region of the United States.
Functions will include: * Evaluation and qualification of customer needs * Demonstration of confocal systems, including configuration and setup * Coordination of installation and initial user training
Necessary qualifications include: * B.A. or B.S. degree in Natural Sciences. Advanced degree desirable. * One year's hands on experience with light microscopy techniques. * Experience and practical knowledge of Confocal microscopy applications. * Experience with computer image analysis and processing preferred. * Sales experience is highly desirable.
The position requires travel of up to 50% within designated sales region.
For consideration, please fax or e-mail your resume in confidence to:
Could some kind, knowledgable person explain why the addition of calcium = to fixatives (buffered aldehydes, biological material, resin embedding and = TEM) is important? =
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org
I am looking for any written materials concerning Reichert MeF = microscope. I have one in mint condition (probably never used), but = without any instructions. The microscope was made after 1954. It is now = disassembled and, although I know more or less how to put it together, I = would rather like to do it according to producer's advices.
I am not sure if my question fits to this mailing list, but I think that = history of microscopy would be of interest for some of us.
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-2 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.2016.0"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} I am looking for any = written=20 materials concerning Reichert MeF microscope. I have one in mint = condition=20 (probably never used), but without any instructions. The microscope was = made=20 after 1954. It is now disassembled and, although I know more or less how = to put=20 it together, I would rather like to do it according to producer's=20 advices. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} {/FONT} {/DIV} {DIV} {FONT face=3DArial size=3D2} I am not sure if my question fits to = this mailing=20 list, but I think that history of microscopy would be of interest = for some=20 of us. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} Thank you {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} Pawel = Karaszkiewicz {/FONT} {/DIV} {DIV} {FONT color=3D#000000 face=3DArial size=3D2} {A=20 href=3D"mailto:zekarasz-at-cyf-kr.edu.pl"} zekarasz-at-cyf-kr.edu.pl {/A} {/FONT} {= /DIV} {DIV} {FONT color=3D#000000 face=3DArial = size=3D2} =20 {/FONT} {/DIV} {/BODY} {/HTML}
There is a Win95/WinNT program called EIKONA3D for volumetric image processing, analysis and visualization, which provides special features and tools for working with image sequences originated from microscopy (e.g. 3D image processing, image alignment, 3D reconstruction, 3D visualization, volume rendering, surface rendering, 3D registration, etc.). One of the tools it provides is 3D segmentation of a 3D data set (image sequence) and labeling/colorization of 3D regions. See the site: http://www.alphatecltd.com/eikona3d.html
Best regards, Nikos Nikopoulos
At 11:10 AM 2/6/99 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have used Adobe PhotoDeluxe which is available (and pretty cheap) for both the Mac and PCs. The program is deceptively simple, but quite usefull if you look into it a bit.
By adding a transparent layer, you can colorize the image without actually modifying it. Once a colored layer is created the colors can be rapidly changed to fit your tastes.
Photoshop will do the same....but for a much higher price, and learning curve.
} } Dear all, } } } } I need to colorize some SEM pictures but I do not know which } } programm to use. If someone can help me, I use PC or Macintosh computer. } } Thanks a lot for your help } } Best regards } } } } Didier } }
-- _______________________________________________ / Michael J. Herron / / U of MN,Medicine/Infectious Diseases / / herro001-at-maroon.tc.umn.edu / / http://128.101.243.213 / /__________________________________________/
I have some samples of muscle tissue in which I would like to determine the distribution of mitochondria within each myocyte.
My life would be made easier, and the sample size bigger, if I could do this at the LM level rather than in the TEM. I figure that the mitochondria will be visible in the LM because they occur in large groups in this tissue, rather than singly. Also, I don't need to see every mitochondrion, just the general pattern.
The question is: does anyone know of a suitable specific stain? I am after a bright-field permanent stain that will work on semi-thin resin sections, not a fluorescent or antibody method. A nice old-fashioned stain!
So far the tissue has been fixed in glutaraldehyde but not processed further. Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department Faculty of Medicine & Health Sciences University of Auckland Private Bag 92019 Auckland New Zealand
I'm studying the transport and localization of some proteins in the cytoplasm of cultured cells. The cells are fixed in paraformaldehyde and stained with a avidin-biotin-fluorescein system. I need to do double staining for this protein and the Golgi, so I intend to use the Bodipy-tr-ceramide probe, as suggested by Molecular Probes .
I would like to know if someone has experience or worked with this specific probe. Particularly I need to know if this probe may be used with fixed cells, that is, after the cells are fixed.
I will be very grateful for any information
Thanks in advance
Francisco Javier Hernandez-Blazquez IARC-WHO Unit of Multistage Carcinogenesis 150 Cours Albert Thomas Lyon - France
I have copies of the operations manuals for both the MeF3 and for its camera system and would be happy to provide you with copies. There would be a slight fee for copying and postage. Please let me know if you are interested.
Best regards,
Barbara Foster
Consortium President
Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118
MME is America's first national consortium providing
customized on-site workshops in all areas of
microscopy, sample preparation, and image analysis.
} } } }
{excerpt}
{fontfamily} {param} Arial {/param} {smaller} I am looking for any written materials concerning Reichert MeF microscope. I have one in mint condition (probably never used), but without any instructions. The microscope was made after 1954. It is now disassembled and, although I know more or less how to put it together, I would rather like to do it according to producer's advices.
{/smaller} {/fontfamily}
{fontfamily} {param} Arial {/param} {smaller} I am not sure if my question fits to this mailing list, but I think that history of microscopy would be of interest for some of us.
{/smaller} {/fontfamily}
{fontfamily} {param} Arial {/param} {smaller} Thank you
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I'm studying the transport and localization of some proteins in the cytoplasm of cultured cells. The cells are fixed in paraformaldehyde and stained with a avidin-biotin-fluorescein system. I need to do double staining for this protein and the Golgi, so I intend to use the Bodipy-tr-ceramide probe, as suggested by Molecular Probes .
I would like to know if someone has experience or worked with this specific probe. Particularly I need to know if this probe may be used with fixed cells, that is, after the cells are fixed.
I will be very grateful for any information
Thanks in advance
Francisco Javier Hernandez-Blazquez IARC-WHO Unit of Multistage Carcinogenesis 150 Cours Albert Thomas Lyon - France
How can you run a lab with the space you are getting? Everytime we are = CAP inspected we are cited (non binding) for lack of space. We have 5 = large rooms that are used for clinical purposes. Fight with the powers = that be on this CAP requirement as opposed to throwing out clinical = specimens.(ie blocks, slides, etc.)
If more room is not posible, how about off site storage? If I had to, I = could store blocks, prints, negs, thicks, etc in a 10 x 10 foot room. = (not much room to move in ) that we have collected in the lab since 1974. = =20
I do not actually now the real requirements that you are asking about, but = check with the histology lab and use those rules as guidelines.
Best of Luck,
Ed
} } } Sharon Drew {drewsh-at-smtpgw2.musc.edu} 02/05 9:45 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html=20=
I am now running a clinical path, diagnostic EM lab. but must reduce my storage. What are the clia and cap regs on how long to keep transmission em blocks, thick section slides, grids and phot mic's? I am being reduced from a 3 room lab to a room and a closet. thanks for you help. S. Drew = = = = = = = = = = =20
I have some samples of muscle tissue in which I would like to=20 determine the distribution of mitochondria within each myocyte.=20
My life would be made easier, and the sample size bigger, if I could=20 do this at the LM level rather than in the TEM. I figure that the=20 mitochondria will be visible in the LM because they occur in large=20 groups in this tissue, rather than singly. Also, I don't need to=20 see every mitochondrion, just the general pattern.
The question is: does anyone know of a suitable specific stain? I am=20 after a bright-field permanent stain that will work on semi-thin resin=20 sections, not a fluorescent or antibody method. A nice old-fashioned=20 stain!
So far the tissue has been fixed in glutaraldehyde but not processed=20 further. Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department Faculty of Medicine & Health Sciences University of Auckland Private Bag 92019 Auckland New Zealand
Hi Paul, During aldehyde fixation the calcium ions tend to leach out, thus the addition of CaCl2 to the buffer solution. Of course there are other important factors to consider such as pH, osmolality, and choice of buffer (PIPES, MOPES, or Cacodylate instead of Phosphate which may result in some percipitation). MgCl2,and KCl are are also sometimes added; depending on what is important to preserve. (Gronblad,M., (1983) Cell Tissue Res. 229,627 and Glauert, A.M., 1975. Fixation, dehydration, and embedding of Biological specimens. Practical Methods in Electron Microscopy. Amer. Elsevier Pub. Co.,Inc., New York 207pp.
At 10:41 PM 2/5/99 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hi all I am wondering if there is any source for SEM images that can be downloaded as "pdf" files. The images are for corroded metals. Regards Mayouf
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Hello all,
I would like to demonstrate the juxtaglomerular cells(=myoepitheloid cells) of kidney on semi-thin sections by using different staining methods. I remember reading a very nice method to show these cells. However, I can not find it now.
I will greatly appreciate if you could send me the suitable staining methods which you used before and also your suggestions about that.
Thanks in advance.
Best regards,
Ranan Gulhan AKTAS, M.D. Trakya University, Faculty of Medicine Pathology Department Edirne 22030 Turkey
Henk, I think that your equation is wrong. The gamma correction is Out = (In) ^(1/gamma). This gives a parabola (upwards turning) for gamma=1/2 which will expand the contrast in the bright region at the expense of the dark regions of the image. For gamma=2, it is a square root function (also a parabola on its side) that will expand the contrast in the dark regions at the expense of the light region. I assume that you would normalize the output range so that it was 0 to 255 with a suitable coefficient in the equation. The curves should go through 0 and 255, so there is no offset constant in the equation. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Hendrik O. Colijn To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hi all,
I was just asked to provide the equation for gamma adjustment to image contrast. I had always assumed that it was just
Output = (Input)^gamma i.e. a simple power law.
Thus for gamma = 0.5 you get a square root curve and for gamma = 2.0 you have a parabola. However, the contrast curves in various books I've checked do not appear to be a power law, but look more like circular arcs. Unfortunately these books only describe gamma but do not provide the equation. Are the curves just artist's license or is my understanding of the gamma correction wrong?
If you include Contrast and Brightness, I would expect the general equation to be
Output = B + C*(Input)^gamma
Where B = Brightness and C = Contrast.
Thanks, Henk Colijn Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu An optimist believes that we live in the best of all possible worlds. A pessimist fears that this is true.
I am looking for public domain images from electron microscopes or anyone in the New York area that might be interested in collaborating with a design firm to produce a high profile television spot. Any help would be appreciated..
Jeff Linnell Liquid Design Group jeff-at-liquidesign.com
My laboratory is equipped with a LEO 912 Omega, the model before the AB if I'm not mistaken, which was purchased 4 years ago. This is an excellent instrument, powerful, versatile and dependable. I use it mostly for conventional and cryo-EM with excellent results. I would reccomend it without hesitation.
For microtome, I have been a satisfied Reichert Ultracut user for years, which is why I bought an Ultracut S for my lab 4 years ago. The fully motorized controls needed getting used to in the beginning but have proven remarkably reliable.
Usual disclaimer: I have no interest or relation to either company other than being a very satisfied customer.
If you have specific questions, don't hesitate to contact me.
Regards, Michel **************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
Never used one of those, so I don't know if these are possible after fixation in glut_de. At least some are possible, acc. to the text, after fixation in "formaldehyde-based fixatives" and "postfixation" in potasium dichromate...
These are all described in Langeron, M: "Precis de microscopie", Masson, Paris, 1934...
Can send you a copy of the protocols if you want (*.tif, *.jpg, whatever...it's in French).
Yvan Lindekens.
---------- } Van: Stephen Edgar {s.edgar-at-auckland.ac.nz} } Aan: Microscopy-at-sparc5.microscopy.com } Onderwerp: LM: A stain for mitochondria? } Datum: maandag 8 februari 1999 5:13
} I have some samples of muscle tissue in which I would like to } determine the distribution of mitochondria within each myocyte. } } The question is: does anyone know of a suitable specific stain? I am } after a bright-field permanent stain that will work on semi-thin resin } sections, not a fluorescent or antibody method. A nice old-fashioned } stain! } } So far the tissue has been fixed in glutaraldehyde but not processed } further. } Regards } } Stephen Edgar
Our group has inherited a Boston-Bradley Adjustable Blade manufactured by Gardener Laboratory of Bethesda, MD. During an on-line search we were able to determine that the company existed in 1952, but no other information was available.
We believe this to be a crude microtome. It has a stainless steel "clamp" and brass inserts that are labeled with exact thickness from 0.001 to 0.101mil. The clamp is a rectangular block with 3/4" on each end 5mm higher than the center section
Profile ____ ____ ____________
Along one side of this center section is an adjustable stainless steel slab with an arrow pointing to the center of one edge. The knurled knobs are on the other side of the center section. This slab may be adjusted such that it is above or below the center height. It may also be loosened from the center area so that it is a distance from the center.
It seems that the only way to cut would be to raise the slab, place a specimen on the center area (top down), hold the specimen in place and cut it off with a razor blade. This seems upside-down to every other microtome I have dealt with.
Has anyone ever used one of these? We need a crude sledge microtome and this may fit our needs if we could just figure out how to use it.
I had always been told that it helped to preserve the membranes. I've never tested that information, however.
Bob Underwood Derm Research Center U of Washington
On 5 Feb 1999, Webster wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } Could some kind, knowledgable person explain why the addition of calcium to fixatives (buffered aldehydes, biological material, resin embedding and TEM) is important? } Regards, } } Paul Webster, Ph.D } House Ear Institute } 2100 West Third Street } Los Angeles, CA 90057 } phone:213 273 8026 } fax: 213 413 6739 } e-mail: pwebster-at-hei.org } } }
} I am looking for public domain images from electron microscopes or } anyone in the New York area that might be interested in collaborating } with a design firm to produce a high profile television spot. Any help } would be appreciated.. } } Jeff Linnell
Jeff -
You'll find a wide variety of images hotlinked at the end of the "Microscopy for Children" bibliography on the Project MICRO web page (URL below). Don't miss the secondary set of links available at "K-12 microscopy resources". Some may be public domain, some are not; you need to check after you've found what you want. There's a stock photo CD-ROM (colorized SEM) available from Corel; it's also in the bibliography .
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
MSA List Recipients, If anyone in possession of a spare Philips CM or EM400 series specimen holder (rod), either regular, low-background, or double-tilt, is willing for a nominal fee or goods and services to part company with said device, please contact me immediately at any of the numbers below. I will be more than appreciative and most remunerative. Thanks. Winston
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W Wiggins, Supervisor 2/9/99 11:25:46 AM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
} Polymer Microscopist - Freeport, Texas and Midland, } Michigan } } Company: The Dow Chemical Company } } Location: Freeport, Texas and Midland, Michigan } } Qualifications (education, certification, language, etc.) and } Experience required: } A candidate with a BS or MS degree in polymer science, material } science or chemistry is preferred with some prior experience in } electron microscopy. } Good written and oral communication skills and the ability to work } both independently and in a team environment are extremely important. } } Job Overview: } } The Surface/Microscopy/X-ray (SMX) Group in Dow's Corporate R&D } Analytical Science Laboratory has two professional level full time } openings for Polymer Microscopists, one position at each of the Dow's } facilities in Midland, MI and Freeport, TX. The primary } responsibilities include working with partners to support research } projects involving new and existing products in Dow's polymer } businesses. } } Key responsibilities will include: } } 1. Extensive problem solving. } 2. Microscopy preparation technique experience including } ultramicrotomy and cryo-ultramicrotomy. } 3. Operation of transmission electron microscope. } 4. Interpretation of images. } 5. Documentation of work. } 6. Compliance with safety and quality programs. } 7. Active member of project and SMX work teams. } } Interested: } Please e-mail or send your resume and cover letter, with reference to } this ad to: } Email: R&D-at-Dow.com or The Dow Chemical Company, Workforce } Planning 98-289, P. O. Box 150, Plaquemine, LA 70765. E-mail } respondents must list Job 98-289 and their last name as the first and } second items on the Subject line. Only those selected for an } interview will be contacted. Only U.S. citizens or aliens who are } authorized to work in the United States will be considered for } employment. } } We are an equal opportunity employer and offer a competitive } compensation and benefits package including 401k, stock purchase, } tuition reimbursement and performance incentives. The Dow Chemical } Company is the fifth largest chemical company in the world with annual } sales of US$20billion. Dow manufactures and supplies chemicals, } plastics and agricultural products for customers in 164 countries and } employs approx. 43,000 people worldwide. For more news and } information about Dow, please visit our web site at www.dow.com. } } Bob Cieslinski } Microscopy & Microanalysis } 1897 E Bldg. } (517) 636-6875 }
} } MSA List Recipients, } If anyone in possession of a spare Philips CM or EM400 series } specimen holder (rod), either regular, low-background, or } double-tilt, is willing for a nominal fee or goods and services } to part company with said device, please contact me immediately } at any of the numbers below. I will be more than appreciative } and most remunerative. Thanks. } Winston } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Winston W Wiggins, Supervisor 2/9/99 11:25:46 AM } CRC-Electron Microscopy Lab. Ofc:704/355-1267 } Carolinas Medical Center Fax:704/355-7648 } P.O. Box 32861 Lab:704/355-7220 } Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ }
-----------------End of Original Message-----------------
can anyone tell me if people still use BSA or bacitracin as 'wetting agents' when negative staining? I cannot find reference to them in any of my texts but I have used them since the dawn of time, so I think I have just lost the references and/or they've gone out of fashion.
If anyone has advice on use, advantages of particular chemicals or references I would be grateful.
thanks
Malcolm
PS our glow discharge unit doesn't work - so I can't use that.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
} } MSA List Recipients, } If anyone in possession of a spare Philips CM or EM400 series } specimen holder (rod), either regular, low-background, or } double-tilt, is willing for a nominal fee or goods and services } to part company with said device, please contact me immediately } at any of the numbers below. I will be more than appreciative } and most remunerative. Thanks. } Winston } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Winston W Wiggins, Supervisor 2/9/99 11:25:46 AM } CRC-Electron Microscopy Lab. Ofc:704/355-1267 } Carolinas Medical Center Fax:704/355-7648 } P.O. Box 32861 Lab:704/355-7220 } Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ }
-----------------End of Original Message-----------------
We have an "Image Galleries" page on the WWW-Virtual Library for microscopy site where you may be able to find something suitable. http://www.ou.edu/research/electron/www-vl/image.shtml
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
Jeff wrote:
} I am looking for public domain images from electron microscopes or } anyone in the New York area that might be interested in collaborating } with a design firm to produce a high profile television spot. Any help } would be appreciated.. } } Jeff Linnell
Jeff -
You'll find a wide variety of images hotlinked at the end of the "Microscopy for Children" bibliography on the Project MICRO web page (URL below). Don't miss the secondary set of links available at "K-12 microscopy resources". Some may be public domain, some are not; you need to check after you've found what you want. There's a stock photo CD-ROM
(colorized SEM) available from Corel; it's also in the bibliography .
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
Hi folks, it has been a while but I am back and of course I need help.
I should submit a proposal to my Dean and Chancellor on a potential centralized morphology core (CMC). They want to see numbers for outlay-cost-income, and I am having trouble coming up with any cost about structure, maintenance.
I will very much appreciate either a brief response to the points below or someone pointing pointing me to a source for this information on the web (phone numbers or email addresses of members with such information will be appreciated). I am envisioning a CMC to provide under one roof: TEM, SEM, Digital Imaging, Confocal, Atomic Force Microscopy and ancillary services such as frozen, histo, immuno-histo and photography. I realize that information for all these may come from different places. Overall, what I need is just an approximate idea for:
1) Cost to operate existing facilities in academic centers?
2) What is the cost for maintaining equipment present at those facilities and to upgrade components?
3) What is the cost to internal and outside users? -Do you charge per case, number of blocks, number of samples, number of cuts?
-How many prints do you provide and at what magnifications for each case?
-Do you retain negatives and when you do not, do you charge extra?
-Charges for embedding cutting and staining frozens, paraffin?
-Charges for plastic processing (JB-4, historesin, etc), sectioning and staining?
-Charges for dupplicating slides, making slides, prints,
-Charges for preparing PhotoShop publication quality composites on color sublimation paper?
-Charges for using microscopy equipment
-Light without and with phase, Normaski, fluorescence, etc.
-Digital confocal optical sectioning, reconstruction, etc.
4) Exceptions.
a) Do junior faculty get service for less than senior and funded faculty?
b) Are there internal mechanisms for covering the costs of promising-emerging faculty, but without active support?
c) How many places have internal mechanisms such as the now extinct NIH BSRG to cover costs?
d) Are any of those costs derived directly or indirectly from grant overheads?
5) Space now housing the facility you describe?
6) How many of the facilities started with external funds?
7) How many of the facilities started with Dean or Chancellor funds?
8) Any other information I miss, but you consider important when considering a CMC?
9) In particular I want to show that most CMC DO NOT make a profit, because of the huge costs for maintenance contract on equipment???
10) Is there anyone out there getting internal support from grant's overhead, and is the money used for CMC considered an incentive-kick-back to funded invetigators?
11) How many of the existing CMC out there offer Cell Sorting (flow) as part of the morphology services?
-Arrangements?
-Maintenance?
-Support?
-Internal and external charges?
Thanks a lot. I will collate and post the results.
*Disclaimer: Whatever... is not Tulane opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web:[ http://www.tmc.tulane.edu/ferminlab/] or [http://www.tmc.tulane.edu/imaging/] Internet: [cfermin-at-mailhost.tcs.tulane.edu] 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
The Michigan Microscopy and Microanalysis (MMM) Society will hold its Spring Meeting on May 7, 1999 at the Genoa Woods Conference Center, 7707 Conference Center Drive, Brighton, MI. The program chair is still soliciting additional student papers for the meeting. If interest or you would like more details on the meeting contact the local affiliate president, Rob Eversole email at eversole-at-wmich.edu.
MSA (the Microscopy Society of America) is making a concerted effort to provide a forum for multiphoton imagers to gather for candid discussions concerning the new technique.
This year's event is condensed to a one day symposium,
#31 Multi-Photon Excitation Microscopy: the Next Generation 2 or 3 AUG 99 in Portland, OR
This year there will be far less time available; however, there are several important goals for this symposium: 1) invite all new speakers for the various applications presentations 2) presentations describing the commercial systems currently available 3) provide a question and answer session forum for technique education
Poster and abstract submissions are welcome, but please hurry since the deadline for abstract inclusion is 15 FEB 99. See http://www.msa.microscopy.com/
The morning session covers a variety of strong applications of (currently quite expensive) ultra-short pulse laser based fluorescence microscopy...
Karel Svoboda, Ph.D Cold Spring Harbor Laboratory "2PELSM for High-Resolution Imaging in Scattering Biological Tissues: Applications to Neuroscience"
Jayne Squirrell University of Wisconsin "2PEFM of Cytoplasmic Organization in Living Mammalian Embryos"
Mary Dickinson, Ph.D. Caltech "Biological Applications of Chromophores With Large Two-Photon Cross-Sections"
Christopher Navara, Ph.D Wayne Hughes Institute "Dynamic Drug Effects Monitoring"
Paul M W French, Ph.D Imperial College of Science, Technology and Medicine "Fluorescence Lifetime Imaging of Biological Tissue"
Afternoon question/answer session panel: to include the morning speakers, plus
Dave Piston, Ph.D Vanderbilt University Warren Zipfel, Ph.D Cornell University Rebbeca Williams Cornell University Vickie Centonze-Frohlich, Ph.D University of Wisconsin John White, Ph.D University of Wisconsin
In addition, we plan to have vendor presentations and handouts for the commercially available two (multi) photon systems.
==============================================
David L. Wokosin Instrumentation Development Engineer Integrated Microscopy Resource University of Wisconsin-Madison 1675 Observatory Drive Madison, WI 53706-1205
******************************************************************************** Victoria Centonze Frohlich, Ph.D. Deputy Director, IMR University of Wisconsin, Madison P(608) 263-6288 F(608) 265-4076 http://www.bocklabs.wisc.edu/imr/home.htm ******************************************************************************** --============_-1293529032==_ma============ Content-Type: text/enriched; charset="us-ascii"
MSA (the Microscopy Society of America)
is making a concerted effort to provide a forum for multiphoton
imagers to gather for candid discussions concerning the new technique.
This year's event is condensed to a one day symposium,
{bold} #31 Multi-Photon Excitation Microscopy: the Next Generation
{/bold} 2 or 3 AUG 99 in Portland, OR
This year there will be far less time available; however, there are several important goals for this symposium:
1) invite all new speakers for the various applications presentations
2) presentations describing the commercial systems currently available
3) provide a question and answer session forum for technique education
Poster and abstract submissions are welcome, but please hurry since the deadline for abstract inclusion is 15 FEB 99. See http://www.msa.microscopy.com/
The morning session covers a variety of strong applications of (currently quite expensive) ultra-short pulse laser based fluorescence microscopy...
Karel Svoboda, Ph.D Cold Spring Harbor Laboratory
"2PELSM for High-Resolution Imaging in Scattering Biological Tissues:
Applications to Neuroscience"
Jayne Squirrell University of Wisconsin
"2PEFM of Cytoplasmic Organization in Living Mammalian Embryos"
Mary Dickinson, Ph.D. Caltech
"Biological Applications of Chromophores With Large Two-Photon Cross-Sections"
Christopher Navara, Ph.D Wayne Hughes Institute
"Dynamic Drug Effects Monitoring"
Paul M W French, Ph.D Imperial College of Science, Technology and Medicine
"Fluorescence Lifetime Imaging of Biological Tissue"
Afternoon question/answer session panel:
to include the morning speakers, plus
Dave Piston, Ph.D Vanderbilt University
Warren Zipfel, Ph.D Cornell University
Rebbeca Williams Cornell University
Vickie Centonze-Frohlich, Ph.D University of Wisconsin
John White, Ph.D University of Wisconsin
In addition, we plan to have vendor presentations and handouts for
the commercially available two (multi) photon systems.
Cesar- you mention what you envision in a CMC what equipment do you currently have? what equipment will you be purchasing? I have used NIH microscopy centers, and I have managed institutional microscopy facilities. The more money you have for staff the better your facility will operate. It is best if you have one staff member per microscope. It gets crazy if your staff is over worked. You will find maintenance costs will be lower, the quality of results will improve, and overall costs will decrease. Maintenace contracts are a killer, we were once paying over $95,000 per year on service contracts for 4 EM scopes. It was driving prices through the roof, try to negotiate with the manfacturers. If you buy all new scopes, maybe buy from one vendor, and see if they will throw in a service engineer as part of the package, or hire a maintenance person yourself. Try to set costs in a per protocol fashion avoid itemizing. If you need any more specifics please feel free to contact me directly. -Mike
On Tue, 9 Feb 1999, Cesar D. Fermin Ph.D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi folks, it has been a while but I am back and of course I need help. } } I should submit a proposal to my Dean and Chancellor on a potential } centralized morphology core (CMC). They want to see numbers for } outlay-cost-income, and I am having trouble coming up with any cost about } structure, maintenance. } } I will very much appreciate either a brief response to the points below } or someone pointing pointing me to a source for this information on the } web (phone numbers or email addresses of members with such information } will be appreciated). I am envisioning a CMC to provide under one roof: } TEM, SEM, Digital Imaging, Confocal, Atomic Force Microscopy and } ancillary services such as frozen, histo, immuno-histo and photography. } I realize that information for all these may come from different places. } Overall, what I need is just an approximate idea for: } } 1) Cost to operate existing facilities in academic centers? } } 2) What is the cost for maintaining equipment present at those facilities } and to upgrade components? } } 3) What is the cost to internal and outside users? } -Do you charge per case, number of blocks, number of samples, number of } cuts? } } -How many prints do you provide and at what magnifications for each } case? } } -Do you retain negatives and when you do not, do you charge extra? } } -Charges for embedding cutting and staining frozens, paraffin? } } -Charges for plastic processing (JB-4, historesin, etc), sectioning and } staining? } } -Charges for dupplicating slides, making slides, prints, } } -Charges for preparing PhotoShop publication quality composites on } color sublimation paper? } } -Charges for using microscopy equipment } } -Light without and with phase, Normaski, fluorescence, etc. } } -Digital confocal optical sectioning, reconstruction, etc. } } 4) Exceptions. } } a) Do junior faculty get service for less than senior and funded } faculty? } } b) Are there internal mechanisms for covering the costs of } promising-emerging faculty, but } without active support? } } } c) How many places have internal mechanisms such as the now extinct NIH } BSRG to cover } costs? } } d) Are any of those costs derived directly or indirectly from grant } overheads? } } 5) Space now housing the facility you describe? } } } 6) How many of the facilities started with external funds? } } } 7) How many of the facilities started with Dean or Chancellor funds? } } } 8) Any other information I miss, but you consider important when } considering a CMC? } } } 9) In particular I want to show that most CMC DO NOT make a profit, } because of the huge costs for maintenance contract on equipment??? } } 10) Is there anyone out there getting internal support from grant's } overhead, and is the money used for CMC considered an incentive-kick-back } to funded invetigators? } } } 11) How many of the existing CMC out there offer Cell Sorting (flow) as } part of the morphology services? } } -Arrangements? } } -Maintenance? } } -Support? } } -Internal and external charges? } } } Thanks a lot. I will collate and post the results. } } *Disclaimer: Whatever... is not Tulane opinion! } Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology } web:[ http://www.tmc.tulane.edu/ferminlab/] or } [http://www.tmc.tulane.edu/imaging/] Internet: } [cfermin-at-mailhost.tcs.tulane.edu] } 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 } Fax 504 587-7389, Voice 584-2521, Main Office 588-5224 } } }
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Our group has inherited a Boston-Bradley Adjustable Blade manufactured by Gardener Laboratory of Bethesda, MD. During an on-line search we were able to determine that the company existed in 1952, but no other information was available. We believe this to be a crude microtome.
It has a stainless steel "clamp" and brass inserts that are labeled with exact thickness from 0.001 to 0.101mil. The clamp is a rectangular block with 3/4" on each end 5mm higher than the center section
Along one side of this center section is an adjustable stainless steel slab with an arrow pointing to the center of one edge. The knurled knobs are on the other side of the center section. This slab maybe adjusted such that it is above or below the center height. It may also be loosened from the center area so that it is a distance from the center. It seems that the only way to cut would be to raise the slab,place a specimen on the center area (top down), hold the specimen in place and cut it off with a razor blade. This seems upside-down to everyother microtome I have dealt with.
Has anyone ever used one of these? We need a crude sledge microtome and this may fit our needs if we could just figure out how to useit.
Thank-you for your help.
Maureen Hunt BP-Amoco p.l.c.
_________________________________________________________ DO YOU YAHOO!? Get your free -at-yahoo.com address at http://mail.yahoo.com
We just replaced the filament in our Jeol 2010, with a Denka filament can anyone tell me who the supplier is in australia, or elsewhere, I'd like to buy another. The filament is a Denka LaB6 Cathode LKSH M3 104051
Thanks George
George Theodossiou Dept Applied Physics RMIT GPO Box 2476V Melbourne 3001 Victoria Australia Ph: +61 3 9925 3394 Fax: +61 3 9925 5290
We just replaced the filament in our Jeol 2010, with a Denka filament can anyone tell me who the supplier is in australia, or elsewhere, I'd like to buy another. The filament is a Denka LaB6 Cathode LKSH M3 104051
Thanks George
George Theodossiou Dept Applied Physics RMIT GPO Box 2476V Melbourne 3001 Victoria Australia Ph: +61 3 9925 3394 Fax: +61 3 9925 5290
Leica Microsystems, Diatome US, and Electron Microscopy Sciences, announce another in a series of TEM Specimen Preparation workshops. This seminar will focus on the hands on participation of the following techniques:
Embedding of industrial samples Specimen trimming
Ultramicrotomy of hard materials Ultramicrotomy of polymers
Collection & handling of sections Staining of polymer sections
Low temperature ultramicrotomy
The format of our workshop is half day lecture and half day bench work in small working groups. Video attachments will be used on the ultramicrotomes in order to maximize the teaching experience for all involved. Samples will be supplied by the course instructors. Participants are encouraged to bring their own samples to work with as time allows.
Course Speakers & Instructors
Dr. Tom Malis Mr. Bob Vastenhout CANMET DOW Chemical Characterization Group Leader Polymer Microscopist Materials Technology Laboratory Analytical Science Department Ottawa, Ontario Terneuzen, The Netherlands
Mr. Helmut Gnagi Product Manager Microtomist Extaordinaire Diatome, Ltd., Switzerland
Hosted by: JoAn Hudson Clemson University Microscopy Facility Clemson, SC
When: June 9-11, 1999
Tuition: $1,400.00
Includes three nights lodging at the lakefront Conference Center & Inn at Clemson University, continental breakfast and lunch daily, one group dinner, course supplies, and lab charges.
Contact: Mike Boykin, Leica Microsystems, Inc. 800-248-0665 X5092
If you are or were using Accu-Scope (3001, 3002, 3018PH etc.) or Wang Biomed (3002, 3004) microscopes - please share your opinion about them and your experience with me.
What did you use it for? Did it prove suitable for the task? Did you have any problems with it? Was its optical/mechanical quality OK for you and for the job?
If you had to use technical support, how helpful/quick were they?
If you used other microscopes too - how do they compare?
Depending on how interesting the answers would be to the other list members, you may either post the replies to the list, or e-mail me directly.
Thank you! -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
Dear Listmembers, Does anybody use or have used ruthenium tetroxide as a fixative for biological material? Any references? Thanks in advance. Chris vd Merwe.
I wonder if anyone knows a good textbook for EDS of minerals? I used to have one in the lab, but it's gone missing, and I don't remember the title or author(s). It had the mineralogical characteristics of most or all of the common minerals, and included typical EDS spectra for each one. A very valuable book for a lab that does a lot of geological work, and I'd really like to replace it (and maybe chain it to the wall this time!)
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B2Y 4A2
Can anyone recommend a good text on EDS of minerals? We used to have a good one in the lab, which described the characteristics of various common minerals, and showed typical EDS spectra for them. Unfortunately, it's disappeared and I cannot remember the title or author, so I guess it's time for a new one.
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B2Y 4A2
Dear microscopists, Our facility needs to replace our print processor. I have looked at several models and wanted some feedback about what models work well. We are limited on the amount of space we can use. The processor can not be any larger then 40" X 38", needs to be easy to clean and durable. We are considering purchasing the ILFORD 2150, but heard it may have lots of problems.
Cheers Malcolm, Yes, the bacitracin wetting technique is still valid as ever. The reference you want is: D.W. Gregory and B.J.S. Pirie, Wetting agents for electron microscopy of biological specimens. Proc. Fifth European Congress on Electron Microscopy, (1972). 234-235. David Gregory recommended using a minimum concentration of 7.5 ug/cm3 for formvar coated grids and 10 ug/cm3 on carbon coated formvar or carbon grids as a wetting agent. All the best, Henry
----------------------------------------------------------------------------- Malcolm wrote: } } Dear all } } can anyone tell me if people still use BSA or bacitracin as 'wetting agents' } when negative staining? I cannot find reference to them in any of my texts } but I have used them since the dawn of time, so I think I have just lost the } references and/or they've gone out of fashion. } } If anyone has advice on use, advantages of particular chemicals or } references I would be grateful. } } thanks } } Malcolm } } PS our glow discharge unit doesn't work - so I can't use that. } } Malcolm Haswell } Electron Microscopy } School of Sciences } Fleming Building } University of Sunderland } SUNDERLAND SR1 3SD } Tyne and Wear } UK } } Tel (0191) 515 2872 } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } Henry Eichelberger, EM Facility Manager Department of Biological Sciences Binghamton, University
I am working with the Zeiss LSM 510 Confocal microscope, and am having trouble achieving 3D image reconstruction using the 3D for LSM program. Do you have any recommendations, or know of the resources available through which I can learn this technique? I'd appreciate any information you have on the topic!
----------------------------------------- Laurie Wallin Technician UCSD Department of Anesthesiology and Neuropathology 9500 Gilman Drive 0629 La Jolla, CA 92093 (619)534-1339 or 822-3271
} Are there any grants available to purchase microscopes for an elementary } school in California?
On a national level, no; but there may be an announcement from a major corporation soon that could help you. Your best current possibility is a local corporation that supports education. You'll need about $1000; you'll find the rationale for that figure and advice on what to buy on the Project MICRO web page (URL below). Although I can't supply the funding, I CAN help you write a proposal, and I can write a supporting letter (but I'll be on vacation 2/18-3/28). Where are you located? California is a big state!
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Dear Chris, Ruthenium tetroxide is slow penetrating, but some structures are better preserved by its use. Examples: Madison, K. C., et. al. J. Invest. Dermatology 88 (1987) 714. and Eichelberger, et. al., Proc. Microscopy & Microanalysis 1994, 270. Best regards, Henry ----------------------------------------------------------------------------- Chris vd Merwe wrote:
} From: "Elektronmikroskopie EM 2075 Lab1-5 NW11" {lab1-at-ccnet.up.ac.za} } Organization: University of Pretoria } To: Microscopy-at-Sparc5.Microscopy.Com } Date: Thu, 10 Feb 1999 12:59:24 CAT-2 } Subject: Ruthenium O4 in biology } Priority: normal } X-mailer: Pegasus Mail for Windows (v2.42a) } } Dear Listmembers, } Does anybody use or have used ruthenium tetroxide as a fixative for } biological material? Any references? } Thanks in advance. } Chris vd Merwe. } } Henry Eichelberger, EM Facility Manager Department of Biological Sciences Binghamton, University
} I wonder if anyone knows a good textbook for EDS of minerals? I used to } have one in the lab, but it's gone missing, and I don't remember the title } or author(s). It had the mineralogical characteristics of most or all of } the common minerals, and included typical EDS spectra for each one. A very } valuable book for a lab that does a lot of geological work, and I'd really } like to replace it (and maybe chain it to the wall this time!)
} F.C. Thomas } MicroAnalysis Facility } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada B2Y 4A2
Perhaps you're thinking of "SEM Petrology Atlas" by Joann E. Welton, Chevron Oil Field Research Company, published by the American Association of Petroleum Geologists, 1984. And no, you can't have my copy! Don't know if it's still in print, but amazon.com will hunt around for a used copy for you (for a small fee, of course).
Cheers, eh?
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Patricia, Of the many print processors I've used, I'd choose the Ilford above them all. The older versions have a problem blowing fuses but that is supposed to have been corrected with the new models. The only difficulty I had was that with heavy usage, it must be cleaned more often which, for me, was more of a nuisance than a problem. It's easy to use and durable, not to mention relatively inexpensive. Go for it. Winston
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W Wiggins, Supervisor 2/10/99 2:53:24 PM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Today I was realesed online free Metallographic Etch's Database. This database is designed to allow you to quick find etchant via powerfull keyword search engine written in Perl. Here are some key features:
1763 records in the database 75 etchants for macro etching 1359 etchants for micro etching 1103 etchants for chemical etching 218 electrolytes for electrolytic etching 75 etchants for physical etching 329 electrolytes for electropolishing
Keyword search engine Browse database by macroetching Browse database by microetching Browse database by chemical etching Browse database by electrolytic etching Browse database by physical etching Browse database by electropolishing Powerfull online help
For more information please see link at microscopy vendors database: http://www.kaker.com/mvd/vendors.html
Henrik Kaker -- SEM-EDS Laboratory Metal Ravne Slovenia http://www2.arnes.si/guest/sgszmera1/index.html
} Can anyone recommend a good text on EDS of minerals? We used to have a good } one in the lab, which described the characteristics of various common } minerals, and showed typical EDS spectra for them. Unfortunately, it's } disappeared and I cannot remember the title or author, so I guess it's time } for a new one. } } F.C. Thomas
Frank,
I think the book you are referring to is SEM Petrology Atlas by Joann E. Welton. It was published in 1984 by The American Association of Petroleum Geologists, Tulsa, OK 74101. ISBN 0-89181-653-4.
Hope this helps. Lou Ross Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777 (573) 882=5458 fax www.missouri.edu/~geosclmr/ebaf.html
Patricia, Our facility has had an Ilford 2150 for 8 years. Though it has had breakdowns, repairs are fairly simple. I feel that the service contract is too expensive for what you get, which is someone walking you through a repair over the phone. I'm no mechanic but I've been able to deal with most of the problems myself and Iford will sell parts if you need to replace something. The one thing that has made the biggest difference in the trouble free operation of the machine is the addition of a dirt/rust filter on the water supply line. After we did that we really have had no problems except for an occassional plumbing leak (also fixable if you can operate a wrench). You'll also find that you don't need to change the chemicals as often as the manufacturer suggests, as they can be expensive. If you need any more information don't hesitate to contact me.
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky Medical Center
--IMA.Boundary.2444868190 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
A colleague and I built a device for doing something similar when I was in graduate school. It consisted of an ultrasonic nebulizer to produce the aerosol at one end of a column, N2 carrier gas (low velocity) to move the aerosol and external heaters to heat the column (along with a few other bells and whistles). Samples were collected on grids at the exit end and the whole thing sat in a hood. The path length was about 2m.
What I'd like to know is if you take any care to get a more uniform aerosol size? Do you work at low enough concentrations to have mostly one (or fewer) particles per drop?
|---------------------------------------------------------------| | Opinions expressed are mine an not those of Rohm and Haas Co. | |---------------------------------------------------------------| | Dr. John R. Reffner | rsrj2r-at-rohmhaas.com | | Rohm and Haas Co. | | | 727 Norristown Road | voice:215-619-5283 | | Spring House, PA 19477 | Fax: 215-619-1607 | |---------------------------------------------------------------|
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Hi
The best way I know of preparing pigments for TEM analysis is by blowing an aerosol of the sample dispersed in alcohol/water solution over a hot plate and collecting the sample on a grid placed at the far end of the hot plate. The company I work for is concerned about the safety issues surrounding this so does anybody know of any less hazardous sample preps. or a simple containment facility for the aerosol spray? I thought about building a glass box for containment but are there any commercially available containment/sample prep. units?
Thanks
Brian Manzor e-mail: brian.manzor-at-grmouth.zeneca.com
Received: from nmho05u.rohmhaas.com ([136.141.252.23]) by ima1.rohmhaas.com with SMTP (IMA Internet Exchange 3.11) id 000E8E24; Wed, 3 Feb 1999 08:41:25 -0500 Received: by nmho05u.rohmhaas.com; id IAA09730; Wed, 3 Feb 1999 08:42:12 -0500 (EST) Received: from sparc5.microscopy.com(206.69.208.10) by nmho05u.rohmhaas.com via smap (3.2) id xma009705; Wed, 3 Feb 99 08:42:01 -0500 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id FAA06651 for dist-Microscopy; Wed, 3 Feb 1999 05:19:30 -0600 Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id FAA06644 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 3 Feb 1999 05:18:59 -0600 Received: from apsn05.zeneca.com (apsn05.zeneca.com [193.132.159.6]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id FAA06637 for {microscopy-at-msa.microscopy.com} ; Wed, 3 Feb 1999 05:18:45 -0600 Received: from apsn15.ukap.zeneca.com by apsn05.zeneca.com (SMI-8.6/SMI-SVR4) id LAA01931; Wed, 3 Feb 1999 11:33:45 GMT Received: by apsn15.ukap.zeneca.com; id LAA29776; Wed, 3 Feb 1999 11:31:39 GMT Received: from ukapzcmsxhub01.ukap.zeneca.com by gbzhp02.ukap.zeneca.com with ESMTP (1.40.112.8/16.2) id AA200221316; Wed, 3 Feb 1999 11:28:36 GMT Received: by ukapzcmsxhub01.ukap.zeneca.com with Internet Mail Service (5.5.2232.9) id {1F6WTK3F} ; Wed, 3 Feb 1999 11:27:36 -0000 Message-Id: {21E55FE938B6D0119A7100805FFEAEC4D0177B-at-UKGR07} } From: Manzor Brian BP {brian.manzor-at-grmouth.zeneca.com} To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-Sparc5.Microscopy.Com}
I'm looking for a gold or graphite/carbon sputter coater for bio specimen preparation.
If you have one to sell, pls let me know details and price.
I'm considering using digital video (DV) for time-lapse/real-time LM. Broadcast/domestic DVcam magazines suggest that DV gives improved resolution over analogue video (S-VHS etc). Added to this, it also seems to give better quality frame by frame editing (+ stills output) when recorded on DV tape (mini or standard) and managed with software like Adobe Premier using Firewire ( aka i.link/IEEE1394) fast serial links.
If anyone has used this technology I'd be interested to hear more about the pros and cons.
TIA
Kevin Jennings -.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-. SmithKline Beecham Pharmaceuticals Analytical Sciences -Microscopy Group New Frontiers Science Park Harlow Essex UK -.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.
I am working on the light reflection of mainly squid iridophores. Are there any groups, preferably in the UK, working with polarising and/or interference microscopes who could give me some advice with this matter?
Thank you very much,
Lydia
PS. Hello Daniele ... back to work !!!
Keith Ryan Marine Biological Association of the UK Citadel Hill Plymouth Devon PL1 2PB England
Gary Gaugler, Ph.D. wrote: =============================================== I'm looking for a gold or graphite/carbon sputter coater for bio specimen preparation.
If you have one to sell, pls let me know details and price. ================================================ There are several very excellent data bases of information of who manufactures what in the microscopy and microanalysis market. You might want to consult some of those listings. Two that I myself make frequent use of are the following:
Microscopy Vendors Data Base http://207.137.96.185/mvd/vendors.html
Microworld Resources and New http://www.mwrn.com/
SPI Supplies is one of the several leading manufacturers of this equipment and all the information you would need our units can be found on our website below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
I can recommend looking at www.bookfinder.com and www.bibliophile.com if you are looking for a used copy of the text (assuming it isn't available new or you are too poor/cheap to pay for it). These are services where you do your own search over a number of used booksellers - I found a couple of technical books I wanted this way and for quite reasonable prices.
Janet Rice MCC Senior Member Technical Staff rice-at-mcc.com 512-338-3266
Yet another lab in search of microtome parts. We have four Leica Ultrotome III ultramicrotomes in very serviceable condition, but a couple need new belts. A supply of spare tubes would also be handy. Is anyone aware of a source for new or used parts for these venerable instruments?
Thanks in advance!
Randy Tindall Electron Microscope Core College of Veterinary Medicine University of Missouri - Columbia Phone: 573-882-8304
Gary: I have a sputter coater for sale. Trouble is I am in Cambridge UK
Patrick Echlin Director, Multi-Imaging Centre School of Biological Sciences Cambridge University
On Wed, 10 Feb 1999, Dr. Gary Gaugler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I'm looking for a gold or graphite/carbon sputter coater for } bio specimen preparation. } } If you have one to sell, pls let me know details and price. } } } } } } Cheers, } Gary Gaugler, Ph.D. } } PGP Public key: } BDC9 8860 AA83 1FB2 66D3 01BB 9EB8 8E6D 671F BEB3 } }
Steve, We actually have an old LKB Microtome we are getting rid of--you are welcome to it, but we are in Boston. If you want to make arrangements to have it shipped to you--it's yours! Peggy Sherwood MGH-Wellman Labs of Photomedicine 224 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-3192 (fax)
Patricia, I didn't see your original e-mail request, but saw the responses! We have a Rapidoprint DD3700 Agfa-Gevaert in our lab for processing EM prints. It's very simple, but like some other processors, it has to be cleaned frequently, especially the water stations, depending upon it's use. Peggy Sherwood MGH-Wellman Labs of Photomedicine
Having worked with minerals of many types, I find that a freeware program for the Mac has been extremely useful in predicting spectra of any mineral. DTSA for either 68K or PPC can be obtained from NIST or possibly from the MSA web site (maybe someone else can confirm this). The program can generate synthetic spectra for just about any detector and will simulate thin or thick specimens. It's a wonderful teaching and research tool that I recommend highly.
Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets Johns Hopkins University Baltimore, Maryland 21218 USA Phone: (410) 516-8342 Fax: (410) 516-7933 e-mail: klivi-at-jhu.edu
} Having worked with minerals of many types, I find that a freeware program } for the Mac has been extremely useful in predicting spectra of any mineral. } DTSA for either 68K or PPC can be obtained from NIST or possibly from the } MSA web site (maybe someone else can confirm this). The program can } generate synthetic spectra for just about any detector and will simulate } thin or thick specimens. It's a wonderful teaching and research tool that I } recommend highly. }
DTSA can be downloaded at http://micro.nist.gov/DTSA/dtsa.html .
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
Summary and compilation: Tissue cultures and coverslips: Thanks to everyone for the tremendous response to my query. The majority of responses suggested the use of Thermonox coverslips. These may be purchased sterile. One side is prepared for cell adhesion, and is labeled. They are compatible with alcohol, acetone and propylene oxide as solvents, and with Epon, Spurr's and araldite mixtures for resin. These coverslips come in sizes which fit well into tissue culture plates. Other investigators have successfully used glass coverslips, membrane inserts and plastic petri plates. Permanox plates were also recommended, cells may be grown, fixed and embedded in the chambers. Most people use a dip into liquid nitrogen to facilitate removal of the coverslip after embedding. Other simply pull the coverslip off while still warm from the embedding oven. If the coverslip is not removed, it may still be sectioned with a diamond knife, however, the coverslip may separate from the resin under the electron beam. If the coverslip if removed, various methods are suggested to position the cells parallel or perpendicular to the bloc face. A compilation of the responses follows, MANY helpful tricks will be found on these pages. -----------------------------------------------------------------------
I use standard glass coverslips - round ones that fit in 24 and 12 well culture dishes. I sterilize them sitting on a clean filter paper in glass petri dishes then transfer them to culture dishes and seed with cells. I fix and process in 20 ml glass scintillation vials, embed in epon (put coverslip cell side up on a slide with a drop of epon, heat 4-8 hrs at 58 C (timing is important), cross-hatch with a razor blade, slowly lower into liquid nitrogen, and little squares of epon pop off with cells attached. re-embed squares in flat molds and section as usual. Thomas E. Phillips, Ph.D. tphillips-at-biosci.mbp.missouri.edu.
We have had good results with thermonox coverslips and even better with aclar provided the cells will grow on it. Process as usual for TEM and IEM. Heat can curl the aclar sometimes. Scott D. Whittaker sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks " Dear Sally, I too have used Thermanox coverslips for TEM of tissue culture cells, but I found they often pulled out from or wrinkled-up my sections. I had much better luck using filter-membrane inserts. The insert fits into the wells (I used 24-well plates but they come in many sizes). You can even polarize the cell line if necessary by placing media with serum in the well under the insert and without serum in the filter-membrane cup. The membrane then can be fixed and processed in the holder and just before embedding, separate the filter, cut it into pieces, and embed. No wrinkles, no pulling out of the resin (I've used both Spurrs and LR White). Granted, they are more expensive, but. . .
"Tina Schwach" tschwach-at-tc.umn.edu
----------------------------------------------------------- I have used Thermanox coverslips extensively for this purpose. (Check your supplier of Lux tissue culture products, as well as your EM suppliers.) They are resistant to acetone (not sure about propylene oxide) and I've used them with Epon, spurr's and an araldite/epon mixture. They are supplied sterile, and peel off easily from a warm polymerised block, leaving the cells embedded in the resin block. I used to UV sterilise them in a laminar flow hood overnight before use. A word of warning though, they are not suitable for light microscopy, and some cells don't adhere very well to them. As I recall, I have used vero cells and other kidney derived epithelial cell lines on them successfully. You can section thermanox, but I think a better result is obtained by peeling it off, and polymerising some fresh resin over the cells. Contact me with problems if you wish. I have no commercial interest in this product. I've just processed heaps of cultured cells. Good luck, Rosey van Driel {Rosemary.van.driel-at-baker.edu.au}
--------------------- We use the Thermanox coverslips that you can buy from EMS. They work really well you just have to be careful to keep the correct side up (Thermanox has a sidedness to it and the cells only grow right on it). ACLAR works well too, though lately the stuff we buy from Ted Pella seems to be thinner than it used to be & has a tendency to curl up when polymerizing. I you have any questions, please feel free to ask. Paula :-) Paula Sicurello UC Berkeley psic-at-uclink4.berkeley.edu http://biology.berkeley.edu/EML --------------------------------------------------------- Besides the Lux coverslips you can also use membrane inserts made by Falcon or Costar. If you would like have the exact protocol let me know and I can send it to you. I have used them both for number of years now without any problems though I prefer costar inserts. Neelima Shah.............. From: Neelima Shah shahn-at-mail.med.upenn.edu http://www.MED.upenn.edu/morphlab/
---------------------------------------------------------- Visit the lab manual on //con-sgi.microbio.emory.edu/eyes-of-the-eagle/index.html For over ten years our lab has floated monolayers of epithelial celllines from the surface of common plastic culture ware with propylene oxide. This is far easier than trying to use coverslips. Call me at 404 727 3508 if you need some coaching on the technology. Regards, Skip } From: L R MELSEN lmelsen-at-emory.edu http://www.MED.upenn.edu/morphlab/
----------------------------------------------------------------- We routinely attach cells to thermanox coverslips (available from EMS). These coverslips can be sterilized, are compatible with conventional embedding methods and can be thin sectioned with a diamond knife. We fix in GA/FA, osmium, dehydrate and embedd in Spurr's with no problem.
If you must section the coverslip itself, try to arrange it diagonally in the block and pick up the sections on formvar, because the coverslip and the resin tend to separate under the beam.
If you are sectioning parallel to the coverslip, embedd this way. Fill a beam capsule with resin and lay the coverslip, cell side down on top. Polymerize; them immediately upon removing the blocks from the oven, pull the coverslip off. If you do this quickly while the blocks are still warm, the cells will remain on the block and you don't have to worry about sectioning the coverslip. Good Luck, Michelle Peiffer 814-865-212 email:mlk101-at-psu.edu
--------------------------------------------------------------------------- For TEM of cultured cells, we grow the cultures on "Thermanox" tissue culture coverslips. From Nalge Nunc INternational, 50 sterile coverslips, 13 mm in diameter is catalog 174950. EMS also sells these thermanox cover slips in a variety of sizes (see page 143 of their cat XIII). The coverslips can be treated with all the same chemistry as tissue including propylene oxide and Spurrs epoxy, which are two components which solubilize polystyrene. These thermanox coverslips can be sunk cell side up in freshly made Spurrs, then following polymerization the coverslips can be removed by first sawing a small area of the epoxy/cell/substrate, then immersing in liquid nitrogen for a few seconds, then prying away the substrate. Now the embedded cells are immediate on the epoxy. Re-embed two fragments of the culture face to face for cross-sections, or cut the block parallel to the face for tangential sections. We particularly like the round 13 mm thermanox coverslips for immunocytochemistry of cultured cells since they can be floated cell side down in a drop of 100 microlitters antibody - gold conjugate, conserving reagents.
If you would like to grow a larger culture, you could also use "permanox" culture dishes, which are equally resistant to chemicals common in TEM processing. These are also available through EMS and other suppliers.
Douglas R. Keene DRK-at-shcc.org
To process cell cultures for grown on round, 13 mm Thermanox tissue culture cover slips for TEM, we use the following hardware:
For fixation, we use porcelain multi-well dishes. These are what most people refer as "staining dishes". They are white, measure about 3.5 x 4.5" and have 12 depressions. We use these for glutaraldehyde, buffer rinses and OsO4. OsO4 can be completely rinsed from the dishes. Once in the last buffer after OsO4 and prior to ethanols and propylene oxide, we transfer the disks to 50 ml polypropylene culture tubes, such as Fisher 05-539-7 (these are sterile, but certainly it is not necessary to pay for sterile containers). There is plenty of room to enter a pipette for fluid exchange without touching the culture disks, and the culture surface will not touch the walls of the cylindrical tube so there is no worry that the cells will be rubbed off. Given the depth of the tube, we do not worry too much that the culture will dry out as fluids are exchanged, since the atmosphere within the tubes is fairly saturated with solvent vapor. Still, fluid exchange is done quickly. The polypropylene tubes are resistant to P.O. and Spurrs. Do not use tubes made from polystyrene as they will dissolve. Once infiltrated with the last change of epoxy, we fill a resin-resistant container with epoxy to a minimum depth of 5 mm, sink the disk so that the cells face up, then polymerize the epoxy. We steal the polypropylene lids from wheaten snap-cap vials (Fisher #0333520E) which are of the appropriate diameter for embedding 13 mm coverslips. Wearing a dust mask, We use a jewelers saw to free small blocks of embedded culture, loosen the cover slip with liquid nitrogen, then remove the disk which exposes the culture to the surface of the block. We then either re-embedded (in some of the same batch of media which was used to infiltrate the culture) face to face for cross sections, or cut the blocks parallel to the culture surface for tangential sections.
Good luck,
Doug Keene EM Facility Shriners Hospital for Children DRK-at-shcc.org
---------------------------------- Here's a few comments to follow from Doug Keene's reply to you celss on coverslips question.
You can process and section Thermanox quite easily with cells grown on them.
Orientation problems can be overcome if you cut the coverslip to fit a BEEM capsule before you innoculate with cells. I have seen good results with various cell types, and if you taper the cut coverslip to fit into the BEEM pot, trimming the block is pretty quick too. If you need, you can also bend up the coverslip at the non-tapered end to indicate which side the cells are growing.
Spurrs resin seems pretty good, but there can be some delamination between the coverslip and resin, but not always! Hope this helps. P.Bond-at-plymouth.ac.uk
--------------------------------------------
Dear Sally, The coverslips that everyone uses is my Thermanox plastic coverslips which are sterile and come in many different sizes depending on your need. They may be found wither in our hard copy catalog or on our website at http://www.emsdiasum.com. Go to our online catalog and click chapter 7. They are listed under Thermanox. In our hard copy catalog XIII they may be found on page 143. Please let me know if I may be of further assistance to you.
Sincerely, Electron Microscopy Sciences 215-646-1566 } From: "SGKCCK-at-aol.com"-at-sparc5.microscopy.com
I used to grow Vero Cells on thermanox cover slips. In Europe we buy such cover slips through our local distributor " Bioblock", but I am quite sure that you can find the same in the EMS catalog ( Nalg Nunc ref.174950 = tissue coverslip, sterile, thermanox size 13 mm in diameter). In fact the thermanox cover slips are sold sterile and the surface treated for cell culture is on the top (where the label is). I grow Vero cells (or others), infect them, fix them before labelling with gold-coupled antibodies. I forgot to say that I put the cover slip on 24 wells (or 4 wells) adapted plates (from Nunc or Falcon ...). Leaving the coverslip on the well, I do all the dehydration and epon embedding in the same plate. Before polymerization I cut the /\ shape off the beem capsule (and put three of these capsules per well (the uncutted side directly over the specimen) over a thin layer of epon. I polymerize for a night and the next day I fill the beem capsules with epon and polymerize further on. When the samples are polymerized you can just remove the capsules and the cells will be adherent on the epon. As usual you trim your block and cut the sections ( the first one will almost be good). We published this method in EmboJournal 1998, 17, 3899-3908.
I hope that this helps, Daniele Spehner From: "Daniele Spehner" {daniele.spehner-at-etss.u-strasbg.fr}
----------------------------------------------------------- Thermonox cover slips (sold by Fisher Scientific, pg. 1793 of the 98/99 catalog) are great for TEM and SEM applications. They are manufactured by Nunc, come in many different shapes and sizes, have one side textured for cell adhesion, and arrive sterile in packs of 50. They are resistant to acetone, alcohol and can be sectioned using diamond knives (I've heard this, but never tried it) without harming the knife.
I've used them with cultured monkey kidney cells, and the cells stayed attached through critical point drying. We've also coated them with poly-L-lysine and settled fish lymphocytes onto them for SEM (critical point drying didn't remove the cells).
Another neat product is the Nunc SonicSeal Slide System (their number 138121, Fisher Cat. No. 12-565-9, Fisher catalog pg. 1792). Because the wells are made of Permanox, we were able to grow cells, then fix, dehydrate, infiltrate and polymerize right in the chambers. After polymerization we separated the slide from the cells by inserting a razor blade into the interface between the slide and resin and twisting it a little. Good luck, Heather Owen, Director Electron Microscope Laboratory University of Wisconsin - Milwaukee (414)229-681 Owenha-at-csd.uwm.edu
--------------------------------------------------- We are working with Caco-2 cells grown in transwell cultures. We use a protocol similar to the one Tom Moninger describes but we embed in Embed 812 (SOLD BY ems AS A REPLACEMENT FOR Epon 812). We encounter severe wrinkling in both thin and semi-thin sections caused by the cells expanding on the surface of the knife boat while the polycarbonate membrane does not expand. We do not yet have aa remedy for this, but I have asked Tom M. to share his experiences with me. Call if you want more detail. Electron Microscopy Laboratories Email: jcoleman-at-bi-pharm.com {jcoleman-at-bi-pharm.com} ------------------------------------------
Hello Sally, you have some experience with TEM polycarbonate filters and cell culture; and you can use acetone and spurr resin with them, there is no problem at all. Good luck Nuria } From: Nuria Cortadellas {nuriac-at-giga.sct.ub.es} ----------------------------------------------- I have processed many filter membranes for TEM and SEM. I follow standard procedures with the following changes;
-dehydrate in EtOH, not acetone (eats the polycarbonate) -embed in Epon 812 or equivalent (I use Ted Pella's Eponate 12) -times can be shortened conciderably (Fix 20 min, wash and dehydrate steps 5 min, resin changes around 30 min.) -do a couple more resin changes than normal
I carry out the entire process (including embedment) in a 24 well plate and cut the filters out with a jeweler's saw after polymerization.
Thomas Moninger moninger-at-emiris.iaf.uiowa.edu http://www.uiowa.edu/~cemrf -------------------------------------------------- Tom } Hi Tom } Thanks for the quick reply!. Part of my sample may be difficult to fix and } embed, I will probably need longer times for this. Did you recommend } shorter times because of the tissue culture, or is this a requirement for } the filter? } Interesting, someone responded that they use Acetone, however I will play } it safe with alcohol. } And do you have trouble with curling of the sections? } Thanks.........SALLY Exactlly right, I use the short times because I am ususally working on monolayers. You might want to test your filters and plates with the acetone but I would bet you will see some immediate hazing and disolving. Feel free to drop me a note if you have any other questions. ----------------- ----------------- ---------------------------- I am fairly new to microscopy so I'm curious why a researcher needs the cells on coverslips to begin with. In the past when I plated cells for a TEM study I would grow them right in the dish or flask with no coverslip and process the cells right in the dish. You can then embed the cells on the dish and remove the dish after it's cured in liquid nitrogen. Unless I was doing a LM study at the same time and I would then split them right before harvesting. I sterilized the cover slips under the hood with a bunson burner and be very quick. All it takes is a fast wave through the flame, any longer and the coverslips warp. After a few seconds they cooled and then you can put them in the dish. If you don't wait they weld to the dish. You might find sterile coverslips at Fisher, Daeger, VWR or Costar. Good luck and drop me a line if you need more info. Steve D'Angelo From: sdangelo-at-batnet.com (steve d'angelo)
______________________________________________--- MSA QUERY: Original Cover slip embedding Colleagues will be growing Vero cells (Green Monkey Kidney Cells) and intestinal epithelial cells on cover slips for subsequent ultrathin sectioning and TEM examination. I am interested to know which cover slips are best to use for this purpose. Can they be purchased sterile? I have checked the Ted Pella and EMS catalogs, they both sell non sterile cellulose acetate cover slips. Are there compatibility issues with resins and solvents? I am interested in any tips or tricks to smooth the way. Thanks, Sally Burns
Sally Burns Center for Electron Optics burnssal-at-pilot.msu.edu
Try Jan Hinsch at the Leica Allendale office: 201-236-5905
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 11:53 AM 2/11/99 -0500, Joseph Passero wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If you are or were using Accu-Scope (3001, 3002, 3018PH etc.) or Wang Biomed (3002, 3004) microscopes - please share your opinion about them and your experience with me.
What did you use it for? Did it prove suitable for the task? Did you have any problems with it? Was its optical/mechanical quality OK for you and for the job?
If you had to use technical support, how helpful/quick were they?
If you used other microscopes too - how do they compare?
[In case it matters, I plan to use it for histology, microbiology and hobby, sometimes pushing resolution to the limit, doing mostly visual, once in a while photomicrography. But please share _your_ experience, regardless whether it's in the areas I mentioned, or not.]
Depending on how interesting in your opinion the answers would be to the other list members, feel free to either post the replies to the list, or e-mail me directly.
Thank you! -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
Vendor Sponsored Short Courses at the UCF/Cirent Materials Characterization Facility
1) Specimen Preparation using the Tripod Polisher (3/12-3/13): sponsored by South Bay Technology, instructor: Ron Anderson, IBM.
2) Specimen Preparation using Focused Ion Beam Milling (3/18-3/19): sponsored by FEI/Philips, instructors: Lucille Giannuzzi, UCF and Fred Stevie, Cirent Semiconductor.
COST $750/person for each course or $1200 for both courses
Registration Fee covers all Program Materials and breakfast and lunch on both days
to register see: http://pegasus.cc.ucf.edu/~ampac
or contact:
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-mail.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
If someone is going to make photocopies, please put me down for a set too, especially for the otholux but any of them would be welcome for the archive. Ed Sharpe
} } } } Dose anybody know where to purchase and/or have, a Service Manual and } Operator Manual for a Leitz Ortholux and a Labolux? } } Originals and or photo copies? } } Thank You } Joseph Passero } jp-at-spacelab.net }
To Randy Tindall: I assume you meant LKB Ultratome III...Try "NJWS-at-aol.com". Norm Woodside deals with used LKB equipment. Bob Santoianni Emory University Hospital Atlanta, GA robert_santoianni-at-emory.org
Topcon/Akashi 002B High-Res TEM for sale. Complete working machine, includes two holders, a double tilt and multi. Air table and Gatan image intensifier system, two complete wehnelts.
Has been under maintainence contract and fully serviced.
Price Negotiable
Contact Mr. Mike Frongillo (617) 253-5092 frong-at-mit.edu
Dr. David C. Bell Room 13-1818 E-mail: dcb-at-MIT.EDU Center for Mat. Sci. and Eng. PH: (617) 253-3317 Massachusetts Institute of Technology FAX: (617) 258-6478 77 Massachusetts Ave, Cambridge, MA 02139
This request comes from a co-worker, who is looking for a software program to plot the scatter of experimental orientation relationship data on a stereographic projection. Please respond to him directly (he is not on the listserver) at mangan-at-anvil.nrl.navy.mil. Thank you. - Richard Fonda
We have generated a considerable amount of data on orientation relationships between two phases using the EBSD/OIM technique on an SEM. I was hoping to plot this up on a stereographic projection, but I have up to 100 data points. I'd like to be able to choose one phase as the reference lattice and then plot three directions in the other phase that correspond to a given orientation relationship between the two. And I'd like to see how far off my experimental data is from the ideal orientation relationship as I define it. Is anyone aware of any software package that could do this for this many data points?
Thanks for your help-
Mike
_____________________________________________________________ Michael A. Mangan Naval Research Laboratory Code 6324 Washington, DC 20375 phone: (202)767-2318 fax: (202)767-2623 _____________________________________________________________
The common American device, originally used for mixing Ag and Hg for dentists, is called a Wig-L-Bug, and is available from many suppliers of infrared sampling devices, e.g.,
Pike Technologies Madison, WI 608-274-2721
There are other devices as well. If the first doesn't work out, mail me directly.
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
Dear Henrik, I have only had success grinding and polishing diamond drill bits with diamond-embedded, brass polishing wheels of various grit sizes. I usually use 45 micron, 20 micron and 6 micron wheels to make a surface suitable for SEM and EDX. You wrote: } Dear All, } } In our lab we need procedure (cutting, grinding and polishing) of } SEM sample with diamond particles in metal matrix. Any suggestions?. } } Henrik } } -- } SEM-EDS Laboratory } Metal Ravne } Slovenia } http://www2.arnes.si/guest/sgszmera1/index.html } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
} } Dear all } } can anyone tell me if people still use BSA
ref...........
Valentine, R.C. (1961) Adv. Virus Res. 8, 287
or bacitracin
ref..............
Gregory. DW and Pirie, BJS, "Wetting Agents for electron microscopy of biological specimens" Proc. 5th European Congress on EM, 1972 (Manchester) p 234
as 'wetting agents'
yes. we still use bacitracin!
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
------------------------------------------------------. } } } Dear Listmembers, } Does anybody use or have used ruthenium tetroxide as a fixative for } biological material? Any references? } Thanks in advance.
Original reference.... Luft JH, 1971 Ruthenium red & violet II ..... Anatomical Record 171 p 369
one of ours.....
CD Garland et al The preservation of surface-associated micro-organisms prepared for SEM...
J. Microscopy 116 pp 227-242
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Can anyone point me to a source of 3mm diameter carbon rods which are likely to be identical with those from my old (} 12 years) box. They were "National" Spectroscopic electrodes, made by Union Carbide Corp Carbon Products Division, 270 Park Av, NY, NY, and were "Carbon 0.12 x 12" L113SP". I have some bought recently from an independent supplier, which are noticeably softer, but they seem to have to get hotter in order to vaporise, so I would like to obtain some as above.
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Ritchie Sims wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Hi } } Can anyone point me to a source of 3mm diameter carbon rods which are } likely to be identical with those from my old (} 12 years) box. } They were "National" Spectroscopic electrodes, made by Union Carbide } Corp Carbon Products Division, 270 Park Av, NY, NY, and were "Carbon } 0.12 x 12" L113SP". } I have some bought recently from an independent supplier, which are } noticeably softer, but they seem to have to get hotter in order to } vaporise, so I would like to obtain some as above. } } thanks } } Ritchie } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
Mr Sims,
Most of the EM supply houses (in the United States at least) sell graphite rods as carbon rods. From your description, we believe that you got graphite this time. We at Ladd sell pure carbon rods. If you are interested we could quote you direct.
Deb Sicard Sales Mgr.
Disclaimer: Ladd Research is a vendor that sells carbon rods. -- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
How do TEM users store spare (i.e. not in use) specimen holders? We have four holders and only one in use at any one time - how to keep the other three 'clean' but readily accessible? The manufacturers' (JEOL) boxes are good but not air-tight and are too large (about 48cm long) for any dessicator I can find in the usual catalogues - Fisher, Jencons, Agar etc.
TIA
Alan Walker
***************************************************************************** Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom Phone: +44-(0)114-2225365 (direct) +44-(0)114-2222000 (switchboard)
What is the best way to store spare (as in, not in use right now) specimen holders? We have four holders, so three are left 'gathering dust' at any one time. The manufacturers' (JEOL) boxes are good but not air-tight, and too long for any dessicator cabinet I can find in UK catalogues.
TIA
Alan Walker
***************************************************************************** Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom Phone: +44-(0)114-2225365 (direct) +44-(0)114-2222000 (switchboard)
We have a Polaron E5100 sputter coater that recently started blowing the high voltage fuse. When I switch to "Set HT" and slowly increase the sputtering current, the current reading doesn't get above ~1 or 2 mA before the Buss GDC, 400 mA, 250 V fuse blows. Has anyone witnessed this before? Any ideas? A web search for Polaron revealed nothing. Does anyone know if they are still in business?
TIA
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
Tim Wakefield ----- / 101 Cary Hall / | \ / Auburn University, AL / --|-- \/ 36849 \ | /\ 334-844-3908 \ | / \ ----- \
On Sun, 14 Feb 1999 katie2468-at-netscape.net-at-sparc5.microscopy.com wrote:
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The Department of Anatomy and Physiology at Kansas State University has a 1977 Phillips Electron Microscope, Model EM400 for sale. This scope was purchased in 1977 by Oral Roberts University for $142,404.00. It was sold to Kansas State University in 1990 for $25,000.00. While here at Kansas State, this microscope has been well cared for and has been under a constant maintenance agreement with FEI. The faculty member who has been using this scope is retiring and no other faculty member has use for it. It is in excellent working condition. Please contact: Dani Goodband 785-532-4538 goodband-at-vet.ksu.edu
I don't know how many microscopists use Photoshop, but the following is text I submitted to the "photoshop list". If you believe in maintaining the integrity of image files, I think you ought to be made aware of Photoshop 5 defaults: (1) to enable ICC profiles and (2) the default color space (sRGB) gamma of 2.2.
~~~~~~~~
I've just installed PS 5 for Windows and have everything calibrated. But I think I'm going to object to the gamma for sRGB and bruceRGB being set to 2.2. I'm objecting for the sake of all my legacy files which were created with monitor compensation near gamma=1.6 which is approximately where 50%blackpoint and 50% whitepoint equals 50% gray ... which I believe (correct me if I'm wrong) is the basis for monitor compensation if you use "Adobe Gamma". A gamma = 2.2 has not been arbitrarily selected for sRGB and bruceRGB, reasons being "most displays", wwweb graphics, and the shadow tonal range. As a test I created with PS4 a RGB image with a flat histogram for all channels, and imported it into PS5 "from" a profile in accordance with my previous compensation and "into" bruceRGB color space. Upon viewing the bruceRGB profiled image it is accurate, HOWEVER if I now examine the histogram it is obvious the translation had done a major number on the pixel values.
I only mention this for the sake of legacy files ... that is, I can understand the profile reasoning from creation to final product. However, I don't know what to suggest. I understand the choice of gamma=2.2, but I believe it assumes most displays will never be compensated and that wwweb graphics will never display in accordance with ICC profiles. If that proves untrue, then the pros will in the future probably suggest a smaller gamma (1.8) and in the mean time we'll have created a lot of fine work profiled for a 2.2 display ... and from what I saw happen to my histogram, I suggest we put out work thru as few profiles as possible.
Can I ask how many of you really do work with sRGB=2.2??
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Maggy Piranian wrote: } } Deb, I saw your response to Ritchie. What is the difference between carbon } and graphite? What are their different virtues and vices? } Maggy Piranian } ***************************************************************** } } Maggy Piranian } Electron Microprobe & X-Ray Diffraction Labs } Dept. of Earth Sciences } Memorial University of Newfoundland } St. John's, Newfoundland Phone (709) 737 8244 } A1B 3X5 Fax (709) 737 2589 } maggy-at-sparky2.esd.mun.ca } *****************************************************************Ladd offers both carbon and graphite but the majority of our customers ask for pure carbon, so that is our standard. Our own lab people feel that carbon provides a superior coating for our substrates.
Both our carbon and graphite are spectroscopic grades (extremely low impurity level). Pros/cons of both are:
Carbon - Harder to cut Needs less amps to heat Preferred by most of our users
Graphite - Softer, easier to cut Needs more amps (heat) to evaporate.
Disclaimer: Ladd Research is a vendor of microscopy supplies.
John Arnott -- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
In a message dated 99-02-15 12:45:15 EST, wise-at-vaxa.cis.uwosh.edu writes:
{ { We have a Polaron E5100 sputter coater that recently started blowing the high voltage fuse. When I switch to "Set HT" and slowly increase the sputtering current, the current reading doesn't get above ~1 or 2 mA before the Buss GDC, 400 mA, 250 V fuse blows. Has anyone witnessed this before? Any ideas? A web search for Polaron revealed nothing. Does anyone know if they are still in business?
TIA
Bob } }
Hi Bob,
Polaron vacuum and coating devices are sold by Energy Beam Sciences in Agawam, MA. You can reach them as follows:
Energy Beam Sciences, Inc. P.O. Box 468 11 Bowles Road Agawam, MA 01001 Tel. (800) 992-9037 Fax (413) 789-2786 www.ebsciences.com
I'm certain they can help you solve your problem.
Cheers,
Bob ****************************** Robert (Bob) Chiovetti, Ph.D. President Microimaging Technologies, Inc. Tucson, Arizona USA Tel./Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems Clinical & Research Microscopy Cytology/Histology/Pathology/EM *******************************
Maggy Piranian wrote: } } Deb, I saw your response to Ritchie. What is the difference between carbon } and graphite? What are their different virtues and vices? } Maggy Piranian}
Sorry, a few lines were missing from our first response:
Ladd offers both carbon and graphite, but the majority of our customers ask for pure carbon so that is our standard. Our own lab people feel that carbon provides a superior coating for our substrates.
Both our carbon and graphite are spectroscopic (extremely low impurity level). Pros/cons:
Carbon - harder to cut needs less amps to heat preferred by most customers
Graphite - softer, easier to cut needs more amps (heat) to evaporate.
disclaimer: Ladd Research is a vendor of microscopy supplies
John Arnott -- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
We have used a 5100 here for many years. I think it has acted up similarly. After having problems reaching vacuum, I pulled it apart and gave it a good cleaning. There was plenty of gold all over the place. I think some of it was even in places where it may have caused a short to the frame. I got over 15 g of gold scraping off what had built up over the years.
Warren
At 11:15 AM 2/15/99 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Just a short heads up. I'll be off-line for a day or so.While I don't expect problems, Murphy says something will go wrong on the List. To minimize problems. I will put Subscriptions and Unscriptions on hold until I reconnect. Please be patient until I get back on the Net.
BTW, yes I did see the XXX mail, and I have added that address to the SPAM filter. However, please remember this is a public posting site and junk mail will occassional get through. The filter, theoretically, permits this only once from a given address, but SPAMer's generally post from different (and faked) addresses all the time. Most of the junk mail is caught, but there will be some that gets through.
Union Carbide sold its line of spectroscopic electrodes to Ultra Carbon in Bay City, Michigan about 15-20 years ago. Shortly thereafter, Union Carbide was fragmented, and UCAR Carbon Company is the remnants of the Union Carbide Carbon Products Division.
The research laboratories at UCAR have probably the only remaining stock of genuine Union Carbide spectroscopic electrodes. We do have a few of the grade and size you want remaining. If you are interested, please contact me.
Helen Mayer UCAR Carbon Company 12900 Snow Road Parma, OH 44130 216-676-2373 FAX 216-676-2276 helen.mayer-at-ucar.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi
Can anyone point me to a source of 3mm diameter carbon rods which are likely to be identical with those from my old (} 12 years) box. They were "National" Spectroscopic electrodes, made by Union Carbide Corp Carbon Products Division, 270 Park Av, NY, NY, and were "Carbon 0.12 x 12" L113SP". I have some bought recently from an independent supplier, which are noticeably softer, but they seem to have to get hotter in order to vaporise, so I would like to obtain some as above.
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
A student in our lab would like to do TEM on Drosophila eggs to look specifically at the micropyle. Our initial fixation was based on a paper on amphibian eggs. We fixed in 3% glutaraldehyde, 2% formaldehyde, 1% acrolein and 2.5% DMSO in 0.1 M cacodylate buffer overnight at room temperature, followed by 2% osmium, 0.2 M sucrose in 0.1M cacodylate buffer, overnight at room temperature. Eggs were then dehydrated in acetone and embedded in EPON.
We observed significant seperation (complete detachment actually) of the chorion from the egg. In fact the chorion appeared to be almost twice the size of the egg. Any advice on why this happened or a better way to fix would be greatly appreciated.
Thanks in advance, Michelle
#################################################### Michelle Peiffer Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:mlk101-at-psu.edu ####################################################
As an accessory to our plasma cleaner we also offer Vacuum Storage Containers. Typically, these are used for short term storage when transferring from the plasma cleaner to the microscope. However, these same storage containers can be fitted with the appropriate valves to evacuate the container, remove water vapor, back fill with dry N2, and store over longer periods of time and be used separate from the plasma cleaner.
We can customize these storage stations to enable you to store multiple holders - even from different manufacturers - in one manifold with individual valves etc. I would be pleased to discuss this system with yo= u further. Please contact me off-line for additional information.
DISCLAIMER: South Bay Technology manufacturers systems for specimen cleaning and storage and therefore I have a vested interest in promoting their use.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by "A.Walker" } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Hi,
How do TEM users store spare (i.e. not in use) specimen holders? We have four holders and only one in use at any one time - how to keep the other three 'clean' but readily accessible? The manufacturers' (JEOL) boxes are good but not air-tight and are too large (about 48cm long) for any dessicator I can find in the usual catalogues - Fisher, Jencons, Agar etc.
I am looking for a commercial source of a good GABA-A beta chain subunit antibody. The one we are using is giving non-specific labeling. Thanking you in advance, Lilith ------------------------------------------------------------------ Lilith Ohannessian-Barry National Research Council Institute of Biological Sciences CANADA Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
There was an excellent article on selecting an RGB working space in the Adobe Photoshop magazine they send free to register users. It was the autumn 1998 issue on p.51- 56. it is probably on their web site in an archive or they will fax it to you. The author concluded that sRGB was the worst possible choice of many. The author thought ColorMatch RGB was a better gamut space to work with. I just read the article this weekend (it is very dense - or maybe i am the dense one but once you get it, it seems very useful. Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Can anyone advise a fax number, an e-mail address, or a website for Agar Aids, UK? Their clocks are 13 hours out of synch with mine, difficult to phone them.
thanks and thanks also to the responders re carbon rods, very interesting to realise the difference between "carbon" rods and "graphite" ones.
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Your comment re: "how many microscopists use Photoshop" is interesting and has a major impact on the scientific quality of microscopy. MME just conducted research at the Cell Biology meeting in December and asked that question. Fifty-four percent of our sample of nearly 500 participants answered this question and 86 % (46% of the total survey population) indicated that they use Photoshop! (And you can quote us on that).
A reminder that this software package was initially designed for a graphics audience which does not need to be concerned with the validity of data carried by each pixel. Clearly, it is important that any issues which affect the scientific content of these images be carefully calibrated and controlled.
Hope this insight is helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 09:48 AM 2/15/99 -0800, shAf wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: SEM "hole and pit" Analysis Trying to update my current software with crrent "automatic or semi-automatic" software and a manual "digitizing Pen" techniques. Because of the edge contrast in SEM, I've use edge detection techniques such as watershed and an old commercial software called "WICS" (which I can't find). Any suggestions? To avoid tieing up the server please respond directly after the first few responses. Thank you. Jeff Day/ "CD" WA5EKH-at-JUNO.COM
___________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com/getjuno.html or call Juno at (800) 654-JUNO [654-5866]
In a message dated 2/16/99 12:07:58 PM, mme-at-map.com writes:
} Your comment re: "how many microscopists use Photoshop" is interesting } and has a major impact on the scientific quality of microscopy. MME just } conducted research at the Cell Biology meeting in December and asked that } question. Fifty-four percent of our sample of nearly 500 participants } answered this question and 86 % (46% of the total survey population) } indicated that they use Photoshop! (And you can quote us on that).
Thanks, Barbara. Although the original point of this thread was the choice of an appropriate color space for imaging, the more general point about Photoshop is one that I believe in as well. That is why we wrote The Image Processing Tool Kit to run with it. This provides the functionality of dedicated image analysis packages costing many thousands of dollars for a few hundred (sorry about the commercial plug on this list, but I think my point here is relevant) by taking advantage of Photoshop as the host. This program supports acquisition with just about every device out there, offers a uniform user interface on Macs and PCs, takes care of storage, printing, virtual memory, etc. etc. (all the system stuff), and (with rev 5) offers a modest amount of automation. It is easily learned and relatively inexpensive. It is also a wonderfully straightforward package for learning about scientific image analysis, with an extensive on-disk tutorial and examples (mostly images from The Image Processing Handbook). Apparently the users agree, because we've sold several thousand copies of the tool kit so far. (http://members.aol.com/ImagProcTK)
Hi Christopher; I have had very good luck staining starch with PAS, followed by counterstain with light green. the starch stains a bright fuschia, and the protein varying intensities of green (depending on type of protein and relative amounts). This was done on GMA sections, btw. If you need more information, feel free to contact me.k cheers shea
Dr. S. Shea Miller Agriculture and Agri-Food Canada Eastern Cereal and Oilseed Research Centre 2068 K.W. Neatby Bldg Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 email: millers-at-em.agr.ca phone: 613-759-1760 fax: 613-759-1701 ! !
Dear Ritchie, I found there was a difference between carbon rods and graphite rods. I prefer the carbon. You may have been supplied the graphite, which, as you say, needs to be heated higher to vaporize. I went back to Carbone of America, Ultra Carbon Div. for spectrographic grade pressed carbon rods. The latest address I have is: ultra carbon 900 Harrison St. Bay City, MI 48708-8244 USA
Good luck, Regards, Mary
At 04:13 PM 15/02/99 GMT+1200, you wrote: } } } Hi } } } } Can anyone point me to a source of 3mm diameter carbon rods which are } } likely to be identical with those from my old (} 12 years) box. } } They were "National" Spectroscopic electrodes, made by Union Carbide } } Corp Carbon Products Division, 270 Park Av, NY, NY, and were "Carbon } } 0.12 x 12" L113SP". } } I have some bought recently from an independent supplier, which are } } noticeably softer, but they seem to have to get hotter in order to } } vaporise, so I would like to obtain some as above. } } } } thanks } } } } Ritchie } } Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Ritchie, The listing in the Kaker.com vendor database shows: Agar Scientific Ltd. 66a Cambridge Road CM24 8DA Stansted UK Tel: +44 1279 81 35 19 Fax: +44 1279 81 51 06 Grids, calibration specimen, tweezers, small tools, light microscope accessories, optical aids, steroscopy, specimen preparation equipment, biological specimen preparation, chemicals, materials science specimen preparation, image storage and analys is, photo materials and equipment, cleaninig materials, safety equipment, publications, starters kits. I was unable to find a Web site listing. At leaast fax will help with the time difference. You wrote:
} Can anyone advise a fax number, an e-mail address, or a website for } Agar Aids, UK? } Their clocks are 13 hours out of synch with mine, difficult to phone } them. } } thanks and thanks also to the responders re carbon rods, very interesting } to realise the difference between "carbon" rods and "graphite" ones. } } cheers } } rtch } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Tom writes ... } } } There was an excellent article on selecting an RGB working } space in the Adobe Photoshop magazine ... The author concluded } that sRGB was the worst possible choice of many. ...
That article was written by Bruce Fraser. I haven't read the article, but I'm reading his and Blatner's "Real World Photoshop 5" which I highly reccommend for unsderstanding the underpinnings of PS's handling of color management. Bruce has created "bruceRGB" which he describes as "social-democratic alternative ..." where sRGB and Color match RGB" are the two ends of a common spectrum. However, where Bruce deviates radically from Color Match is his choice of gamma=2.2 for the working space gamma (cmRGB gamma=1.8). I understand his point of view if your display gamma is indeed 2.2 .. but if you read (for example) a spacial distribution of an elemental composition into this color space (where the gamma is considered unity), the PS conversion (bruceRGB or sRGB) will indeed do a number on the image map's histogram. I do understand Photoshop is used within our community primarily for presentation and not really as a data tool ... I only mention this to those who may be considering the PS5 upgrade and as a word of caution.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I agree. I purchased a copy of the toolkit and have found it a very handy addition to Photoshop. You get boolean math, background adjustment, fft stuff, thresholding, auto-counting capabilities and much more. It appears to me to do just about everything that my (orders of magnitude) more expensive image processing software does with the exception of macro writing capabilities. I've been using it lots for semi-automatic counting. I add a grid to my acquired image and then use the mouse to mark intersections with the grid. The program will count the number of marks you put on with your mouse. If you are using Photoshop it is a worthwhile addition to your lab software even if you have a full blown image processing package. If you don't have a full-blown image processing program (and can't afford one) then it would be EXTREMELY helpful. John also has several useful books about image processing. All very practical, knowledgable and useful in my opinion.....
Robin Griffin - who has no financial interest in John's stuff!
-----Original Message----- } From: "DrJohnRuss-at-aol.com"-at-Sparc5.Microscopy.Com [mailto:"DrJohnRuss-at-aol.com"-at-Sparc5.Microscopy.Com] Sent: Tuesday, February 16, 1999 7:50 AM To: mme-at-map.com; Microscopy-at-Sparc5.Microscopy.Com
In a message dated 2/16/99 12:07:58 PM, mme-at-map.com writes:
} Your comment re: "how many microscopists use Photoshop" is interesting } and has a major impact on the scientific quality of microscopy. MME just } conducted research at the Cell Biology meeting in December and asked that } question. Fifty-four percent of our sample of nearly 500 participants } answered this question and 86 % (46% of the total survey population) } indicated that they use Photoshop! (And you can quote us on that).
Thanks, Barbara. Although the original point of this thread was the choice of an appropriate color space for imaging, the more general point about Photoshop is one that I believe in as well. That is why we wrote The Image Processing Tool Kit to run with it. This provides the functionality of dedicated image analysis packages costing many thousands of dollars for a few hundred (sorry about the commercial plug on this list, but I think my point here is relevant) by taking advantage of Photoshop as the host. This program supports acquisition with just about every device out there, offers a uniform user interface on Macs and PCs, takes care of storage, printing, virtual memory, etc. etc. (all the system stuff), and (with rev 5) offers a modest amount of automation. It is easily learned and relatively inexpensive. It is also a wonderfully straightforward package for learning about scientific image analysis, with an extensive on-disk tutorial and examples (mostly images from The Image Processing Handbook). Apparently the users agree, because we've sold several thousand copies of the tool kit so far. (http://members.aol.com/ImagProcTK)
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
---------- Forwarded message ----------
Tuesday, February 16, 1999
InvestingNow.com Wrote
http://www.InvestingNow.com
At 8 AM stocks appear to be set for a big move this morning. The S&P 500 futures were up 7.2, more than 11 above fair value and cueing the market for a good jump upward at the open. At 8:10 a.m. EST, the 30-year Treasury bond was up 28/32 to 98 10/32, dropping the yield to 5.36%.
Japan's Nikkei added 177.92 to 14,232.64. Hong Kong's stock market was closed for the Chinese New Year holiday. Europe's major averages were all higher. In Frankfurt, the Dax was up 15.24 to 4894.79. In Paris, the CAC was up 10.33 to 4075.52. In London, the FTSE was up 93.2, or 1.5%, to 6116.4.
Dell (DELL:NYSE) and Hewlett-Packard (HWP:NYSE) will report earnings after the close. American Airlines expects nearly all its flight schedule to be back to normal today. American is a unit of AMR (AMR:NYSE).
Volvo (VOLVY:Nasdaq ADR) of Sweden is considering a takeover of Navistar (NAV:NYSE), the U.S. truck and engine manufacturer, in a move that could more than double Volvo's share of the North American truck market, the Financial Times reported. Earlier this month, Volvo agreed to sell its car division to Ford (F:NYSE). Chicago-based Navistar has a market capitalization of $2.3 billion, meaning that a buyer would probably have to pay at least $3 billion to take full control of the company, the newspaper said. If Volvo, however, were only to buy Navistar's heavy truck operations, the price could be substantially lower, the newspaper said.
The Dow Jones Industrial Average (DJIA) should maintain its trading range of 9,000 to 9,100 on the bottom end and 9,375 to 9,500 on the top end. These markets have been fantastic for trading the last week. As can be seen from our calls, for short-term calls they have been predictable. The wild card continues to be the technology sector. The nervousness Friday concerning DELL will continue today as we await the earning announcement this afternoon. Overall we are positive on the DJIA and broad market, and continue to dislike technology stocks. For the long term we remain BULLISH.
Did you receive our e-mail execution to cover our short in Microsoft on Friday? We covered this short in MSFT at $158 at the close on Friday afternoon. This short was established on February 3, 1999 at $168. Be sure to check out our trading suggestions and portfolio recommendations regularly. And get on our mailing list!
At 8 AM stocks appear to be set for a big move this morning. The S&P 500 futures were up 7.2, more than 11 above fair value and cueing the market for a good jump upward at the open. At 8:10 a.m. EST, the 30-year Treasury bond was up 28/32 to 98 10/32, dropping the yield to 5.36%.
Japan's Nikkei added 177.92 to 14,232.64. Hong Kong's stock market was closed for the Chinese New Year holiday. Europe's major averages were all higher. In Frankfurt, the Dax was up 15.24 to 4894.79. In Paris, the CAC was up 10.33 to 4075.52. In London, the FTSE was up 93.2, or 1.5%, to 6116.4.
Dell (DELL:NYSE) and Hewlett-Packard (HWP:NYSE) will report earnings after the close. American Airlines expects nearly all its flight schedule to be back to normal today. American is a unit of AMR (AMR:NYSE).
Volvo (VOLVY:Nasdaq ADR) of Sweden is considering a takeover of Navistar (NAV:NYSE), the U.S. truck and engine manufacturer, in a move that could more than double Volvo's share of the North American truck market, the Financial Times reported. Earlier this month, Volvo agreed to sell its car division to Ford (F:NYSE). Chicago-based Navistar has a market capitalization of $2.3 billion, meaning that a buyer would probably have to pay at least $3 billion to take full control of the company, the newspaper said. If Volvo, however, were only to buy Navistar's heavy truck operations, the price could be substantially lower, the newspaper said.
The Dow Jones Industrial Average (DJIA) should maintain its trading range of 9,000 to 9,100 on the bottom end and 9,375 to 9,500 on the top end. These markets have been fantastic for trading the last week. As can be seen from our calls, for short-term calls they have been predictable. The wild card continues to be the technology sector. The nervousness Friday concerning DELL will continue today as we await the earning announcement this afternoon. Overall we are positive on the DJIA and broad market, and continue to dislike technology stocks. For the long term we remain BULLISH.
Did you receive our e-mail execution to cover our short in Microsoft on Friday? We covered this short in MSFT at $158 at the close on Friday afternoon. This short was established on February 3, 1999 at $168. Be sure to check out our trading suggestions and portfolio recommendations regularly. And get on our mailing list!
I have an instruction booklet(no service manual) for the Ortholux, as well as booklets for attachments called Ultropak and Interference Contrast Device T. Someone give me an address to send copies to.
Julie Gross UCONN Health Center jgross-at-neuron.uchc.edu
Another plus with respect to using Image Processing Toolkit Version 2.5 is that John has included his text on Stereology on the CD-ROM. -Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
---------- } From: "DrJohnRuss-at-aol.com"-at-Sparc5.Microscopy.Com To: mme-at-map.com; Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
In a message dated 2/16/99 12:07:58 PM, mme-at-map.com writes:
} Your comment re: "how many microscopists use Photoshop" is interesting } and has a major impact on the scientific quality of microscopy. MME just } conducted research at the Cell Biology meeting in December and asked that } question. Fifty-four percent of our sample of nearly 500 participants } answered this question and 86 % (46% of the total survey population) } indicated that they use Photoshop! (And you can quote us on that).
Thanks, Barbara. Although the original point of this thread was the choice of an appropriate color space for imaging, the more general point about Photoshop is one that I believe in as well. That is why we wrote The Image Processing Tool Kit to run with it. This provides the functionality of dedicated image analysis packages costing many thousands of dollars for a few hundred (sorry about the commercial plug on this list, but I think my point here is relevant) by taking advantage of Photoshop as the host. This program supports acquisition with just about every device out there, offers a uniform user interface on Macs and PCs, takes care of storage, printing, virtual memory, etc. etc. (all the system stuff), and (with rev 5) offers a modest amount of automation. It is easily learned and relatively inexpensive. It is also a wonderfully straightforward package for learning about scientific image analysis, with an extensive on-disk tutorial and examples (mostly images from The Image Processing Handbook). Apparently the users agree, because we've sold several thousand copies of the tool kit so far. (http://members.aol.com/ImagProcTK)
At 8 AM stocks appear to be set for a big move this morning. The S&P 500 futures were up 7.2, more than 11 above fair value and cueing the market for a good jump upward at the open. At 8:10 a.m. EST, the 30-year Treasury bond was up 28/32 to 98 10/32, dropping the yield to 5.36%.
Japan's Nikkei added 177.92 to 14,232.64. Hong Kong's stock market was closed for the Chinese New Year holiday. Europe's major averages were all higher. In Frankfurt, the Dax was up 15.24 to 4894.79. In Paris, the CAC was up 10.33 to 4075.52. In London, the FTSE was up 93.2, or 1.5%, to 6116.4.
Dell (DELL:NYSE) and Hewlett-Packard (HWP:NYSE) will report earnings after the close. American Airlines expects nearly all its flight schedule to be back to normal today. American is a unit of AMR (AMR:NYSE).
Volvo (VOLVY:Nasdaq ADR) of Sweden is considering a takeover of Navistar (NAV:NYSE), the U.S. truck and engine manufacturer, in a move that could more than double Volvo's share of the North American truck market, the Financial Times reported. Earlier this month, Volvo agreed to sell its car division to Ford (F:NYSE). Chicago-based Navistar has a market capitalization of $2.3 billion, meaning that a buyer would probably have to pay at least $3 billion to take full control of the company, the newspaper said. If Volvo, however, were only to buy Navistar's heavy truck operations, the price could be substantially lower, the newspaper said.
The Dow Jones Industrial Average (DJIA) should maintain its trading range of 9,000 to 9,100 on the bottom end and 9,375 to 9,500 on the top end. These markets have been fantastic for trading the last week. As can be seen from our calls, for short-term calls they have been predictable. The wild card continues to be the technology sector. The nervousness Friday concerning DELL will continue today as we await the earning announcement this afternoon. Overall we are positive on the DJIA and broad market, and continue to dislike technology stocks. For the long term we remain BULLISH.
Did you receive our e-mail execution to cover our short in Microsoft on Friday? We covered this short in MSFT at $158 at the close on Friday afternoon. This short was established on February 3, 1999 at $168. Be sure to check out our trading suggestions and portfolio recommendations regularly. And get on our mailing list!
} What are operating temperatures of the two Diffusion pumps in a JEOL } 100-CX, and can I replace the oil with Santovac 5. } I do not know ofhand what the operating temperature is but it is using a 200V 600W heater. We have been running the 100S TEM and 100C TEM as well as JSM 840 Scanner on Santovac 5 without anny problems.
Hope this is some help.
Mr. S H Coetzee Electron Microscope Unit Private bag X3 Wits Johannesburg 2050 Tell: +27 11 716 2419 Fax : +27 11 339 3407 E-mail stephan-at-gecko.biol.wits.ac.za
To Laurence Marks and Amy Yarbrough, There is an excellent history of the Electron Microscope, "EMSA and Its People--the First Fifty Years" by Sterling P. Newberry. It was published in 1992 by EMSA (new MSA).
A group here in the medical school needs to do some immunoEM and will require cryoultramicrotomy. Are there any labs in the Boston area which have this capability and would make it available perhaps on a fee for service basis? Thanks. Don Gantz Boston Univ Med School gantz-at-med-biophd.bu.edu 617-638-4017
A colleague here asked me whether I knew a simple stain for bacteria that would selectively demonstrate the presence or absence of lipid. The idea would be to stain E. coli before or after lipid extraction, and see in a light microscope whether the lipid had been removed.
I know nothing about microbial stains. Any thoughts on whether this is possible or how to go about this experiment?
Gary Radice 804-289-8107 Department of Biology 804-289-8233 (FAX) University of Richmond gradice-at-richmond.edu Richmond VA 23173
As a supplier of items for microscopy we may be able to help you with your storage problem. If our standard desiccator cabinet is too small we can, (at extra cost sadly), probably provide a custom unit. I will send details of our stock item.
All commercial disclaimers apply
Terry Cooper TAAB Laboratories Equipment Ltd 3 Minerva House, Calleva Park Aldermaston, Berks, RG7 8NA, UK
Dr. E. McCain at Muhlenberg College, Allentown, PA, is looking for an aperture drive for a Carl Zeiss Transmission Microscope EM109. If anyone out there can help her, please contact her directly at mccain-at-muhlenberg.edu. Thanks in advance. Pete Dondl
I recently attended a two day course on adhesion. The topic of the different types of cleaning came up and one that was mentioned was UV/ozone. I am familiar with that and I thought that someone had told me that they stored their TEM holders in a UV light box. The problem is that it takes two wavelengths of UV for this type of cleaning to work (One that creates the ozone and one that dissociates it to form nascent oxygen). Are there any systems out there that have ports for holders could be inserted in order for them to maintain their cleanliness while not being used? You would not want a system that the whole holder gets into because the UV might degrade the plastic parts and O-ring seals.
Just curious.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This is about the Image Processing Tool Kit available from Dr. John Russ = and Chris Russ, which are plug ins for Photoshop and NIH Image.
Just wanted to add that not only is it a great addition for Photoshop and = NIH Image as everyone has already mentioned, it also includes a nice = tutorial that students can use to learn how to use the filters. It = further helps them develop their skills in using Photoshop. We use it = for part of our Digital Imaging Course. It really makes Photoshop a much = more powerful program than it already is.
I have no financial interest in the product, but I do love it.
Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
__________________________________________________________________________= _____ } From: Walck. Scott D. on Tue, Feb 16, 1999 6:14 PM
Another plus with respect to using Image Processing Toolkit Version 2.5 = is that John has included his text on Stereology on the CD-ROM. -Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
---------- } From: "DrJohnRuss-at-aol.com"-at-Sparc5.Microscopy.Com To: mme-at-map.com; Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
In a message dated 2/16/99 12:07:58 PM, mme-at-map.com writes:
} Your comment re: "how many microscopists use Photoshop" is interesting } and has a major impact on the scientific quality of microscopy. MME = just } conducted research at the Cell Biology meeting in December and asked = that } question. Fifty-four percent of our sample of nearly 500 participants } answered this question and 86 % (46% of the total survey population) } indicated that they use Photoshop! (And you can quote us on that).
Thanks, Barbara. Although the original point of this thread was the = choice of an appropriate color space for imaging, the more general point about Photoshop is one that I believe in as well. That is why we wrote The Image = Processing Tool Kit to run with it. This provides the functionality of dedicated = image analysis packages costing many thousands of dollars for a few hundred = (sorry about the commercial plug on this list, but I think my point here is relevant) by taking advantage of Photoshop as the host. This program supports acquisition with just about every device out there, offers a uniform user interface on Macs and PCs, takes care of storage, printing, virtual = memory, etc. etc. (all the system stuff), and (with rev 5) offers a modest amount = of automation. It is easily learned and relatively inexpensive. It is also a wonderfully straightforward package for learning about scientific image analysis, with an extensive on-disk tutorial and examples (mostly images from The Image Processing Handbook). Apparently the users agree, because we've sold several thousand copies of the tool kit so far. (http://members.aol.com/ImagProcTK)
John Russ
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I'm looking for a used unit in working condition. This 1600T has the "universal" stage....small round model. Need the detector and amplifier with BNC connector interface to signal selector switch.
I'm looking for a used unit in working condition. This 1600T has the "universal" stage....small round model. Need the detector and amplifier with BNC connector interface to signal selector switch.
This is about the Image Processing Tool Kit available from Dr. John Russ and Chris Russ, which are plug ins for Photoshop and NIH Image.
Just wanted to add that not only is it a great addition for Photoshop and NIH Image as everyone has already mentioned, it also includes a nice tutorial that students can use to learn how to use the filters. It further helps them develop their skills in using Photoshop. We use it for part of our Digital Imaging Course. It really makes Photoshop a much more powerful program than it already is.
I have no financial interest in the product, but I do love it.
Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
_______________________________________________________________________________ } From: Walck. Scott D. on Tue, Feb 16, 1999 6:14 PM
Another plus with respect to using Image Processing Toolkit Version 2.5 is that John has included his text on Stereology on the CD-ROM. -Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
---------- } From: "DrJohnRuss-at-aol.com"-at-Sparc5.Microscopy.Com To: mme-at-map.com; Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
In a message dated 2/16/99 12:07:58 PM, mme-at-map.com writes:
} Your comment re: "how many microscopists use Photoshop" is interesting } and has a major impact on the scientific quality of microscopy. MME just } conducted research at the Cell Biology meeting in December and asked that } question. Fifty-four percent of our sample of nearly 500 participants } answered this question and 86 % (46% of the total survey population) } indicated that they use Photoshop! (And you can quote us on that).
Thanks, Barbara. Although the original point of this thread was the choice of an appropriate color space for imaging, the more general point about Photoshop is one that I believe in as well. That is why we wrote The Image Processing Tool Kit to run with it. This provides the functionality of dedicated image analysis packages costing many thousands of dollars for a few hundred (sorry about the commercial plug on this list, but I think my point here is relevant) by taking advantage of Photoshop as the host. This program supports acquisition with just about every device out there, offers a uniform user interface on Macs and PCs, takes care of storage, printing, virtual memory, etc. etc. (all the system stuff), and (with rev 5) offers a modest amount of automation. It is easily learned and relatively inexpensive. It is also a wonderfully straightforward package for learning about scientific image analysis, with an extensive on-disk tutorial and examples (mostly images from The Image Processing Handbook). Apparently the users agree, because we've sold several thousand copies of the tool kit so far. (http://members.aol.com/ImagProcTK)
John Russ
------------------ RFC822 Header Follows ------------------ Received: by sjdccd.cc.ca.us with ADMIN;16 Feb 1999 18:14:18 -0800 Received: from Sparc5.Microscopy.Com ([206.69.208.10]) by ent2.sjdccd.cc.ca.us (Netscape Messaging Server 3.6) with SMTP id 296 for {jmurphy-at-sjdccd.cc.ca.us} ; Tue, 16 Feb 1999 18:13:57 -0800 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id PAA22384 for dist-Microscopy; Tue, 16 Feb 1999 15:18:10 -0600 Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id PAA22370 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 16 Feb 1999 15:17:39 -0600 Received: from gateway.ppg.com (gateway.ppg.com [199.221.65.2]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id PAA22358 for {microscopy-at-sparc5.microscopy.com} ; Tue, 16 Feb 1999 15:17:25 -0600 Received: by gateway.ppg.com id QAA25744 (SMTP Gateway for microscopy-at-sparc5.microscopy.com); Tue, 16 Feb 1999 16:33:07 -0500 Message-Id: {199902162133.QAA25744-at-gateway.ppg.com} Received: by gateway.ppg.com (Protected-side Proxy Mail Agent-1); Tue, 16 Feb 1999 16:33:07 -0500 } From: "Walck. Scott D." {walck-at-ppg.com} To: Micro {microscopy-at-Sparc5.Microscopy.Com}
} From: Administrator_at_RLT013-at-ccmailgw.mcgawpark.baxter.com } X-SMTP: helo gaugler.com from administrator_at_rlt013-at-ccmailgw.mcgawpark.baxter.com server -at-server14.aitcom.net ip 208.234.0.15 } Date: Wed, 17 Feb 1999 20:41:46 -0600 } To: "Dr. Gary Gaugler" {gary-at-gaugler.com} } Subject: Message not deliverable } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Chih-Hung, You could put them in a grid box and hand carry them to avoid any rough handling. Another option is to use membrane boxes which are boxes with PE membranes stretched across the openings. When sandwiched together they hold the grid in place suspended between the two membranes. They are commercially available. Russ
-----Original Message----- } From: "chanch-at-ufl.edu"-at-sparc5.microscopy.com [mailto:"chanch-at-ufl.edu"-at-sparc5.microscopy.com] Sent: Wednesday, February 17, 1999 6:50 PM To: microscopy-at-sparc5.microscopy.com
Dear Professional friends,
I have to use a distant microscope. What's the best way to transport (from FL to CO) fragile TEM samples?
best regards,
chih-hung chang Dept. of Chemical Engineering University of Florida Gainesville,FL 32611
I use membrane boxes, one sample per box. I've used them with III-V, II-VI, and now glass samples. You can buy them from a variety of EM supply houses. I use the one inch wide white Post-it tape to label the samples. When I'm done with the sample, I reuse the box and tear off the label. They stack nicely and you can pack a bunch into small boxes that are easily transportable. They also make great pill boxes. -Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: "chanch-at-ufl.edu"-at-Sparc5.Microscopy.Com To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Dear Professional friends,
I have to use a distant microscope. What's the best way to transport (from FL to CO) fragile TEM samples?
best regards,
chih-hung chang Dept. of Chemical Engineering University of Florida Gainesville,FL 32611
Chih-Hung, Transporting specimens is routinely undertaken by many people, the best method depends on your specimens and what you are going to do with them. Are they very fragile, air sensitive, contaminate easily or do they change with time? They can be transported by hand or mail. They can be sealed in an inert, dry, gas in a grid box, stored in alcohol filled tubes or packed in velin tissue and filter paper in a small box. Undoubtably there are many other ways as well. If you have particular requirements for your specimens then let us know and we will see if we can help with your specific problem,
Good luck, Ron
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
I'm hoping that someone will be able to help me out with this as I am primarily a biological science person.
1- Is there someone in the RTP area that has an SEM with an environmental chamber?
2- Any suggestions on a plastic polymer that could poured onto a bunch of particals (eg: sand) which after polymerization could be fractured and the fractured surface examined by an SEM so that partical size and intrastitial space measurments could be made using image analysis?
3- To complicate matters, it would be nice to seperate (again via SEM) the organics from the minerals vissually. We need to see if the organic is chemically attached to the mineral or if it forms aggregates between them.
best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**Suppose you were an idiot... And suppose you were a member of Congress .. But I repeat myself.-Mark Twain**
I'm hoping that someone will be able to help me out with this as I am a biological science person (give me a cell any time).
1- Is there someone in the RTP area that has an SEM with an environmental chamber?
2- Any suggestions on a plastic polymer that could poured onto a bunch of particals (eg: sand) which after polymerization could be fractured and the fractured surface examined by an SEM so that partical size and intrastitial space measurments could be made using image analysis?
3- To complicate matters, it would be nice to seperate (again via SEM) the organics from the minerals vissually. We need to see if the organic is chemically attached to the mineral or if it forms aggregates between them.
best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
I'm hoping that someone will be able to help me out with this as I am a biological science person (give me a cell any time).
1- Is there someone in the RTP area that has an SEM with an environmental chamber?
2- Any suggestions on a plastic polymer that could poured onto a bunch of particals (eg: sand) which after polymerization could be fractured and the fractured surface examined by an SEM so that partical size and intrastitial space measurments could be made using image analysis?
3- To complicate matters, it would be nice to seperate (again via SEM) the organics from the minerals vissually. We need to see if the organic is chemically attached to the mineral or if it forms aggregates between them.
best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**Suppose you were an idiot... And suppose you were a member of Congress .. But I repeat myself.-Mark Twain**
One nice carrier I used in the past consisted of a two-part plastic container, where a plastic foil was suspended over the opening of both halves. You would place the sample on the foil in the lower half of the container and close it with the upper half of the container. The sample is then fixed between the two foils which hang suspended in the center of the box.
I don't know who manufacturs this box, but I can find out if you want to. Perhaps somebody else here has a source for this?
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
} ---------- } From: } "chanch-at-ufl.edu"-at-sparc5.microscopy.com[SMTP:"chanch-at-ufl.edu"-at-sparc5.mi } croscopy.com] } Sent: Wednesday, February 17, 1999 4:50 PM } To: microscopy-at-sparc5.microscopy.com } Subject: TEM sample transport } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Dear Professional friends, } } I have to use a distant microscope. } What's the best way to transport (from FL to CO) fragile TEM samples? } } best regards, } } chih-hung chang } Dept. of Chemical Engineering } University of Florida } Gainesville,FL 32611 } } }
} Phil, } } I'm having a major problem posting to the Microscopy ListServ would you } post the following for me please. The MSA filter seems to think that I } shouldnt be allowed to post. } } I'm hoping that someone will be able to help me out with this as I am } a biological science person (give me a cell any time). } } 1- Is there someone in the RTP area that has an SEM with an } environmental chamber? } } 2- Any suggestions on a plastic polymer that could poured onto a bunch } of particals (eg: sand) which after polymerization could be fractured } and the fractured surface examined by an SEM so that partical size and } intrastitial space measurments could be made using image analysis? } } 3- To complicate matters, it would be nice to seperate (again via SEM) } the organics from the minerals vissually. We need to see if the organic } is chemically attached to the mineral or if it forms aggregates between } them. } Bob Schoonhoven } rschoonh-at-imap.sph.unc.edu
I am a graduate student at the university of Toronto in the department = of Laboratory medicine . My thesis is a comparative study of the anterior and posterior = cingulate gyrus in Alzheimer's disease and Lewy body dementia . I will = have to count plaques (accumulation of amyloid) and tangles and Lewy = bodies(intracytoplasmic inclusion bodies ) in both the anterior and = posterior portion of the cingulate gyrus which is part of the cortex . = I use immunohistochemisty techniques to stain plaques tangles and Lewy = bodies . I need information (softwares, books,papers, Web sites) to learn about = stereology avoid bias in my count, and take into consideration all the = tissue factors that could affect my count. Thank you for your help. My E.Mail address is mervat.kahil-at-utoronto.ca
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT size=3D2} I am a graduate student at the university of Toronto = in the=20 department of Laboratory medicine . {/FONT} {/DIV} {DIV} {FONT size=3D2} My thesis is a comparative study of the = anterior and=20 posterior cingulate gyrus in Alzheimer's disease and Lewy body dementia = . I will=20 have to count plaques (accumulation of amyloid) and tangles and Lewy=20 bodies(intracytoplasmic inclusion bodies ) in both the anterior and = posterior=20 portion of the cingulate gyrus which is part of the cortex . I use =
immunohistochemisty techniques to stain plaques tangles and Lewy bodies=20 . {/FONT} {/DIV} {DIV} {FONT size=3D2} I need information (softwares, books,papers, Web = sites) to=20 learn about stereology avoid bias in my count, and take into = consideration all=20 the tissue factors that could affect my count. {/FONT} {/DIV} {DIV} {FONT size=3D2} Thank you for your help. {/FONT} {/DIV} {DIV} {FONT size=3D2} My E.Mail address is {A=20 href=3D"mailto:mervat.kahil-at-utoronto.ca"} mervat.kahil-at-utoronto.ca {/A} {/FO= NT} {/DIV} {/BODY} {/HTML}
I wish to examine the morphology of Desmids (single celled freshwater algae) using SEM. Can anyone give me suitable references for their preparation. Please send them to michaeld-at-amsg.austmus.gov.au
I wish to examine the morphology of Desmids (single celled freshwater algae) using SEM. Can anyone give me suitable references for their preparation. Please send them to michaeld-at-amsg.austmus.gov.au
There have been several recent postings on the use of membrane boxes for the transport and storage of prepared grids. We too are advocates of these boxes, however, with one caveat: You want to be careful that there is nothing on your sample that could react with the thin polyethylene stretched film (membrane).
When even the slightest reaction occurs, the grids are effectively destroyed in terms of their usefulness. Or that is what was described to us by an unhappy customer.
There has been at least one instance where the specimen wanted to stick better to the membrane than to the grid, with the end result that the specimen was lost (stuck) on the membrane and the user was left with a pristine grid sans sample.
We recommend a "test" before storing important samples this way. Some might call it "over kill" but to us it seems like good advice: Take a typical sample and heat age it at 40 deg. C. What ever might happen at room temperature, this test should speed it up about 100 times faster. If a test time about equal to the expected storage or transport time is done, and if on examination, there are no signs of any reaction with the membrane, then you can be reasonably certain that the grid and sample are going to be inert and not changed because of the close proximity to the membrane.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Dear all, We are imaging pumice sections on glass slides, using BSE detector at 25kv, condenser lens 4, using a Robinson BSE detector on a Hitachi FE4000 The idea is to obtain highly contrasted images to transform into black and white binary images. The problem is we are getting horizontal banding which looks like a charging problem, although the samples are well coated and earthed. Could it be due to anything else? The problem has recently developed and was also apparent on a more conducting sample of some TEM grids. Attached are some images. Any comments would be helpful.
Thank { {horiz_bd.tif} } { {18299_29.tif} } s David Wild
Hi! Does anyone know where I can get grid freezing device for cryo-EM? I am looking for commercially available apparatus that can plunge EM grid into liquid ethane and prepare biological samples in vitrified ice. Thank you very much for your help!
Best regards, Sam Li ******************************************************************************* 213 Wintrobe Building Biochemistry Department University of Utah Salt Lake City, UT 84132 phone: 801-585-5490, fax: 801-581-7959 sam.li-at-m.cc.utah.edu *******************************************************************************
Pardon this belated contribution to the carbon discussion. After returning from the excellent New Zealand Microscopy Conference I found an email were the correspondent wants to exchange graphite for carbon rods. I think that the discussion on those rods was a little incomplete and would like to add this:
Pure (amorphous) carbon rods without graphite (crystalline carbon) would not work; they are essentially non-conductors. If carbon was to volatilise at a much lower temperature than graphite, graphite would be deposited as particulate matter. Clearly that is not so - both carbon and graphite give smooth films under the right conditions. Graphite rods simply have a higher graphite content than carbon rods and purity of analysis is quite another topic.
Higher graphite content rods are more easily shaped or sharpened and since they conduct better than "carbon" rods, they require more current to heat to sublimation point. Of course the diameter of the rods at contact point is another important variable.
I have found that most carbon rod evaporation difficulties are due to wrong voltage selection. Graphite, let alone carbon rods are near impossible to deal with at 10 volts. I used 30 volts and about 60 amps to evaporate from graphite rods. A slightly higher voltage may be preferable for carbon rods. Incidentally 10 volts gives better control for metal evaporation. Clearly maximum amps and an easily selectable range of voltages are a major consideration when purchasing an evaporator.
I am of the persuasion that a strong preference for carbon versus graphite rods is largely a matter of belief. It's too easy to mistake technique and equipment parameters as proving that one or the other rod is "better". I rather state my belief that: Graphite rods are less brittle and carbon cord, because of a larger source area, gives greater uniformity in thickness.
Disclaimer: ProSciTech like all EM suppliers supplies carbon and graphite rods and we have no preference which are purchased. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
Are the images you posted digitally acquired, or from photographs? The bands in your images look just like banding we have had in the past on our Hitachi S-800 FESEM. It drove us nuts for many long months! It appeared in both SE and BSE images, and in all our photographed images. Service personnel could not find the problem. Finally I realized that it really *only* appeared on the recording CRT, and not on the visual CRTs. I hauled out a really old SEM manual from the late 1960s that dealt with all kinds of weirdness. It showed banding on a CRT that was a result of dirty or deficient high voltage to the CRT. I gingerly cleaned the HV contacts to the CRT and the problem disappeared. It reappears every few years, and I imagine it's our humid and somewhat volcanic air that corrodes these contacts. Cleaning them up works. Hitachi remains dubious about this, by the way.
If the bands appear on digital images, I guess you still need to track down where your signal is coming from and see if you have weak contacts.
Good luck! I know how frustrated we were.
Aloha, Tina
On Fri, 19 Feb 1999, D.Wild wrote:
} Dear all, } We are imaging pumice sections on glass slides, using BSE detector at 25kv, } condenser lens 4, using a Robinson BSE detector on a Hitachi FE4000 The } idea is to obtain highly contrasted images to transform into black and } white binary images. The problem is we are getting horizontal banding which } looks like a charging problem, although the samples are well coated and } earthed. Could it be due to anything else? The problem has recently } developed and was also apparent on a more conducting sample of some TEM } grids. Attached are some images. } Any comments would be helpful.
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
It sure does look like charging (dark) and discharging (bright), but ther= e are other problems which show up in a similar fashion to this when you us= e backscatter.
All this assumes that you do have a good earth on your specimen stage? =
Although BSE see charge far less than SE remember you tend to use much larger currents for BSE observation and you may be trying to cope with th= e large current when you have a very poor earth?
If it is charging one would expect the problem to change in character if you changed the beam current, or more dramatic, change the kV. Lower the=
beam current or the kV and the problem should decrease. Still not convinced then take the sample out and look at a clean stub. No problems then your problem is specimen oriented. If you prove it is charge and t= he stage earth is good -
1. try a faster scan speed for your photography 2. try to work with smaller specimens. 3. try to work at a lower kV (10 or 15) and increase the emission curre= nt (I think a 4000 will go up to 20uA via one of the F keys)
If the above do not markedly change the image form then the problem is probably due to the processing within the backscattered detector system. =
Some BSE detectors allow you to change the response of the amplifier, the= y have viewing and photographic processing options. If you do not have the= se options try taking a photograph at a totally different scan speed?
Good luck
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
======================================================== FOCUS ON MICROSCOPY 1999
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
We wish to announce an important change in the abstract submission and modification deadline. Because of an improved schedule for the printing services of the abstract booklet, we are able to extend the deadline. However, we encourage you to submit and finalise your abstract as quickly as possible since we are starting to arrange the talks into the different sessions.
The new abstract submission and modification deadline is now:
!!! Monday, 8 March 1999 --- 15.00 hours Heidelberg Time !!!
The "abstract submission part" of the database will definitely be closed at 15.00 hours, so that it is not possible to submit further abstracts or make any further modifications to the abstracts after this date.
The "registration only part" of the database will remain open until 9 April 1999!!!
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meeting point for developers and users working in these rapidly evolving fields and play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic developments in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
We have a Hitachi model S-570 that may be available for sale (80 % sure we will sell, 20 % chance it will be relocated at another Rodel location.). This dual detector tungsten system is in good shape and accepts a 6 in diameter sample. The system will be sold as is. If interested, please e-mail me for more details. Michael Ingram Rodel, Inc Polishing Lab 451 Bellevue Rd Newark, DE 19713 (302) 366-0500, ext.: 2545
I have a question that is not directly related to microscopy but maybe someone out there can help me. One of our customers has asked our company to source and supply a resin used for injection into the vascular system to display the blood vessels (after removing organic material) for teaching, research and museum work.
I have a vague memory of a Japanese resin fulfilling these requirements but can find no immediate reference. Any help gratefully received,
Regards
Terry Cooper TAAB Laboratories Equipment Ltd 3 Minerva House, Calleva Park Aldermaston, Berks, RG7 8NA, UK
Rodel, Inc. had a Hitachi model S-570 that may be available for sale (80 % sure we will sell, 20 % chance it will be relocated at another Rodel location.). This dual detector tungsten system is in good shape and accepts a 6 in diameter sample. The system will be sold as is. If interested, please e-mail me for more details.
Michael Ingram Rodel, Inc Polishing Lab 451 Bellevue Rd Newark, DE 19713 (302) 366-0500, ext.: 2545
"Microwave Techniques for Clinical Histopathology"
There will be a microwave session at Scanning 99 entitled "Microwave Techniques for Clinical Histopathology". Scanning 99 is being held at the Hyatt Regency O'Hare in Rosemont, IL from April 11-14, 1999. The "Microwave Techniques for Clinical Histopathology" session is set for Tuesday, April 13, 1999, in the morning and will consist of invited and contributed presentations. The details of the conference and deadlines/forms for abstract submissions can be found at http://www.scanning.org {http://www.scanning.org} or details can be requested from Mary Sullivan (e-mail: scanning-at-fams.org {mailto:scanning-at-fams.org} , tel: 201-818-1010).
A preliminary list of invited speakers and the titles for the presentation is shown below.
Co-Chairs:
Beverly Giammara - Virtek Vision Inc.
Title "Simple Microwave Methods for Electron Microscopy"
Steven E. Slap - Energy Beam Sciences, Inc.
Title "Introduction to Laboratory Histoprocessing Techniques"
Speakers:
Richard W. Dapson, Ph.D - Anatech Ltd
Title "Microwave Fixation for Histopathology"
D. Denise Hardy - ARUP
Title "Routine Microwave Processing in the Clinical Laboratory"
Linda M. Chicoine - Cognetix/Viatech Imaging, Inc.
Title "Microwave Techniques for Immuno Labeling in Electron Microscopy"
Nathan T. Brinn - Leica Microsystems
Title "Microwave Staining Techniques in Histopathology"
Dear Bob, } } I'm hoping that someone will be able to help me out with this as I am } primarily a biological science person. } } 3- To complicate matters, it would be nice to seperate (again via SEM) } the organics from the minerals vissually. We need to see if the organic } is chemically attached to the mineral or if it forms aggregates between } them. } There are many possibilities for how the organic material could be attached along the continuum from physically adjacent to covalently bonded; however, that said, suspending the specimen in water and applying varying amounts of agitation should distinguish among some of the possi- bilities. If there is only the loosest association between the minerals and the organics, I would expect the former to sink while the latter should float (perhaps you may need to add CsCl to the water if the organ- ics have a specific gravity slightly } 1). If the organics remain attached when the specimen is sonicated, that would signify a tight association. This assumes that neither the minerals nor the organics are water-soluble. Good luck. Yours, Bill Tivol
Our Denton Vaccum Evaporator EV502 won't sputter. We can glow discharge and carbon coat without any trouble but no gold sputtering.
All leads and wiring on the cathode have been changed. We have a new gold target. The cathode was cleaned. =20 The magnets were hopefully put back in the right order. Bioengineering says the electrical system is OK. The vacuum appears OK.
Any suggestions or ideas to get the sputter to sputter=20 would be appreciated.
George Lawton Chief Electron Microscopist Microscopy and Imaging Service Center UT Southwestern Medical Center at Dallas Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
Terry Cooper wrote: ============================================ I have a question that is not directly related to microscopy but maybe someone out there can help me. One of our customers has asked our company to source and supply a resin used for injection into the vascular system to display the blood vessels (after removing organic material) for teaching, research and museum work.
I have a vague memory of a Japanese resin fulfilling these requirements but can find no immediate reference. Any help gratefully received, ================================================== I am 99% certain that you thinking about the Mercox Resin System which is used pretty universally for corrosion casting studies. It is available from SPI Supplies and some of the other main suppliers of consumables for light and electron microscopy.
You can find out more about the Mercox system from our website URL http: //www.2spi.com/catalog/chem/embed1.html
If you have kept your previous issues of MICROSCOPY TODAY ( Issue 98-7, Sept . 1998), there was an excellent publication by Fred E. Hossler. If you did not save your old issues, there is a link to the electronic version of that publication from the above mentioned URL.
Disclaimer: SPI Supplies distributes the Mercox resin system and therefore we are interested in promoting its use.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Our Denton Vacuum Evaporator DV502 won't sputter. We can glow discharge and carbon coat without any=20 trouble but no gold sputtering. All the leads and wiring on the cathode have been changed. We have a new gold target. The cathode was cleaned. Magnets were put back in the right order. Bioengineering says the electrical system is OK. The vacuum appears OK. =20
Any suggestions or ideas to get the sputter to sputter would be appreciated.
George Lawton Chief Electron Microscopist Microscopy and Imaging Service Center UT Southwestern Medical Center at Dallas Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
I can tell you from painful experience to avoid acetone. If you do an acetone rinse on your sample prior to putting it in the box, make sure that all of the acetone has evaporated. If not the sample will stick or you will get a nice hole in the membrane. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Garber, Charles A. To: MICROSCOPY BB -----------------------------------------------------------------------.
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
There have been several recent postings on the use of membrane boxes for the transport and storage of prepared grids. We too are advocates of these boxes, however, with one caveat: You want to be careful that there is nothing on your sample that could react with the thin polyethylene stretched film (membrane).
When even the slightest reaction occurs, the grids are effectively destroyed in terms of their usefulness. Or that is what was described to us by an unhappy customer.
There has been at least one instance where the specimen wanted to stick better to the membrane than to the grid, with the end result that the specimen was lost (stuck) on the membrane and the user was left with a pristine grid sans sample.
We recommend a "test" before storing important samples this way. Some might call it "over kill" but to us it seems like good advice: Take a typical sample and heat age it at 40 deg. C. What ever might happen at room temperature, this test should speed it up about 100 times faster. If a test time about equal to the expected storage or transport time is done, and if on examination, there are no signs of any reaction with the membrane, then you can be reasonably certain that the grid and sample are going to be inert and not changed because of the close proximity to the membrane.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
The latest edition of the Testing Buyers Guide, compiled by the editors of ASM Internationals Advanced Materials & Processes magazine, is now available online at http://www.asm-intl.org/testing
If you are looking for a SEM supplier, or for a firm to conduct some failure analysis for you, then the place to look is the Equipment/Supplies/Services Listing. Click on the category of interest, and you'll be taken to a listing of companies providing that service. To find the contact details for a particular company, simply click on its name. If you already know the name of a company, then you can proceed directly to the Company Directory. Here you will find an alphabetical listing of the more than 400 companies listed in this directory.
I hope you find this a useful resource.
Leslie H. Chom LChom-at-po.asm-intl.org Manager, Online Services ASM International ph. 440.338.5151 Ext. 5510 fx. 440.338.4634
Need property, composition, and processing data? Alloy Digest Webfaxx --- http://www.asm-intl.org
At 16:23 17/02/99 -0400, you wrote: } Content-Type: text/plain; } charset=3D"utf-8" } X-MIME-Autoconverted: from 8bit to quoted-printable by electra.ciens.ucv.ve id EAA17539 } } Estimados colegas: Para facilitar la difusi=C3=B3n de la informaci=C3=B3n= acerca del } V Congreso Interamericano de Microscop=C3=ADa Electr=C3=B3nica , se la = estamos } enviando bajo tres distintas modalidades (direcci=C3=B3n de la p=C3=A1gina= web, texto } en este correo y un documento anexo en formato Word). Agradecer=C3=ADamos= su } valiosa colaboraci=C3=B3n en ayudarnos a distribuirla. Con un saludo= cordial. } Miren Gonz=C3=A1lez-Elorriaga } } Dear colleagues: In order to facilitate the spreading of the information } relative to the Vth Interamerican Electron Microscopy Congress we are } sending it under three different formats ( web site address, text in this } e-mail, and as an attachment document in Word). We will appreciate very= much } your collaboration in delivering it to other colleagues that could be } interested in attending the Congress. Sincerely yours. Miren } Gonz=C3=A1lez-Elorriaga. } } http://electra.ciens.ucv.ve/~svme/ } } } } SOCIEDAD VENEZOLANA DE MICROSCOPIA ELECTRONICA } UNIVERSIDAD CENTRAL DE VENEZUELA } FACULTAD DE CIENCIAS } CENTRO DE MICROSCOPIA ELECTR=C3=93NICA } } } V INTERAMERICAN } ELECTRON MICROSCOPY } CONGRESS } } VIII VENEZUELAN ELECTRON } MICROSCOPY CONGRESS } } ISLA DE MARGARITA } VENEZUELA } OCTOBER 24 - 28, 1999 } } } } GENERAL INFORMATION } } The forthcoming Interamerican Electron Microscopy Congress is the fifth of= a } series which had its origin in M=C3=A9rida, Venezuela, in 1986. } The scientific program will begin on sunday October the 24th, running } through Thursday October the 28th. The program will feature Simposia, } Invited Talks, Oral Presentations and Posters Sessions related to Electron } Microscopy and its applications to Medicine, Biology and Materials. } We look forward to the participation of colleagues from Latin-America, } North America, Europe and Asia. Given the quality of the participants we } want this event to be very fruitful for both undergraduate and postgraduate } students from the specialities mentioned above. } As usual, we also count with the participation of important Commercial } Companies. } From now we give all participants a warm welcome, wishing them a } nice stay in our beautiful Venezuela. } } } INSTRUCTIONS FOR AUTHORS } } All papers must have a two-page extent. The first page must include only } text; the first 19 lines of the second page must contain the rest of the } text and could also be used to write the References. The remaining area= must } be dedicated to the Figures: graphs, tables and photographs. } } } } We request very specially to follow the format mentioned above. } } Papers must be submitted according to the following guidelines: } Title (Times New Roman, capitals, boldface, 12 points, centered). Leave one } blank line and in the following line write the names of authors,= underlining } the name of the author in charge of the presentation. On the following line } write the institutional affiliation of the authors. Leave two blank lines } and then start writing the text. The text must contain a brief= introduction, } objectives, experimental methods, results, discussion and conclusions.= These } points will be taken into account for the selection of the submitted= works. } Languages: Spanish, English or Portuguese. } Paper size: Letter or A4. } Type font: Times New Roman, 10 points, 1.5 spacing. } Margins: Top (2 cm), Bottom (3.5 cm), Left (3 cm), Right (2 cm). } Please, mail one original and two copies of each paper, together with the } registration form and fee. } Reception of papers will be acknowledged promptly. } } Please, do not send your payment separated from the paper and the } registration form. } } } } } } } } } } REGISTRATION FEES } } SVME / IFSM Members } Until On site } 04/30/99 } } Professionals US$ 200 US$ 230 } Students* US$ 100 US$ 130 } } Non Members of SVME / IFSM } } Until On site } 04/30/99 } } Professionals US$ 250 US$ 280 } Students* US$ 125 US$ 155 } } * Students must include a certificate from their institution stating their } status. } } The registration fee entitles the participant the submission of one paper. } Additional papers will pay a fee of US$ 30. } The registration fee includes a copy of the } proceedings and the participation in all } cultural and social events. } } The registration form, paper and payment must be sent to the following } address: } } V INTERAMERICAN ELECTRON } MICROSCOPY CONGRESS. } Universidad Central de Venezuela } Facultad de Ciencias } Centro de Microscop=C3=ADa Electr=C3=B3nica } Avenida Los Ilustres, Los Chaguaramos, } Caracas, VENEZUELA } } Additional information : } } Tel: 58-2-6052174 } Tel-Fax: 58-2-6930694 } e-mail: } svme-at-electra.ciens.ucv.ve } curbina-at-electra.ciens.ucv.ve } alcastel-at-telcel.net.ve } } PAPERS DATA FORM } This completed form must accompany all papers } } Title: } } } } } } } } } Author (s): } } } } } } Three Keywords: } } } } Poster : } } Space recerved for the poster is } 150 cm.-high by 90cm.-wide. } } } Please return this form with your paper (original and two copies) and the } payment. } } } If the paper is not accepted the author will be refunded a 70% of the } cancelled fee. } } } } } } REGISTRATION FORM } Please fill in Capital letters } } Name: } } Institution: } } Complete Address: } } Phone: FAX: } } e-mail: } } Member of SVME/IFSM: } YES =EF=81=AF NO =EF=81=AF } } Student* =EF=81=AF Professional =EF=81=AF } } } REGISTRATION FEES } } SVME/IFSM Members } Until On Site } 04/30/99 } } Professional US$ 200 US$ 230 } Student* US$ 100 US$ 130 } } SVME/IFSM Non-Members } } Until On Site } 04/30/99 } } Professional US$ 250 US$ 280 } Student* US$ 125 US$ 155 } } * Students must include a copy of their official student I.D. } } } PAPERS DATA FORM } This completed form must accompany all papers } } } } } } } } ORGANIZING COMMITTEE } } President: Dr. Alan Castellano } General Secretary: Dr. Caribay Urbina } Scientific Secretary: Dr. Gema Gonz=C3=A1lez } M.Sc. Sonia Camero. } Dr. Carlos Rojas. } Dr. H=C3=A9ctor Finol. } Dr. Miren Gonz=C3=A1lez } M.Sc. Fredy S=C3=A1nchez } Secretary of Special } Events: Dr. Blanca M=C3=BCller } Treasurer: Dr. Humberto Rojas } Technical Support: Dr. Pedro Rodr=C3=ADguez } } } NATIONAL ADVISORY COMMITTEE } } } } INTEVEP M.Sc.Margarita Navas } IVIC Dr. Ra=C3=BAl Padr=C3=B3n } UCV Dr. Fracehuli Dagger } UCV Dr. Mariana Staia } ULA Dr. Mauro Brice=C3=B1o } LUZ Dr. Orlando Castej=C3=B3n } UNEFM Dr. Auristela de Mirt } IDEA Dr. Gloria Villegas } UDO Dr. Oscar Gonz=C3=A1lez } UDO Dr. Benjam=C3=ADn Hidalgo } USB Dr. Augusto Ruiz } USR Dr. Antonio Breta=C3=B1a } IUT M.Sc. Freddy Arenas } } } } } } } } } } } } COURSES } During October the 24th and the 28th a variety of courses related to } techniques of sample preparation, observation and } microanalysis will be offered. } } Further details about these courses will be in our web site: } http://electra.ciens.ucv.ve/~svme } } COMMERCIAL EXHIBITION } } The Congress will include an exhibition presented by companies that } trade products related to Electron Microscopy. The exhibition area will be } located at the entrance to the halls of conference and close to the posters } area. } } HOTELS } Margarita Hilton International Hotel (Congress Headquarters), use } e- mail or Fax to reserve. 58-95-620810 } http://www.hilton.com } } Details about accomodation will be } informed in our website: } http://electra.ciens.ucv.ve/~svme } } } CALL FOR PAPERS } Deadline: May the 15th 1999 } Notification of acceptance: } from July the 31st 1999 } } } } } } } } Attachment Converted: "C:\FREDDY\Attach\V Interamerican Electron Microscopy Congress.url" } } Attachment Converted: "C:\FREDDY\Attach\VCngIME.doc" }
The latest edition of the Testing Buyers Guide, compiled by the editors of ASM Internationals Advanced Materials & Processes magazine, is now available online at http://www.asm-intl.org/testing
If you are looking for a SEM supplier, or for a firm to conduct some failure analysis for you, then the place to look is the Equipment/Supplies/Services Listing. Click on the category of interest, and you'll be taken to a listing of companies providing that service. To find the contact details for a particular company, simply click on its name. If you already know the name of a company, then you can proceed directly to the Company Directory. Here you will find an alphabetical listing of the more than 400 companies listed in this directory.
I hope you find this a useful resource.
Leslie H. Chom LChom-at-po.asm-intl.org Manager, Online Services ASM International ph. 440.338.5151 Ext. 5510 fx. 440.338.4634
Need property, composition, and processing data? Alloy Digest Webfaxx --- http://www.asm-intl.org
Leslie H. Chom LChom-at-po.asm-intl.org Manager, Online Services ASM International ph. 440.338.5151 Ext. 5510 fx. 440.338.4634
Need property, composition, and processing data? Alloy Digest Webfaxx --- http://www.asm-intl.org
does anybody know the correct setup of the two RS232 remote interfaces on a LEO982. the manual is pretty sketchy on how to actually send and receive data (or i'm just doing something terribly wrong).
i've configured a comm port on a PC to meet the stated specifications of either or both of the RS232 ports on the scope, and i can send data (as seen on a cable monitor) but i don't get any response from the scope. i've used both null modem and standard cable configurations. it seems the ports just are not listening.....
also, does the 982 data screen do anything in response to the remote interface being activated?
any ideas?
thx! brian
____________________________________________________________________ Brian McIntyre mailto:mcintyre-at-optics.rochester.edu Univ. of Rochester-EM Lab 716-275-3058 voice The Institute of Optics 716-244-4936 fax Rochester, NY 14627 http://www.optics.rochester.edu
For some reason I did not see your original message and attachment images that you posted. Therefore I have only seen the responses. I'll throw in my two cents anyway. You mentioned that you have seen the problem using a TEM grid. I would use something like this that is very conductive with strong contrast if you can readily reproduce the problem. If you can reproduce the banding or flashing only with the samples on glass. It is probably your sample. Unless in increasing the contrast(gain in the Robinson BSE), you inadvertently create some "crosstalk" from the voltage for the preamp to a video line.
First of all to eliminate some things a few questions need to be answered (Assuming you have reproduced the banding with a conductive sample). Make sure the grounding plug on the stage near the X Y controls is plugged in correctly. Think back to when the banding started. WAs anybody inside the instrument? Was anything added? Has the BSE detector been adjusted, moved? A wise old service engineer once told me that 80% of all problems are created by someone fiddling with something. After years of service myself he was absolutely right. Do you see the same banding in normal SE mode as well as BSE? If so the Robinson BSE detector is probably not the culprit. Do you see the problem on the view CRT? If so start at a fast raster then step down to slower and slower raster speeds. Take note if the amounts of banding or flashing increases. If so it is probably charging. Photo speeds will show the most charging. If you only see the problem on your photos in SE and BSE then I would suspect the High Voltage to the photo CRT as someone has already suggested. Make sure you know what your doing before messing with this. The anode on the CRT has 8 to 10KV on it. Even after turning the power off it'll hold a considerable shock. If both view and photo CRT's show banding, the problem is visible in SE and BSE, and you have eliminated charging; the high voltage circuit in the high voltage tank has been known to have problems. This is rare however.
} From what little I know I strongly suspect the Robinson has developed a problem. Usually a ground has developoed a poor connection. It has been awhile but I recall a little gold or silver tab that makes contact with the main shaft on the main housing. I don't remember whether you can get to it without taking the housing apart. Make sure this and any other grounds you find have good connections. If you go into the housing make sure the voltage or power cables are seperate from the video cables. I've rambled and can think of nothing else right now. Good luck.
Joel McClintock EM Specialist U of Kentucky (606)257-1242
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Our Denton Vacuum Evaporator DV502 won't sputter. We can glow discharge and carbon coat without any trouble but no gold sputtering. All the leads and wiring on the cathode have been changed. We have a new gold target. The cathode was cleaned. Magnets were put back in the right order. Bioengineering says the electrical system is OK. The vacuum appears OK.
Any suggestions or ideas to get the sputter to sputter would be appreciated.
George Lawton Chief Electron Microscopist Microscopy and Imaging Service Center UT Southwestern Medical Center at Dallas Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
(InterMail v03.02.07 118 124) with SMTP id {19990219212714.GINO7957-at-oldserver} for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 19 Feb 1999 21:27:14 +0000 Message-ID: {36CDD78A.C0C-at-worldnet.att.net}
Terry Cooper wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Dear fellow Microscopists, } } I have a question that is not directly related to microscopy but maybe } someone out there can help me. One of our customers has asked our company } to source and supply a resin used for injection into the vascular system to } display the blood vessels (after removing organic material) for teaching, } research and museum work. } } I have a vague memory of a Japanese resin fulfilling these requirements but } can find no immediate reference. Any help gratefully received, } } Regards } } Terry Cooper } TAAB Laboratories Equipment Ltd } 3 Minerva House, Calleva Park } Aldermaston, Berks, RG7 8NA, UK
Dear Terry Cooper,
The product I believe you are referring to is Mercox. It is available from Ladd in Red (#21245), Blue (#21246), and Clear (#21247).
John Arnott Chairman
-- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
I would like to reiterate what Joel McClintock mentioned in his post, and which I neglected to point out. The high voltage supply to your CRT is 10 kV! Be careful! The cleaning I have done has so far not involved actually polishing contacts, but merely blowing compressed clean, dry air over the contacts, making sure they are seated, cleaning the surrounding areas, and carefully closing everything up again.
There are a number of circuits in most instruments that are still energized or retain a charge when the instrument is OFF or even DISCONNECTED.
I hate having a perfectly good, sunny, warm, Hawaiian winter day with great surf ruined by getting thrown up against the wall and electrocuted.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Dear Bob, I cannot help you with the location of the variable-pressure SEM, but the sample prep I can. 2. The usual practice is to mount the sample in a quick-setting epoxy resin, with most of the sample in the bottom. I use silicon rubber cups one inch in diameter, about 1/2 inch deep. The set resin is then ground on a graded series of sandpaper disks, then polished on diamond suspension to cross-section the particles. The sample is then gold or carbon coated. 3. organic and minerals are easily separated by backscattered imaging (BSE). The organics, being composed of light elements, will show darker than the mineral, which is composed of heavier elements. A photo of SEM and BSE of the same area is often helpful. Particle size measurements may be better done on a sample of grains sprinkled on a sticky tab and iamged by BSE. Hope this helps. You wrote: } I'm hoping that someone will be able to help me out with this as I am } primarily a biological science person. } } 1- Is there someone in the RTP area that has an SEM with an } environmental chamber? } } 2- Any suggestions on a plastic polymer that could poured onto a bunch } of particals (eg: sand) which after polymerization could be fractured } and the fractured surface examined by an SEM so that partical size and } intrastitial space measurments could be made using image analysis? } } 3- To complicate matters, it would be nice to seperate (again via SEM) } the organics from the minerals vissually. We need to see if the organic } is chemically attached to the mineral or if it forms aggregates between } them. } } best regards, } Bob } Robert Schoonhoven } Laboratory of Molecular Carcinogenesis and Mutagenesis } Dept. of Environmental Sciences and Engineering } University of North Carolina } CB#7400 } Chapel Hill, NC 27599 } Phone } office 919-966-6343 } Lab 919-966-6140 } Fax 919-966-6123 } } **Suppose you were an idiot... And suppose you were a member of Congress } .. } But I repeat myself.-Mark Twain** } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
} Date: Fri, 19 Feb 1999 15:48:43 -0800 } To: "rschoonh-at-sph.unc.edu"-at-Sparc5.Microscopy.Com, Microscopy } From: Mary Mager {mager-at-interchange.ubc.ca} } Subject: Re: material sciences question } } Dear Bob, } I cannot help you with the location of the variable-pressure SEM, but the sample prep I can. } 2. The usual practice is to mount the sample in a quick-setting epoxy resin, with most of the sample in the bottom. I use silicon rubber cups one inch in diameter, about 1/2 inch deep. The set resin is then ground on a graded series of sandpaper disks, then polished on diamond suspension to cross-section the particles. The sample is then gold or carbon coated. } 3. organic and minerals are easily separated by backscattered imaging (BSE). The organics, being composed of light elements, will show darker than the mineral, which is composed of heavier elements. A photo of SEM and BSE of the same area is often helpful. } Particle size measurements may be better done on a sample of grains sprinkled on a sticky tab and iamged by BSE. } Hope this helps. } You wrote: } } I'm hoping that someone will be able to help me out with this as I am } } primarily a biological science person. } } } } 1- Is there someone in the RTP area that has an SEM with an } } environmental chamber? } } } } 2- Any suggestions on a plastic polymer that could poured onto a bunch } } of particals (eg: sand) which after polymerization could be fractured } } and the fractured surface examined by an SEM so that partical size and } } intrastitial space measurments could be made using image analysis? } } } } 3- To complicate matters, it would be nice to seperate (again via SEM) } } the organics from the minerals vissually. We need to see if the organic } } is chemically attached to the mineral or if it forms aggregates between } } them. } } } } best regards, } } Bob } } Robert Schoonhoven } } Laboratory of Molecular Carcinogenesis and Mutagenesis } } Dept. of Environmental Sciences and Engineering } } University of North Carolina } } CB#7400 } } Chapel Hill, NC 27599 } } Phone } } office 919-966-6343 } } Lab 919-966-6140 } } Fax 919-966-6123 } } } } **Suppose you were an idiot... And suppose you were a member of Congress } } .. } } But I repeat myself.-Mark Twain** } } } Regards, } Mary } Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
With a little more time I have re read your mail and the difficulty of coating your specimen suddenly hits me in the eye!
At the kV you are using (25) with the complicated surface of your specime= n you would have to go to some pretty complex sputtering techniques to remo= ve the possibilities of charging.
If it was me I would conduct the empty stub test I mentioned in an earlie= r =
mail and if this was OK I would then look at the size and shape of the specimen and the coating method.
Sputter coaters are not as good as people like to think they get beaten b= y complex surfaces when a conducting path to earth is very very difficult t= o achieve.
Try this
1. Coat the specimen for a minute or so at 45 degrees 2. Repeat with it tilted 45 degrees in exactly the opposite directi= on 3. Get the best vacuum that you can, by turning off the sputter gas,=
and try to sputter again.
See if this works as it is my standard method for very difficult specimen= s. I find this is the only way to bury the metal deep into the specimen por= es but you should also lower the kV!
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
I would also add my two cents to Mary Mager response to adding epoxy to sand/organic mixture. To eliminate voids, we would evacuate the powder sample and then add the liquid via a tube from the outside. The liquid had been previously degassed by either heating or evacuation. The apparatus is simple to construct. Our high resolution density measurements indicated almost complete filling of all voids limited only by the molecular size of the liquid.
Another technique is to simply put the powder and epoxy together in a vaccuum oven and then very slowly evacuate. Make sure there is plenty of room for the bubbles in the sample holder.
J. Roy Nelson, PhD Material Testing Laboratory Pennington, NJ (609) 730-0575 jrnelson-at-nj1.aae.com
-----Original Message----- } From: Tina Carvalho [SMTP:tina-at-pbrc.hawaii.edu] Sent: Saturday, February 20, 1999 12:22 AM To: Joel McClintock Cc: microscopy-at-Sparc5.Microscopy.Com
Hi, all-
A researcher has approached me with confocal images of part of an insect's nervous system that has cells that particularly light up with FITC. They don't know what these cells are, and they want to be able to locate these cells in the TEM. They are looking for something in particular, but I am not allowed to be more specific.
Their idea is to have HRP-conjugated anti-FITC, hoping for a dark reaction product that I will be able to find in the TEM.
I haven't done any HRP staining in (yikes!) decades, and so do not have a protocol handy. Could someone suggest one or, perhaps better, an alternative plan? I am very open to suggestions! Since all they know about these cells is that they fluoresce with FITC, I haven't come up with any other labelling ideas. When I asked if they could microinject the cells, I got blank and then fearful stares.
Thanks in advance!
Mahalo, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Before taking my new position as an educator who uses the SEM to improve high school curriculum, the SEM affected my life in incidental ways. For example, the SEM allowed scientists to study and improve their understanding of biology, medicine. Material manufacturing and the aerospace industry used scanning electron microscopes to improve manufacturing processes and to maintain the global superiority of the United States Air Force. I was aware of its use and power, but did not have much contact with the technology.
I was privileged, as a teacher, to take classes on field trips to local industry to use their SEM. Once, a class engaged in a project to determine the damage done by painting the ceiling tiles in our high school's cafeteria. We did sound spectrum studies and painted acoustical tile with unaltered tile under the scanning electron microscope. As you can imagine, the paint had filled in most of the sound-absorbing gaps, reducing the effectiveness of the tiles.
More recently, I have been using a "Personal Scanning Electron Microscope" to improve the curriculum in Dayton area schools. We have used the SEM to examine the adaptations of stream invertebrates to various functions in their ecosystem, to compare the structure of red blood cells (eukaryotes) to blue- green algae (prokaryotes), to examine the pollen comb, brush, and pocket on the hind legs of honeybees, and to measure microscopically.
We have also described the crystalline forms of rocks and minerals and studied microfossils found in diatomaceous earth. I am currently at the Museum of Discovery in Dayton, Ohio, where we are photographing tiny ants and snails that have never been seen under an SEM before. Some of these images will be published in scientific journals to advance our understanding of the diversity of our environment while others will be archived along with the actual specimens for the benefit of future generations.
Even if I had never had the opportunity to use an SEM, it would have been improving my life indirectly through the contributions of scientists who do use one. Having the opportunity to use one several days a month is enriching my life as I am able to work and contribute on the border between science and art, using both sides of my brain!
Good luck with your report, Amy!
Sincerely,
Carlton Bowers Alliance for Education TECH TREK Mobile Research Laboratory Curriculum & Technology Specialist (937) 222-2934
George Lawton wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Our Denton Vaccum Evaporator EV502 won't sputter. } We can glow discharge and carbon coat without any } trouble but no gold sputtering. } } All leads and wiring on the cathode have been changed. } We have a new gold target. The cathode was cleaned. } The magnets were hopefully put back in the right order. } Bioengineering says the electrical system is OK. The } vacuum appears OK. } } Any suggestions or ideas to get the sputter to sputter } would be appreciated. } } George Lawton } Chief Electron Microscopist } Microscopy and Imaging Service Center } UT Southwestern Medical Center at Dallas } Phone: 214-648-7291 } eMail: George.Lawton-at-email.swmed.edu
George, I had this happen on my Polaron 5100. Try going to full voltage at a very poor vacuum and see if you can make it start. It may have something to do with contamination.
Ken Converse owner Quality Images third party SEM service Delta, PA
Bob, At the risk of running on- I'd just like to add my two cents worth to the good advise that you have received from Mary Mager.
Particle Size We have had the best results determining particles size by the "keep it simple" rule for SEM preparations. We usually use the sticky tab method. One of the commercially available, double sided carbon adhesives such as SPI Carbon Tape or Ted Pella Carbon Conductive Tabs work well. SEM secondary electron imaging (SEI) usually does great, but you will probably have great backscattered electron contrast as well (BEI). This simple prep works best (for us) for the particle size aspect of the problem, because you needn't worry yourself with the problem of recognizing small particle fragments due to the fracturing or grinding/polishing.
Composition/ Organic and inorganic As Mary recommends, you will want to embed and cross-section (by grinding /polishing) to expose a topography free surface. SEM-BEI will show the spatial correlation between the mineral and carbonaceous components. Also don't overlook PLM. If you have access to a good optical microscope, you may learn much about the structure here.
Intrastitial and interstitial spaces Again the cross-section prep is your best shot. I use Buehler's EPO-COLOR fast cure epoxy for these application. It is a nice bright red color that will likely be different in color fron any other components in your sample. Use a vacuum infiltration before curing. This allows examination by optical techniques with differentiation between the epoxy embedding material and the other organics in the specimen - works great for identifying porosity. Then on to the SEM for a closer look and identification of compositions.
Hopefully SEM has the resolution to examine the intrastitial space that your interested in. Otherwise its on to the TEM. isn't it fun?! Regards, Brad Huggins
Mary Mager Wrote: Dear Bob, I cannot help you with the location of the variable-pressure SEM, but the sample prep I can. 2. The usual practice is to mount the sample in a quick-setting epoxy resin, with most of the sample in the bottom. I use silicon rubber cups one inch in diameter, about 1/2 inch deep. The set resin is then ground on a graded series of sandpaper disks, then polished on diamond suspension to cross-section the particles. The sample is then gold or carbon coated. 3. organic and minerals are easily separated by backscattered imaging (BSE). The organics, being composed of light elements, will show darker than the mineral, which is composed of heavier elements. A photo of SEM and BSE of the same area is often helpful. Particle size measurements may be better done on a sample of grains sprinkled on a sticky tab and iamged by BSE.
You wrote: } I'm hoping that someone will be able to help me out with this as I am } primarily a biological science person. } } 1- Is there someone in the RTP area that has an SEM with an } environmental chamber? } } 2- Any suggestions on a plastic polymer that could poured onto a bunch } of particals (eg: sand) which after polymerization could be fractured } and the fractured surface examined by an SEM so that partical size and } intrastitial space measurments could be made using image analysis? } } 3- To complicate matters, it would be nice to seperate (again via SEM) } the organics from the minerals vissually. We need to see if the organic } is chemically attached to the mineral or if it forms aggregates between } them. } } best regards, } Bob } Robert Schoonhoven
Hi all, I have recently finished my third year project in electron microscopic observations of the entry and exit of influenza virus. In it I used Thermanox coverslips to culture the cells on, one of the problems that I had was delamination of the coverslip from the resin (Spurrs) and wondered if this had any thing to do with the different densities of the Thermanox and resin. If anyone knows their densities I would be most grateful.
I'm back on-line, albeit remotely. I have noticed several recent postings this week (of course) in which users have appended images to their mailings.
Please remember this is not accepted practice on this listserver. Too many people have low bandwidth or expensive or slow connections. Fortunately for me, my software is set to ignore large downloads while I'm on the road and operating via a modem. Many users do not have this luxury. This is, of course, detailed in the rules of listserver usage which you all have received when you subscribed.
If you wish to share images with list users place them on a WWW site and just provide a URL in the body of your message..
I am putting the final touches on a presentation on the state of microscopy education for a presentation this coming Thursday (New York Microscopical Society) and would love to include some good "up-to-date" stats about what's going on in the field. While I have a lot of material on courses at Delta San Joaquin, Woods Hole, Lehigh, and through MSA, I could use some info "from the field". If you are teaching a course at your company/university, could you please send (email or snail mail) a quick synopsis? Also of interest: if you are including a microscopy unit as part of another course.
Many thanks! Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
Michael: I worked with Desmids more than a quarter of a century ago. Cannot lay may hands on those reference now but the important things learned about their preparation for SEM would not have changed!
These cells are very sensitive to osmotic shock. We used the buffer quite dilute - 0.01M at the most and dehydration has to be slower than normal, have more gradual steps and begin with about 20% ethanol, when most other specimens can tolerate dehydration from 70%. Since these are large cells you can observe osmotic effects under a light microscope.
I think that we used our improvised freeze drying method back then, but solvent drying or critical point drying too, if you can retain the cells should give good results. Call me if you have further questions. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Friday, February 19, 1999 1:24 PM, MichaelD [SMTP:michaeld-at-amsg.austmus.gov.au] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } I wish to examine the morphology of Desmids (single } celled freshwater } algae) using SEM. Can anyone give me suitable } references for their } preparation. } Please send them to michaeld-at-amsg.austmus.gov.au } } Thanking you in anticipation } } } Mike Dingley.
Does anyone have a reference or recipe for post-fixing with acetate-veronal (AV)-buffered OsO4, and/or en bloc staining with AV-buffered UA?? I believe Palade authored the technique, probably others have modified it.
Thanks, Ann Lehman Trinity College Hartford, CT v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-exchange.cc.trincoll.edu
We have a new semester course in EM. It covers theory and application of both SEEM and TEM. It is an undergraduate course, at the 5000 level (senior, graduate school acceptable). Students prepare and view their own samples on the Hitachi S4000 SEEM and on the Zeus 902 TEM. Digital images are generated as TIFF files and the students produce portfolios with PhotoShop. Final prints are made with an Alps color printer. We are establishing a web site, and student work will be posted on this site.
William McManus, MS Lab Supervisor Electron Microscope Facility Department of Biology Utah State University Logan UT 854322-5305
435-797-1920
-----Original Message----- } From: Barbara Foster [mailto:mme-at-map.com] Sent: Sunday, February 21, 1999 5:42 PM To: Confocal Microscopy List; microscopy-at-Sparc5.Microscopy.Com
Dear list-listeners,
I am putting the final touches on a presentation on the state of microscopy education for a presentation this coming Thursday (New York Microscopical Society) and would love to include some good "up-to-date" stats about what's going on in the field. While I have a lot of material on courses at Delta San Joaquin, Woods Hole, Lehigh, and through MSA, I could use some info "from the field". If you are teaching a course at your company/university, could you please send (email or snail mail) a quick synopsis? Also of interest: if you are including a microscopy unit as part of another course.
Many thanks! Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
Tina Carvalho wrote: } } A researcher has approached me with confocal images of part of an insect's } nervous system that has cells that particularly light up with FITC. } They don't know what these cells are, and they want to be able to locate } these cells in the TEM. They are looking for something in particular, but } I am not allowed to be more specific. } } Their idea is to have HRP-conjugated anti-FITC, hoping for a dark reaction } product that I will be able to find in the TEM. } } I haven't done any HRP staining in (yikes!) decades, and so do not have a } protocol handy. Could someone suggest one or, perhaps better, an } alternative plan? I am very open to suggestions! Since all they know } about these cells is that they fluoresce with FITC, I haven't come up with } any other labelling ideas. When I asked if they could microinject the } cells, I got blank and then fearful stares. } Dear Tina, Jim Turner has done a lot of corellative confocal LM and EM. Try some of his publications for a protocol. 10 kV would ruin one of your wonderful Hawaiian days; we just use the shock to recover from Albany winter :-). Yours, Bill Tivol
This is a multi-part message in MIME format. --------------3B1287CCAA802B61D5BACA63 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
Dear Dr. Carvalho
An alternative tehcnique to anti-FITC antibodies in electron microcopy may be the photoconversion of DAB by the irradiation of the FITC-marked cells using a laser or a fluorescent microscope. I never used this technique but it appears to be relatively easy. This technique is described by Pagano and Martin (1997) in Cell Biology: A Laboratory Handbook, 2nd ed., Julio E Celis, Ed: 505-510, Academic Press, San Diego.
I hope it works Good Luck!
Francisco Javier Hernandez-Blazquez IARC-WHO Unit of Multistage Carcinogenesis 150 Cours Albert Thomas Lyon - France
fhernandez-at-iarc.fr
Tina Carvalho wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, all- } } A researcher has approached me with confocal images of part of an insect's } nervous system that has cells that particularly light up with FITC. } They don't know what these cells are, and they want to be able to locate } these cells in the TEM. They are looking for something in particular, but } I am not allowed to be more specific. } } Their idea is to have HRP-conjugated anti-FITC, hoping for a dark reaction } product that I will be able to find in the TEM. } } I haven't done any HRP staining in (yikes!) decades, and so do not have a } protocol handy. Could someone suggest one or, perhaps better, an } alternative plan? I am very open to suggestions! Since all they know } about these cells is that they fluoresce with FITC, I haven't come up with } any other labelling ideas. When I asked if they could microinject the } cells, I got blank and then fearful stares. } } Thanks in advance! } } Mahalo, } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
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Hi Barbara, In brief: We teach TEM, SEM, and Electron Diffraction courses open to gradurates and undergraduates. The TEM course is a 6 hr. credit course (2hrs/lecture, 4hrs/lab) that involves producing a report which is 50% of their grade. The main challenge is producing good sections with a tempermental 21 year old ultramicrotome. The SEM course is for 2 hrs. credit again is with 50% of the grade on their report. Here we are at mercy of our 25 year old ETEC behaving well. The EM Diffraction course is taught only every few years. But every year we offer a one afternoon EM Diffraction Lab as part of a Solid State Chemistry Techniques course. In addition we conduct highschool tours, and have involved high school students in special summerschool projects here on the University campus. On occasion we have trained researchers from industry in EM instrumentation techniques. The microscopy courses serve important functions in that some of my former students have gone on to direct EM or microscopy imaging facilities in industry and Universities, and others in their capacity as teachers or researchers will be able to share and expand the microscopy knowledge with others. Our concerns: 1.) Because of the small class size (6-10 students on average) there is pressure to drop the courses. Yet it is usually the students who end up doing the research for faculty. 2.) Departments faced with budget concerns want to cut service contracts. This is especially determental to TEM performance and the production of quality results. 3.) Lack of interest in replacing, repairing and/or upgrading equipment, and support for training courses for staff in new microscopy techniques. Suggestion: For MSA or local Microscopy Societies to produce a brochure, or better, a series of brochures on different microscopy and imaging techniques and their relevance and applications to research and commerce aimed at high school level science students. If these were available to hand out for high school tours or accessable to high school teachers, it would do much in furthering the interest and understanding of microscopy.
Barbara Foster wrote: } Dear list-listeners, } I am putting the final touches on a presentation on the state of microscopy } education for a presentation this coming Thursday (New York Microscopical } Society) and would love to include some good "up-to-date" stats about } what's going on in the field. While I have a lot of material on courses at } Delta San Joaquin, Woods Hole, Lehigh, and through MSA, I could use some } info "from the field". If you are teaching a course at your } company/university, could you please send (email or snail mail) a quick } synopsis? Also of interest: if you are including a microscopy unit as part } of another course. } } Many thanks! } Barbara Foster } Consortium President } Microscopy/Microscopy Education ...Educating microscopists for greater } productivity. } } 125 Paridon Street Suite 102 Springfield, MA 01118 } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } Visit our web site {http://www.MME-Microscopy.com/education} } ****************************************************** } MME is America's first national consortium providing } customized on-site workshops in all areas of } microscopy, sample preparation, and image analysis.
Henry Eichelberger, EM Facility Manager Department of Biological Sciences Binghamton, University
I have been giving a course since 1993 to biomedical graduate students here at the University of Michigan entitled "Morphology for Molecular Biologists." You can get an idea of the course and see a schedule in this URL:
http://www.umich.edu/~mmb533/
Unfortunately, you won't be able to view the lectures, which have to be restricted to the University of Michigan, because they contain some copyrighted material.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www.umich.edu/~akc/
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear list-listeners, } } I am putting the final touches on a presentation on the state of microscopy } education for a presentation this coming Thursday (New York Microscopical } Society) and would love to include some good "up-to-date" stats about } what's going on in the field. While I have a lot of material on courses at } Delta San Joaquin, Woods Hole, Lehigh, and through MSA, I could use some } info "from the field". If you are teaching a course at your } company/university, could you please send (email or snail mail) a quick } synopsis? Also of interest: if you are including a microscopy unit as part } of another course. } } Many thanks! } Barbara Foster } Consortium President } Microscopy/Microscopy Education ...Educating microscopists for greater } productivity. } } 125 Paridon Street Suite 102 Springfield, MA 01118 } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } Visit our web site {http://www.MME-Microscopy.com/education} } ****************************************************** } MME is America's first national consortium providing } customized on-site workshops in all areas of } microscopy, sample preparation, and image analysis. } }
Since I am the person who, I think, started this discussion, I'd like to explain why I am developing a preference (purely personal) for "carbon" over "graphite". It may help others.
My coater, an old Edwards 306, uses a 3mm sharpened rod which is caused, by a spring, to bear on to a flat-ended 0.25" carbon rod (which I call the anvil). The 3mm (consumable) rod is held in a hole, about 8mm long, drilled into the end of another piece of 0.25" rod (pity about the ban on images).
When I was using my previous supply of "National" spectroscopic carbon rods, the anvil and the holder used to last for about a year. When I switched (somewhat unwittingly) to "graphite" rods, the higher temperature (or maybe it was the higher current) caused both the holder and the anvil to get so hot that the former lasted for three coatings and the latter for five.
I could, of course, modify my holder and anvil set-up, but it seems to me that it would be easier just to try to obtain some rods similar to those which I had before.
I'm sure there must be other coaters can cope with the graphite rods better than mine can.
It's not just "belief", Jim, but rather, as you also say, different operating and instrument parameters.
Disclaimer: As a hard-pressed University employee, I just want things which work satisfactorily to continue to do so with the minimum hassle factor.
cheers
Ritchie
} From: Jim J Darley {jim-at-proscitech.com.au} } Reply-to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-sparc5.microscopy.com} } Subject: Carbon versus graphite rods } Date: Fri, 19 Feb 1999 14:52:38 +1100 } Organization: proscitech
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Pardon this belated contribution to the carbon discussion. } After returning from the excellent New Zealand Microscopy } Conference I found an email were the correspondent wants to } exchange graphite for carbon rods. I think that the } discussion on those rods was a little incomplete and would } like to add this: } } Pure (amorphous) carbon rods without graphite (crystalline } carbon) would not work; they are essentially } non-conductors. If carbon was to volatilise at a much lower } temperature than graphite, graphite would be deposited as } particulate matter. Clearly that is not so - both carbon } and graphite give smooth films under the right conditions. } Graphite rods simply have a higher graphite content than } carbon rods and purity of analysis is quite another topic. } } Higher graphite content rods are more easily shaped or } sharpened and since they conduct better than "carbon" rods, } they require more current to heat to sublimation point. Of } course the diameter of the rods at contact point is another } important variable. } } I have found that most carbon rod evaporation difficulties } are due to wrong voltage selection. Graphite, let alone } carbon rods are near impossible to deal with at 10 volts. I } used 30 volts and about 60 amps to evaporate from graphite } rods. A slightly higher voltage may be preferable for } carbon rods. Incidentally 10 volts gives better control for } metal evaporation. Clearly maximum amps and an easily } selectable range of voltages are a major consideration when } purchasing an evaporator. } } I am of the persuasion that a strong preference for carbon } versus graphite rods is largely a matter of belief. It's } too easy to mistake technique and equipment parameters as } proving that one or the other rod is "better". I rather } state my belief that: Graphite rods are less brittle and } carbon cord, because of a larger source area, gives greater } uniformity in thickness. } } Disclaimer: ProSciTech like all EM suppliers supplies } carbon and graphite rods and we have no preference which } are purchased. } Cheers } Jim Darley } ProSciTech Microscopy } PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 7 4774 0370 Fax: +61 7 4789 2313 } Great microscopy catalogue, 500 Links, MSDS, User Notes } ********************** www.proscitech.com.au ***** }
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
With regard to your quest for information on training organisations.
Protrain, based in Oxford England, and its sister company SSEM, based in California, have been running "in house" electron microscopy training courses around the world for the past eighteen years.
Our staff are made up of professional electron microscopists who have bee= n in the business for at least 33 years. We all started out as service engineers in the days when you literally built the instruments on the customers site. Our paths then moved toward those of application specialists, combining instrument demonstration and customer training, these roles with a number of the most well known manufacturers. Since we=
moved into running our own training organisations we have run courses in university and industrial organisations covering all the natural English speaking countries in the world and some where English was not their firs= t language.
Our training courses are run on customer demand in the clients laboratory= , but we are often employed by universities to run specific courses for the= m within their own E.M. Units. Many of our courses will be similar to othe= rs that you may hear about in your survey, but I believe that we are the onl= y organisation in the world than run courses for technical staff on the maintenance of electron microscopes. Our latest move has been to make th= e courses available with sound on CD ROM, taking information to those that are unable to afford the hands on training.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
If it is not a violation of state laws or institutional policy to provide the information, I am interested in determining the extent to which laboratories in U. S. academic institutions regulate their chemical waste and provide radiation safety protection and laser safety protection policies and procedures for their employees. Are appropriate OSHA and other regulatory agency rules mandated by your institutions? Have some been granted extensions in order to set up appropriate procedures?
I would specifically like to know if anyone has direct knowledge of laboratory safety inspections by the EPA (Environmental Protection Agency) or OSHA (Occupational Safety and Health Administration) or any other state or federal safety regulating agency. If so, what has been the outcome of the inspections? Have fines have been levied against institutions for improper chemical storage or waste disposal procedure violations? Have any fines been imposed for radiation or laser safety violations? Obviously, this listserver is primarily microscopy lab directed, however, it would be useful to know if laboratories other than electron microscope labs have been inspected as well.
The purpose of my request is to gather data for a talk I will be giving. I would like to know how rigorously regulatory agency rules are inforced for academic institutions compared to industry enforcement.
Our institution has a strict waste disposal policy. No chemical is flushed down the sink. All waste is put in appropriate containers and is removed by university Risk Management people as needed. State inspectors have inspected laser labs. This resulted in a request for better beam stop protection. I am not aware of any fines. To date, there have been no OSHA or EPA inspections. Our Radiation Protection Office oversees radiation and laser safety on campus and provides excellent protection resources.
All answers will be reported generically. If any information is collected and if a summary is requested, no institution will be listed in my talk or in a summary to this listserver. Respondents may wish to reply to me directly.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
We offer a two year Associate's Degree in Electron Microscopy at Madison Area Technical College in Madison, Wisconsin. The four semester program deals with SEM, FESEM, TEM, AFM, sample prep for materials and biologic, and basic EM maintenance. Check out our web site for more info: http:// Electron-Microscopy.madison.tec.wi.us
Bill Carmichael EM Program MATC 3550 Anderson St. Madison, WI 53704 billc-at-jvlnet.com
Barbara Foster wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear list-listeners, } } I am putting the final touches on a presentation on the state of microscopy } education for a presentation this coming Thursday (New York Microscopical } Society) and would love to include some good "up-to-date" stats about } what's going on in the field. While I have a lot of material on courses at } Delta San Joaquin, Woods Hole, Lehigh, and through MSA, I could use some } info "from the field". If you are teaching a course at your } company/university, could you please send (email or snail mail) a quick } synopsis? Also of interest: if you are including a microscopy unit as part } of another course. } } Many thanks! } Barbara Foster } Consortium President } Microscopy/Microscopy Education ...Educating microscopists for greater } productivity. } } 125 Paridon Street Suite 102 Springfield, MA 01118 } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } Visit our web site {http://www.MME-Microscopy.com/education} } ****************************************************** } MME is America's first national consortium providing } customized on-site workshops in all areas of } microscopy, sample preparation, and image analysis.
We offer a two year Associate's Degree in Electron Microscopy at Madison Area Technical College in Madison, Wisconsin. The four semester program deals with SEM, FESEM, TEM, AFM, sample prep for materials and biologic, and basic EM maintenance. Check out our web site for more info:
http://Electron-Microscopy.madison.tec.wi.us
Bill Carmichael EM Program MATC 3550 Anderson St. Madison, WI 53704 billc-at-jvlnet.com
Barbara Foster wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear list-listeners, } } I am putting the final touches on a presentation on the state of microscopy } education for a presentation this coming Thursday (New York Microscopical } Society) and would love to include some good "up-to-date" stats about } what's going on in the field. While I have a lot of material on courses at } Delta San Joaquin, Woods Hole, Lehigh, and through MSA, I could use some } info "from the field". If you are teaching a course at your } company/university, could you please send (email or snail mail) a quick } synopsis? Also of interest: if you are including a microscopy unit as part } of another course. } } Many thanks! } Barbara Foster } Consortium President } Microscopy/Microscopy Education ...Educating microscopists for greater } productivity. } } 125 Paridon Street Suite 102 Springfield, MA 01118 } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } Visit our web site {http://www.MME-Microscopy.com/education} } ****************************************************** } MME is America's first national consortium providing } customized on-site workshops in all areas of } microscopy, sample preparation, and image analysis.
A very big hello to everyone that I haven't spoken to since leaving the Microscopy List some 2 years ago. My new vocation leads me to re-subscribing to ask you all a very big favour.
***Have you done any EM work (imaging or analytical) for archaeologists? If so, would you be so kind as to drop me a line to let me know the aim and nature of the work? Reasons follow;
I left the Microscopy Labs of the Australian Museum to follow my wife's work in Brussels, Belgium, where I'm doing a Ph.D. in Archaeology (in 3D modelling). The undergraduate courses in our faculty do not cover many scientific techniques, so my fellow students have badgered me into giving a seminar on EM's and their applications in archaeology, and a demo on the uni's SEM. I have plenty of examples from my work in zoology, but obviously I need examples from archaeology. I have some references to work with but I figure that the BEST references are the people on this List.
Please understand that my motivations are honest. My only use of your information will be this once-off seminar for my fellow students. They are undergrads who are not in a position to poach any research for their own advantage. All contributors will be acknowledged. And I will be personally very grateful for your help.
We recently purchased Pixera Professional digital camera (for optical microscopy). Nice little thing, but:
We want to have the camera on PC (Pentium II, 333MHz, 128MB RAM) with Win NT 4.0 operating system. We successfully installed the NT driver Pixdrv.drv and Pixera Visual Communication Suite (from a CD). After starting Studio Viewfinder (a software for picture acquisition), there was just fine b/w raster (noise) instead of picture. Using other software and Twain interface led us to the same disappointment. We repeated experiments with both service packs for NT 4.0, namely 3 and 4.
We also tried to install the same camera on another computer (Pentium MMX 200MHz, 64MB RAM ), running on Win 98. New hardware was installed without a problem, but when opening the Pixera VC Suite and running Viewfinder program a message "Can not initialize the camera" appeared.
I would be very grateful for any suggestion.
Best regards,
Goran Drazic
------------------------------
Dr. Goran Drazic J. Stefan Institute Jamova 39 SI-1001 Ljubljana Slovenia
{!doctype html public "-//w3c//dtd html 4.0 transitional//en"} {html} I have a surplus 3.5 mm {b} never been used {/b} Diatome diamond knife old style boat for sale. {/html}
In a message dated 99-02-23 08:57:29 EST, Goran.Drazic-at-ijs.si writes:
{ { We also tried to install the same camera on another computer (Pentium MMX 200MHz, 64MB RAM ), running on Win 98. New hardware was installed without a problem, but when opening the Pixera VC Suite and running Viewfinder program a message "Can not initialize the camera" appeared.
} }
Dr. Drazic,
I had the same problem just yesterday on a new Pixera installation. Pixera told me this is a problem with some high-speed computers.
We switched out the interface card to a "Cirrus Logic PCIC compatible PCI to PCMCIA Bridge," and I had to load a new set of drivers from a floppy disk that was included with the PCI card from Pixera. I also found out that I did not have "32 bit support" enabled on the computer. With the new card and 32 bit support enabled, the camera now works beautifully.
If your local representing agency can not help, I suggest you contact Pixera directly at:
www.pixera.com
} From there you can contact Technical Support. The new set of drivers are two fairly small files. If you can get the correct PCI board, I am sure Pixera could send them to you via e-mail.
Good luck, I hope this helps!
Best regards,
Bob ****************************** Robert (Bob) Chiovetti, Ph.D. President Microimaging Technologies, Inc. Tucson, Arizona USA Tel./Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems Clinical & Research Microscopy Cytology/Histology/Pathology/EM *******************************
Any suggestion on proper etchant procedure to use in order to do NiTi and NiTiCu Memory Shape Alloys SEM images? We have tried several polishing procedure (cutting speed, different clothes and a chemical etch with 50:50 mixture of HF acid and HNO3 acid) but no grain borders can be seen (but some pinning if you use acid for too long!) Thank You in advance, Edoardo
{!DOCTYPE HTML PUBLIC "-//W3C//DTD HTML 3.2//EN"} {HTML} {HEAD} {META HTTP-EQUIV=3D"Content-Type" CONTENT=3D"text/html; = charset=3Dus-ascii"} {META NAME=3D"Generator" CONTENT=3D"MS Exchange Server version = 5.5.1960.3"} {TITLE} SEM preparation of SMA {/TITLE} {/HEAD} {BODY}
{P} {FONT SIZE=3D2 FACE=3D"Courier New"} Any suggestion on proper etchant = procedure to use in order to do NiTi and NiTiCu Memory Shape Alloys SEM = images? {/FONT} {BR} {FONT SIZE=3D2 FACE=3D"Courier New"} We have tried several polishing = procedure (cutting speed, different clothes and a chemical etch with = 50:50 mixture of HF acid and {/FONT} {/P}
{P} {FONT SIZE=3D2 FACE=3D"Courier New"} HNO3 acid) but no grain borders = can be seen (but some pinning if you use acid for too long!) {/FONT} {BR} {FONT SIZE=3D2 FACE=3D"Courier New"} Thank You in advance, = Edoardo {/FONT} {/P}
I am looking for software/hardware that will help me count the number amount , as well as note the area of the particle, off a negative (print) image that I have produced using a TEM. I could do it by hand, but the time neccessary is not available. I know that there has to be something out there that will help me, and any help would be greatly appreiciated.
Joseph Montoya Physics Department New Mexico State University
I am unsure about your imaging goals, but can you use BSE channeling contrast instead of an agressive etch? I haven't consulted the phase diagrams, but it is often possible to see grains (orientation related contrast) if the Z differences don't produce a poor "signal to noise ratio". Noise in this case being any undesired image signal.
Any suggestion on proper etchant procedure to use in order to do NiTi and NiTiCu Memory Shape Alloys SEM images? We have tried several polishing procedure (cutting speed, different clothes and a chemical etch with 50:50 mixture of HF acid and
HNO3 acid) but no grain borders can be seen (but some pinning if you use acid for too long!) Thank You in advance, Edoardo
Does anyone out there know of a source of single crystal LiF substrates or other similar single crystal fluorides, chlorides, etc? If not, would you perhaps know of an entrepreneurial crystal grower who might be tempted? Thanks in advance.
Cheers John
John P. McCaffrey Institute for Microstructural Sciences National Research Council of Canada M-50 Montreal Rd. Ottawa, Ontario K1A 0R6 Canada
As a result of a recent trade, I have a nice clean LKB-Huxley Ultra Microtome - seems to be in good working condition and has the original accessory box with a variety of chucks and adapters. And a new - unused - EdgeCraft 2.6mm diamond knife. All of my work is LM, so I need to get this stuff sold or traded (hopefully in the greater Baltimore-DC area for the Huxley, since I don't want to crate it). Money would be nice (my wife thinks my self-supported research costs way too much), but also open to interesting trades for useful LM equipment - I am Leitz 170mm based, but open to other possibilities. Also would consider selling/trading the Huxley and the diamond knife as seperate items. Mainly, I need to get the Huxley out of my wife's laundry room and don't have room for it in my lab - OK, I know ultra microtomes in the laundry room is not a common problem, but we are a little strange. And, this is clearly not a commercial enterprise.
Geoff Avern wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } A very big hello to everyone that I haven't spoken to since leaving the } Microscopy List some 2 years ago. My new vocation leads me to } re-subscribing to ask you all a very big favour. } } ***Have you done any EM work (imaging or analytical) for archaeologists? } If so, would you be so kind as to drop me a line to let me know the aim } and nature of the work? Reasons follow; } } I left the Microscopy Labs of the Australian Museum to follow my wife's } work in Brussels, Belgium, where I'm doing a Ph.D. in Archaeology (in 3D } modelling). The undergraduate courses in our faculty do not cover many } scientific techniques, so my fellow students have badgered me into giving } a seminar on EM's and their applications in archaeology, and a demo on the } uni's SEM. I have plenty of examples from my work in zoology, but } obviously I need examples from archaeology. I have some references to } work with but I figure that the BEST references are the people on this } List. } } Please understand that my motivations are honest. My only use of your } information will be this once-off seminar for my fellow students. They } are undergrads who are not in a position to poach any research for their } own advantage. All contributors will be acknowledged. And I will be } personally very grateful for your help. } } Hoping to hear from you, } } Geoff Avern } Brussels, Belgium
Geoff One person you might want to try and chase down is Dr. Alan Walker. He used to operate an SEM at Johns Hopkins University in Baltimore, MD, but I don't believe he is there at the present time. I beleive it was the Lucy find in Africa that he was associated with and he might be able to give a lot of input into the archeological uses of the SEM. Hope this helps.
Ken Converse owner Quality Images third party SEM service
What you want to do is relatively straightforward. You do have to be careful that the area fraction that you measure for the particle will not be the volume fraction because you have to take account for the thickness of the sample in transmission. For relatively inexpensive solutions, you can use NIH image on a MAC or Photoshop with John Russ' Image Processing Toolkit Plug-ins for both Mac and PC. IPTK is about $250 and comes with a very good tutorial and a primer on stereology measurements. It will also work with other programs. The latest version has digital darkroom on it. I also think that his plug-ins work with NIH image, but I have never tried them with it. IPTK also goes very well with his book, The Image Processing handbook. Visit their web site: http://members.aol.com/ImagProcTK/index.htm
John Russ monitors the Listserver and he is very helpful to people with quantitative microscopy problems as he was with me just over the past couple of days. Thanks again, John!
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Rudy To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
I am looking for software/hardware that will help me count the number amount , as well as note the area of the particle, off a negative (print) image that I have produced using a TEM. I could do it by hand, but the time neccessary is not available. I know that there has to be something out there that will help me, and any help would be greatly appreiciated.
Joseph Montoya Physics Department New Mexico State University
Hello,=20 Can somebody help me about the preparation of the Sato's Lead Citrate? = What's the chemicals and its proportion? Thank's in advance. M.Sc. Rinaldo Pires dos Santos Lab. of Plant Anatomy - Dept. of Botany - UFRGS Porto Alegre - RS - Brazil
Hello, Can somebody help me about the preparation of the Sato's Lead Citrate? What's the chemicals and its proportion? Thank's in advance. M.Sc. Rinaldo Pires dos Santos Lab. of Plant Anatomy - Dept. of Botany - UFRGS Porto Alegre - RS - Brazil
We did some work on materials investigation of a 300-year old Virgin Mary stone (Indo-Christian style). We picked up small pieces of samples from different positions of the stone, e.g., coating, and stone itself. The results are interesting. We analyed mineral constituents and chemical compositions and finally we were able to determine the most possible locality where the stone was produced.
The paper was published at MRS Proceedings: M.S. Barger and W.L. Gong, La Virgincita: Technical study of an Indo-Christian Statue, Mat. Res. Soc. Proc. Symp. 462, 381 (1997).
I would appreciate comments or opinions regarding this software. I saw the presentation and I was quite impressed. Howver before I can make any recommendation I would like to hear from users of this software especially those using the Confocal Imaging module and Cell Tracking.
My two cents worth is to throw that mental switch into your head in the analytical mode. As you know your EM, and my Pol Scope, do analysis as well as make WOW! pictures. To not assume, I will refer you to the Journal of Archaeological Sciences for many articles using the EM.
You have a "tool" that can analysis micro quantities, which archaeologist are not used to working with. You need to educate them, not what IS done, but the types of things can be done. Archaeological problems of the "what the .... is this" tend to come on a more or less dig by dig basis. Build an easy bridge for them to walk accross to your analystical tool. Problem solving is best done with the interest of the various are openly and freely discussed.
While not archaeology, don't forget conservation and restoration as you consider your course.
Good luck and Shalom from Jerusalem, Azriel
} } Brussels, Belgium } } Geoff } One person you might want to try and chase down is Dr. Alan } Walker. He used to operate an SEM at Johns Hopkins University in } Baltimore, MD, but I don't believe he is there at the present time. I } beleive it was the Lucy find in Africa that he was associated with and } he might be able to give a lot of input into the archeological uses of } the SEM. } Hope this helps. } } Ken Converse } owner } Quality Images } third party SEM service } } +-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+ Azriel Gorski azrielg-at-cc.huji.ac.il ICQ User ID No. - 1750739
CHOICE - The enchanted blade, with an edge that shapes lifetimes. Richard Bach RUNNING FROM SAFETY
A friend is someone who knows the song in your heart, and can sing it back to you when you have forgotten the words. +-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+
We run in-house courses on EDS analysis, WDS analysis, combined EDS / WDS analysis and Quantitative X-ray Mapping. These courses are all 'hands on' using ETEC and JEOL Microprobes. We offer these courses to all University and Industry Personnel.
Moran Scientific Pty Ltd P.O. Box 651 Goulburn NSW 2580 Australia Tel 02 4844 4234 (International, 61 2 48444234) Fax 02 4844 4291 (International, 61 2 48444291) Email {kmoran-at-goulburn.net.au} Web Page http://www.goulburn.net.au/~kmoran "Tell me and I'll forget, show me and I may remember, involve me and I'll understand" - an Old Chinese Proverb
Dear all, has anybody experience or can give me tips for the preparation/examination of liposomes ( {100nm diameter) for TEM or SEM . We want to analyse the homogeneity of suspensions and to estimate the single volume of the liposome particles? Unfortunately we don't have any unit for cryomicroscopical observations! Any suggestions are welcome... best regards, Bernward Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit„tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
Hello: We are considering doing TEM to look at the microstructure around surface pits in molded stainless steel parts. We are trying to understand the source of these pits. So far SEM and OM have provided us with no relevant clues. The pits are barely visible by eye and they are only very few of them on a given part. We are thinking of doing tripod polishing , but I would appreciate any comments/suggestions on how one might best approach this.
Rinaldo, Here's the recipe for Sato's Triple Lead stain that we use:
0.5 gm Pb nitrate 0.5 gm Pb acetate 0.5 gm Pb citrate 1.0 gm Na citrate 41 ml boiled,distilled water
-Mix in covered tri-pour beaker for at least 2 hours (all day is better) -Add 9.0 ml 1N NaOH to clear(changes milky-white color to clear)
If you have any questions or comments, don't hesitate to contact me. Winston
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W Wiggins, Supervisor 2/24/99 8:43:32 AM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I recall reading a few years ago a description of the Hilbert (spelling?) raster pattern generator consisting of interlocking H's in a fractal regression of scale. As I recall, the advantages touted for this raster were equivalent (on overage) probe deflection speeds in the x & y directions, a continuous path for the raster probe, and a low sensitivity to scan coil saturation at rapid scan rates.
I have an "opportunity" to replace my scan generator with a digital device, and would like to give the Hilbert a try. The problem is that I misplaced my copy of the article. Does anyone recall the source of this reference?
Thanks,
valdemar-at-fast.net or rwafu-at-bsco.com Valdemar Furdanowicz Homer Research Labs G-165 Bethlehem Steel Co. Bethlehem, PA 18016
Dear Rinaldo: Here is the protocol for Sato's Lead Citrate:
Anhydrous lead citrate Pb(C6H5O7)2 0.20 g Lead nitrate Pb(NO3) 0.15 g Lead acetate Pb(CH3COO)2.3H2O 0.15 g Sodium citrate Na3(C6H5O7).2H2O 1.00 g Distilled water 41.0 ml
The above reagents placed in a 50 ml voletric falsk were mixed well to make a yellowish milky solution. Then the solution was added 9.0 ml of 1 N NaOH and mixed well until the solution became transparent with light yellowish color. KEEP IT IN REFRGERATOR, IT COULD BE GOOD OVER ONE YEAR.
Here is some useful references: Sato, T: Electron Microsc, 17: 158, 1968 Watson, ML: J Biophys. Biochem Cytol, 4:727, 1958 *Hanaichi T, Sato T, Hoshino M and Mizuno N: Proc XIth Int Cong on Electron Microscopy, Kyoto, 1986 -- This is where "my" recipe came from. Good Luck.
Zhaojie Zhang ********************************* Zhaojie Zhang Department of Botany and Microbiology University of Oklahoma Norman, OK 73019 Phone: 405-325-6234 http://students.ou.edu/Z/Zhaojie.Zhang-1/ *********************************
Rinaldo Pires dos Santos wrote:
} Hello, } Can somebody help me about the preparation of the Sato's Lead Citrate? } What's the chemicals and its proportion? } Thank's in advance. } M.Sc. Rinaldo Pires dos Santos } Lab. of Plant Anatomy - Dept. of Botany - UFRGS } Porto Alegre - RS - Brazil
there are several software packages available that do exactly what you want: threshold the particle, analyze them and give you particle results such as area, max. radius, etc. In my opinion you should look for software that:
a) calculates all the parameters you need b) allows you to define your own parameters (for future extension) c) provides an upgrade path if you need more processing at a later time d) reads and writes standard files for data exchange e) provides features for particle separation (watershed or other) f) allows you to set up Regions of Interest (connected and non-connected) g) provides density measurements h) deals correctly with edge particles i) allows user defined classifications
Please check out our website for more information.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com web: http://www.soft-imaging.com
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
DATE: Tue, 23 Feb 1999 13:06:56 } From: Rudy {mojoseph-at-NMSU.Edu} To: microscopy-at-Sparc5.Microscopy.Com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for software/hardware that will help me count the number amount , as well as note the area of the particle, off a negative (print) image that I have produced using a TEM. I could do it by hand, but the time neccessary is not available. I know that there has to be something out there that will help me, and any help would be greatly appreiciated.
Joseph Montoya Physics Department New Mexico State University
Sato lead citrate was published by T. Sato on the Journal of Electron Microscopy vol.17,No.2, 158-19, 1968.
The recipe is as follows:
Lead nitrate 1 g Lead acetate 1 g Lead citrate 1 g sodium citrate 2 g
in 82 ml of distilled water, then add 4% NaOH 18 ml. pH at 12.
There is another recipe in J. Electron Micro., 116, 133 (1976).=20
Best regards,
Ming
On Tue, 23 Feb 1999, Rinaldo Pires dos Santos wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America= =20 } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Hello,=20 } Can somebody help me about the preparation of the Sato's Lead Citrate? } What's=A0 the chemicals and its proportion? } Thank's in advance. } M.Sc. Rinaldo Pires dos Santos } Lab. of Plant Anatomy - Dept. of Botany - UFRGS } Porto Alegre - RS - Brazil } =20 } =20
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * #1074B Dentistry Pharmacy Building * * University Of Alberta. * * Edmonton, Alberta, Canada T6G 2N8 *=20 * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Dear Rinaldo: Here are two references that may be helpful: 1) Takagi, I., etal, 1990. Penetration and stainability of Modified Sato's Lead Staining Solution. J. Electron Microsc. 39:67-68.
2) Hanaichi, T.,etal.. 1986. A Stable Lead by Modification of Sato's Method. J. Electron Microsc. 35:304-306.
The original Sato reference is J. Electron Microsc 17:158 (1968).
Hmmm.. I guess that's three. If you don't have access to these, contact me directly.
Don Gantz Boston Univ Med School gantz-at-med-biophd.bu.edu
On Tue, 23 Feb 1999, Rinaldo Pires dos Santos wrote:
} Date: Tue, 23 Feb 1999 21:14:06 -0300 } From: Rinaldo Pires dos Santos {rinaldop-at-botanica.ufrgs.br} } To: Microscopy-at-sparc5.microscopy.com } Subject: Sato Lead Citrate } =20 } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America= =20 } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Hello,=20 } Can somebody help me about the preparation of the Sato's Lead Citrate? } What's=A0 the chemicals and its proportion? } Thank's in advance. } M.Sc. Rinaldo Pires dos Santos } Lab. of Plant Anatomy - Dept. of Botany - UFRGS } Porto Alegre - RS - Brazil } =20 There are several recipes for something called "Sato lead;" I don't know=20 what the original was. Here is the one we use:
1g lead nitrate 1g lead citrate 1g lead acetate 2g sodium citrate 82 ml distilled water
Shake 1 min. Sol'n will look milky. Then add 18 ml freshly prepared (from pellets) 4% NaOH (1N). Sol'n will=20 clear, except for large grains in bottom. Filter rapidly and store=20 sealed in dark bottle (plastic, not glass; can wrap with Al foil). Keep li= d=20 on. If white precipitate forms on top, remove stain from area away from=20 ppt. Can microfuge just before use to remove ppt. Lead carbonate ppt=20 on the grid looks like little "Pacmans" in the scope--very black circles=20 (100-200 nm +/-) with one side usually rough or indented. (Uranyl acetate= =20 precipitate is finer-grained--looks like pepper. Phosphate buffers can=20 also look like pepper, but grains are usually a little darker. UA=20 deposits are frequently ON subcellular structures--membranes, ribosomes,=20 etc; PO4 deposits are usually in the cytosol--where you would expect water.= )
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710=20 Ph: 919 684-3452 FAX: 919 684-8735
McCaffrey, John (IMS) wrote: } } Does anyone out there know of a source of single crystal LiF } substrates or other similar single crystal fluorides, chlorides, etc? } Dear John, I got mine from Ted Pella. I am not affiliated with them ex- cept as a customer. Yours, Bill
Dear Bernward Laube: A relatively simple method to evaluate liposomes is to fix with osmium tetroxide to make them more rigid and less likely to flatten (effectiveness will depend upon composition of liposome) and then negative stain them on a grid. If you want to discuss in detail, contact me directly. Don Gantz Boston Univ Med School gantz-at-med-biophd.bu.edu
I am searching for any information regarding electrochemical etching of {110} Iridium to form a very sharp tip suitable for FIM use. Any information or direction towards information would be appreciated. Thanks, Randy
Try negative staining with a 2% ammonium molybdate (aq). Take a coated grid (carbon, formvar, etc.), put a drop of liposomes on top of the grid for a minute. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the ammonium molybdate. After a minute, draw off drop as before and allow the grid(s) to dry. Take it to the TEM.
The above procedure is only a starting point. The concentration, stain, and times can all be varied to achieve optimum results. You might check out some EM texts on negative staining for other ideas. Good luck.
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Dear all, has anybody experience or can give me tips for the preparation/examination of liposomes ( {100nm diameter) for TEM or SEM . We want to analyse the homogeneity of suspensions and to estimate the single volume of the liposome particles? Unfortunately we don't have any unit for cryomicroscopical observations! Any suggestions are welcome... best regards, Bernward Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit?tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
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John Hren's book on FIM has it. Send me a message at home to remind me and I will look it up for you. (SKAB-Walck-at-worldnet.att.net) Another option is to call Alan Melmed at John Hopkins-He'll know for sure. -Scott
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---------- } From: Randy Percival To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
I am searching for any information regarding electrochemical etching of {110} Iridium to form a very sharp tip suitable for FIM use. Any information or direction towards information would be appreciated. Thanks, Randy
We have a manual metalographic grinding/polishing wheel. The sample being prepared is held in the hand as it is pressed against the rotating wheel. Our safety people have asked us to provide finger protection for this device. Does anyone have any solution/suggestion? Everett Ramer
Jordi, Polishing is only going to contaminate your pit with all sorts of junk. The best way is to microtome the pit either normal or preferably in cross section. A diamond knife would make this quite easy although we have done good work with glass but this is a slow process as the knife must be repositioned every few cuts. One hint is to keep your cutting area very small, like a fraction of a mm. Good Luck, Russ Gillmeister
-----Original Message----- } From: Marti, Jordi [mailto:Jordi.Marti-at-alliedsignal.com] Sent: Wednesday, February 24, 1999 8:43 AM To: Microscopy
Hello: We are considering doing TEM to look at the microstructure around surface pits in molded stainless steel parts. We are trying to understand the source of these pits. So far SEM and OM have provided us with no relevant clues. The pits are barely visible by eye and they are only very few of them on a given part. We are thinking of doing tripod polishing , but I would appreciate any comments/suggestions on how one might best approach this.
Thanks to all who responded to my problem. But I still have the problem. I got a new cylinder of Argon. I checked the hoses to make sure they were not clogged. I double checked the polarity. My vacuum is good. But when I=20 turn on my DSM-5 I get a blue arc around the=20 middle of the cathode. After 3 minutes, I shut the sputter down but I have no coating on my sample. I checked the inside of the DSM-5 and found no loose wires, and the fuses were good. Any other suggestions would be greatly appreciated.
George Lawton Chief Electron Microscopist Microscopy and Imaging Service Center UT Southwestern Medical Center at Dallas Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
Have you tried SWLI (scanning white light interferometry) or AFM?
SWLI has XY resolution comparable to OM (since you can see these pits by naked eye, that should suffice) but can have Z resolution as fine as about 5 nm. I have a Xerox copy of an older article available on this technology if you are interested (Amer Lab, Nov 1994) or you can visit the web sites of several of the manufacturers: Zygo, Wyko (now part of Veeco) and Phase Shift Technology. This approach would not only allow you to image but measure parameters such as depth, profile immediately around the pit, etc.
Caveat: I have no financial interest in sales derived from this message.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 06:43 AM 2/24/99 -0700, Marti, Jordi wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Summer School on Computing in Electron Microscopy slotted for August 9-13, 1999 , Berkeley, California
(Berkeley, CA) The seventh annual Summer School on Computer-Interactive HRTEM Image Acquisition, Processing and Simulation will be held at the National Center for Electron Microscopy (NCEM), Lawrence Berkeley National Laboratory, University of California, Berkeley from August 9 through August 13, 1999.
The curriculum will focus on training participants in techniques of computer-assisted acquisition and interpretation of high-resolution electron microscope images, including remote-control microscopy. Participants will learn general principles and apply them to specific cases. Instruction on use of computer assistance to obtain images on NCEM microscopes will be followed by training in the use of specific application programs for image interpretation by image processing and simulation.
Participants who wish to apply newly acquired techniques to their own projects are encouraged to extend their visit at NCEM into the next week. Please note: this type of arrangement requires advance submission of a proposal. Projects may involve prepared specimens for microscopy, images and diffraction patterns for processing, or crystal and defect data for simulations. The fee of $375 will cover all materials, instruction, continental breakfast daily, two lunches and an evening reception. Deadline for applications is June 15, 1999. For more information and downloadable application materials contact:
What kind of resolution can be achieved when imaging noble metal particles in catalyst? The catalyst comprises binder and zeolite. I image the system using a STEM/ADF and have been able to see {1 nm particles on binder.
However, I have not been able to see the ultra-fine particles (5-10 --0__=Su6dP4Po6DV5U1roKslOtYgCgwxd5t1scAJybnVta3mV5BnNAUWoo8fA Content-type: text/plain; charset=iso-8859-1 Content-Disposition: inline Content-transfer-encoding: quoted-printable
=C5) in the zeolite. Has anyone been able to image 5-10=C5 metal particles in zeolites? I am trying to reconcile the difference between images (which don't sho= w {1nm particles on zeolites) and chemisorption technique which indicate that the metal is very highly dispersed.
Thank you, Sincerely, Mohan Kalyanaraman
Sr. Staff Material Scientist Catalyst Characterization Catalyst Technology Group Mobil Technology Company PO Box 480 Paulsboro, NJ 08066 609-224-3989 (ph#) 609-224-3608 (fax) =
A couple of weeks ago I posted a query, looking for a specific LM stain for mitochondria. There were several responses from people wanting to know the same thing so here are the results.
First of all, thanks to everyone who replied. Only some of you got a personal thankyou because I lost track of who I had thanked and who I hadn't.
If you remember, I had some tissue already fixed in glutaraldehyde but not processed further. Ideally I wanted a LM stain that would work on semithin resin sections and preferably be compatible with TEM processing.
The only practical method anyone suggested was to do what I do now: process the tissue as normal for TEM and stain semi-thins with toluidine blue at high pH. The only problem with this is that nearly everything stains with tol. blue, and you have to identify the mitochondria by the intensity of their staining. So it is not a specific stain, but it is easy to do and fits in with the TEM processing.
Other respondents suggested Janus Green, and Mitotracker from Molecular Probes, but both of these only stain mitochondria in living cells and my cells are already fixed. According to the Molecular Probes catalogue, Mitotracker is retained well in stained cells after fixation (but the cells need to be alive when they are stained).
The only other suggestions were to try the methods developed several decades ago. I had read all these before posting my query and rejected them because none of them use the type of aldehyde fixation we use today.
So that's it. No ideal methods, and I haven't the time to play around and develop my own. You know how it is.
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department Faculty of Medicine & Health Sciences University of Auckland Private Bag 92019 Auckland New Zealand
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(1) Reflection Optical Microscopy in NOMARSKI (Differential Interference Contrast) mode. This has similar high resolution in the Z direction as SWLI (Barbara Foster reply), but it does have the disadvantage, that it only works for nearly flat surfaces: if your steel objects have curved surfaces then the next might be useful:
(2) for TEM it is very easy to make replicas of steel surfaces using cellulose acetate, followed by shadow / carboning the first stage replica. We have used this on all sorts of specimens. It is very easy if your surface is flat or even cylindrical (as long as the curvature is not too great) but for spherical surfaces you can either (i) replicate a square part with only a small solid angle - say from the N.Pole to 80^ north; or (ii) reliplicate a thin nearly cylindrical strip along a latitude or longitude.
There are books which discuss extraction replicas, which not only give you the surface but can pull off tiny inclusions for electron diffraction, etc.
On Wed, 24 Feb 1999, Marti, Jordi wrote:
} We are considering doing TEM to look at the microstructure around } surface pits in molded stainless steel parts. We are trying to understand } the source of these pits. So far SEM and OM have provided us with no } relevant clues. The pits are barely visible by eye and they are only very } few of them on a given part. We are thinking of doing tripod polishing , but } I would appreciate any comments/suggestions on how one might best approach } this.
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Online registration is available also at the same site.
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Hmmm.... Then you may want to check/replace the target. Blue arc may indicate you have the correct power supply and vacuum. But you should make a double check. -cy TEMist, Batta Labs, Delaware
On Wed, 24 Feb 1999, George Lawton wrote: } Thanks to all who responded to my problem. } But I still have the problem. I got a new cylinder } of Argon. I checked the hoses to make sure they } were not clogged. I double checked } the polarity. My vacuum is good. But when I } turn on my DSM-5 I get a blue arc around the } middle of the cathode. After 3 minutes, I shut } the sputter down but I have no coating on my } sample. I checked the inside of the DSM-5 and } found no loose wires, and the fuses were good. } Any other suggestions would be greatly } appreciated. } } George Lawton } Chief Electron Microscopist } Microscopy and Imaging Service Center } UT Southwestern Medical Center at Dallas } Phone: 214-648-7291 } eMail: George.Lawton-at-email.swmed.edu
I did not see the original tread. There is a list of "mite experts" who did a lot of TEM, SEm on mites. Don Griffin in the UK, Enrico de Lillo in Italy Carin Kameric in SA just to name a few. These individuals will be more than happy to help, give references etc. We are active in South Africa as well. I did a bit in confocal and SEM.
Hope this does help.
Mr. S H Coetzee Electron Microscope Unit Private bag X3 Wits Johannesburg 2050 Tell: +27 11 716 2419 Fax : +27 11 339 3407 E-mail stephan-at-gecko.biol.wits.ac.za
Mohan Rice and Treacy were able to image an individual Pt atom in a zeolite using high angle annular dark field microscopy with a HB501. Imaging an individual atom on a catalyst support or zeolite is quite difficult because of the problem in differentiating whether the intensity is a result of tip noise or scatter from a higher atomic number atom. The smallest cluster in a zeolite that I have been able to image and then reimage to demonstrate that it is scattering is about 0.4nm. One also has to be careful so as not to agglomerate the metal because of the beam energy and structural collapse of the zeolite. Pennycook has shown some work with the HB603 that an individual Pt atom on alumina can be imaged. The key to performing this type of work is utilizing a very thin sample so that scattering from the catalyst support is minimized. Orienting along a zone axis is often helpful. Sample thickness may be the reason you can image clusters on the binder and not in the zeolite. On the other hand, all of the Pt may be on the binder and not in the zeolite. Don't always believe that the impregnation and subsequent finishing accomplished the desired metals location.
Steve Bradley UOP sabradle-at-uop.com (847) 391-3321
} -----Original Message----- } From: Mohan Kalyanaraman [SMTP:mohan_kalyanaraman-at-EMail.mobil.com] } Sent: Wednesday, February 24, 1999 3:57 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Imaging of ultra-fine metal particles } } Fellow Microscopists, } } What kind of resolution can be achieved when imaging noble metal particles } in catalyst? } The catalyst comprises binder and zeolite. I image the system using a } STEM/ADF and have been } able to see {1 nm particles on binder. } } However, I have not been able to see the ultra-fine particles (5-10 { { } File: ATT17711.txt } }
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Hi all, I have a new problem that I believe hinges on fixation at very low pH. The problem is this: I want to fix virus crystals that appear to be very sensitive to pH change...i.e. they dissolve when put into any solution having a pH above about 4.6. They are stable in a stabilization buffer (40% PEG MME 2000 and 10mM CaCl2 in 0.1M sodium acetate pH4.6).
We have been successful fixing virus crystals in the past by first fixing in 0.1M glutaraldahyde in a stabilization buffer at pH 6.0 (47% saturated ammonium sulfate, 0.1M MES) for 72 hrs. Then we fixed briefly in 0.1 Glut in 0.67M PO4 buffer (pH 6.8) and finally in 1% OsO4 in 0.67 PO4 buffer for 1 hr. However, this was all done at a much higher pH than the present crystals can tolerate.
We have tried fixing the crystals by first adding glutaraldehyde to the stabilization buffer to a final concentration of 0.1% and leaving the crystals in this for 72 hrs. The crystals are fine. If moved to fix in higher pH buffer, the crystals dissolve. Therefore I then rinsed with stabilization buffer and added aqueous OsO4 directly to the buffer to a final concentration of 1% OsO4. This was followed by washes with buffer. The crystals still seemed to be intact. Next was to try to dehydrate by gently and gradually adding ETOH to the buffer-crystal prep. As soon as I started adding the ETOH, which both diluted the stabilization buffer and affected the pH, the crystals started to dissolve.
I suspect that the fixatives did not crosslink at the low pH and that the crystals were still so unstable that they disintegrated in the dilute ETOH/buffer mix. It did not surprise me that the glut would not work at the low 4.6 pH but I did think the OsO4 would work. By the way, the crystals did not turn brown with the OsO4.
Does anyone know what is the minimum pH that 0.1% glutaraldehyde and 1% Osmium will crosslink? Has anyone else had a similar fixation problem and come up with a solution? These crystals are very difficult to make and I hate to keep dissolving them.
Looking forward eagerly to suggestions. Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
for {Microscopy-at-sparc5.microscopy.com} Paris Thu, 25 Feb 1999 15:01:09 +0100 (MET) Message-ID: {36D55860.979FBCD4-at-wanadoo.fr}
Materials for sale (well cared and under maintenance) : 1/- Ultramicrotome MT-1 /SORVALL-PORTER BLUMM (manual type), weight 12
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We have a manual metalographic grinding/polishing wheel. The sample being prepared is held in the hand as it is pressed against the rotating wheel. Our safety people have asked us to provide finger protection for this device. Does anyone have any solution/suggestion? Everett Ramer
Could you elaborate on how you would fix liposomes w/ Osmium tetroxide.= We have tried the negative stain approach with some success but have to li= ve with the obvious artifacts such as flattening. There are likely other artifacts caused by ionic or chemical changes of the stain. The chemic= al fixation sounds promising but I'm not sure how this could be done on a = liquid sample.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com =
Position Open: Research Electron Microscopist, to start July, 1999. (http://www.burnham-inst.org).
Highly motivated, conscientious individuals, especially with experience in negative staining, frozen hydrated, or other high resolution biological specimen preparation techniques, are encouraged to apply. Qualifications include a BA/BS or equivalent experience, preferably in one of the biological or physical sciences, or in bioengineering. Knowledge of the operation of a high resolution TEM is required. An ability to work independently as well as in a small group environment is important. The candidate will not be solely responsible for microscope maintenance, but will assist with such, as well as with operating/testing/developing instrumentation (e.g., transmission electron microscopes, cryo-holders, freeze-etch machine, cryo-microtome). Previous experience with and aptitude for such activities is desirable, although some on-the-job training is possible. Additional duties include routine and publication-quality photographic work and general assistance in examining biological specimens for members of the group. Applications should include a statement of experience and goals plus three letters of reference. Salary is competitive and commensurate with experience. EOE. Send applications to: The Burnham Institute, 10901 N. Torrey Pines Rd., La Jolla, Ca 92037 Attn: Dr. Dorit Hanein c/o Sherri Marinovich Director of Human Resources or e-mail www.humanresources-at-burnham-inst.org. REFERENCE JOB CODE DH.
"Sequential treatment by phosphotungstic acid and uranyl acetate enhances the adherence of lipid membranes and membrane proteins to hydrophobic EM grids", A. N. Barnakov, Journal of Microscopy. Vol 175, Pt 2, August 1994, pp. 171-174.
I've not actually tried the above, just collected the paper last year for future reference.
Gib Ahlstrand
----------------------------------------------------------------------------- } Try negative staining with a 2% ammonium molybdate (aq). Take a coated grid (carbon, formvar, etc.), put a drop of liposomes on top of the grid for a minute. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the ammonium molybdate. After a minute, draw off drop as before and allow the grid(s) to dry. Take it to the TEM.
} The above procedure is only a starting point. The concentration, stain, and times can all be varied to achieve optimum results. You might check out some EM texts on negative staining for other ideas. Good luck.
} Chuck Butterick Engineered Carbons, Inc.
______________________________ Reply Separator _________________________________ } Subject: TEM/SEM of liposomes Author: {"B.Laube-at-biologie.uni-bielefeld.de"-at-sparc5.microscopy.com} at
} } Dear all, has anybody experience or can give me tips for the preparation/examination of liposomes ( {100nm diameter) for TEM or SEM . We want to analyse the homogeneity of suspensions and to estimate the single volume of the liposome particles? Unfortunately we don't have any unit for cryomicroscopical observations! Any suggestions are welcome... best regards, Bernward } } Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit?tsstrasse 25
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Dear Everett, Grinding your fingers down is the first rite of passage for any Metallurgist worth his/her salt. No good metallographer has any feeling left in thumb or forefinger. Will you mess with tradition? Seriously, any glove thick enough to provide protection will hamper the fine control necessary to get a flat surface. The only thing I can think of is some sort of clamp or vise to hold the sample. I have never seen one, however. Now you know why automatic polishers are so popular. You wrote: } } We have a manual metalographic grinding/polishing wheel. The sample } being prepared is held in the hand as it is pressed against the rotating wheel. } Our safety people have asked us to provide finger protection for this device. } Does anyone have any solution/suggestion? } Everett Ramer } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Mohan, I have imaged the catalyst used by Petro Canada in my 200 kV TEM and seen the larger particles you mentioned at 40,000X, but I wasn't looking for the small ones and I no longer have the sample. Have you tried conventional TEM at 100 or 200 kV? The size you are looking for is difficult, since they are clusters of only a few atoms, so they have very little electron stopping power. The problem is not resolution, but contrast. Have you tried conventional darkfield? You wrote: } Fellow Microscopists, } } What kind of resolution can be achieved when imaging noble metal particles } in catalyst? } The catalyst comprises binder and zeolite. I image the system using a } STEM/ADF and have been } able to see {1 nm particles on binder. } } However, I have not been able to see the ultra-fine particles (5-10 } =C5) in } the zeolite. } Has anyone been able to image 5-10=C5 metal particles in zeolites? } I am trying to reconcile the difference between images (which don't show } {1nm particles on zeolites) and } chemisorption technique which indicate that the metal is very highly } dispersed. } } Thank you, } Sincerely, } Mohan Kalyanaraman Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
What is the crystal made of (what is the virus? is it in a matrix inside the cell? what is on the surface of the virus or crystal?). Maybe it's something that doesn't get cross-linked by either glut or Os. How about water sol resins to avoid organic solvents? What are the research questions you want to ans by sectioning the virus?
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
O.k., using Methylmethcrylate and Butyl methacrylate the instructions say " degas with nitrogen" does anyone know what that actually means? Does it mean simply bubbling N2 gas through the resin mixture? Any one have any specific instructions / recommendations?
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
Widely used as a routine stain for ultrathin sections, lead solutions are frequently employed after uranyl staining to produce high contrast and stain many cellular and tissue components. Sato's method of lead staining(1967) is one of several variations of lead staining since its introduction by Watson(1958), among which are two methods by Karnovsky(1961), one by Millonig(1961), Reynolds (1963), Venable and Coggeshall(1965), and Fahmy(1967).
We have recently begun using a refinement of Sato's method introduced by Hanaichi et al.(1986) to produce a long-term-storage lead solution that seems to give virtually no -CO3 precipitate and claims to be good for one year at room temperature. We'll see.
Indicated references: - Karnovsky MJ.(1961), Simple methods for 'staining with lead' at high pH in electron microscopy, J.biochem.biophys. Cytol.11,729.
- Millonig G.(1961). A modified procedure for lead staining of thin sections, J.biophys.biochem Cytol 11,736.
- Reynolds ES.(1963). The use of lead citrate at high pH as an electron-opaque stain in electron microscopy.J.Cell Biol.17,208.
- Venable JH, and Coggeshall R.(1965). A simplified lead citrate stain for use in electron microscopy. J.Cell Biol.25,407.
- Fahmy A.(1967). An extemporaneous lead citrate stain for electron microscopy, Proc. 25th Ann. Conf. EMSA,p.148.
- Sato T.(1967). A modified method for lead staining of thin sections.J. Electron Microsc., 16:133.
- Hanaichi T.,et al.,(1986). A stable lead by modification of Sato's method. J.Electron Microsc.,35:304.
General references: - Staining Methods for Sectioned Material, Lewis PR;Knight DP, Practical Methods in Electron Microscopy, Ed:Glauert AM.
- Principles and Techniques of Electron Microscopy,Biological Applications,3rd ed.,Hayat MA.
- Electron Microscopy, Principles and Techniques for Biologists, Bozzola JJ,Russell LD.
- Technical Tips II, Electron Microscopy Sciences
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W Wiggins, Supervisor 2/25/99 2:44:49 PM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I need some advice about sample prep for TEM. I have several amorphous silicon thin films that have been deposited on glass substrates from which I would like to make samples for TEM. The films are ~200 A thick, and it is important that they not be folded over or significantly thinned during the preparation. I would generally prefer a chemical process, as I worry that mechanical processes or ion milling could significantly change the microstructure of the films.
I have been told that it is possible to prepare the samples I want to first scoring the surface of the film, then immersing it and the substrate in a weak solution of HF. A little gentle scraping and the film is supposed to float free of the substrate and I can then pick it up on a copper mesh grid. Does anyone have any experience with this technique? Any advice on how weak a "weak" solution might be, or how long I need to let the film+substrate soak before scraping?
I would also be very glad to hear of any alternative means to prepare the samples I need that don't involve HF.
Thanks,
Paul Voyles
Paul Voyles (609) 951-2627 voyles-at-research.nj.nec.com NEC Research Institute, 4 Independence Way, Princeton, NJ 08540
I need to evaporate nickel in my Balzers BAF 301 Freeze Etcher. Everything is ready to go except I can't find what the density of nickel is in grams/cc so that I can calibrate my Quartz Crystal monitor. No, this information is not in the Merck Index, a disappointment indeed! Any help would be appreciated.
Richard: I do this by attaching a glass pasteur pipet to a rubber tube on a N2 tank and very slowly (barely perceptible flow against my cheek) flow the gas. I immerse the tip into the resin mix (in the fume hood) and bubble it for about 5-10 min. It is worth the effort - we have done the expt. Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Hi all, I have recently finished my third year project in electron microscopic observations of the entry and exit of influenza virus. In it I used Thermanox coverslips to culture the cells on, one of the problems that I had was delamination of the coverslip from the resin (Spurrs) and wondered if this had any thing to do with the different densities of the Thermanox and resin. If anyone knows their densities I would be most grateful.
I need to purchase a liquid nitrogen filling device for the Reichert KF-80. This consists of the 35 liter dewar and the filling head that goes into the dewar. I have contacted Leica/Reichert and they are unable to supply me with the unit because I needed the older model version. The new one available does not have the same circuitry and therefore will not be not compatible.
This dewar and filling head are of the same vintage of the F-C through D series of the cryo attachment for their ultramicrotome. So if any of you purchased the cryo attachment for your Reichert Ultramicrotome and are no longer doing cryo-microscopy and would want to part with at least half of the equipment, please get in touch with me.
Thank you very much.
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
We are looking for a used, but late model "research grade" SEM. A large chamber is imperative. Chamber porting for EDS, WDS, and BSE is also required. The system must be configured to provide a clean, high vacuum (heavy element carbides & similar work). VP systems will certainly qualify and are desirable if the clean high vacuum criteria can also be met. Digital (=} 1024x1024) or analog is OK. I do prefer manual/knobs over automatic/windows mode. Software may set things up correctly on the average, but we don't do average work. Specimens are metallics/oxides/carbides etc., elements range from boron through uranium in almost any possible combination.
For a point of reference, We currently have an Etec Autoscan which has been modified to use a mag-lev turbo pump. It is also fitted with a windowless EDS detector, Microspec 2A WDS, and GW Electronics BSE. Although the Etec is a 70s vintage SEM, it is an excellent instrument. It only falls short in the realm of very low kV / high resolution imaging, high resolution at hefty WDS beam currents (somewhat of a conflict) and chamber size.
Woody White McDermott Technology, Inc. Lynchburg Research Center P.O. Box 11165 Lynchburg, VA 24506-11165
} } We have a manual metalographic grinding/polishing wheel. The sample } } being prepared is held in the hand as it is pressed against the rotatin= g wheel. } } Our safety people have asked us to provide finger protection for this device. } } Does anyone have any solution/suggestion? } } Everett Ramer
Everett, =
Mary Mager's response, while funny, is actually quite accurate. There ar= e not too many ways to protect fingers while properly holding a small (1-1/4", typically) =
sample for grinding/polishing. =
I preface my next remarks by pointing out that I work for a manufacturer = of
metallographic equipment and consumables, and therefore have a financial interest in solving your problem:
We at BUEHLER, do offer a simple grinding fixture which might help. This=
fixture is a squat, stainless steel, hollow cylinder with a carbide ring around it's base. The sample is clamped within another hollow cylinder seated within the first. =
The two cylinders are threaded, so that the inner can be raised or lowere= d =
with respect to the carbide 'stop' of the outer. Engraved markings allow= =
material removal in increments as fine as 20microns. While this is not actually =
finger protection, per se, it will allow you to grasp something larger so=
that your =
fingers are not in such close proximity to the grinding wheel. We also offer =
a motor system which allows the fixture to rotate, in place, on the wheel=
My manual for our Blazers quartz crystal film thickness monitor QSG 301 says the density of nickel is 8.9
Aloha, Tina
} I need to evaporate nickel in my Balzers BAF 301 Freeze Etcher. Everything } is ready to go except I can't find what the density of nickel is in grams/cc } so that I can calibrate my Quartz Crystal monitor. No, this information is } not in the Merck Index, a disappointment indeed! Any help would be } appreciated.
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
We have such a device, but are not willing to part with it. The reason I replied is that I can assure you that you can still use your KF80. The pump unit sometimes drives me nuts, so when I plunge freeze I merely fill the unit by hand. Take care to cover the cryogen tube, then pour LN2 in the working area with a small Dewar. I've done this with a blown fuse, but if your electronics work, it's easy to keep an eye on the LN2 level on the front panel. I find I have to add LN2 after every plunge (level goes down one light), but it just becomes part of the rhythm of freezing. Freeze, remove sample, cover tube, pour in nitrogen, uncover tube, freeze.
I have not tried this with impact freezing ("slamming").
Good luck!
Tina
} I need to purchase a liquid nitrogen filling device for the } Reichert KF-80. This consists of the 35 liter dewar and the filling head } that goes into the dewar. I have contacted Leica/Reichert and they are } unable to supply me with the unit because I needed the older model version. } The new one available does not have the same circuitry and therefore will } not be not compatible. } } This dewar and filling head are of the same vintage of the F-C } through D series of the cryo attachment for their ultramicrotome. So if any } of you purchased the cryo attachment for your Reichert Ultramicrotome and } are no } longer doing cryo-microscopy and would want to part with at least half of } the equipment, please get in touch } with me.
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
If you know of anyone that may be interested in (and suited for) the "exciting world of industrial research" please pass them the following job posting. The position has been advertised at a technical level but we now have the opportunity to expand the position to a research scientist (Ph.D.) position. We have just installed a state-of -the art FEI XL830 DualBeam FIB and I am looking for someone to be the "FIB expert".
I am looking for a person with extensive focused ion beam (FIB) experience and preferably some transmission electron microscopy (TEM) experience to fill a position in the Materials Analysis and Characterization Group at the IBM Almaden Research Center. Much of the work will involve the operation of a dual beam FIB in collaboration with other scientists in the study of small structures, thin films, and magnetic structures. Areas of application include photoresists, magnetic thin films, and tips for atomic force microscopes. The scientist is expected to take responsibility for the operation of the FIB, assist in the training of other users, and collaborate with a wide variety of applications of the that device. In addition, the scientist will apply TEM techniques to related problems. Experience with FIB instruments and TEM is extremely important.
Experience with TEM sample prep - especially tripod polishing and micromanipulation, as well as TEM and SEM would be a plus.
IBM is an Equal Opportunity Employer, Women and Minorities are encouraged to apply.
Please include the names of three references with your resume.
Please contact me directly. Submit your resume to:
Dr. Philip M. Rice K19/D1 IBM Almaden Research Center 650 Harry Road San Jose, CA 95120-6099
Dear Listers, Just a question about the MSA standard for x-ray spectrum file format: do any of you use it and under what circumstances? I am looking to upgrade my old x-ray analyser computer soon and want to know whether this standard is ever used and when. Please reply to me off-list and I will summarize to the list if I get any results.
Thank you all, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
The density of nickel is 8.902 g/cc according to the American Society for Metals, Metal Handbook.
Mark Robertson National Research Council Integrated Manufacturing Technology Institute Vancouver, B.C. Canada Phone (604) 221-3073 Fax (604) 221-3088 mark.robertson-at-nrc.ca http://www.nrc.ca/imti/vanc_e.html
} -----Original Message----- } From: Grazul, John [SMTP:Grazul-at-nel-exchange.Rutgers.EDU] } Sent: Thursday, February 25, 1999 12:46 PM } To: 'Microscopy-at-sparc5.microscopy.com' } Subject: density of nickel for evaporation } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Evaporators All, } } I need to evaporate nickel in my Balzers BAF 301 Freeze Etcher. } Everything } is ready to go except I can't find what the density of nickel is in } grams/cc } so that I can calibrate my Quartz Crystal monitor. No, this information } is } not in the Merck Index, a disappointment indeed! Any help would be } appreciated. } } TIA, } }
In a message dated 99-02-25 14:56:29 EST, edelmare-at-casmail.muohio.edu-at-sparc5.microscopy.com writes:
{ { O.k., using Methylmethcrylate and Butyl methacrylate the instructions say " degas with nitrogen" does anyone know what that actually means? Does it mean simply bubbling N2 gas through the resin mixture? Any one have any specific instructions / recommendations? } }
Hi Richard,
Yes, this is correct. Simply bubble dry nitrogen gas at a *very low* flow rate through a pipette that's immersed in the mixture. Regarding how long to do this, we used to bubble with nitrogen for 15 minutes as a matter of routine. If the mixture contains benzoyl peroxide or a similar granular or dry catalyst, just bubble until the catalyst is completely dissolved.
Bob ****************************** Robert (Bob) Chiovetti, Ph.D. President Microimaging Technologies, Inc. Tucson, Arizona USA Tel./Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems Clinical & Research Microscopy Cytology/Histology/Pathology/EM *******************************
Huge thanks to you all - your responses to my query on EM & Archaeology has given me a stack of leads to chase up for my seminar. I will be contacting many of you privately in the course of the next week or so - s= o expect a call!
Geoff Avern Universit=E9 Libre de Bruxelles
P.S. It was nice to get responses from familiar names - it made me realise that I'm missing working in EM more than I thought!
Hi Paul, I looked at a similar sample recently, although in my case the Si was laser annealed and was crystalline. Here is the protocol I used. I don't think you will be able to pick up the Si from the etch in your case as the layer is too thin.
1) Grind the sample to a thickness of ~100 um by mounting a ~1 cm square piece face down on a glass slide using thermoplastic wax. By placing grade zero cover slips (~120 um thick) on either side of the sample you don't have to worry about grinding the sample away.
(By the way, I think that hand protection whilst using a grinding wheel is just not necessary. It is not a chainsaw! I have never hurt myself when grinding a sample which is properly mounted on a proper (large enough) stub - maybe you could put a big flange on your stubs to keep your decidedly overzealous safety officer happy.)
2) Take the sample off the glass slide and put it in a small amount of conc. HF (48%). This will dissolve the glass completely in less than half an hour. The Si film will probably break up into many very small fragments (biggest being a few hundred um across).
3) Dilute the HF at least 20 times with DI water.
4) Filter the solution (preferably using a smooth filter paper).
5) Wash out any remaining HF from the filter paper with DI water.
6) Let the filter dry.
7) Take a 200 mesh TEM grid and 5-minute epoxy. Make up a small blob of epoxy and dunk the grid in it. Put the grid between two pieces of filter paper and remove most of the glue. Do this two or three times more so that you have a grid coated with a very thin layer of tacky glue.
8) Holding the grid in tweezers and working under a low mag optical microscope, pick up Si fragments by touching them with the grid.
9) If you're really in a hurry, you can stick the grid on a hot plate at ~170C for 30 secs (after the epoxy has cured) to outgas it.
It took me less than 90 minutes from getting the sample to having it in the microscope.
} I need some advice about sample prep for TEM. I have several amorphous } silicon thin films that have been deposited on glass substrates from which } I would like to make samples for TEM. The films are ~200 A thick, and it } is important that they not be folded over or significantly thinned during } the preparation. I would generally prefer a chemical process, as I worry } that mechanical processes or ion milling could significantly change the } microstructure of the films. } } I have been told that it is possible to prepare the samples I want to } first scoring the surface of the film, then immersing it and the substrate } in a weak solution of HF. A little gentle scraping and the film is } supposed to float free of the substrate and I can then pick it up on a } copper mesh grid. Does anyone have any experience with this technique? } Any advice on how weak a "weak" solution might be, or how long I need to } let the film+substrate soak before scraping? } } I would also be very glad to hear of any alternative means to prepare } the samples I need that don't involve HF. } } } Thanks, } } Paul Voyles } } Paul Voyles (609) 951-2627 voyles-at-research.nj.nec.com } NEC Research Institute, 4 Independence Way, Princeton, NJ 08540
Its not clear to me what your trying to measure here, but if it is the surface profile (ie the physical form, depth etc.) of these corrosion pits, then how about conoscopic holography. Uniscan manufacture a non- contact optical surface profiler which can use a conoscopic head in order to measure surface profiles on the sub - micronic scale, without touching the surface (and down holes too! - not too many optical techniques can do this).
Much less involved than TEM - or SEM for that matter.
If it the dynamics of pitting you want to measure then we also manufacture various scanning probe electrochemistry systems (SRET,SVET,LEIS) which will measure the localised pitting currents, and hence the corrosion rate, dynamically whilst the thing is pitting.
Let me know if you want more details - or see www.uniscan.co.uk.
Good luck.
Graham Johnson.
In message {Pine.GSO.3.96.990225093315.6816A- 100000-at-suma3.reading.ac.uk} , Robert H. Olley {R.H.Olley-at-reading.ac.uk} writes } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Graham R. Johnson PhD. CPhys. MInstP. ________________________________________________________________________
I asked my metalsmith wife this question, as she's often grinding little bits of metal and such, and has a fondness for her fingers. Her recommendations are: 1) *Do not* wear gloves! They are a safety hazard and can be caught in the grinding wheel. Then you won't have to worry about your fingers. 2) Try a dop stick. This is a wooden dowel with sealing wax on the end in which the sample is held. Krazy glue can also be used to attach the sample. These sticks allow fine control. Gem facetters use dop sticks, for instance. 3) If you prefer to hold the pieces in your fingers, use leather finger tips. They cover just the tips of the fingers and since they come off easily are no hazard. They may not allow you the fine control you want.
Check your local college's art department or any art supply or lapidary store for these supplies (dop sticks are just dowels, so get them at a cheap hardware store--the wax may have to be gotten elsewhere).
Phil
} We have a manual metalographic grinding/polishing wheel. The sample } being prepared is held in the hand as it is pressed against the rotating wheel. } Our safety people have asked us to provide finger protection for this device. } Does anyone have any solution/suggestion? } Everett Ramer
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 Voice: (608) 833-2885 Fax: (608) 836-1969 (please make sure my name is on any fax) oshel-at-terracom.net
I want to thank everyone for all the responses and suggestions I received on looking at surface pits in molded stainless steel parts.
The suggestions ranged from doing microtomy to FIB , SEM (and EDS) as well as replication . Polishing seems to be the less desirable because it could introduce artifacts and contaminants that would complicate matters.
Currently we have two reconditioned microscopes available for purchase and can provide service contracts. Please contact me at this address for more details.
Mark W. Rigler, Ph.D. Vice President MAS, Inc. Suwanee, GA
Holy Cow! Talk of your rapid and accurate responce! I guess I either have to buy the CRC, or take up Liechtensteinian {so that I can translate my manuals}. It does turn out that the Quartz Crystal control manual is the one manual that I do not have; the others are in German, at least the pictures are really cool.
Polishing (crossections) can give you very good results if you apply low viscosity epoxy on surface in advance - to protect edge and debris in pits. You can get good sample for SEM/EDS investigation - use diamond and do not use soft polishing wheels, or you can prepare thin sections for STEM/EDS - do not use electrolytic thinning, ion mill is better for samples as yours. I do not believe you will get sufficient results for your investigation using TEM.
Vladimir Dusevich Electron Microscope Lab Manager UMKC
On Fri, 26 Feb 1999, Marti, Jordi wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I want to thank everyone for all the responses and suggestions I received } on looking at surface pits in molded stainless steel parts. } } The suggestions ranged from doing microtomy to FIB , SEM (and EDS) as } well as replication . Polishing seems to be the less desirable because it } could introduce artifacts and contaminants that would complicate matters. } } } } Thanks again, } } Jordi Marti } } }
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I'm sorry for trouble, but I have missed the discussion about = free(share)ware software for frame averaging a month ago and would like = to ask someone who initiated the discussion to send me a summary. Net is increadibly slow at my place and I can't search the archive. TIA Andrew
Dear Woody, I saw a notice last week in this listserver for a Hitachi S-570 for sale. This is what I use to do my EDX, WDX (Microspec WD-3PC) and BSE. It is very reliable, has incredible beam stability, high resolution and very high beam current capabilities. With an optioal low-kV gun it can do great low kV work. I bought a new S-570 years ago to replace my old Etec Autoscan. I recently purchased a used S-570 for less than $10,000 for EBSP work. The S-2500 is the later model of this with a larger chamber and it is also very good. You wrote: } We are looking for a used, but late model "research grade" SEM. A } large chamber is imperative. Chamber porting for EDS, WDS, and BSE } is also required. The system must be configured to provide a } clean, high vacuum (heavy element carbides & similar work). VP } systems will certainly qualify and are desirable if the clean high } vacuum criteria can also be met. Digital (=} 1024x1024) or analog } is OK. I do prefer manual/knobs over automatic/windows mode. } Software may set things up correctly on the average, but we don't } do average work. Specimens are metallics/oxides/carbides etc., } elements range from boron through uranium in almost any possible } combination. } } } For a point of reference, We currently have an Etec Autoscan which } has been modified to use a mag-lev turbo pump. It is also fitted } with a windowless EDS detector, Microspec 2A WDS, and GW } Electronics BSE. Although the Etec is a 70s vintage SEM, it is an } excellent instrument. It only falls short in the realm of very low } kV / high resolution imaging, high resolution at hefty WDS beam } currents (somewhat of a conflict) and chamber size. } } Woody White } McDermott Technology, Inc. } Lynchburg Research Center } P.O. Box 11165 } Lynchburg, VA 24506-11165 Good luck, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
I have a Kevex 7700 XRF unit and have found out recently that the = printer interface card isn't working. Does anyone know where I can get a = spare other than Kevex (Kevex is on the expensive side). Thanks.
______________________ Roberto Garcia Senior Analyst, Metallography North Carolina State University Analytical Instrumentation Facility Box 7531, Room 303 EGRC Raleigh, NC 27695-7531 rgarcia-at-unity.ncsu.com http://spm.aif.ncsu.edu/aif
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD} {META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {STYLE} {/STYLE}
{META content=3D'"MSHTML 5.00.0910.1309"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT size=3D3} I have a Kevex 7700 XRF unit and have found out = recently that=20 the printer interface card isn't working. Does anyone know where I can = get a=20 spare other than Kevex (Kevex is on the expensive side). = Thanks. {/FONT} {/DIV} {DIV} {/DIV} {DIV} ______________________ {BR} Roberto Garcia {BR} Senior Analyst,=20 Metallography {BR} North Carolina State University {BR} Analytical = Instrumentation=20 Facility {BR} Box 7531, Room 303 EGRC {BR} Raleigh, NC 27695-7531 {BR} {A=20 href=3D"mailto:rgarcia-at-unity.ncsu.com"} rgarcia-at-unity.ncsu.com {/A} {BR} {A=20 href=3D"http://spm.aif.ncsu.edu/aif"} http://spm.aif.ncsu.edu/aif {/A} {/DIV= } {/BODY} {/HTML}
1. EELS imaging was recommended for catalyst particles; 2. You need to worry about contrast more than resolution; 3. Finer the probe size in the STEM, the resolution is going to be better 4. Difficulty in seein g these due to the fact they are only a few atom clusters; may be try to compare contrast between zeolite with Pt atoms vs. no Pt atoms 5. Zeolites are prone to damage in beam - computer acquisition helpful to lowering current and acquiring metal particle images 6. Tracey and Rice have imaged indivudual Pt particles in zeolite L; agglomeration of metal under beam due to zeolite structure damage is a problem; thin samples are better 7. Energy filtered TEM may work, but try STM (scanning tunneling microscope) 8. Size of probe and Thickness of zeolite important - affects signal to noise ratio 9. Thin sectioning will help - microtoming
Various people have been able to image single atoms on alumina.
Thanks to all those that responded.
As always, it is great that we have a forum to share the collective experience. I have a detailed summary of postings that I can mail it to anyone who is interested.
Mohan Kalyanaraman
Sr. Staff Material Scientist Catalyst Characterization Catalyst Technology Group Mobil Technology Company PO Box 480 Paulsboro, NJ 08066 609-224-3989 (ph#) 609-224-3608 (fax)
I'm in search of a TIFF image that was taken using an Philips XL series SEM that is running with a 50Hz mains. The image detail is unimportant as is linescan time. It just needs to be taken with the standard XL TIFF export function. This is for an internal project that we are doing. You can e-mail it to me at the below address (watch doing a reply as it might go to the list server and we don't want that).
Thanks Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
I was under the impression that this server was NOT the place for posting items for sale. If that is the case then ignore this message. If I have been wrong or the rules have changed then fellow readers I have some equipment that is available:
edelmare-at-casmail.muohio.edu-at-sparc5.microscopy.com wrote: } } O.k., using Methylmethcrylate and Butyl methacrylate the instructions say " degas with } nitrogen" does anyone know what that actually means? Does it mean simply bubbling N2 } gas through the resin mixture? Any one have any specific instructions / } recommendations? } Dear Richard, Bubbling N2 through the resin will displace disolved O2, so that would be an indicated procedure if you don't want O2 in the resin. After this, however, I'd pull a vacuum on the mix (using house vac) to get rid of the N2. The latter will prevent bubble formation during polymerization. Yours, Bill Tivol
I will offer this initial reply on-list, then suggest that we take the discussion off-line.
We have used a Pixera for a couple years now and have no problems with it. Software updates did much to improve the quality of the viewfinder.
Are you sure that you rebooted after installing the Pixera drivers and software? (That may require a full reboot rather than just a suspend/resume thing like many new PCs support.) We recently moved our Pixera to a Pentium II 450 from a Pentium 200, and I think I got the "can not initialize camera" message after I loaded the software but before I rebooted. Unfortunately, I cannot tell you about the NT behavior. Although I had NT on our Pentium 200, I never tried installing the Pixera software for NT. Maybe I can.
If you still need help, perhaps you can reply directly. I also have a contact at Pixera if they need to step in.
At 02:08 PM 2/23/99 +0100, you wrote: } Dear Colleagues, } } We recently purchased Pixera Professional digital camera (for optical } microscopy). Nice little thing, but: } } We want to have the camera on PC (Pentium II, 333MHz, 128MB RAM) with Win NT } 4.0 operating system. } We successfully installed the NT driver Pixdrv.drv and Pixera Visual } Communication Suite (from a CD). After starting Studio Viewfinder (a } software for picture acquisition), there was just fine b/w raster (noise) } instead of picture. Using other software and Twain interface led us to the } same disappointment. } We repeated experiments with both service packs for NT 4.0, namely 3 and 4. } } We also tried to install the same camera on another computer (Pentium MMX } 200MHz, 64MB RAM ), running on Win 98. New hardware was installed without a } problem, but when opening the Pixera VC Suite and running Viewfinder } program a message "Can not initialize the camera" appeared. } } I would be very grateful for any suggestion. } ------------------------------ } Dr. Goran Drazic } J. Stefan Institute } Jamova 39 } SI-1001 Ljubljana } Slovenia
I like to have some way to pull the x-ray spectra from our EDS for use elsewhere. Once in a while we get an adventurous grad student who thinks they want a spectrum in an Excel file so they can do their own processing. Other times, we want to overlay spectra from our two, disimilar x-ray systems. In those cases, we want to be able to export the data to something that a spreadsheet can read. The exact format doesn't matter that much to me. It could be MSA format or plain text, preferably in a single column or in two columns of energy and intensity.
MSA format is one answer to the problem, but I would probably relax the constraints to simply something readable by a spreadsheet.
Warren
At 02:39 PM 2/25/99 -0800, you wrote: } } Dear Listers, } Just a question about the MSA standard for x-ray spectrum file format: do } any of you use it and under what circumstances? I am looking to upgrade my } old x-ray analyser computer soon and want to know whether this standard is } ever used and when. Please reply to me off-list and I will summarize to the } list if I get any results. } } Thank you all, } Mary } } Mary Mager } Electron Microscopist } Metals and Materials Engineering } University of British Columbia
Let me first describe the reason for this request. As a professor in a Faculty of Engineering, I co-ordinate the teaching of a basic course in Materials Engineering. This course is annually given to nearly one thousand students. Presently, I am preparing an interactive CD-ROM which will be a complement for a book already published (in French) and used as a reference manual in this course. Today, there is the opportunity to include video-clips, short movies, animations, sounds on a media such as CD-ROM with the pedagogical benefits of such tools. In Materials Science, some topics such as the glide of dislocations on a slip plane, cross-slip of dislocations, pile-ups of dislocations along grain boundaries, Frank-Read sources, Orowan’s mechanism are difficult to teach with only the help of traditional 2D figures in a book. Video-clips with sound track and movies can now easily be included on a CD-ROM and will surely help the students to better understand these topics.
So, my request is the following: I am looking for videos or movies showing the movement of dislocations or any of the dislocation mechanisms described above as directly seen in thin foils observed in TEM. If any of you has already on his shelves such video or movies (or knows somebody who has this type of audiovisuals), please get in touch directly with me at this e-mail address jbailon-at-email.phys.polymtl.ca ). Arrangements will be made for the duplication of the documents and for their copyright. Obviously, due credits will be given in the CD-ROM for the authors of these documents. Many thanks in advance.
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD} {META HTTP-EQUIV=3D"Content-Type" CONTENT=3D"text/html; = charset=3Diso-8859-1"}
{META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {P align=3Djustify} {FONT size=3D2} Let me first describe the reason for = this request.=20 As a professor in a Faculty of Engineering, I co-ordinate the teaching = of a=20 basic course in Materials Engineering. This course is annually given to = nearly=20 one thousand students. Presently, I am preparing an interactive CD-ROM = which=20 will be a complement for a book already published (in French) and used = as a=20 reference manual in this course. Today, there is the opportunity to = include=20 video-clips, short movies, animations, sounds on a media such as CD-ROM = with the=20 pedagogical benefits of such tools. In Materials Science, some topics = such as=20 the glide of dislocations on a slip plane, cross-slip of dislocations, = pile-ups=20 of dislocations along grain boundaries, Frank-Read sources, = Orowan’s=20 mechanism are difficult to teach with only the help of traditional 2D = figures in=20 a book. Video-clips with sound track and movies can now easily be = included on a=20 CD-ROM and will surely help the students to better understand these=20 topics. {/FONT} {/P} {P} {FONT size=3D2} So, my request is the following: I am looking for = videos or=20 movies showing the movement of dislocations or any of the dislocation = mechanisms=20 described above as directly seen in thin foils observed in TEM. If any = of you=20 has already on his shelves such video or movies (or knows somebody who = has this=20 type of audiovisuals), please get in touch directly with me at this = e-mail=20 address ( {A=20 href=3D"mailto:jbailon-at-email.phys.polymtl.ca"} jbailon-at-email.phys.polymtl.= ca {/A} ).=20 Arrangements will be made for the duplication of the documents and for = their=20 copyright. Obviously, due credits will be given in the CD-ROM for the = authors of=20 these documents. Many thanks in advance. {/FONT} {/P} {/DIV} {DIV} {SPAN class=3D300292522-26021999} {FONT color=3D#000000 face=3DArial =
I am trying to return an older microscope to service. It is a Reichert, I think of 1960's or 1970's vintage, and has been "in storage" ( ie. collecting dirt) for about 15 years, so its moving parts are frozen. It has only a serial number for identification; Nr. 236 628. It is a big black microscope with trinocular head and camera. The light source is rear-mounted, the lamp housing making a right angle with a tube containing a focuser for the bulb filament, a diaphragm, and two filter holder slides. It provides sub-stage illumination through the base, from an external transformer. There are "toggles" on either side of the base for switching to "Epi" or "Mix" illumination. The condenser looks like a dedicated phase type, Nr. 13 259. The objectives are Plan 4:1 (brightfield), Ph. 20:1, Ph. 44:1, and 100.1 (also appears to contain a phase ring, and has an internal diaphragm). Does this description ring bells for anyone? I used the same model microscope briefly back in the early 70's, but this one has a different condenser. I can't recall how to set it up properly as I have not used phase contrast since then. I seem to recall that something called a "phase telescope" (??) was used in alignment. Of course I can't find any other parts or the manual!
Could anyone offer advice on how to set it up correctly? Or know how I could get a manual? The lamp assembly is puzzling; the bulb is so large that it is touching the housing. I wonder if it is the correct bulb - any ideas?
To all who desired more info on fixing and staining of liposomes-- I apologize for the delay in my response. We had a major snowstorm and I lost a day. The initial query by Bernward Laube was about evaluating homogeneity of liposome suspensions ( {100nm diam) and estimating particle volumes. We have been using a osmium fix/negative stain technique for a number of years to look at size distribution of VLDL, chylomicrons, and synthetic lipid emulsions. The osmium fix firms up these particles nicely so that flattening is greatly reduced. In fact, based upon metal shadowing exps. the slight enlargement of diameter due to flattening after fixation is negated by slight shrinkage of particles during processing. The result is an excellent estimate of diameter and volume.
In regard to lipid vesicles/liposomes, our experience with smaller ones of 50nm or so prepared by sonication and processed with only negative stain is that they are preserved pretty well. We would expect some flattening although we have not shadowed these. However, with larger liposomes of 100nm+ diam, we believe OsO4 will aid in preservation and stability and presumably reduce flattening.
If one requires differentiation of lamellar structure (uni vs multi) among particles within a population, negative staining is unreliable because of variability of stain penetration. For this purpose, we use vitreous ice cryomicroscopy as we are fortunate to have it available.
In general our fixation/negative staining procedure is as follows:
1) Add 1 volume of 2% OsO4 in Cacodylate buffer to 2 volumes of emulsion or liposome prep. Fix for 30 mins.+
2) Place small aliquot on freshly flow-discharged carbon formvar-coated grid for a few seconds.
3) Remove and blot off excess fluid and immediately stain with 1% NaPT.
4) After a few seconds, remove and blot excess fluid. Air dry.
Optimum concentration is trial and error. Fixed suspension can be diluted as needed. Hope this helps.
Don Gantz Biophysics Dept Boston Univ Med School gantz-at-med-biophd.bu.edu
As the applications and inquiries about the UBC Live-cell Course have come in, it has become clear to me that some of the descriptions of it may have given some wrong impressions.
Two of the common misconceptions seem to be:
1. That you will not be welcome unless you have a live-cell project.
2. That you should have a very high level of confocal experience before attending.
In fact, many students participate fully in the course even though they do not come with their own live-cell project. This is because they are paired in small groups with others who do have such projects. We stress the "live-cell" emphasis in the course for two reasons:
- We think of it as the most difficult type of 3D microscopy to do. Therefore, learning to do it, will teach the best techniques in other areas.
- We think that the ability of LM techniques to perform live-cell studies is one of their greatest advantages: one that should not be thrown away when one moves to 3D.
However, many excellent images are produced each year from specimens that have been fixed and stained. (Try http://corn.eng.buffalo.edu/19983d.htm and http://confo.pulmonary.ubc.ca/~dietrich for some examples)
On the second point, it is true that many who have attended in the past have run major microscopy facilities for years, it is also true that others have had only slight prior experience.
We start with Kohler illumination (taught by Ernst Keller from Carl Zeiss) and go on from there. In addition, this year we will add a one-day pre-course meeting with Ernst for those but who really haven't thought seriously about microscopes before, although they may have used them.
The best news is that, because of a misprint in one of our announcements, the deadline for applications has been extended to March 15.
Hope to see you there,
Jim Pawley
PS: For more info: see http://www.cs.ubc.ca/spider/ladic/course/bulletin.html
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
Dear Microcopists: In my recent study of Si dislocations caused by MeV ion implantation and annealing (long time), I found that many linear dislocations in the implanted area. In the attached file (observed along [110] of (001) Si), there two kinds of dislocations. One on {111} planes and running along {112} directions and the other on (001) plan and enlongated along {100} . I am sure there are not {311} dislocations either the Frank loops. But I can't give a defination of these dislocation with a Burgers vector, because my HRTEM is not suitable to do that defect analysis. Hope some one can give me some suggestions about this dislocations (how to call them and what's their possible B vector and they are partial or perfect dislocations). I realy appreciate any information about it. Thank you very much.
{ {dislocation draw.doc} } DAI Jiyan IMRE Singapore
The microscope is (almost) certainly a Reichert Zetopan. This famous stand was made in the Reichert factory in Vienna, Austria and was discontinued in1975. Reichert is now part of the Leica group. I can send you a copy of the manual (multiple page *.tif file, zipped, about 1M).
I have included a (small) picture of the Zetopan from the manual for identification purposes...
Consider yourself lucky: this is one of the best microscopes ever made!
You may want to double check and see if Ar really flows into the chamber 'cause lighter ions MAY NOT be able to knock off atoms off your target. -cy
On Wed, 24 Feb 1999, George Lawton wrote: } Thanks to all who responded to my problem. } But I still have the problem. I got a new cylinder } of Argon. I checked the hoses to make sure they } were not clogged. I double checked } the polarity. My vacuum is good. But when I } turn on my DSM-5 I get a blue arc around the } middle of the cathode. After 3 minutes, I shut } the sputter down but I have no coating on my } sample. I checked the inside of the DSM-5 and } found no loose wires, and the fuses were good. } Any other suggestions would be greatly } appreciated. } } George Lawton } Chief Electron Microscopist } Microscopy and Imaging Service Center } UT Southwestern Medical Center at Dallas } Phone: 214-648-7291 } eMail: George.Lawton-at-email.swmed.edu
In a message dated 2/27/99 12:13:30 AM, jbailon-at-phys-server.phys.polymtl.c= a writes:
} ...In Materials Science, some topics such as the glide of dislocations on } a slip plane, cross-slip of dislocations, pile-ups of dislocations } along grain boundaries, Frank-Read sources, Orowan=92s mechanism are } difficult to teach with only the help of traditional 2D figures in a } book...
I couldn't agree more. Have you seen the diagrams, animations and video cl= ips in our ViMS (Visualizations in Materials Science) CD? (http://www.pws.com/ge/russ.html, PWS Publishing Co) The CD is available f= or both Mac (ISBN 0-534-95052-3) and Windows (ISBN 0-534-95736-6) users. The = CD is in use along with or in some cases in place of a traditional textbook a= t more than 40 universities, worldwide. When our own classes are not in sess= ion at N. C. State University, you can access them via the web at http://vims.ncsu.edu
I have been trying to find the fax or email of Leitz Microscopes. I have been unable to find it after a lot of efforts on the web. Can anybody pass me the relevant info so that I can get in touch with their sales people.
{html} {head} {/head} {BODY bgcolor=3D"#FFFFFF"} {p} {font size=3D2 = color=3D"#000000" face=3D"Arial"} I have been trying to find the fax or = email of Leitz Microscopes. I have been unable to find it after a lot of = efforts on the web. Can anybody pass me the relevant info so that I can = get in touch with their sales people. {br} {br} Thanks {br} {br} Dr.Usman = Rafi {/p} {/font} {/body} {/html} ------=_NextPart_000_01BE628A.433AF9E0--
In a message dated 99-02-27 10:13:27 EST, pulse-at-shoa.net writes:
{ { I have been trying to find the fax or email of Leitz Microscopes. I have been unable to find it after a lot of efforts on the web. Can anybody pass me the relevant info so that I can get in touch with their sales people. } }
Dr. Rafi,
Leitz is now part of the Leica group (along with Reichert-Jung, American Optical, Bausch & Lomb and Cambridge Instruments). The address for Leica will depend on what country you are in, but the easiest way to contact them is at their website:
www.leica.com
} From there you can fill out a request form, and it will be passed to the group which represents Leica in your area.
Best regards,
Bob ****************************** Robert (Bob) Chiovetti, Ph.D. President Microimaging Technologies, Inc. Tucson, Arizona USA Tel./Fax (520) 546-4986 rchiovetti-at-aol.com Manufacturers' Representatives Systems Integrators Analog & Digital Imaging Systems Clinical & Research Microscopy Cytology/Histology/Pathology/EM *******************************
It sounds as though you have a very treasured microscope called a Zetopan. If you can take a picture (even a polaroid) and send it to me, I can confirm that.
If, indeed, this is the instrument you have, I can supply supportive documentation (manual, brochure with decription of parts, etc.). Since this microscope was changed to a dramatically different design in the early 60's (the Polyvar), I suspect that it may be older than you think.
Setting up phase is simple: (I know that this looks like a large number of steps, but once you get the hang of it, it should take less than a minute) 1. Put a well-behaved sample (thin section or cheek smear) on the stage and bring it into focus using the 20x Phase Objective. Set the condenser to brightfield (either BF or H). 2. Close the field iris (on the light port) until you can see it in the field of view. Set up regular Koehler illumination. (If you need a review: a. Focus the image of the field iris using the focus control on the condenser mount b. Center the image of the field iris using the centration screws at about 5 o'clock and 7 o'clock) When you are done, open the field iris so that it is just outside the field of view. 3. Replace one of the eyepieces with the "phase telescope". If you can't find the phase telescope, just remove one of the eyepieces and look way down the tube (this plane is called the Back Focal Plane of the Objective and what goes on there is critical to good imaging). You will see a smokey ring (the phase plate). 4. Rotate the condenser so that the "PH20" aligns with the white marker on the condenser. Look back down the tube/phase telescope. You should see a bright ring. Align the bright ring so that it sits completely and exactly under the smokey ring. I don't have either my manuals or my microscope here at the office, so I am not sure where these alignment controls are. Frequently they are sliders on the rotating plate and/or secondary centering screws on the condenser rather than on its mount. TROUBLE-SHOOTING: If the ring is not the same size, rotate through all the options until you find the best match. If it is not exactly in focus as you look down the Back Focal Plane, use the condenser focus to rack the condenser up or down slightly. If you cannot see the ring, check to make sure that both the field iris and, especially, the condenser aperture iris are open. Most condensers are designed so that this second iris is out of the way when you rotate to the Phase position, but not all. Finally, Phase works best if you insert a good quality green (546nm) filter over the light port. However, the illuminator for the Zetopan may not be powerful enough to supply adequate illumination. In this case you have two options: (a) work without the green filter or (b) contact me privately. I am having my Zetopan illuminator "modernized" shortly and may have some alternatives for you.
For further information on how Phase Contrast works and how to optimize it, see our book "Optimizing Light Microscopy". Details are on our website: MME-Microscopy.com/education
Hope this was helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 06:07 PM 2/26/99 -0500, Sharon Godkin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Finally AFM that compliments the SEM lab.=20 =20 AFM designed for imaging in fluids and under controlled environmental = condition. =20 Capable of force spectroscopy measurements down to the piconeuton of = force (protein folding or antibody - antigen binding force = characterization) with the same instrument that can measure very soft = biological samples at 37C with better that 1 manometer resolution.
Some images http://www.molec.com/biology/index.html
Hands on workshop (bring you own samples) Detail will be posted on = www.molec.com in about a week. In Situ Surface Chemistry 4/27 to 4/28 In Vitro Biology 4/28 to 4/30
If two peaks are overlapped, and you want to distinguish between the two, you can increase the acceleration voltage, right? But how does this help to distinguish between the elements?
My next question concerns Energy Dispersive X-ray Spectroscopy (EDS). The baseline on the EDS spectra is influenced by inelasic scattering of the incident electron beam by the atomic nuclei of the sample, which results in a peaked background. This is called the bremsstrahlung effect. What causes this???
I don't get your first question. You are probably referring to the EDS peak overlap. If so, I don't think increased acceleration voltage can increase the EDS resolution directly. Instead, higher voltage may generate/promote peaks with higher characteristic energy values. These peaks may not overlap and therefore can be distinguished.
A simple answer to your second question is that X-ray photons are not only, though mainly from the sample, but also from everywhere in the chamber because of the elastic/inelastic scattering of electrons.
Hope the above helps.
-cy
On Sat, 27 Feb 1999 Seadoohog-at-aol.com-at-sparc5.microscopy.com wrote:
} I just have a couple of questions regarding SEM. } } If two peaks are overlapped, and you want to distinguish between the two, you } can increase the acceleration voltage, right? But how does this help to } distinguish between the elements? } } My next question concerns Energy Dispersive X-ray Spectroscopy (EDS). The } baseline on the EDS spectra is influenced by inelasic scattering of the } incident electron beam by the atomic nuclei of the sample, which results in a } peaked background. This is called the bremsstrahlung effect. What causes } this??? } } Thanks! } } } }
Does any one have any experience with the $279.00 eyepiece camera that www.imicrovision.com sells? It is a CMOS camera that is read through the printer port.
It claims a 704 X 576 max resolution.=20
I know that CMOS cameras are noisy. But it looks like it might be an adequate illustration tool. If I want resolution I can use the 4X5 camera and do 3 color separations.
Thanks Gordon
Gordon Couger gcouger-at-couger.com Owner PRAG-L PRactical AGriculture List www.couger.com/prag-l Stillwater, OK 405 624-2855 GMT -6:00
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 = HTML//EN"} {!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D4} Does any one have any experience = with the=20 $279.00 eyepiece {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D4} camera that {A=20 href=3D"http://www.imicrovision.com"} www.imicrovision.com {/A} sells? It = is a CMOS=20 camera {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D4} that is read through the printer=20 port. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D4} {/FONT} {/DIV} {DIV} {FONT size=3D4} It claims a 704 X 576 max resolution. {/FONT} {/DIV} {DIV} {FONT size=3D4} {/FONT} {/DIV} {DIV} {FONT size=3D4} I know that CMOS cameras are noisy. But it looks = like it=20 might {/FONT} {/DIV} {DIV} {FONT size=3D4} be an adequate illustration tool. If I want = resolution I can=20 use the {/FONT} {/DIV} {DIV} {FONT size=3D4} 4X5 camera and do 3 color separations. {/FONT} {/DIV} {DIV} {FONT size=3D4} {/FONT} {/DIV} {DIV} {FONT size=3D4} Thanks {/FONT} {/DIV} {DIV} {FONT size=3D4} Gordon {/FONT} {/DIV} {DIV} {FONT size=3D4} {/FONT} {/DIV} {DIV} {FONT size=3D4} Gordon Couger {A=20 href=3D"mailto:gcouger-at-couger.com"} gcouger-at-couger.com {/A} {BR} Owner = PRAG-L=20 PRactical AGriculture List {A=20 href=3D"http://www.couger.com/prag-l"} www.couger.com/prag-l {/A} {BR} Stillw= ater,=20 OK 405 624-2855 = GMT=20 -6:00 {/FONT} {/DIV} {/BODY} {/HTML}
---------- } Van: Barbara Foster {mme-at-map.com} } Aan: Sharon Godkin {GodkinS-at-em.agr.ca} ; microscopy-at-sparc5.microscopy.com } Onderwerp: Re: LM - old Reichert; need manual } Datum: zaterdag 27 februari 1999 19:02
Hi all,
According to Mr. Kappl, head of the service department of the Reichert-factory in Vienna, Austria (now Leica-Reichert), the Zetopan was discontinued in 1975 (personal communication, 1995).
There were two models of the Zetopan: the (older) Zetopan with black finishing, and the (more recent) one with grey finishing. Essentialy these two are the same except that some spare parts have other dimensions (i.e. the UV-block filterholder).
Some spare parts (new/second hand) are still availlable trough some German and Austrian dealers (bought some time ago a new micro flash unit for Zetopan and some filter holders).
Second-hand Zetopans aren't to expensive (if you can find one). Judging from the prices on some online auctions, Zetopan is far less expensive than comparable instruments made by Zeiss and Leitz (Leica). This puzzles me as the Zetopan is IMHO far better than those, spare parts are (more or less: with luck and time) availlable at decent prices (at least in Europe).
One problem is, that the bulbs used in the illuminator "LUX FNI" are difficult to find. It's probably interesting to know that most European microscopes are using either Osram-bulbs or some kind of modified Osram-bulb. This is also the case for the lightsource "LUX FNI". The bulb used is essentialy an Osram nr. 8100 with E14 foot (BTW: this is the same bulb as the one used in another famous microscope: the Leitz Ortholux...) The bulb is soldered in some kind of a "Reichert adapter", thus providing a "precentered and preadjusted" bulb... It's perfectly possible to remove the old bulb, clean up this adapter and fit a regular Osram 8100 in it. That bulb is also far less expensive than the "Reichert bulb"...
At least its true that electrons scatter like light and X-rays with the characteristics of the last reflecting surface (sufficient oomph permitting) are generated. However, EDS detectors have a collimator tube which limits most of the scattered X-rays. In any case the percentage of scattered X-rays is small - to wit: this type of analyses is very location specific and on polished specimen can give very accurate results.
Bremsstrahlung (literally "Braking Radiation") is also called continuum. Peaks sit on top of this background radiation. Lower kV results in a higher continuum as does an average low atomic number specimen. Continuum is the product of electron interaction with the nucleus and not other electrons.
Remember that most EDS/WDS analysis is done at 15kV. KV is varied to identify problem peaks. Perhaps to see another higher peaks or obtain better resolution in the low end of the spectrum. Change from 15kV is there is a good reason only. Disclaimer: ProSciTech has no bremsstrahlung but plenty of continuum on offer. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Sunday, February 28, 1999 12:05 PM, Chao-ying Ni; Office 312 SPL; Phone 1013 [SMTP:ni-at-me.udel.edu] wrote: } } } I don't get your first question. You are probably } referring to the EDS } peak overlap. If so, I don't think increased acceleration } voltage can } increase the EDS resolution directly. Instead, higher } voltage may } generate/promote peaks with higher characteristic energy } values. These } peaks may not overlap and therefore can be distinguished. } } A simple answer to your second question is that X-ray } photons are not } only, though mainly from the sample, but also from } everywhere in the } chamber because of the elastic/inelastic scattering of } electrons. } } Hope the above helps. } } -cy } } } } On Sat, 27 Feb 1999 } Seadoohog-at-aol.com-at-sparc5.microscopy.com wrote: } } } I just have a couple of questions regarding SEM. } } } } If two peaks are overlapped, and you want to distinguish } } between the two, you } } can increase the acceleration voltage, right? But how } } does this help to } } distinguish between the elements? } } } } My next question concerns Energy Dispersive X-ray } } Spectroscopy (EDS). The } } baseline on the EDS spectra is influenced by inelasic } } scattering of the } } incident electron beam by the atomic nuclei of the } } sample, which results in a } } peaked background. This is called the bremsstrahlung } } effect. What causes } } this??? } } } } Thanks! } } } } } } } } }
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We had a similar problem. Eventually the horizontal banding became so bad we thought we needed a new detector or at least a new photomultiplier tube in the detector. The banding was absent when we switched to our Robinson detector. It turned out to be a loose grounding wire in the detector unit itself. See if wiggling the wires coming out of your detector effect your image (e.g. increased banding, lost of contrast, bright white screen), if it does, then the solution is simple.
Roy Nelson Material Testing Laboratory Pennington, NJ 08534 jrnelson-at-nj1.aae.com
P.S. It took forever for your images to load. --------------FFDDB42C033F1E52239010A3 Content-Type: text/x-vcard; charset=us-ascii; name="jrnelson.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for jrnelson Content-Disposition: attachment; filename="jrnelson.vcf"
Does the banding appear in your secondary images? If so there may be a contaminant in the column or on the BSED itself. Slide the BSED out and see if the SED image improves. (I believe Robinson detectors can be moved in or out). If the banding in your SED imaging goes away with the BSED out, there is probably a piece of "dirt" on the forks of the BSED. Try to clean the forks with a blast of nitrogen or other aerosol. Careful if you have to clean the forks of the BSED by wiping them, there is a thin conductive coating on the Lucite forks which comes off easily. Also check the metal strip on top of the BSED forks with an ohmeter to make sure it makes good electrical contact with the detector tube. (As long as you have an ohmeter out, make sure your sample is indeed grounded by measuring the resistance from your coated sample to ground.) Also inspect the bottom of the objective lens for any contaminants. If the banding happens in both SE and BSE modes, any contaminant in the column, or even an ungrounded aperture in the beam path is suspect. Also, make sure the beam emission current is stable. Hope this helps!
Bill Carmichael
"D.Wild" wrote:
} Dear all, } We are imaging pumice sections on glass slides, using BSE detector at 25kv, } condenser lens 4, using a Robinson BSE detector on a Hitachi FE4000 The } idea is to obtain highly contrasted images to transform into black and } white binary images. The problem is we are getting horizontal banding which } looks like a charging problem, although the samples are well coated and } earthed. Could it be due to anything else? The problem has recently } developed and was also apparent on a more conducting sample of some TEM } grids. Attached are some images. } Any comments would be helpful. } } Thank { {horiz_bd.tif} } { {18299_29.tif} } s David Wild } } ------------------------------------------------------------------------ } Name: horiz_bd.tif } horiz_bd.tif Type: TIFF Image (image/tiff) } Encoding: base64 } } Name: 18299_29.tif } 18299_29.tif Type: TIFF Image (image/tiff) } Encoding: base64
Actually the bremsstrahlung, (I think is German for "braking radiation") is from the beam/specimen interaction, and infact, can tell you much about the specimen surface. As I understand it, the bremsstrahlung is the result of the inelastic scattering events that occur when the high energy beam electrons interact with the specimen. As the primary electron (beam) approaches the specimen, it is affected by the electrons and nuclei of the specimen atoms, and it loses energy. The energy lost is released in the form of a photon with a given energy. These events are quite random and the energies of the photons are also quite random, with photon energies ranging from as high as the beam energy, down to small fractions of the beam energy. These photons when detected by the EDS system give a continuous background of energy counts from nearly zero to the accelerating voltage energy. By the way this is a good way for determining the actual high energy (accelerating voltage) that is being emitted by your SEMs "electron gun".
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I just have a couple of questions regarding SEM.
If two peaks are overlapped, and you want to distinguish between the two, you can increase the acceleration voltage, right? But how does this help to distinguish between the elements?
My next question concerns Energy Dispersive X-ray Spectroscopy (EDS). The baseline on the EDS spectra is influenced by inelasic scattering of the incident electron beam by the atomic nuclei of the sample, which results in a peaked background. This is called the bremsstrahlung effect. What causes this???
by mailhost.auckland.ac.nz (8.9.2/8.9.2/8.9.2-ua) with ESMTP id IAA20889 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 1 Mar 1999 08:57:21 +1300 (NZDT) Received: from GLGNOV2/SpoolDir by glgnov2.auckland.ac.nz (Mercury 1.21); 1 Mar 99 08:54:57 +1200 Received: from SpoolDir by GLGNOV2 (Mercury 1.21); 1 Mar 99 08:54:30 +1200
Dear All
My vote would be that ads for the disposal of used equipment by the owner are OK but that ads by dealers are not. I hope that this is the rule.
Ritchie
} I was under the impression that this server was NOT the place for posting } items for sale. } If that is the case then ignore this message. } If I have been wrong or the rules have changed then fellow readers I have } some equipment that is available: } } Sorvall MT-2 ultramicrotomes(2). } Mettler analytical balance H78AR } TMC Micro-G pneumatic table 24x40 } AFM probes } GAST compressor/vacuum pump 60psi. } } All are in excellent working order and are being offered at substancially } reduced prices. } I need the cash. } } Contact; } Steve D'Angelo, Pacifica California, 650.738.2699, 650.400.8063 or email } sdangelo-at-batnet.com
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Hi there listers, Someone in our lab is about to start a project on microvascular casting of varicose veins for SEM viewing. We were wondering what most peoples preference was for some of the commercial polymers and kits available? Your ideas of pro's and con's for these products would be nice too!
Look forward to hearing from you,
Rich.
----------------------------------------------------------------------- Richard Lander Electron Microscopist South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254 mailto:richard.lander-at-stonebow.otago.ac.nz "Southernmost EM Unit in the World!" ------------------------------------------------------------------------
-----Original Message----- } From: "Seadoohog-at-aol.com"-at-sparc5.microscopy.com {"Seadoohog-at-aol.com"-at-sparc5.microscopy.com} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
I am wanting to label some ultra thin sections of marrow embedded in LR White for alkaline phosphatase for viewing on the TEM. Can any-one help me with a method, I have heard that the use of lead citrate is the way to go. My specimens were originally fixed in either 4% paraformaldehyde, or 2% paraformaldehyde and 0.05% glutaraldehyde, then decalcified in 10% EDTA. Do I need to reactivate the enzyme with Mg2+ treatment? If so how? Thanks, Susie Nilsson PhD. Peter MacCallum Cancer Institute Melbourne, Australia s.nilsson-at-pmci.unimelb.edu.au
I appreciate FS posts. For those that do not, it would be easy for the poster to preface their post in the Subject block with FS: xxxxx that way, those that do not want to see FS items can filter them out.
Dear All This is all fun. Lost a bit of skin myself. Latex gloves does help a bit. Home made clamping devices I pressume will help, but I prefer to have a "hands on" onto the sample. For large amounts of samples automation is a option.
} Everett Ramer wrote: } } } } We have a manual metalographic grinding/polishing wheel. The sample } } } being prepared is held in the hand as it is pressed against the rotating } wheel. } } } Our safety people have asked us to provide finger protection for this } device. } } } Does anyone have any solution/suggestion? } } } Everett Ramer } } Everett, } Mary Mager's response, while funny, is actually quite accurate. There are } not too } many ways to protect fingers while properly holding a small (1-1/4", } typically) } sample for grinding/polishing. } } I preface my next remarks by pointing out that I work for a manufacturer of } } metallographic equipment and consumables, and therefore have a financial } interest in solving your problem: } } We at BUEHLER, do offer a simple grinding fixture which might help. This } fixture } is a squat, stainless steel, hollow cylinder with a carbide ring around } it's base. } The sample is clamped within another hollow cylinder seated within the } first. } The two cylinders are threaded, so that the inner can be raised or lowered } with respect to the carbide 'stop' of the outer. Engraved markings allow } material removal in increments as fine as 20microns. While this is not } actually } finger protection, per se, it will allow you to grasp something larger so } that your } fingers are not in such close proximity to the grinding wheel. We also } offer } a motor system which allows the fixture to rotate, in place, on the wheel} Mr. S H Coetzee Electron Microscope Unit Private bag X3 Wits Johannesburg 2050 Tell: +27 11 716 2419 Fax : +27 11 339 3407 E-mail stephan-at-gecko.biol.wits.ac.za
I am curious if anyone is using an automatic section stainer for TEM. If so, what brand are you using. We are using an LKB section stainer (15 years old now). I just wonder if there is anything else in the market. Thanks,
Cora Bucana ******************************************************* Corazon D. Bucana, Ph.D. Department of Cancer Biology U.T. M.D. Anderson Cancer Center 1515 Holcombe Blvd. Box 173 Houston, Texas 77030 Phone: (713) 792-8106 Email:bucana-at-audumla.mdacc.tmc.edu FAX: (713) 792-8747
May 20-22 and May 24-26, 1999 North Carolina State University Raleigh, North Carolina, USA
and
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For detailed information and registration contact Cindy Allen, Dept. of Continuing and Professional Education, N. C. State University, Raleigh, NC 27695-7401, 919-515-8171, fax 919-515-7614, email: Cindy_Allen-at-NCSU.edu
Information is available on-line at the following sites:
Seadoohog-at-aol.com-at-sparc5.microscopy.com wrote: } } My next question concerns Energy Dispersive X-ray Spectroscopy (EDS). The } baseline on the EDS spectra is influenced by inelasic scattering of the } incident electron beam by the atomic nuclei of the sample, which results in a } peaked background. This is called the bremsstrahlung effect. What causes } this??? } } Thanks!
Dear Seadoohog, Brehmsstrahlung, or "braking radiation" is caused by the ac- celeration of the electron by a large mass (nucleus). Both Maxwell's equations and quantum mechanics predict that accelerated charges will give off electromagnetic radiation. The large mass is necessary so that conservation of both energy and momentum can be satisfied. Yours, Bill Tivol
There are some second hand equipment outlets here in the US who occasionally get bits and pieces for Zetopans. One is John Oren, in VT.
Re: illuminators - OptiQuip has suggested some interesting alternatives for an upgrade path. We will be working on this in Apr/May. Anyone interested is welcome to email privately for further info.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 01:41 PM 2/28/99 +0100, Yvan Lindekens wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In the process of trying to produce some Ag coated samples, I instead obtained some silver beads, which would be dandy if that's what I was after ;^ { ...
The specifics were: Denton DV502A evaporator, 4cm of 0.2mm dia Ag wire wrapped around ~1mm dia section of a standard carbon rod, 5x10^-7 Torr. Melting of the wire occured abruptly at well under 20A.
Can anyone suggest conditions or parameters to that will yield evaporation rather than melting???
Later I expect to coat with Al, which like Ag melts and boils at much lower temperatures than Pd and Pt (which are no problem with the above...) so if anyone can provide similar information regarding Al, that would also be helpful.
(This is probably pushing my luck, but if anyone knows any rule of thumb for how thick the films of the above are as a function of conditions & time, that would be super to hear....)
Thanks.
_______________________________________________________ Get your free, private email at http://mail.excite.com/
O.k., I've lost my cheat sheet which had the sizes of Tobbaco Mosaic Virus components, and I can't locate the information presently. Can anyone help me out here? I know the particles are 18.0 nm in diameter "300 nm" in length (o.k. some of them break). What is the diameter of the central core?
What is the spacing between the sprialing sub units?
Thanks!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
Silver has a melting point of 961C. Its vapor pressure is 847C = 1e-8 torr 958C = 1e-6 torr 1105C = 1e-4 torr
An equilibrium vapor pressure of 1e-6 will give ~1 monolayer per second deposition rate (using a kinetic theory of gas model with a sticking coefiicient of 1). Since this will undoubtedly not be an equilibrium situation, you can this value to be an upper limit. So, at the melting point of silver, you will get much less than 0.2nm/sec deposition rate near the sample. Farther away, it will drop as 1/r^2.
Aluminum too, will melt long before you get much evaporation 660C = melting point 677C = 1e-8 torr 821C = 1e-6 torr 1010C = 1e-4 torr
Also, thermal evaporation of Ag, Au, Al will tend to produce metal islands on the sample which can interfere with high mag imaging. Sputtering often will give a smaller grain size.
Cheers, Henk
At 08:40 AM 3/1/99 -0800, you wrote: } {snip} } } Can anyone suggest conditions or parameters to that will yield evaporation } rather than melting??? } } Later I expect to coat with Al, which like Ag melts and boils at much lower } temperatures than Pd and Pt (which are no problem with the above...) so if } anyone can provide similar information regarding Al, that would also be } helpful. } } (This is probably pushing my luck, but if anyone knows any rule of thumb for } how thick the films of the above are as a function of conditions & time, } that would be super to hear....) } } Thanks.
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu An optimist believes that we live in the best of all possible worlds. A pessimist fears that this is true.
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There was a request recently for information on a new EM techniques book. = The book is scheduled to be published in mid March so there is no = information yet other than the chapter titles. I post them here.
Electron microscopy methods and protocols / edited by M.A. Nasser Hajibagheri. (Methods in molecular biology ; v. 117) ISBN 0-89603-640-5 Expected publication date is mid March.
Contents: 1 General Preparation of Material and Staining of Sections ................= ... 1 Heather A. Davies 2 Negative Staining of Thinly Spread Biological Particulates ..............= . 13 J. Robin Harris 3 Preparation of Thin-Film Frozen-Hydrated/Vitrified Biological Specimens for Cryoelectron Microscopy .....................................= .. 31 J. Robin Harris and Marc Adrian 4 The Production of Cryosections Through Fixed and Cryoprotected Biological Material and Their Use in Immunocytochemistry .......... 49 Paul Webster 5 High-Pressure Freezing for Preservation of High Resolution Fine Structure and Antigenicity for Immunolabelling ............................= . 77 Kent McDonald 6 The Application of LR Gold Resin for Immunogold Labeling .............. = 99 J. R. Thorpe 7 Low-Temperature Embedding in Acrylic Resins .............................= . 111 Pierre Gounon 8 Quantitative Aspects of Immunogold Labeling in Embedded and Nonembedded Sections...................................................= ..... 125 Catherine Rabouille 9 Microwave Processing Techniques for Electron Microscopy: A Four-Hour Protocol ......................................................= ............. 145 Rick T. Giberson and Richard S. Demaree, Jr. 10 Electron Microscopic Enzyme Cytochemistry...............................= .... 159 Nobukazu Araki and Tanenori Hatae 11 In Situ Molecular Hybridization Techniques for Ultrathin Sections ....................................................= ................ 167 Jean-Guy Fournier and Fran=E7oise Escaig-Haye 12 Preparation of the Fission Yeast Schizosaccharomyces pombe for Ultrastructural and Immunocytochemical Study ..................... 183 M. A. Nasser Hajibagheri, Kenneth Sawin, Steve Gschmeissner, Ken Blight, and Carol Upton 13 Preparation of Double/Single-Stranded DNA and RNA Molecules for Electron Microscopy ...................................................= ............ 209 M. A. Nasser Hajibagheri 14 Applications of Electron Microscopy for Studying Protein-DNA Complexes..................................................................= .................. 229 Maria Schnos and Ross B. Inman 15 X-Ray Microanalysis Techniques .........................................= ............. 245 A. John Morgan, Carole Winters, and Stephen St=FCrzenbaum
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/apw.htm
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Please note the following job announcement:
ELECTRON MICROSCOPE TECHNICIAN
Lehigh University seeks an Electron Microscope Technician to perform technical duties in support of the Electron Microscopy Laboratory of the Materials Science and Engineering Department. Technician will instruct students in the operation of microscopes and other equipment; maintain and repair instruments; oversee upkeep of the lab; support research professors and students; analyze samples; give tours and demonstrations; supervise students; maintain a safe environment; and other assigned duties. Bachelors degree in physical science and/or 4+ years related work experience required. Candidates should be familiar with electron microscopes, mechanical and electronic equipment, vacuum systems, PC and/or MAC and EDS/WDS systems. Good communication and interpersonal skills are essential.
Lehigh University offers excellent benefits and vacation package. Interested candidates should forward resume to: Deanne Hoenscheid, Materials Science and Engineering, Lehigh University, 5 E. Packer Ave., Bethlehem, PA 18015. EOE. M/F/D/V.
On Mon, 1 Mar 1999 edelmare-at-casmail.muohio.edu-at-sparc5.microscopy.com wrote:
} Date: Mon, 1 Mar 1999 11:42:21 -0500 } From: edelmare-at-casmail.muohio.edu-at-sparc5.microscopy.com } To: microscopy-at-sparc5.microscopy.com } Subject: Size of TMV? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } O.k., I've lost my cheat sheet which had the sizes of Tobbaco Mosaic Virus components, } and I can't locate the information presently. Can anyone help me out here? I know the } particles are 18.0 nm in diameter "300 nm" in length (o.k. some of them break). What } is the diameter of the central core? } } What is the spacing between the sprialing sub units? } } Thanks! } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." }
TMV:
Pitch = 2.3 nm
Don't know about inner diameter of core; try: http://www.ncbi.nlm.nik.gov/ICTVdb/welcome.htm
Let me know the diameter if you find it. S
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I've used W baskets to produce thin films of Au,Cr,Al and Cu in similar evaporators. I would think it would work with Ag too. The liquid metal seems to "hang" in the basket by what, I would guess, are surface tension effects. Any of the major EM houses should have these baskets.
I've also attempted to calculate, a priori, what the film thickness should be and never been too thrilled at the agreement with the result. A quartz crystal thickness monitor can be calibrated for most metals but if you don't have one, you might try using the change in resistance across a glass coverslip (Steere approach) or cross sectioning a series of thin films generated under known evaporation conditions and looking at them in a TEM.
cheers, John John Heckman Michigan State University
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radsci-at-excite.com writes ... } } } In the process of trying to produce some Ag coated samples, I instead } obtained some silver beads, which would be dandy if that's } what I was after } ... } The specifics were: Denton DV502A evaporator, 4cm of 0.2mm } dia Ag wire wrapped around section of a standard carbon rod, } ...
I might suggest, rather than carbon rod, you wrap the Ag wire around tungsten wire which will be wetted. I imagine the Ag on carbon was like water on a duck's back.
BTW, why did a simple reply to this post want to send a message to {"radsci-at-excite.com"-at-sparc5.microscopy.com} ???
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Everett, When I have to do a quick grind on something small that I don't want to put into a mount, I use the "rubber fingers" that secretaries wear. I have found these to be much better than finger cots, which don't last one spin of the wheel. I don't know if this solution will satisfy the "safety" people but I still have finger prints. Dorrance
} ---------- } From: EVERETT RAMER } Sent: Thursday, February 1999 8:23 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Finger Protection during Sample Prep } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have a manual metalographic grinding/polishing wheel. The sample } being prepared is held in the hand as it is pressed against the rotating } wheel. } Our safety people have asked us to provide finger protection for this } device. } Does anyone have any solution/suggestion? } Everett Ramer } }
We have been asked this question by a customer: =========================================== We are looking for a holder for the Wehnelt cap for filament adjustment. AMRAY had one when we went up there but they do not sell one nor could they tell us where to buy one. It looks like a custom job. Do you know what we are talking about? =========================================== I am sure that someone has this hidden away somewhere in their catalog but I have not been able to find it. Any help would be appreciated.
Chuck
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I run a very old probe in a small Geology department. I want to buy a JEOL 733 or later (eg 840A). I would be very pleased to hear from anyone who has one currently for sale or who is planning to sell one this year.
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Dear all We do evaporation of various metals on a old Edwards evaporator. First the Tungstan (W) wire gets heated up under vacuum to clean it from all dirt. Then the requiered metal wire gets wraped around a V bent in the W wire. when heated up the wire will melt and form a melted globule. this will evaporate only after futher heating. }
} } The specifics were: Denton DV502A evaporator, 4cm of 0.2mm dia Ag wire } wrapped around ~1mm dia section of a standard carbon rod, 5x10^-7 Torr. } Melting of the wire occured abruptly at well under 20A. } } Can anyone suggest conditions or parameters to that will yield evaporation } rather than melting??? }
} } (This is probably pushing my luck, but if anyone knows any rule of thumb for } how thick the films of the above are as a function of conditions & time, } that would be super to hear....) } No it does exist.
p = density of the of the coating material in g. ( cm^-3) M = weight of coating material a source (g) R = source samle seperation, (cm) dx = film thickness.
(4 pie R^2) dx p = M
But 1) efficiency factor of evaporative method method is rarely better than 75%. thuss: (4 pie R^2) dx p = M.3/4 this will only work if the sample is directly under the evaporated source
If it is at a angle theta,
3/4.M = (4 pie R^2 dx p)/ sin theta Where theta is the angle between the gounr and the tip of the V.
since Em people do like a really thin coating (nm) for M in g/cm^3 R in cm and dx in nm
M = 4/3. ( 4pie R^2 dx p)/sin theta . 10^-7
For coating material in wire form for a diameter d cm and l lenth (cm0
M = (pie d^2 lp)/4
length of wire required for a coatint thickness of dx cm 1 = 64/3 .[( R^2 dx)/d^2 sin theta)]
hope this does help
Mr. S H Coetzee Electron Microscope Unit Private bag X3 Wits Johannesburg 2050 Tell: +27 11 716 2419 Fax : +27 11 339 3407 E-mail stephan-at-gecko.biol.wits.ac.za
Good day Listmembers A while ago there was a discussion on this list on the preparation of insect eggs for ultra-thin sectioning. Without re-opening the discussion, can somebody that collected all the replies and other experts in this regard please supply me with a general protocol if there is one?
TIA Alan N Hall Laboratory for Microscopy and Micro-Analysis NWII Building University of Pretoria Pretoria 0002 Republic of South Africa Tel: +27-12-420 3896(Office) +27-12-420 2075(Laboratory) Fax: +27-12-362 5150
Dear all, In the evaporation of metal from tungsten wire it is important not to raise the temperature of the wire too fast. If you do this, the metal will melt and drop off in a blob. The thing to do is to raise the temperature until the metal just starts to melt. Then wait until the wire is wetted - you'll see the liquid creeping up the sides of the V. The bulk of the metal will remain as a blob at the apex of the V but will not drop off. Then you can turn the heat up more until the evaporation takes place.
Eric
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
At 03:54 PM 3/1/99, shAf wrote: } } BTW, why did a simple reply to this post want to send a } message to {"radsci-at-excite.com"-at-sparc5.microscopy.com} ??? } I am not sure about the why, but I see that too for various posts. I think it has something to do with the formating of the address, perhaps the use of the quotes.
You know, it could almost be beneficial. You can reply to the sender or the whole list by removing the unwanted part. However, about half the time, I forget to do the editing and my posts get kecked back for me to fix them.
Open Position: Microscopy Characterization Scientist
Essential Functions:
Provide polymer morphology, particle and aesthetics characterization support at GE Plastics in Mt. Vernon, IN. In particular, TEM, SEM, optical microscopy, AFM, particle sizing, surface and other more specialized techniques are used. Leadership in effective problem solving approaches is a must.
Developing Six Sigma quality methods, leveraging information technology to improve problem solving/productivity, and establishing world class techniques appropriate for supporting the businesses are key components to success. This position will have as an area of responsibility the interface role between Analytical Technology and the High Performance Polymer business. The role requires development of effective test plans to support customer issue resolution, Technology Design for Six Sigma programs and manufacturing issues. Broad analytical problem solving skills or the ability to develop these skills.
Qualifications/Requirements:
Ph.D. in Polymer Science, Analytical Chemistry or related areas with expertise in Microscopy characterization of materials. Excellent problem solving and innovation skills are required. Six Sigma Quality Training or the ability to learn these skills. Good interpersonal , teamwork, and communication skills. High energy.
Interested candidates should send their resume to:
Matt Harthcock GE Plastics Global Leader Materials Characterization and Analytical Technology Building 1 1 Lexan Lane Mt. Vernon, Indiana 47620 Phone: 812-831-4776 FAX 812-831-4917
The C.M.I./A.A.B. Descriptions of Plant Viruses No. 151 (Oct. 1975) on tobacco mosaic virus (type strain) gives the following information:
..cylindrical canal of radius c. 2 nm.....
As Sarah Miller states, a pitch of c. 2.3 nm is given.
More infromatiom about particle structure is given: If anyone would like a complete copy of this bulletin, I can mail you a copy. Dont' know if the fax machine will accept this document the way it is folded.
Maureen Petersen
************************************************************************ Maureen Petersen Department of Plant Pathology 1453 Fifield Hall University of Florida
My grandson in the states recently and proudly wrote to his Grand-Dad that he NOW has a microscope and could I send him some samples to look at. He told me that .....
"the microscope is EDU-SCIENCE and the magnifications are 100x 300x 600x."
Since my grandson is 10 years old and egar, I don't want to ask him questions like - Are the objectives aprochromats?. Is anyone on the list familiar with this microscope? I specifically want to know if it's optics are good, and if they are color corrected.
Once I know if he has a decent microscope or a toy that makes things biger and the user blind or crazy, I will better know how to encourage him, and what type of samples and reading matter to send.
Thanks for you help, from a land far away.
Shalom from Jerusalem, Azriel
+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+ Azriel Gorski, Head Fibers and Polymers Laboratory Division of Identification and Forensic Science Israel National Police Jerusalem
azrielg-at-cc.huji.ac.il ICQ User ID No. - 1750739
CHOICE - The enchanted blade, with an edge that shapes lifetimes. Richard Bach RUNNING FROM SAFETY
A friend is someone who knows the song in your heart, and can sing it back to you when you have forgotten the words. +-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+
Hi! I have a reference to two stains/methods for nerves: Koelle's acetyl cholinesterase technique and Winklemann's Silver Impregnation method. I have not been able to find either one. I found a "general" reference to acetyl cholinesterase from the University of Nottingham web page in which they recommend using 'DPK' mounting resin. I have never heard of it--can anyone help me out? Thanks!
Peggy Sherwood Wellman Labs of Photomedicine (224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-3192 (fax)
} -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } We have been asked this question by a customer: } =========================================== } We are looking for a holder for the Wehnelt cap for filament adjustment. } AMRAY had one when we went up there but they do not sell one nor could they } tell us where to buy one. It looks like a custom job. Do you know what we } are talking about? } =========================================== } I am sure that someone has this hidden away somewhere in their catalog but I } have not been able to find it. Any help would be appreciated. } } Chuck
Chuck:
I believe these Wehnelt cap holders are available from Energy Beam Sciences.
Ken Bart
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323 USA kbart-at-hamilton.edu (315) 859-4715
} O.k., I've lost my cheat sheet which had the sizes of Tobbaco Mosaic Virus components, } and I can't locate the information presently. Can anyone help me out here? I know the } particles are 18.0 nm in diameter "300 nm" in length (o.k. some of them break). What } is the diameter of the central core?
The diameter of the central core is 40 A, it is packed with the nucleic acid (RNA). The protein subunits are arranged in a helix containing 16 1/3 subunits per turn ( or 49 subunits per three turns). Each subunit consists of 158 amino acids in a constant sequence.
} What is the spacing between the sprialing sub units? } It is the nucleic acid.
Best regards,
Ming
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * #1074B Dentistry Pharmacy Building * * University Of Alberta. * * Edmonton, Alberta, Canada T6G 2N8 * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Charles writes ... } } We have been asked this question by a customer: } =========================================== } We are looking for a holder for the Wehnelt cap for filament } adjustment. } AMRAY had one when we went up there but they do not sell one } nor could they tell us where to buy one. ...
Just to clarify ... this sounds like a "jig" strictly for holding the wehnelt on the workbench(??) From personal experience I've never seen one of these as a necessary component for the adjustment ... or if it were, it came with the instrument, or was available from the manufacturer. Usually, its primary use is as a heat sink so that the wehnelt assy can be removed while it is hot and to allow it to cool down in a minimum amount of time. Since wehnelts vary in design and geometry I can't imagine this being available thru any other source other than the manufacturer ... which leaves you with what would seem to be a very simple job for a machine shop. (... why do I think this isn't much help? ...)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
} From: Mriglermas-at-aol.com Return-path: {Mriglermas-at-aol.com} To: Woody.N.White-at-mcdermott.com Cc: Mriglermas-at-aol.com
I wish to purchase a picoammeter for specimen current measurement (x-ray analysis work) and am requesting information on models, prices and vendors. Please contact me directly. Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Sonneborn, T.M. (1975), "The Paramecium aurelia complex of fourteen sibling species", Transactions of the American Microscopical Society 94: 155-178.
This is to support a high school student doing a very interesting project on individual variation in protozoa. The local library ordered a copy but estimates 3 weeks delivery.
Can someone out there help me to get it quickly? I will be happy to reimburse reasonable expenses.
Posted by our evaporater, dating from who-knows-when, but certainly long ago, is a sheet of recommended procedures for evaporation different metals. For silver it recommends using a molybdenum or tantalum wire as the filament. It says to wrap a fine wire of silver around the Ta or Mo filament, and then simply evaporate. I recently did a number of successful silver evaporations by fashioning (by hand) a Mo spiral basket and putting a small bead of Ag in it. (I had some Mo wire and Ag beads). I then evaporated the Ag just as I would have evaporated Au from a W basket. It worked fine, but, as always with evaporating from a basket in this way, the thickness uniformity over distances of 2-3 cm. was not very good.
My favorite form of fingertip protection has always (20+ years) been long fingernails. Not talons by any means, but longer than fingertip length. You can feel when a nail is touching without actually losing any skin. Granted, you sometimes generate a strange-looking manicure but an emery board or some 600 grit paper will soon put that to rights. One thing to watch out for: don't let the water to your wheel get so cold as to cause numbness in your fingers. If the water's too cold, you won't know you've ground off your fingerprints until you notice blood on the wheel. And it will really smart when the feeling comes back!
Stephan Coetzee wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear All } This is all fun. Lost a bit of skin myself. Latex gloves does help a } bit. Home made clamping devices I pressume will help, but I prefer } to have a "hands on" onto the sample. For large amounts of samples } automation is a option. } } } } Everett Ramer wrote: } } } } } } We have a manual metalographic grinding/polishing wheel. The sample } } } } being prepared is held in the hand as it is pressed against the rotating } } wheel. } } } } Our safety people have asked us to provide finger protection for this } } device. } } } } Does anyone have any solution/suggestion? } } } } Everett Ramer } } } } Everett, } } Mary Mager's response, while funny, is actually quite accurate. There are } } not too } } many ways to protect fingers while properly holding a small (1-1/4", } } typically) } } sample for grinding/polishing. } } } } I preface my next remarks by pointing out that I work for a manufacturer of } } } } metallographic equipment and consumables, and therefore have a financial } } interest in solving your problem: } } } } We at BUEHLER, do offer a simple grinding fixture which might help. This } } fixture } } is a squat, stainless steel, hollow cylinder with a carbide ring around } } it's base. } } The sample is clamped within another hollow cylinder seated within the } } first. } } The two cylinders are threaded, so that the inner can be raised or lowered } } with respect to the carbide 'stop' of the outer. Engraved markings allow } } material removal in increments as fine as 20microns. While this is not } } actually } } finger protection, per se, it will allow you to grasp something larger so } } that your } } fingers are not in such close proximity to the grinding wheel. We also } } offer } } a motor system which allows the fixture to rotate, in place, on the wheel} } Mr. S H Coetzee } Electron Microscope Unit } Private bag X3 } Wits } Johannesburg } 2050 } Tell: +27 11 716 2419 } Fax : +27 11 339 3407 } E-mail stephan-at-gecko.biol.wits.ac.za
Help- We are having a problem with getting nuclear autofluorescence of tissue culture cells fixed (Paraformaldehyde) and prepped for immunofluorescence. If you fix and mount the cells in glycerol-phenylene diamine with no antibody treatment, they usually look blank immediately, then over time they develop nuclear fluorescence. The nuclear stain is primarily in the fluorescein window and somewhat variable but seems to come up over time. Is it possible that this is coming from a breakdown product of the phenylene diamine we are using as an antifade? An apparently unrelated problem is nuclear background with secondary antibodies alone. If we treat with secondary and look immediately (before the other problem seems to arise), we also get nuclear staining. This seems to vary with the batch of secondary antibody so appears to be non-specific binding of the secondary. It is not blocked by serum etc. Any ideas appreciated. THanks- Dave
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax)
I am trying to dissolve Brefeldin A in Dulbecco's MEM for use in a cell = culture experiment for TEM study. However the drug does not seem to be = readily soluble in the MEM.=20 Can someone give me some help?
Dr. A.P. Alves de Matos Curry Cabral Hospital TEM Unit Portugal
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} I am trying to dissolve Brefeldin A = in=20 Dulbecco's MEM for use in a cell culture experiment for TEM study. = However the=20 drug does not seem to be readily soluble in the MEM. {BR} Can someone = give me=20 some help? {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Dr. A.P. Alves de Matos {BR} Curry = Cabral=20 Hospital {BR} TEM Unit {BR} Portugal {/FONT} {/DIV} {/BODY} {/HTML}
I have been watching what has been going on in relation to your sputter coating problems. I did not look too hard at the early postings so what = I have set out below may be covering old ground, but I hope it may help? M= ay I say that there have been many who have tried so hard to help you, my comments below in no way are critical of their stirling efforts.
Sputter coaters need a RP vacuum of just to the left of dead centre on yo= ur coater's vacuum gauge. It does not matter what the gauge has as its calibration; this setting should work for gold, gold-palladium or platinu= m.
There are three styles of sputter coater low voltage {600volts, medium voltage 1,000 to 1,500volts and a high voltage coater at 1,000 to 3,000volts.
The low voltage coaters do not have a voltage control, simply a depositio= n control which you should set at it's mid point. Set the voltage controls=
for the other two coaters in their mid range too.
As a test specimen use a 2" square piece of white paper weighted down wit= h a stub. Set the target to specimen distance at 5cms. Set the time at tw= o minutes (much too long for normal coating but fine for this test).
Pump the system down YOU DO NOT NEED ARGON! We often run with air, the difference is that argon gas gives a constant coating rate but air, wet o= r dry, gives different coatings. If you are using argon, flush to full lef= t scale once or twice to clean out the system during the pump down.
When you have a vacuum greater than 60% of the scale bleed the gas into t= he system until the pressure holds on the position mentioned above; just to the left of dead centre. When the vacuum is at the correct level, switch=
on the plasma and increase the voltage to the correct level. You should have a plasma and by adjustment of the voltage, deposition, or gas, adjus= t the current to ~20mA. Let the system run for the two minutes.
When the system completes its cycle open it up and take a look at the paper. Using a gold target - very light blue, its working but not very efficiently, pale blue, better but still not good enough, darker blue, still not good enough, blue-grey much more like it, grey to gold tint, it= s working fine.
If you do not have any of these colours the target material may be your problem. Is it one of the metals mentioned below? Even if the target is=
gold on brass and all the gold has gone you will still sputter brass! =
Chromium, aluminium and carbon will not sputter with the very simple systems used for conventional SEM. Much higher specification systems, i= n relation to current and vacuum, are required.
Well I do hope that helps? Please remember that sputter coaters are not 100% efficient when it comes to coating non-conducting specimens. Even a=
good coater will have trouble with rough and porous specimens. You shoul= d never expect to run above 15kV if you have coated this type of specimen.
Well that's it I do hope we can sort out your problem?
Regards
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
If anyone is disposing of this Amray scope, I would appreciate knowing about it so that I could purchase some spare parts and assemblies from it before it is dumped. I am particularly interested in the main 30KV power supply and the CRT power supply, photomult assembly, and any of the circuit boards. (Specific model is 1600T without the digital controls and pushbuttons. All components and assemblies are of interest.)
I am also looking for a Robinson detector for this scope. Pls let me know if you have one and want to dispose of it.
I also have an EDAX 9100 with detector and LSI-11 console that I do not need. suggestions on its disposition are welcome.
Slightly off subject, but I am sure that someone has encountered this obstacle. We have ASCII data files on 8" floppies from various sort of data acquisision hardware/software and are seeking out a way to transfer the data to a useable media 3.5" would suffice, a 8" drive on a system w/internet access would be great, etc. etc.
Does anyone know of someone I could obtain assistance from?
__ _-==-=_,-. /--`' \_-at---at-.-- { Tim (TJ) LaFave Jr. `--'\ \ {___/. Department of Physics \ \\ " / University of North Carolina, Charlotte } =\\_/` { Charlotte, NC 28223 ____ /= | \_|/ _' `\ _/=== \___/ (704)547-3244 `___/ //\./=/~\====\ (704)509-6622 [Hm] \ // / | ===: http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ \/ \\ \\`--| / \\ ---------- +*+ ---------- | _ \\: /==:-\ `.__' `-____/ |--|==: Such that the future be theirs \ \ ===\ :==:`-' to shape and direct. _} \ ===\ /==/ -----------------------------------------------------------------
Hi there there seems to be 2 versions of this book, one was pub in 1988 and one in 1995, what is the diff in them besides the ISBN number?
I have the older one I just got it today and it is a lot of fun was wondering what the newer one might offer..... The one I have has all black and white pictures in it. thanks Ed Sharpe
We are interested in quantifying the amount of sulphur in wool via EDXS analysis in the TEM.
Wool has approximately 5% sulphur within the fibre which varies for different components of the fibre. We would like to be able to mount the sample and standard in the same block and microtome them together. It is not necessary for the material to be fibrous but that would help!
To date we have tried a couple of methods without much success.
In the first instance we attempted to mix sulphur compounds in with the resin to obtain a homogenous material but found that no matter what we did there was always significant variability in the amount of sulphur throughout the resin.
The next attempt used a commercially available fibre which had a known amount of sulphur in it (0.25%) which we embedded in the resin and microtomed. This "sort of" worked OK but the level of sulphur was not quite high enough for our purpose and the uncertainties in our measurements were too high.
So, if anyone has any ideas or a source of materials we could use, I (and the rest of the team) would be most grateful.
Thanks very much in anticipation.
Colin Veitch
Instrumentation Scientist Fibre Structure & Function Group CSIRO Wool Technology PO Box 21 BELMONT Vic 3216 Australia
Tel: +61 (0)3 5246 4000 Fax: +61 (0)3 5246 4811
The information contained in this email message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Wool Technology on +61 3 5246 4000.
If anyone is disposing of this Amray scope, I would appreciate knowing about it so that I could purchase some spare parts and assemblies from it before it is dumped. I am particularly interested in the main 30KV power supply and the CRT power supply, photomult assembly, and any of the circuit boards. (Specific model is 1600T without the digital controls and pushbuttons. All components and assemblies are of interest.)
I am also looking for a Robinson detector for this scope. Pls let me know if you have one and want to dispose of it.
I also have an EDAX 9100 with detector and LSI-11 console that I do not need. suggestions on its disposition are welcome.
I need information about SOPHIA process for aluminium castings ?
Best regards
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager of Structural and Mechanical Research Laboratory str. Zakopianska 73 Call (*48 12) 2618356 (after 8th march) 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
Hi Lesley S. Bechtold, =20 I have read with interest your contribution from 1998-11-27 where you=20 described your treatment of mouse sperm prior to SEM. I want to=20 prepare thrombocytes in a similar way and wonder whether you always=20 apply 1% solutions of poly-L-lysine to render coverslips adhesive. As=20 I understand 0.01% solutions are suitable (and much less expensive...)= =20 for other applications. I deal with moderate amounts of "precious"=20 thrombocytes from knockout mice so I can not play around with=20 conditions. You certainly have extensive experience with this.=20 Any comments would be highly appreciated! =20 All the best, Matthias
I think DPK is a misprint for DPX which is a mounting medium consisting of polystyrene and plasticizers dissolved in xylene and should be widely available from most microscopy suppliers
Dan Hill Biochemistry Department Cambridge University Cambridge UK
On Tue, 2 Mar 1999, Peggy Sherwood wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi! } I have a reference to two stains/methods for nerves: Koelle's acetyl } cholinesterase technique and } Winklemann's Silver Impregnation method. I have not been able to find } either one. I found a "general" reference to acetyl cholinesterase from } the University of Nottingham web page in which they } recommend using 'DPK' mounting resin. I have never heard of it--can } anyone help me out? Thanks! } } Peggy Sherwood } Wellman Labs of Photomedicine (224) } Massachusetts General Hospital } 50 Blossom Street } Boston, MA 02114 } 617-724-4839 (voice mail) } 617-726-3192 (fax) } } } }
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There are 3-4 sulfur standards for the carbon black industry, ASTM D-1619. The concentrations I know about are 0.82%, 1.54%, and 1.93%. These are carbon black powders so combining them in block with your fibers would be a real challenge. I do know a source for these standards if you want to try them.
Chuck Butterick Engineered Carbons, Inc. ______________________________ Reply Separator _________________________________
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Hi All,
We are interested in quantifying the amount of sulphur in wool via EDXS analysis in the TEM.
Wool has approximately 5% sulphur within the fibre which varies for different components of the fibre. We would like to be able to mount the sample and standard in the same block and microtome them together. It is not necessary for the material to be fibrous but that would help!
To date we have tried a couple of methods without much success.
In the first instance we attempted to mix sulphur compounds in with the resin to obtain a homogenous material but found that no matter what we did there was always significant variability in the amount of sulphur throughout the resin.
The next attempt used a commercially available fibre which had a known amount of sulphur in it (0.25%) which we embedded in the resin and microtomed. This "sort of" worked OK but the level of sulphur was not quite high enough for our purpose and the uncertainties in our measurements were too high.
So, if anyone has any ideas or a source of materials we could use, I (and the rest of the team) would be most grateful.
Thanks very much in anticipation.
Colin Veitch
Instrumentation Scientist Fibre Structure & Function Group CSIRO Wool Technology PO Box 21 BELMONT Vic 3216 Australia
Tel: +61 (0)3 5246 4000 Fax: +61 (0)3 5246 4811
The information contained in this email message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Wool Technology on +61 3 5246 4000.
by smtp.uky.edu (8.8.8/8.8.8) with ESMTP id JAA09170 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 Mar 1999 09:50:05 -0500 (EST) Received: from nc.gws.uky.edu ([128.163.193.169]) by pop.uky.edu (8.8.8/8.8.8) with SMTP id JAA13068 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 Mar 1999 09:50:05 -0500 (EST) Message-Id: {2.2.32.19990303145459.006bcec0-at-pop.uky.edu} X-Sender: jmcclin-at-pop.uky.edu X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Woody,
We have a JEOL 35CF for sale that was made in '82. Excellent condition and has backskatter detector. Dual supply Haskris water chiller also available that cooled a TEM as well as this SEM.
We have a stock of spare parts, vacuum tubes, manuals, tools, etc., for an old RCA EMU-3G microscope. Since the scope itself is long gone, these items are destined for that mysterious place where old scope parts go to die. UNLESS someone wants them.
Preserve history! Keep your EMU going! Read the manuals! Now's your chance. All yours for the mere cost of shipping.
Randy Tindall Electron Microscope Core College of Veterinary Medicine University of Missouri - Columbia Phone: 573-882-8304
I made a makeshift but useable holder for filament adjustment for a Amray 1830. I took one of the cardboard boxes that hold the coaters for Polaroid film (about 9"X2"X3/4") and glued two metal washers(1" outer diameter and ~7/16 inner diameter) on top of one another and to the middle of the box. The height of the washers was about 1/8". I then positioned the posts of the filament in the center of the washers to mark the spots. Then I used a needle to poke holes in the cardboard. The Wehnhelt can then be upright and it is easier to center the filament.
Mike Baxter Lehman College mykkb-at-juno.com
___________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com/getjuno.html or call Juno at (800) 654-JUNO [654-5866]
If this is what I think it is it should be available from E Fjeld C., Inc. 978-667-1416. CAA-FB Filament Alignment Base is item discription in my 1985 catalog.
I am posting this on behalf of nearby hospital colleagues:
1. JEOL 1200EX TEM for sale, serviced twice yearly since installation in 1985. In excellent condition. Very light use of up to 65 cases per year.
2. Reichert (now Leica) Ultracut S ultramicrotome, new in 1993, with table, drawers plus LKB Knifemaker 7800. Hardly used and in storage over the last three years.
For information, telephone Mike Sale on England (0)1872 255006. Cash offers to Keith Atkinson on England (0)1872 255006. An e-mail address: tracey.leigh-at-rcht.swest.nhs.uk
Please do not "Reply to sender" - although I would pass replies on to the correct contact!
Keith Ryan Marine Biological Association of the UK Plymouth, UK
I have been asked to research the latest methods for colloidal gold labelling in biological tissues. The researchers would first like to do some diaminobenzidine peroxidase staining on kidney tubules embedded in paraffin to see if the cell membrane antigen and distribution can be detected at the LM-peroxidase level.
Then the project would progress to preparation, sectioning, and labelling of kidney tissue at the TEM level. I have been asked to provide a beginning point of up-to-date protocols that are presently being done.
I am currently scanning Medline and Ovid search engines. Any suggestions would be greatly appreciated.
Sincerely,
Ginger Baker EM Lab Manager OSU-College of Osteopathic Medicine 918-561-8232 lizard-at-osu-com.okstate.edu
I just wanted to say a wholehearted thank you to all of you on this listserver. With your suggestions I have managed to inject an antibody which was directly conjugated to ultrasmall gold particles (thanks to Dr. Jan Leunissen and Electron Microscopy Sciences), perfuse the mouse, and embed the tissues with the specific orientation that I wanted in airtight flat embedding molds to polymerize under UV (thanks to so many people for different aspects of the embedding that I don't have space to name them all), then cut, silver enhance and examine by TEM!
The antibody in question had failed to label anything when the tissue was fixed with ANY fixative but these results were unquestionable and the morphology great!!
My most sincere thanks to you all - I'm off to try new things as our little research lab is closing down. Thanks and Good-bye!
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-med.mcgill.ca
hi, Brefeldin A does not dissolve in media , you need first to dissolve it in either Methanol or DMSO. We dissolve the Brefeldin A in Methanol at a conc of 5mgs/ml and use this as our stock to then be diluted into our buffer or media.
I am currently looking to upgrade my EDS system. I work in a semiconductor manufacturing facility(CMOS) and I currently have a Cambridge S200 SEM with Kevex Delta II.
I have an interest in Oxford, EDAX and a company called EVEX (which I am not familiar with)? Does anyone have experience with these companies - Good or Bad?
I am looking for a light element detector with possibly a WDS for Boron quantification.
Thanks in advance.
Jim Arnold Microelectronics and Technology Center AlliedSignal Electronics and Avionics Systems 9140 Old Annapolis Road Columbia, MD 21045
I have a requirement to process liquid and agar-bound pathogenic bacteria for SEM & TEM. I've been collecting them in cacodylate-buffered 3% glutaraldehyde where the final concentration has been 1.5% for the liquid suspended bacteria. These have been kept in the refrigerator. Does anyone have a protocol(s) that would lead me to SEM and TEM specimens from this point? Have I made a mistake already? My instruments are a Philips EM-400 and JEOL 6300V.
We have an immediate opening for a field service technician in the Los Angeles area. Responsibilities include the repair and maintenance of Scanning Electron Microscopes of a number of manufacturers including JEOL, Hitachi, Leo.
Candidate should have a minimum of three years electronic or instrumentation experience. Please e-mail or fax me at:
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Hello,
I have been asked to research the latest methods for colloidal gold labelling in biological tissues. The researchers would first like to do some diaminobenzidine peroxidase staining on kidney tubules embedded in paraffin to see if the cell membrane antigen and distribution can be detected at the LM-peroxidase level.
Then the project would progress to preparation, sectioning, and labelling of kidney tissue at the TEM level. I have been asked to provide a beginning point of up-to-date protocols that are presently being done.
I am currently scanning Medline and Ovid search engines. Any suggestions would be greatly appreciated.
Sincerely,
Ginger Baker EM Lab Manager OSU-College of Osteopathic Medicine 918-561-8232 lizard-at-osu-com.okstate.edu
We bought some of those chain meshed type gloves that protect fingers pretty well when hand grinding. Our metallographers say you can easily hold the samples and mounts. They look and feel rather clumsy to me tho. But if they work....alrighty then.
There are two special issues of journals on immunogold labeling which appeared recently. Cell Vision, Vol. 4, No. 6 (1997), and Microscopy Research and Technique, Vol. 42, No. 1 (1998). If you have trouble finding a hard copy of that issue of Cell Vision (I think it was from before it was indexed by Medline), you can download all the articles from the journal's web site - if you go to
this is the table of contents for this issue, and it links to the full text articles in pdf format.
Disclaimer: we supply immunogold reagents, and both these special issues contain several articles which feature results obtained with our products (Nanogold).
A good recent reference on post-embedding immunogold labeling is "Colloidal Gold Post-Embedding Immunocytochemistry" (Progress in Histochemistry and Cytochemistry, Vol 29, No 4) by Moise Bendayan (Gustav Fischer, 1995).
Hope this helps,
Rick Powell Nanoprobes, Incorporated.
} } Hello, } } I have been asked to research the latest methods for colloidal gold } labelling in biological tissues. The researchers would first like to do } some diaminobenzidine peroxidase staining on kidney tubules embedded in } paraffin to see if the cell membrane antigen and distribution can be } detected at the LM-peroxidase level. } } Then the project would progress to preparation, sectioning, and labelling } of kidney tissue at the TEM level. I have been asked to provide a } beginning point of up-to-date protocols that are presently being done. } } I am currently scanning Medline and Ovid search engines. Any suggestions } would be greatly appreciated. } } Sincerely, } } Ginger Baker } EM Lab Manager } OSU-College of Osteopathic Medicine } 918-561-8232 } lizard-at-osu-com.okstate.edu
****************************************************************** * NANOPROBES, Incorporated | US Toll-free: (877) 808-2101 * * 25 East Loop Road, Suite 113 | Tel: (516) 444-8815 * * Stony Brook, NY 11790-3350, | Fax: (516) 444-8816 * * USA | rpowell-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
Hello Jim., Since you mentioned the word evex, I suspect you missed the past discussions on this list server regarding them. It centered around their registering URLs with the trade names of all competitors so that any web search by manufacture's name produced the evex web page. Reports were that they were willing to sell the URLs to the principles for 10s of K$, perhaps more. Seems they (someone) did try a defense with an unsigned response. Personally wouldn't have confidence in any claim they made. You can probably find these discussion in the list server archives, ~a year ago.
Bruce Brinson Rice U.
Disclaimer: I use EDS & WDS on occasion. I have absolutely no financial interest & know no one with financial interest in the sales of equipment in this industry & no qualms about bringing this issue back to light.
Arnold, Jim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am currently looking to upgrade my EDS system. I work in a semiconductor } manufacturing facility(CMOS) and I currently have a Cambridge S200 SEM with } Kevex Delta II. } } I have an interest in Oxford, EDAX and a company called EVEX (which I am not } familiar with)? Does anyone have experience with these companies - Good or } Bad? } } I am looking for a light element detector with possibly a WDS for Boron } quantification. } } Thanks in advance. } } Jim Arnold } Microelectronics and Technology Center } AlliedSignal Electronics and Avionics Systems } 9140 Old Annapolis Road } Columbia, MD 21045 } } email: Jim.arnold-at-alliedsignal.com } voice: (410) 964-4118 } fax: (410) 964-5046
I have to prepare pure Al foils for TEM investigation. What I need is (as usual :) a large thin area. What are the best electrolytes for the thinning of Al? Is there anything else to consider (e.g. temperature, ...)? As you see, I have not a lot of experience with this technique. Could anybody recommend a book concerning electrolytical thinning?
TIA,
Petra -------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu Visit our WWW site! http://www.crpcu.lu/~wahlbrin
Don't forget our annual meeting in Gainesville Florida April 8-10
Dealines are fast approaching
For details see the latest issue of the Beam or our web site at :
http://www.biotech.ufl.edu/cbr/sems/sems.html
or cal or fax me below.
Greg Erdos Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
Hi, All We are looking for a Wenhelt Block Filament for filament ( SEM Philips 505 or 501), If you got one we're interested in it ( Free, or for sale , reasonable price)) , please contact us.
Thanks
***************************************************************** Mohamed Belhaj UFR SCIENCES Laboratoir d'Analyse des Solides Surfaces et Interfaces DTI/LASSI EP CNRS 120 BP 1039 Reims 51687 Cedex 2 Tel : 03 26 05 33 27 ou 03 26 05 33 14 Fax : 03 26 05 33 12 ******************************************************************
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Hi,
I have a usual question, which was posted before and, I believe, will be posted again and again. It is about printers for SEM images. So:
- What is the best printer for monochrome SEM/STEM images? - What is the best inexpensive printer for the same images? - What is the best printer for EDS output (color maps, maps+images)?
Thank you,
J.Meen
____________________________________________________________________ Get free e-mail and a permanent address at http://www.netaddress.com/?N=3D= 1
======================================================== FOCUS ON MICROSCOPY 1999
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
We wish to remind you about the upcoming deadline for abstract submission and modification.
The abstract submission and modification deadline is:
!!! Monday, 8 March 1999 --- 15.00 hours Heidelberg Time !!!
The "abstract submission part" of the database will definitely be closed at 15.00 hours, so that it is not possible to submit further abstracts or make any further modifications to the abstracts after this date.
The "registration only part" of the database will remain open until 9 April 1999!!!
Our website now also includes informations about some highlights of the commercial exhibition. This is an extract:
Leica Microsystems GmbH: Two-Photon Confocal System Leica TCS MP L.O.T. Oriel GmbH & Co KG: Nanopositioning Unit and Fiber Laser Molecular Probes: New fluorescent reagents for Cell Biology and Imaging highly - Fluorescent Alexa dyes Olympus Optical Co. GmbH: Confocal Laser-Scanning-Microscope with two-photon excitation Omicron Vakuumphysik GmbH: Scanning Near Field Optical Microscope (SNOM) Visitron Systems GmbH: 2D/3D Fluorescence Imaging System based on Cooled Digital Camera System Wallac Distribution GmbH: Confocal Microscope
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meeting point for developers and users working in these rapidly evolving fields and play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic developments in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
I have never done any material sciences work - I've only ever done biological specimens for EM. Our engineering department has asked me to look at some ball bearings that are failing, as a favour. I know I don't need to fix or dehydrate but do I simply clean them, mount them (using double-sided tape?) and coat them as usual? Or is coating unnecessary? What whould I clean these with? I'm assuming there is grease somewhere that is not good for my vacuum!
Any help would be appreciated!! Thanks in advance.....
Lesley Bechtold
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
Ginger R Baker {lizard-at-osu-com.okstate.edu} CC: Bruce A Benjamin {benjamb-at-osu-com.okstate.edu} , Connie A Hebert {hconnie-at-osu-com.okstate.edu} , William D Meek {meekwd-at-osu-com.okstate.edu} X-Mailer: QuickMail Pro 1.5.3 (Mac) X-Priority: 3 MIME-Version: 1.0 Reply-To: Paul Webster {pwebster-at-mailhouse.hei.org} Content-Type: multipart/alternative; boundary="====50515355485450534849===1"
Reply to: RE: LM and TEM Need help locating the latest = Colloidal Gold T Dear Ginger,
The methods you are looking for can be slit into two sections; (i) = specimen preparation and (ii) colloidal gold labeling. For both, you have = many choices that will be determined by your lab equipment, lab skills = and budget. A good book to have next to you is "Fine structure = Immunocytochemistry" by G. Griffiths (published 1993 by Springer Verlag) = because it will give you advice on all your choices. First, specimen preparation: Your aim is to prepare samples that have = good morphology but also have good accessibility to antigen by the = antibodies and colloidal gold. The best way to achieve this is to obtain = sections. To section, you can either embed in resin (Lowicryl, LR White/= Gold, Unicryl, Monostep), or use cryosections. If you decide to use resin,= will you embed using the PLT method or by freeze substitution? Whatever = your choice, if you are to test the system first by light microscopy then = I would highly recommend using the same system for LM and EM. Do not try = to compare paraffin-embedded, HRP-DAB labeled material with aldehyde-fixed,= sectioned material. =
The gold labeling is also filled with many choices. It is clear that = sequential labeling (ie adding the primary antibody followed by a gold-= labeled secondary) is the best choice. However, what you choose as the = gold-labeled secondary may affect your labeleing pattern. If a = quantitative result is your aim (do you want a signal that reflects the = amount of antigen present?) then you must try not to use signal = amplification. For this either protein A-gold or immunoglobulin-gold as a = single secondary step is best. If you want to get a "strong" signal and = are not too worried about quantitative result you may want to try silver = enhancement of the gold marker.
If I were doing this study, I would fix the kidney, by perfusion, in 4% = formaldehyde (buffered in phosphate buffer). I would then cryoprotect = with sucrose, freeze in liquid nitrogen and embed part of it in Lowicryl = resin through freeze substitution. Other pieces of the cryoprotected, = frozen tissue, I would then section in a cryoultramicrotome. Both the = Lowicryl and cryo sections can be mounted onto glass substrate and labeled = with antibody and fluorescent secondary antibody. You can also label the = sections with the gold conjugated secondary you choose to use for EM and = visualize this for LM by silver enhancement. The advantage of using the = same reagents and tissues for LM is that you can easily test your system = before moving onto EM preparation. Once you have the conditions worked = out, the same blocks you used for LM can be sectioned again for EM (just = mount the sections on grids) and labeled with the same (or similar) = protocols you used for LM.
The protocols you need are all covered in the Griffiths book. Some of the = newer advances have been covered in the a book that had the contents = posted recently.
Contact me if you want more details, resources or references.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm --====50515355485450534849===1 Content-Type: text/html; charset="US-Ascii" Content-Transfer-Encoding: quoted-printable
{HTML} {HEAD} {/HEAD} {BODY} {PRE WIDTH=3D"132"} Reply to: RE: LM and TEM Need help locating the latest = Colloidal Gold T
{/PRE} {FONT FACE=3D"Geneva" SIZE=3D3 = COLOR=3D"#000000"} Dear Ginger, {BR} {BR} The methods you = are looking for can be slit into two sections; = (i) specimen preparation and (ii) colloidal = gold labeling. For both, you have many = choices that will be determined by your = lab equipment, lab skills and budget. A = good book to have next to you is "Fine = structure Immunocytochemistry" by G. = Griffiths (published 1993 by Springer Verlag) = because it will give you advice on all your = choices. {BR} First, specimen preparation: = Your aim is to prepare samples that have = good morphology but also have good accessibility = to antigen by the antibodies and colloidal = gold. The best way to achieve this is to = obtain sections. To section, you can either = embed in resin (Lowicryl, LR White/Gold, = Unicryl, Monostep), or use cryosections. = If you decide to use resin, will you embed = using the PLT method or by freeze substitution? = Whatever your choice, if you are to test = the system first by light microscopy then = I would highly recommend using the same = system for LM and EM. Do not try to compare = paraffin-embedded, HRP-DAB labeled material = with aldehyde-fixed, sectioned material. = {BR} {BR} The gold labeling is also filled = with many choices. It is clear that sequential = labeling (ie adding the primary antibody = followed by a gold-labeled secondary) is = the best choice. However, what you choose = as the gold-labeled secondary may affect = your labeleing pattern. If a quantitative = result is your aim (do you want a signal = that reflects the amount of antigen present?) = then you must try not to use signal amplification. = For this either protein A-gold or immunoglobulin-gold = as a single secondary step is best. If = you want to get a "strong" signal = and are not too worried about quantitative = result you may want to try silver enhancement = of the gold marker. {BR} {BR} If I were doing = this study, I would fix the kidney, by perfusion, = in 4% formaldehyde (buffered in phosphate = buffer). I would then cryoprotect with = sucrose, freeze in liquid nitrogen and embed = part of it in Lowicryl resin through freeze = substitution. Other pieces of the cryoprotected, = frozen tissue, I would then section in a = cryoultramicrotome. Both the Lowicryl and = cryo sections can be mounted onto glass = substrate and labeled with antibody and fluorescent = secondary antibody. You can also label = the sections with the gold conjugated secondary = you choose to use for EM and visualize this = for LM by silver enhancement. The advantage = of using the same reagents and tissues for = LM is that you can easily test your system = before moving onto EM preparation. Once = you have the conditions worked out, the = same blocks you used for LM can be sectioned = again for EM (just mount the sections on = grids) and labeled with the same (or similar) = protocols you used for LM. {BR} {BR} The protocols = you need are all covered in the Griffiths = book. Some of the newer advances have been = covered in the a book that had the contents = posted recently. {BR} {BR} Contact me if you = want more details, resources or references. {/FONT} {FONT = FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#000000"} {BR} {BR} Paul Webster, Ph.D {BR} House = Ear Institute {BR} 2100 West Third Street {BR} Los = Angeles, CA 90057 {BR} phone:213 273 8026 {BR} fax: = 213 413 6739 {BR} e-mail: pwebster-at-hei.org {BR} http://www.hei.org/htm/aemi.htm {/FONT} {/BODY} {/HTML} --====50515355485450534849===1--
Has anyone had any experience with these knives and would you like to comment on their performance relative to "standard" diamond knives?
Tom
Thomas Moninger moninger-at-emiris.iaf.uiowa.edu University of Iowa Central Microscopy Research Facility http://www.uiowa.edu/~cemrf Views expressed are mine.
On Wed, 3 Mar 1999 14:32:22 -0700, jwright-at-dugway-emh3.army.mil wrote... } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
For the TEM samples, fixation in small 1.5 mL eppendorf microfuge tubes works well since you can pellet the samples between solution changes if necessary. From the point you are at right now, you should rinse in your cacodylate-buffer (I used 0.1M cacodylate with 7.5% sucrose), then move on to post-fixation with 1% OsO4 in 0.1M cacodylate (no sucrose) until pellets turn dark brown or black (usually 30-min at room temp). If the pellets are really big, separate them before osmium treatment to make sure the entire pellet is fixed. Rinse in cacodylate buffer (no sucrose), dehydrate in an acetone or ethanol series. I use ethanol because I like to embed in LR White. After 2 changes in 100%, separate the SEM run (for critical point drying and coating) from the TEM samples- move on to resin infiltration right in the eppendorf tubes. I usually do 3:1 (solvent:resin) on a rotator for 1 hr., 1:1, 1:3, 100% times two and cure overnight. Good luck.
As promised, here is a summary of the replies I received to my enquiry about who uses the MSA (formerly EMSA) spectra file format.
Of eight replies, two didn't care for the format or never used it. "My personal preference is not to use MSA format for these purposes. It's way overcomplicated".
The rest of the replies were either reluctant or enthusiastic users who use the format to transfer spectra from one EDX analyser to another, or to the DTSA program, or into a desktop computer. Three would be just as happy to have any method to convert the spectra into Excel format, but three were quite enthusiastic about the ability to exchange spectra between different labs, different EDX programs on commercial systems, even different analysers from the same company of differnet versions.
Thanks to all who replied. I can send the text of all replies to anyone who wishes to see them.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
The best quality will probably always be from a dye sublimation printer. There are Kodak and Codonics machines at the high end for full page output at a cost of about $10K. Kodak also makes a small format (4x5) printer that does 3-color for about $600 and under $1/print. I don't know that they yet offer a black ribbon for true monochrome prints.
For general purpose work, most any of the inkjets will do decent work for both color and monochrome. You may just want to try some at your local computer stores to see what resolution you can get and how fast. We use an Epson 600 ( {$200) for pretty good work at maybe 2 minutes per full page print.
1200 dpi laser printers do a pretty good job for monochrome. We have a Lexmark Optra R. I have seen similar good results from an HP LaserJet 5. It takes a while to transmit the data to the printer, but then they whip out prints fast.
In all cases, the paper can do a lot to improve appearance. You will want some good stuff on had for the best prints.
None of what I have said is peculiar to EM work. You may want to enlist a few others in your area to get a good printer. Happy hunting.
Warren S.
At 09:13 AM 3/4/99 -0600, you wrote: } Hi, } } I have a usual question, which was posted before and, I believe, will be } posted again and again. It is about printers for SEM images. So: } } - What is the best printer for monochrome SEM/STEM images? } - What is the best inexpensive printer for the same images? } - What is the best printer for EDS output (color maps, maps+images)? } } Thank you, } } J.Meen
I am more of a TEM eprson, but I have a suggestion from watching my colleagues. Take an aluminium stub, which would normally go into an SEM, and using a 5 mm drill bit just drill a small , 1-2 mm deep, hole in top so that the ball bearing will sit in it. Before you mount the bearing, wash the bearing in organic solvents first, say two acetone washes and then two ethanol washes, and ultrasonically clean the bearing in an ultrasound bath with both solvents.
Dry it off by putting it into an oven at say 150 C for ten minutes and then mount it onto the stub using silver-dag mounting paint (used by almost everyone here) to fix the bearing onto the stub. If you have access to vacuum coating system with a Radio Frequency inductive plasma ring attachment or anything that can produce an Argon plasma, then put the stub in for ten minutes so that there are no organic residues left. Once it is finished you can put it straight into the SEM knowing that there should be a clean surface to look at. The silver dag paint should earth the bearing to the stub.
I hope this helps.
Jon
-- ***************************************** Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
Yes, that's right - if you are somewhere between Baltimore, Maryland and Sarasota, Florida (with a detour to visit my son in Chapel Hill, NC) I can deliver this lovely item to you in early April. I still don't want to crate it for shipment, but I am driving down to visit my mother and can drop it off on the way if it is not too far off my path. The item in question is an LKB-Huxley Ultra Microtome (model 9801-1) in nice clean condition with the knife holder and a set of chucks and collets. I have about $500 tied up in it which I would like to recover - if you can use it, get in touch and perhaps we can work something out. It just does not fit the work I am doing now, and I need both the $$ and space.
Dear Petra, We have had good success with Al alloys with the Struers Tenupol Twin-jet polisher, using their A-7 electrolyte at about -5 degrees C and 12V. The A-7 is: 100 ml. perchloric acid 700 ml. methanol 200 ml. gycerine The usual precautions with perchloric acid solutions apply. Another I have had recommended to me is: 100 ml. perchloric acid 900 ml. ethanol The solution must be kept at less than 10 degrees C. while polishing, and a voltage range of 7.5V to 20 V, depending on the alloy. I have no experience with pure Al, only commercial Al alloys. A book I recommend for these things is: "Metallography Principles and Practices" by Vander Voort, but I don't know if it is still in print.
You wrote: } } Hello Everyone, } } I have to prepare pure Al foils for TEM investigation. What I need is (as } usual :) a large thin area. What are the best electrolytes for the thinning } of Al? Is there anything else to consider (e.g. temperature, ...)? } As you see, I have not a lot of experience with this technique. Could } anybody recommend a book concerning electrolytical thinning? } } TIA, } } Petra } -------------------------------------------------------------- } Dr. Petra Wahlbring Best of luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
We've been very pleased with the Epson Color Stylus 740 (approx. $280) and Epson Stylus Photo 700 (approx. $250) using Epson's Photo Quality Glossy Film. Image and text quality is very good to excellent.
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center
Research Scientist, Chemistry Williams College
On Thu, 4 Mar 1999, Warren E Straszheim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } The best quality will probably always be from a dye sublimation printer. } There are Kodak and Codonics machines at the high end for full page output } at a cost of about $10K. Kodak also makes a small format (4x5) printer that } does 3-color for about $600 and under $1/print. I don't know that they yet } offer a black ribbon for true monochrome prints. } } For general purpose work, most any of the inkjets will do decent work for } both color and monochrome. You may just want to try some at your local } computer stores to see what resolution you can get and how fast. We use an } Epson 600 ( {$200) for pretty good work at maybe 2 minutes per full page print. } } 1200 dpi laser printers do a pretty good job for monochrome. We have a } Lexmark Optra R. I have seen similar good results from an HP LaserJet 5. It } takes a while to transmit the data to the printer, but then they whip out } prints fast. } } In all cases, the paper can do a lot to improve appearance. You will want } some good stuff on had for the best prints. } } None of what I have said is peculiar to EM work. You may want to enlist a } few others in your area to get a good printer. Happy hunting. } } Warren S. } } At 09:13 AM 3/4/99 -0600, you wrote: } } Hi, } } } } I have a usual question, which was posted before and, I believe, will be } } posted again and again. It is about printers for SEM images. So: } } } } - What is the best printer for monochrome SEM/STEM images? } } - What is the best inexpensive printer for the same images? } } - What is the best printer for EDS output (color maps, maps+images)? } } } } Thank you, } } } } J.Meen } } }
Depending on the suspected failure mechanism, just give them a good soaking in acetone or some kind of degreasing detergent (local auto store) followed by a good rinse and dry. That should remove any grease. I would however talk to the Eng. folks to make sure that your cleaning would not disturb the failure mecahnism or worse.... enhance it. Yes, I would give them a light coat of Au or AuPd. Mounting on any kind of tape (carbon or otherwise) might present some drifting problems in the SEM.......perhaps Ag or C paint would do the trick....that's our solution around here anyway. Hope this helps.
John Staman LSI Logic, Process Analysis Lab, Colorado Springs, CO. 719-573-3282
} -----Original Message----- } From: Lesley S. Bechtold [SMTP:lsb-at-aretha.jax.org] } Sent: Thursday, March 04, 1999 8:53 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Biologist needs help from Material Scientists!! } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } I have never done any material sciences work - I've only ever done } biological specimens for EM. Our engineering department has asked me to } look at some ball bearings that are failing, as a favour. I know I don't } need to fix or dehydrate but do I simply clean them, mount them (using } double-sided tape?) and coat them as usual? Or is coating unnecessary? } What whould I clean these with? I'm assuming there is grease somewhere } that is not good for my vacuum! } } Any help would be appreciated!! Thanks in advance..... } } Lesley Bechtold } } } Lesley S. Bechtold } Supervisor, Biological Imaging } The Jackson Laboratory } 600 Main St. } Bar Harbor, ME 04609 } 207-288-6191 }
Assuming that you want to look at the bearings in an SEM and that the balls are made of steel, the solution is very simple:
Clean the balls to get rid of any grease. A few years back we used Trichlorethylene to degrease materials, but that is environmentally unsafe. There are other possibilities for safer degreasing.
Then you could just use a simple specimen holder and glue the balls to the holder with silver paint or carbon paint, mainly to fix them in place. I am not sure what you mean by double-sided sticky tape, but I would not use that. Who knows what the tape might do when the chamber is pumped down. Since they are steel, you don't have to worry about conductivity, just put them in the chamber, pump down and enjoy.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
} ---------- } From: Lesley S. Bechtold[SMTP:lsb-at-aretha.jax.org] } Sent: Thursday, March 04, 1999 8:53 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Biologist needs help from Material Scientists!! } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Hi, } } I have never done any material sciences work - I've only ever } done } biological specimens for EM. Our engineering department has asked me } to } look at some ball bearings that are failing, as a favour. I know I } don't } need to fix or dehydrate but do I simply clean them, mount them (using } double-sided tape?) and coat them as usual? Or is coating } unnecessary? } What whould I clean these with? I'm assuming there is grease } somewhere } that is not good for my vacuum! } } Any help would be appreciated!! Thanks in advance..... } } Lesley Bechtold } } } Lesley S. Bechtold } Supervisor, Biological Imaging } The Jackson Laboratory } 600 Main St. } Bar Harbor, ME 04609 } 207-288-6191 } }
I need a razor blade holder to use in an old A/O CryoCut vintage 1978. It's a weird one, the blade is mounted to the far right of the holder to accommodate the knife mounting mechanism of the cryostat.
I could also use a razor blade holder for use in an old A/O rotary microtome. This is pretty standard, with the blade in the center of the holder.
I think they are available new, but are pretty expensive given the scope of my project. Anything old but serviceable would work for me. If you have something, maybe we can workout a deal.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I have a few technical papers that deal with electropolishing aluminum using our Model 550 Single Vertical Jet Electropolisher which may be of interest:
1)" Characterization of the Microstructure, Fracture, Morphology and Toughness in Particulate and Short Fibre Reinforced Aluminum Matrix Composites", M.R. Krishnadev et al
2) "Preparation of Iron and Aluminum Samples for Ion Implantation and TEM=
Examination" J.M. McDonald
and the "Bible" of Jet Polishing:
3) "Polishing Methods for Metallic and Ceramic Transmission Electron Microscopy Specimens", B.J. Kestel
Kestel report is 60+ pages and give preparation guidelines for dozens of materials. He has recipes for pure aluminum as well as several aluminum alloys. I would recommend this report for anyone doing electropolishing.=
I have copies of all 3 of these reports as well as a bibliography of over=
200 technical reports mostly dealing with specimen preparation. I would = be pleased to mail you a copy of any or all of these if you think they will = be of use. Please let me know.
DISCLAIMER: South Bay Technology does manufacture the Model 550 Single Vertical Jet ELectropolisher and has a vested interest in promoting its use.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy. Message text written by Petra Wahlbring } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Hello Everyone,
I have to prepare pure Al foils for TEM investigation. What I need is (as=
usual :) a large thin area. What are the best electrolytes for the thinni= ng of Al? Is there anything else to consider (e.g. temperature, ...)? As you see, I have not a lot of experience with this technique. Could anybody recommend a book concerning electrolytical thinning?
J. Meen asks ... } } } ... } } - What is the best printer for monochrome SEM/STEM images?
A dye-sublimation printer with its optional gray scale media.
} - What is the best inexpensive printer for the same images?
A good 600dpi laser printer configured for random dithering, although the next printer would work well too. I would opt for $$ being spent up front on the printer ... it will still cost pennies per page.
} - What is the best printer for EDS output (color maps, maps+images)?
A photographic quality ink jet would work well here ... inexpensive and could as well serve your second purpose, 'cept for speed.
If you're concerned with archiving, fading can be a problem with color ink jets ... moreso than with dye-subs ... so the dye-sub can serve dual purposes, gray scale and color. But, it is a hassle to constantly be changing its purposes from monochrome to color. The dye-sub's color media can be used to print gray scale, but it almost always has a tint because CMY is used, therefore I would not classify a color mode dye-sub as the best printer for your first need.
my $0.02 and cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Lesley writes ... } } } ... Our engineering department has asked me to } look at some ball bearings that are failing, as a favour. I } know I don't } need to fix or dehydrate but do I simply clean them, mount them (using } double-sided tape?) and coat them as usual? Or is coating } unnecessary? } What whould I clean these with? I'm assuming there is grease } somewhere } that is not good for my vacuum! } } ...
I would clean them first with acetone and then with clean ETOH and then dry to remove the small amount of absorbed water. They shouldn't need coating, but I'd mount them with conductive double-stick tape. Should be as easy as that :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I am using a Drukker Diamond Knive at 3 years. It is a excellent knive. I did perfect sections without compression. I cut plant tissues and pollen grains with a hard cell wall.
Rinaldo Pires dos Santos Dept of Botany - Plant Anatomy Laboratory - UFRGS Porto Alegre - RS - Brazil
A student of ours is going to the Washington, D.C. area this summer. I've already told her to stay away from the White House. What she really wants to know is: are any EM/Imaging facilities in the area? She would like to use one to help her with her work while she's there. So drop me a line and I'll pass the info on to her.
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Warren E Straszheim says: } The best quality will probably always be from a dye sublimation printer. } There are Kodak and Codonics machines at the high end for full page output } at a cost of about $10K. Kodak also makes a small format (4x5) printer that } does 3-color for about $600 and under $1/print. I don't know that they yet } offer a black ribbon for true monochrome prints.
1. Several other manufacturers (JVC, Sony, Panasonic) offer dye sublimation printers, marketed for catching/printing video frames. [I'm planning to buy one of those, probably Sony.]
2. I doubt that a term "ribbon" can be applied to such a printer...
} For general purpose work, most any of the inkjets will do decent work for } both color and monochrome. You may just want to try some at your local } computer stores to see what resolution you can get and how fast. We use an } Epson 600 ( {$200) for pretty good work at maybe 2 minutes per full page } print.
Interesting. For monochrome of course inkjets hardly cut it. But for color - it suddenly becomes cost-effective and attractive.
} 1200 dpi laser printers do a pretty good job for monochrome. We have a } Lexmark Optra R. I have seen similar good results from an HP LaserJet 5. It } takes a while to transmit the data to the printer, but then they whip out } prints fast.
1. I have yet to see a laser printer (within $2K price range) that comes close to Optra in reproducing photographs. LaserJet-5 is good, but not that good.
2. I wonder what kind of printer connection you use, and how large your files are. My printers are on Ethernet, and 16MB files "fly in" in less than half-a-minute.
} In all cases, the paper can do a lot to improve appearance. You will want } some good stuff on head for the best prints.
(:-) -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
Additional to the useful replies relating to the preparation of ball bearings, I would like to add a little regarding the microscopy: Small, clean and undamaged ball bearings are a good instructive aid to explain SEM functioning. This may be of some use to Jonathan when viewing those bearings and to anybody trying to explain some SEM effects. 1 At high kV (say above 20) you may have trouble seeing anything, because fine surface structure and dirt will be invisible. 2 At low kV (say 10 and below) such surface structures will be visible. 3 At low powers, regardless of kV, the ball bearing will appear like a disk, but the outer part of the disk is brighter. This nicely shows that in secondary mode, brightness almost entirely is increased with the angle of incidence. Being a sphere, tilt has no effect on the brightness distribution over that image. 4 A BS detector mounted at an angle (whereas the Robinson and some others are vertical) will make the distinction that the specimen is not a disk but a sphere, because the BS electrons directed away from the detector leave a shadow. 5 A similar effect is produced when the bias current of the secondary detector is turned off and the condenser current is turned down. This floods the secondary detector's scintillator with backscattered electrons and produces a BS image quite suitable for low powers.
Nice teaching exercise, but its useful to know about these effects when actually looking at those bearings. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Friday, March 05, 1999 6:11 AM, Jonathan Barnard [SMTP:J.Barnard-at-bristol.ac.uk] wrote: } } I am more of a TEM eprson, but I have a suggestion from } watching my } colleagues. } Take an aluminium stub, which would normally go into an } SEM, and using a } 5 mm drill bit just drill a small , 1-2 mm deep, hole in } top so that the } ball bearing will sit in it. Before you mount the bearing, } wash the } bearing in organic solvents first, say two acetone washes } and then two } ethanol washes, and ultrasonically clean the bearing in an } ultrasound } bath with both solvents. } } Dry it off by putting it into an oven at say 150 C for ten } minutes and } then mount it onto the stub using silver-dag mounting } paint (used by } almost everyone here) to fix the bearing onto the stub. If } you have } access to vacuum coating system with a Radio Frequency } inductive plasma } ring attachment or anything that can produce an Argon } plasma, then put } the stub in for ten minutes so that there are no organic } residues left. } Once it is finished you can put it straight into the SEM } knowing that } there should be a clean surface to look at. The silver dag } paint should } earth the bearing to the stub. } } I hope this helps. } } Jon } } -- } ***************************************** } Jonathan Barnard } } Microstructural Physics, } H.H.Wills Physics Laboratory, } University of Bristol, } Tyndall Avenue, } Bristol BS8 1TL. } } 0117 928 9000 ext 8750 } } ***************************************** } }
The string concerning preparation of bacteria touched on delayed second fixation. This is worth a separate discussion on delayed second (usually Os) fixation: Sabatini, first to publish GA as a fixative, also published that Os fixation could be delayed by several months. That seems true for some tissues, which show no ill-effects when compared with the usual, immediate double fixation. However, we found in the lab that other tissues are sensitive to that delay. I used to run a couple of busy service labs and cannot remember specifically which tissues and what structures were affected. It would be interesting to know when delayed double fixation is acceptable and others may have experience to share. I believe that specimen preparation for SEM is never affected by delayed second fixation. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Thursday, March 04, 1999 11:14 PM, Tina Schwach [SMTP:tschwach-at-tc.umn.edu] wrote: } } On Wed, 3 Mar 1999 14:32:22 -0700, } jwright-at-dugway-emh3.army.mil wrote... } } I have a requirement to process liquid and agar-bound } } pathogenic } } bacteria for SEM & TEM. I've been collecting them in } } cacodylate-buffered 3% glutaraldehyde where the final } } concentration has } } been 1.5% for the liquid suspended bacteria. These have } } been kept in } } the refrigerator. Does anyone have a protocol(s) that } } would lead me to } } SEM and TEM specimens from this point? Have I made a } } mistake already? My } } instruments are a Philips EM-400 and JEOL 6300V. } } } } John Wright } } Microbiologist } } } } West Desert Test Center } } Dugway, UT } } } } } } John, } I have stored samples in primary fixative (glut-para- } ruthenium red in } cacodylate) for several weeks, even months and they appear } to be fine. } For SEM, you may want to place (dry) your samples on some } kind of surface, } ie stainless steel chips, so you'll be able to view them. } You'll have to } do this at the end anyway. You can even place them on } nucleopore filter } membranes. Depending on what you want to see, the agar } strands can get in } the way. } } For the TEM samples, fixation in small 1.5 mL eppendorf } microfuge tubes } works well since you can pellet the samples between } solution changes if } necessary. From the point you are at right now, you } should rinse in your } cacodylate-buffer (I used 0.1M cacodylate with 7.5% } sucrose), then move on } to post-fixation with 1% OsO4 in 0.1M cacodylate (no } sucrose) until pellets } turn dark brown or black (usually 30-min at room temp). } If the pellets are } really big, separate them before osmium treatment to make } sure the entire } pellet is fixed. Rinse in cacodylate buffer (no sucrose), } dehydrate in an } acetone or ethanol series. I use ethanol because I like } to embed in LR } White. After 2 changes in 100%, separate the SEM run (for } critical point } drying and coating) from the TEM samples- move on to resin } infiltration } right in the eppendorf tubes. I usually do 3:1 } (solvent:resin) on a } rotator for 1 hr., 1:1, 1:3, 100% times two and cure } overnight. } Good luck. } } } } }
The peaks have suddenly started to move around a bit, although the resolution stays good. The problem comes and goes, in its good times the standard deviation of the position of the Co Ka peak is about 0.4 eV (10 determinations), but sometimes it's about 10 or even 20 eV. My thinking is that if it were the detector, the resolution would be degrading, but it's not, so maybe the culprit is the (analog) pulse-processor. Anyone got any thoughts on how to pin it down as being either a the detector b the subsequent signal-processing stuff, eg pulse-proc? Could I successfully test the pulse-proc with a ramp from a standard signal generator, or would that signal, being relatively clean compared with that from a detector, not really check it out rigorously enough?
cheers (well, I try)
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
======================================================== FOCUS ON MICROSCOPY 1999
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
We wish to remind you about the upcoming deadline for abstract submission and modification.
The abstract submission and modification deadline is:
!!! Monday, 8 March 1999 --- 15.00 hours Heidelberg Time !!!
The "abstract submission part" of the database will definitely be closed at 15.00 hours, so that it is not possible to submit further abstracts or make any further modifications to the abstracts after this date.
The "registration only part" of the database will remain open until 9 April 1999!!!
Our website now also includes informations about some highlights of the commercial exhibition. This is an extract:
Leica Microsystems GmbH: Two-Photon Confocal System Leica TCS MP L.O.T. Oriel GmbH & Co KG: Nanopositioning Unit and Fiber Laser Molecular Probes: New fluorescent reagents for Cell Biology and Imaging highly - Fluorescent Alexa dyes Olympus Optical Co. GmbH: Confocal Laser-Scanning-Microscope with two-photon excitation Omicron Vakuumphysik GmbH: Scanning Near Field Optical Microscope (SNOM) Visitron Systems GmbH: 2D/3D Fluorescence Imaging System based on Cooled Digital Camera System Wallac Distribution GmbH: Confocal Microscope
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meeting point for developers and users working in these rapidly evolving fields and play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic developments in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
Others have offered their suggestions but may I add a friendly warning, o= r offer a bit of fun?
Ball bearings are a great eye opener for the novice SEM operator. Fix a small ball bearing to a stub with a NON conducting adhesive. Run the microscope at 20 to 25kV for a few minutes and the ball will charge like=
mad. let it charge { one of the very few reasons for using such high kV := -) }. NOW drop to 2kV and wonder of wonders you will see the inside of the=
specimen chamber. Move the ball around and change the magnification and you will be able to view the chamber in detail. Take a look at the aperture of the final lens, the EDX detector face, or the backscattered detector surface. Great fun if you know what is happening, totally confusing if you did not do it deliberately.
This "problem" may occur in other circumstances where, after using high k= V and charging a specimen, moving to a low kV the charge does not go away b= ut simply acts as a reflector of the beam. Double sided tape which is non conducting is the biggest culprit. It should never be used in SEM where = we now have available carbon double sided media which is great for a quick f= ix with lighter weight specimens.
Have fun?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
As an assistance to people who have day to day problems with their equipment the listserver provides a very valuable service and I am often pointing people in this direction to solve their problems.
My area of knowledge is in the operation and maintenance of scanning and transmission electron microscopes so I take an interest in and discussion= s in this area.
Of late I have noticed a tendency for replies to become very complicated,=
so much so that the person asking the question would in my mind only beco= me bemused by the reply. Should we not build the knowledge base step by ste= p rather than leap in with solutions far distant from the actual question/problem?
As a made-up example -
Q - What do I do if I do not have a good quality image on the screen in m= y TEM?
A - Change the phosphor.
Have you seen answers like this?
I like many of you reading this I would not like to inhibit anyone sendin= g in a reply to a question, but should they not place their reply on a leve= l?
Example
A - Assuming that you have prepared the specimen correctly, have the correct alignment, kV and apertures, have the condenser lenses set correctly and are in a well darkened room and have become dark adapted - = if you notice your screen is not as bright as a colleagues TEM perhaps you should consider changing the screen as they fade due to beam damage and contamination?
I look forward to being put in my place! =
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
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The string concerning preparation of bacteria touched on delayed second fixation. This is worth a separate discussion on delayed second (usually Os) fixation: Sabatini, first to publish GA as a fixative, also published that Os fixation could be delayed by several months. That seems true for some tissues, which show no ill-effects when compared with the usual, immediate double fixation. However, we found in the lab that other tissues are sensitive to that delay. I used to run a couple of busy service labs and cannot remember specifically which tissues and what structures were affected. It would be interesting to know when delayed double fixation is acceptable and others may have experience to share. I believe that specimen preparation for SEM is never affected by delayed second fixation. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Thursday, March 04, 1999 11:14 PM, Tina Schwach [SMTP:tschwach-at-tc.umn.edu] wrote: } } On Wed, 3 Mar 1999 14:32:22 -0700, } jwright-at-dugway-emh3.army.mil wrote... } } I have a requirement to process liquid and agar-bound } } pathogenic } } bacteria for SEM & TEM. I've been collecting them in } } cacodylate-buffered 3% glutaraldehyde where the final } } concentration has } } been 1.5% for the liquid suspended bacteria. These have } } been kept in } } the refrigerator. Does anyone have a protocol(s) that } } would lead me to } } SEM and TEM specimens from this point? Have I made a } } mistake already? My } } instruments are a Philips EM-400 and JEOL 6300V. } } } } John Wright } } Microbiologist } } } } West Desert Test Center } } Dugway, UT } } } } } } John, } I have stored samples in primary fixative (glut-para- } ruthenium red in } cacodylate) for several weeks, even months and they appear } to be fine. } For SEM, you may want to place (dry) your samples on some } kind of surface, } ie stainless steel chips, so you'll be able to view them. } You'll have to } do this at the end anyway. You can even place them on } nucleopore filter } membranes. Depending on what you want to see, the agar } strands can get in } the way. } } For the TEM samples, fixation in small 1.5 mL eppendorf } microfuge tubes } works well since you can pellet the samples between } solution changes if } necessary. From the point you are at right now, you } should rinse in your } cacodylate-buffer (I used 0.1M cacodylate with 7.5% } sucrose), then move on } to post-fixation with 1% OsO4 in 0.1M cacodylate (no } sucrose) until pellets } turn dark brown or black (usually 30-min at room temp). } If the pellets are } really big, separate them before osmium treatment to make } sure the entire } pellet is fixed. Rinse in cacodylate buffer (no sucrose), } dehydrate in an } acetone or ethanol series. I use ethanol because I like } to embed in LR } White. After 2 changes in 100%, separate the SEM run (for } critical point } drying and coating) from the TEM samples- move on to resin } infiltration } right in the eppendorf tubes. I usually do 3:1 } (solvent:resin) on a } rotator for 1 hr., 1:1, 1:3, 100% times two and cure } overnight. } Good luck. } } } } }
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Additional to the useful replies relating to the preparation of ball bearings, I would like to add a little regarding the microscopy: Small, clean and undamaged ball bearings are a good instructive aid to explain SEM functioning. This may be of some use to Jonathan when viewing those bearings and to anybody trying to explain some SEM effects. 1 At high kV (say above 20) you may have trouble seeing anything, because fine surface structure and dirt will be invisible. 2 At low kV (say 10 and below) such surface structures will be visible. 3 At low powers, regardless of kV, the ball bearing will appear like a disk, but the outer part of the disk is brighter. This nicely shows that in secondary mode, brightness almost entirely is increased with the angle of incidence. Being a sphere, tilt has no effect on the brightness distribution over that image. 4 A BS detector mounted at an angle (whereas the Robinson and some others are vertical) will make the distinction that the specimen is not a disk but a sphere, because the BS electrons directed away from the detector leave a shadow. 5 A similar effect is produced when the bias current of the secondary detector is turned off and the condenser current is turned down. This floods the secondary detector's scintillator with backscattered electrons and produces a BS image quite suitable for low powers.
Nice teaching exercise, but its useful to know about these effects when actually looking at those bearings. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Friday, March 05, 1999 6:11 AM, Jonathan Barnard [SMTP:J.Barnard-at-bristol.ac.uk] wrote: } } I am more of a TEM eprson, but I have a suggestion from } watching my } colleagues. } Take an aluminium stub, which would normally go into an } SEM, and using a } 5 mm drill bit just drill a small , 1-2 mm deep, hole in } top so that the } ball bearing will sit in it. Before you mount the bearing, } wash the } bearing in organic solvents first, say two acetone washes } and then two } ethanol washes, and ultrasonically clean the bearing in an } ultrasound } bath with both solvents. } } Dry it off by putting it into an oven at say 150 C for ten } minutes and } then mount it onto the stub using silver-dag mounting } paint (used by } almost everyone here) to fix the bearing onto the stub. If } you have } access to vacuum coating system with a Radio Frequency } inductive plasma } ring attachment or anything that can produce an Argon } plasma, then put } the stub in for ten minutes so that there are no organic } residues left. } Once it is finished you can put it straight into the SEM } knowing that } there should be a clean surface to look at. The silver dag } paint should } earth the bearing to the stub. } } I hope this helps. } } Jon } } -- } ***************************************** } Jonathan Barnard } } Microstructural Physics, } H.H.Wills Physics Laboratory, } University of Bristol, } Tyndall Avenue, } Bristol BS8 1TL. } } 0117 928 9000 ext 8750 } } ***************************************** } }
We lost our Ar/Kr laser very early in its life. (it was still under warranty) A bit of advice was given to us from Zeiss regarding our Ar/Kr laser. That is to run it at 50% power rather than full blast all the time. It seems to be much better now.
} Just to give an opposite example about the laser, } } I'm running an LSM 410 equipped with an Omnichrome Ar/Kr 488/568/647 laser } since 5 years (1750 hours) and i had no problem yet ! Maybe it's just luck } though... } } } At 05:01 PM 25/2/99 -0800, you wrote: } } As for my own experience, I found the 568nm Kr laser sadly unreliable, we } had to } change it twice in the past year. } } Mr. S H Coetzee Electron Microscope Unit Private bag X3 Wits Johannesburg 2050 Tell: +27 11 716 2419 Fax : +27 11 339 3407 E-mail stephan-at-gecko.biol.wits.ac.za
A number of suggestions have been made about the preparation and observation of failed ballbearings. These will certainly give good images and may well be enough to solve the problem, but if the fault lies in the alloy rather than in the physical state of the balls, it will not show up. You might have to consider embedding the bearing in resin and polishing a flat on it. A thin coat of carbon is needed unless you use a conductive resin. Then the BSE image (and EDX if possible) will reveal the internal structure of the alloy.
Eric
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
} The peaks have suddenly started to move around a bit, although the } resolution stays good. } The problem comes and goes, in its good times the standard deviation } of the position of the Co Ka peak is about 0.4 eV (10 } determinations), but sometimes it's about 10 or even 20 eV. } My thinking is that if it were the detector, the resolution would be } degrading, but it's not, so maybe the culprit is the } (analog) pulse-processor. Anyone got any thoughts on how to pin it } down as being either } a the detector } b the subsequent signal-processing stuff, eg pulse-proc? } Could I successfully test the pulse-proc with a ramp from a standard } signal generator, or would that signal, being relatively clean } compared with that from a detector, not really check it out } rigorously enough?
One thing to note is if your movement is due to gain or offset drifts. If offset drift then both a low peak and high peak will move the same amount. If gain the low will move a little, high will move much more.
Sometimes the gain and offset pots on the pulse processor get "dirty". Run them up and down a few times or clean with electronic cleaner spray.
Disconnect and reconnect every connector. The contacts can get "dirty" over time. The physical active of disconnecting/reconnecting will clean the contacts.
You can test your MCA by using a sliding pulser. We use a Berkley Nucleonics Corp (BNC) GL-3 pulse generator to test every MCA that we ship. It's the only way to really know how well the MCA is working. It's about $5k, not cheap. You really need a very good pulser to test linearity. 10 to 20eV is a really small voltage difference.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
Is it OK to (thin) section the Falcon cell culture inserts in cross section, and would anyone who's done this please contact me directly? I have a question or two about the best way to embed to maximize cell numbers. (I think this has been discussed before.) Thank you. Grace
In the spirit recently suggested by Steve Chapman, try disturbing the cables and processor box. If this affects the peak positions or resolution, there's a good chance you have a loose or dirty connector. Try reseating the PC boards in the processor box.
Larry Thomas Washington State University
---------- From: Ritchie Sims Sent: Friday, March 5, 1999 1:28 PM To: microscopy-at-Sparc5.Microscopy.Com Subject: EDS trouble-shooting
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Dear Experts
The peaks have suddenly started to move around a bit, although the resolution stays good. The problem comes and goes, in its good times the standard deviation of the position of the Co Ka peak is about 0.4 eV (10 determinations), but sometimes it's about 10 or even 20 eV. My thinking is that if it were the detector, the resolution would be degrading, but it's not, so maybe the culprit is the (analog) pulse-processor. Anyone got any thoughts on how to pin it down as being either a the detector b the subsequent signal-processing stuff, eg pulse-proc? Could I successfully test the pulse-proc with a ramp from a standard signal generator, or would that signal, being relatively clean compared with that from a detector, not really check it out rigorously enough?
cheers (well, I try)
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
American Soc List {microscopy-at-Sparc5.Microscopy.Com} X-Mailer: QuickMail Pro 1.5.3 (Mac) X-Priority: 3 MIME-Version: 1.0 Reply-To: Paul Webster {pwebster-at-mailhouse.hei.org} Content-Type: multipart/alternative; boundary="====56495250495150514856===1"
Reply to: RE: Complications With regard to this comment By Steve Chapman, I agree that the discussion = aspect of the listserver is slowly being erroded and would sometimes like = to see the more detailed, carfully considered answers being posted again. = I use the server as a source of assistance but also find that by reading = discussions on subjects I know little about, I can actually learn = something. This sort of passive learning is useful in a busy lab environme= nt.
I am not sure we can hold the solution providers totally responsible for = this errosion either. It is sometimes difficult to know exactly the = problem, the skill of the poster or the environmentally limiting factors = from such brief postings as in the example given by Steve ("What do I do = if I do not have a good quality image on the screen in my TEM?"). Why is = it that some questions are posted in almost annotated form from posters = who do not even provide us with an identification? =
The tendency for discussions to be carried out "off-line" in more detail = is not very beneficial either. It is true that some of the really = detailed discussions, and perhaps some of the comments about particular = products, are best covered in a more private setting, but it would be = great to see edited summaries and final results.
I know writing is tough and it takes some time to write but by putting = more effort into this skill we could improve our list. I am not getting = at non-English writers here but at anyone who writes messages which are so = brief that relevant information is omitted. The more we practice our = writing, the better we get (hopefully). =
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm http://www.hei.org/htm/apw.htm
Steve Chapman wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
{HTML} {HEAD} {/HEAD} {BODY} {PRE WIDTH=3D"132"} Reply to: RE: Complications
{/PRE} {FONT FACE=3D"Geneva" SIZE=3D3 = COLOR=3D"#000000"} With regard to this comment By Steve = Chapman, I agree that the discussion aspect = of the listserver is slowly being erroded = and would sometimes like to see the more = detailed, carfully considered answers being = posted again. I use the server as a source = of assistance but also find that by reading = discussions on subjects I know little about, = I can actually learn something. This sort = of passive learning is useful in a busy = lab environment. {BR} {BR} I am not sure we can = hold the solution providers totally responsible = for this errosion either. It is sometimes = difficult to know exactly the problem, the = skill of the poster or the environmentally = limiting factors from such brief postings = as in the example given by Steve ("What = do I do if I do not have a good quality = image on the screen in my TEM?"). Why = is it that some questions are posted in = almost annotated form from posters who do = not even provide us with an identification? = {BR} {BR} The tendency for discussions to be = carried out "off-line" in more = detail is not very beneficial either. It = is true that some of the really detailed = discussions, and perhaps some of the comments = about particular products, are best covered = in a more private setting, but it would = be great to see edited summaries and final = results. {BR} {BR} I know writing is tough and = it takes some time to write but by putting = more effort into this skill we could improve = our list. I am not getting at non-English = writers here but at anyone who writes messages = which are so brief that relevant information = is omitted. The more we practice our writing, = the better we get (hopefully). {BR} {BR} Regards, {BR} {/FONT} {FONT = FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#000000"} {BR} {/FONT} {FONT FACE=3D"Geneva" SIZE=3D3 COLOR=3D"#000000"} Paul = Webster, Ph.D {BR} House Ear Institute {BR} 2100 = West Third Street {BR} Los Angeles, CA 90057 {BR} phone:213 = 273 8026 {BR} fax: 213 413 6739 {BR} e-mail: pwebster-at-hei.org {BR} http://www.hei.org/htm/aemi.htm {BR} http://www.hei.org/htm/apw.htm {/FONT} {FONT = FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#000000"} {BR} {BR} {BR} {/FONT} {FONT FACE=3D"Geneva" = SIZE=3D3 COLOR=3D"#000000"} Steve Chapman wrote: {/FONT} {FONT FACE=3D"Geneva"= = SIZE=3D1 COLOR=3D"#000000"} {BR} >-----------------------------------------------------------------------= - {BR} >The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America {BR}
id xma019702; Fri, 5 Mar 99 13:37:30 -0500 Received: by eastman.com id AA1958416 (5.67b/IDA-1.5 for microscopy-at-Sparc5.Microscopy.Com); Fri, 5 Mar 1999 13:37:57 -0500 Received: from ntmcon02.emn.com by eastman.com with SMTP id AA2032906 (5.67b/SMI-4.1 for {microscopy-at-Sparc5.Microscopy.Com} ); Fri, 5 Mar 1999 13:37:57 -0500 Received: by ntmcon02.emn.com with Internet Mail Service (5.5.2232.9) id {FZ4RM9PM} ; Fri, 5 Mar 1999 13:38:06 -0500 Message-Id: {E788D998AAC8D1119EC10000F8CD1E8A5B42BA-at-ntmail23.emn.com}
Regarding moving peaks in EDS, we once experienced a problem caused by the video monitor on the EDS unit being too close to the wires coming from the detector. Our problem became quite obvious when the resolution began to degrade. Moving the wires fixed the problem. Perhaps your problem is being caused by electromagnetic fields.
Dennis B. Barr (dennbarr-at-eastman.com) Physical Chemistry Research Laboratory Physical & Analytical Chemistry Research Division Eastman Chemical Company Kingsport, TN 37662-5150
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} The peaks have suddenly started to move around a bit, although the } resolution stays good. } The problem comes and goes, in its good times the standard deviation } of the position of the Co Ka peak is about 0.4 eV (10 } determinations), but sometimes it's about 10 or even 20 eV. } My thinking is that if it were the detector, the resolution would be } degrading, but it's not, so maybe the culprit is the } (analog) pulse-processor. Anyone got any thoughts on how to pin it } down as being either } a the detector } b the subsequent signal-processing stuff, eg pulse-proc? } Could I successfully test the pulse-proc with a ramp from a standard } signal generator, or would that signal, being relatively clean } compared with that from a detector, not really check it out } rigorously enough?
One thing to note is if your movement is due to gain or offset drifts. If offset drift then both a low peak and high peak will move the same amount. If gain the low will move a little, high will move much more.
Sometimes the gain and offset pots on the pulse processor get "dirty". Run them up and down a few times or clean with electronic cleaner spray.
Disconnect and reconnect every connector. The contacts can get "dirty" over time. The physical active of disconnecting/reconnecting will clean the contacts.
You can test your MCA by using a sliding pulser. We use a Berkley Nucleonics Corp (BNC) GL-3 pulse generator to test every MCA that we ship. It's the only way to really know how well the MCA is working. It's about $5k, not cheap. You really need a very good pulser to test linearity. 10 to 20eV is a really small voltage difference.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
I have a couple of questions w/r to EELS output in other formats. I am acquiring EELS data on a GIF at another institution, but would like to take the files back to my lab. I saved the files as ASCII X,Y, tagged ASCII, and EMSA/MAS format. I can plot the X,Y data with no problem. However, in the tagged ASCII and the EMSA/MAS format, the data are written in 5 wide rows that wrap the data. I typically use Excel to plot stuff like this, but I don't know how to unwrap the data. What I am doing is going into a Wordprocessor and getting rid of the hard returns and commas and then resaving as a text file. I can automate it a little with macros, but it still takes time.
1. Are there ways to unwrap the data directly into Excel?
2. Is there a public domain EELS program that will plot the data and perhaps do some work with the EMSA/MAS format that will run on a PC or a Mac? (I would prefer PC because our Macs are old and there is not much chance of getting new ones here.)
Thanks in advance.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Lesley S. Bechtold wrote: ============================================ I have never done any material sciences work - I've only ever done biological specimens for EM. Our engineering department has asked me to look at some ball bearings that are failing, as a favour. I know I don't need to fix or dehydrate but do I simply clean them, mount them (using double-sided tape?) and coat them as usual? Or is coating unnecessary? What whould I clean these with? I'm assuming there is grease somewhere that is not good for my vacuum! ================================================== It is interesting how so many different persons, skilled in the art, would approach the same kind of problem so many different ways. Of course, not much information was given either so we could all be those proverbial blind men feeling a different part of the elephant.
We ourselves approach bearing failures somewhat differently. For one thing, at the onset, it is rarely clear whether it is a straight SEM job. If corrosion is involved, for example, the ability to analyze corrosion product is lost if vigorous washing/cleaning procedures are used.
No mention was made of the examination of the raceway, but that is also something of importance in any kind of a failure analysis of this type.
Our approach is to liquid wash in cold solvent, such as acetone and/or heptane, perhaps even xylene, but you don't want a solvent that is "too good ." You want to leave some organic layer on the metal surfaces. We then remove the last vestiges of the lubricant system with exposure to an oxygen plasma such as in our Plasma Prep™ II plasma etcher. This is a dry process approach, corrosion product is left in situ in place, right where it is, and with EDS, sometimes complimented with Auger, one can learn information about the failure mechanism that would not be learned by straight topographical examination.
For the mounting of the bearing "balls", depending on size, we would always use one of the conductive double sided adhesive products, either carbon sheets or if larger, then Tempfix™.
We ourselves believe one should use caution before washing away the information that could be contained in corrosion product.
Disclaimer: SPI Supplies manufactures the Plasma Prep II plasma etcher and supplies the carbon based adhesives. Our Structure Probe services laboratory performs failure analysis as a service on these kinds of samples.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Ritchie Sims wrote: } Dear Experts } } The peaks have suddenly started to move around a bit, although the } resolution stays good. } The problem comes and goes, in its good times the standard deviation } of the position of the Co Ka peak is about 0.4 eV (10 } determinations), but sometimes it's about 10 or even 20 eV. } My thinking is that if it were the detector, the resolution would be } degrading, but it's not, so maybe the culprit is the } (analog) pulse-processor. Anyone got any thoughts on how to pin it } down as being either } a the detector } b the subsequent signal-processing stuff, eg pulse-proc? } Could I successfully test the pulse-proc with a ramp from a standard } signal generator, or would that signal, being relatively clean } compared with that from a detector, not really check it out } rigorously enough? } } cheers (well, I try) } } Ritchie } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } }
Dear Ritchie! In similar cases the first I do is the following: I take off the signal cable from preamplifier and processor and measure its resistance, which should be near zero, and then check stability of this resistance during curving this cable. Two times in my practice I have met the case when after several working years the copper core of the cable was corroded as result of chemical interaction with material of cable insulation, and the resistance between many wires of core became rather big and unstable. After change to cable with Teflon insulation the peaks became very (!) stable. Regards.
I have run across the same problems with EMSA format and with other files that store multiple points of data per row.
I have developed macros that insert the necessary number of blank lines (5 for EMSA) following a given row, I select the data in the row, Copy-Special using the transpose function into the blank rows, delete the original row, and move down to the next row, and repeat. It isn't pretty, but it gets the job done.
If I was doing it regularly, I think I would write a program (standalone or Word Basic) that would incorporate the smarts enough to do it all automatically given the file name. As of yet, I have not been so driven. But maybe someone else has already done one.
Good luck.
At 03:54 PM 3/5/99 -0500, you wrote: } I have a couple of questions w/r to EELS output in other formats. I am } acquiring EELS data on a GIF at another institution, but would like to take } the files back to my lab. I saved the files as ASCII X,Y, tagged ASCII, and } EMSA/MAS format. I can plot the X,Y data with no problem. However, in the } tagged ASCII and the EMSA/MAS format, the data are written in 5 wide rows } that wrap the data. I typically use Excel to plot stuff like this, but I } don't know how to unwrap the data. What I am doing is going into a } Wordprocessor and getting rid of the hard returns and commas and then } resaving as a text file. I can automate it a little with macros, but it } still takes time. } } 1. Are there ways to unwrap the data directly into Excel? } } 2. Is there a public domain EELS program that will plot the data and } perhaps do some work with the EMSA/MAS format that will run on a PC or a } Mac? (I would prefer PC because our Macs are old and there is not much } chance of getting new ones here.) } } Thanks in advance. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc.
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
On Fri, 5 Mar 1999, Walck. Scott D. wrote: | |2. Is there a public domain EELS program that will plot the data and |perhaps do some work with the EMSA/MAS format that will run on a PC or a |Mac? (I would prefer PC because our Macs are old and there is not much |chance of getting new ones here.) |
I've been using XlispStat {http://www.xlispstat.org/} for both plotting and analysis ( Mostly EDS but some EELS ).
It's a free, cross-platform (Mac,Windows,Unix) version of Lisp enhanced with vector and matrix arithmetic, math and statistical function, linear and non-linear regression, smoothing, FFTs, and simple object oriented graphics.
It's extensible with function written either in Lisp or C: I've written functions to read & write EMSA/MAS format (roughly -- the specs are somewhat ambiguous), read spectra from a 4pi SpectraEngine on a Mac, convolution and digital filtering and other utilities. ( Haven't quite gotten it all wrapped up into a complete EDS/EELS package yet. )
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| University of Virginia Health Sciences Center |--- ---| P.O. Box 10011 Charlottesville, VA 22906-0011 |---
Caldera Open Linux: "Powerful and easy to use!" -- Microsoft(*) (*) {http://www.pathfinder.com/fortune/1999/03/01/mic.html}
Dear Lim (S'pore), There are many infrared camera in the market. You might need to specify = what is the usage are for. By the way, how is Singapore nowadays? Still M= oney no enough?
Thank you for the replies so far. I should have included the info that it's a gain problem ie the zero peak stays in the right place. I guess it boils down to:
given that the resolution stays good, does anyone think that it could be a detector problem, or does everyone think that it's the pulse-processor?
rtch
} From: Self {GLGNOV2/RSIMS} } To: microscopy-at-sparc5.microscopy.com } Subject: EDS trouble-shooting } Date: Fri, 5 Mar 1999 21:28:27 GMT+1200
} Dear Experts } } The peaks have suddenly started to move around a bit, although the } resolution stays good. } The problem comes and goes, in its good times the standard deviation } of the position of the Co Ka peak is about 0.4 eV (10 } determinations), but sometimes it's about 10 or even 20 eV. } My thinking is that if it were the detector, the resolution would be } degrading, but it's not, so maybe the culprit is the } (analog) pulse-processor. Anyone got any thoughts on how to pin it } down as being either } a the detector } b the subsequent signal-processing stuff, eg pulse-proc? } Could I successfully test the pulse-proc with a ramp from a standard } signal generator, or would that signal, being relatively clean } compared with that from a detector, not really check it out } rigorously enough? } } cheers (well, I try) } } Ritchie }
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
As one of the committee members of the MSA/MAS Format I'm perplexed about your comment on the 5 wide rows. That sounds very odd indeed and I recall nothing in the format that calls out that type of coding. Perhaps there is an error somewhere or a misinterpretation of the spectral file format. .
Send me a "private" copy of the 3 files at my ANL address (Zaluzec-at-aaem.amc.anl.gov) and I'll have a look at them when I get back to the US.
Nestor
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I think that the main problem here is that we have a defined M&M=20 =46ormat (MSA/MAS in old parlance, let's face it we are a Microscopy=20 and Microanalysis community and the sooner we realize it the=20 better!), but the XEDS and EELS manufacturers don't fully support it.=20 They either write it but don't read it or read it but don't write it.=20 I think the format should be refined so that the manufacturers will=20 use it and let them have tags in the format similar to the TIFF.
Just my two cents worth (probably start a war, but just my opinion)
Jfm.
John Mansfield PhD CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
Ritchie Sims wrote: } Dear Experts } } The peaks have suddenly started to move around a bit, although the } resolution stays good. } The problem comes and goes, in its good times the standard deviation } of the position of the Co Ka peak is about 0.4 eV (10 } determinations), but sometimes it's about 10 or even 20 eV. } My thinking is that if it were the detector, the resolution would be } degrading, but it's not, so maybe the culprit is the } (analog) pulse-processor. Anyone got any thoughts on how to pin it } down as being either } a the detector } b the subsequent signal-processing stuff, eg pulse-proc? } Could I successfully test the pulse-proc with a ramp from a standard } signal generator, or would that signal, being relatively clean } compared with that from a detector, not really check it out } rigorously enough? } } cheers (well, I try) } } Ritchie } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } }
Dear Ritchie! In similar cases the first I do is the following: I take off the signal cable from preamplifier and processor and measure its resistance, which should be near zero, and then check stability of this resistance during curving this cable. Two times in my practice I have met the case when after several working years the copper core of the cable was corroded as result of chemical interaction with material of cable insulation, and the resistance between many wires of core became rather big and unstable. After change to cable with Teflon insulation the peaks became very (!) stable. Regards.
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Hello
Ritchie Sims wrote: } Dear Experts } } The peaks have suddenly started to move around a bit, although the } resolution stays good. } The problem comes and goes, in its good times the standard deviation } of the position of the Co Ka peak is about 0.4 eV (10 } determinations), but sometimes it's about 10 or even 20 eV. } My thinking is that if it were the detector, the resolution would be } degrading, but it's not, so maybe the culprit is the } (analog) pulse-processor. Anyone got any thoughts on how to pin it } down as being either } a the detector } b the subsequent signal-processing stuff, eg pulse-proc? } Could I successfully test the pulse-proc with a ramp from a standard } signal generator, or would that signal, being relatively clean } compared with that from a detector, not really check it out } rigorously enough? } } cheers (well, I try) } } Ritchie } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } }
Dear Ritchie! In similar cases the first I do is the following: I take off the signal cable from preamplifier and processor and measure its resistance, which should be near zero, and then check stability of this resistance during curving this cable. Two times in my practice I have met the case when after several working years the copper core of the cable was corroded as result of chemical interaction with material of cable insulation, and the resistance between many wires of core became rather big and unstable. After change to cable with Teflon insulation the peaks became very (!) stable. Regards.
Arnold, Jim wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am currently looking to upgrade my EDS system. I work in a semiconductor } manufacturing facility(CMOS) and I currently have a Cambridge S200 SEM with } Kevex Delta II. } } I have an interest in Oxford, EDAX and a company called EVEX (which I am not } familiar with)? Does anyone have experience with these companies - Good or } Bad? } } I am looking for a light element detector with possibly a WDS for Boron } quantification. } } Thanks in advance. } } Jim Arnold } Microelectronics and Technology Center } AlliedSignal Electronics and Avionics Systems } 9140 Old Annapolis Road } Columbia, MD 21045 } } email: Jim.arnold-at-alliedsignal.com } voice: (410) 964-4118 } fax: (410) 964-5046
Jim, One of my customers sent their Kevex detector to Evex for repair. They didn't complete the repair, they bent the dewar, and they never returned. A very bad bet to do business with them.
Ken Converse owner Quality Images third party SEM service Delta, PA
Lesley S. Bechtold wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } } I have never done any material sciences work - I've only ever done } biological specimens for EM. Our engineering department has asked me to } look at some ball bearings that are failing, as a favour. I know I don't } need to fix or dehydrate but do I simply clean them, mount them (using } double-sided tape?) and coat them as usual? Or is coating unnecessary? } What whould I clean these with? I'm assuming there is grease somewhere } that is not good for my vacuum! } } Any help would be appreciated!! Thanks in advance..... } } Lesley Bechtold } } Lesley S. Bechtold } Supervisor, Biological Imaging } The Jackson Laboratory } 600 Main St. } Bar Harbor, ME 04609 } 207-288-6191
Lesley, All of the suggestions have been pretty much on the mark. Just two things 1) Get a degausser so you can demagnetize the sample, because if you don't, your resolution will have you swearing at your microscope for its poor performance, 2) a ball bearing that has a good surface may require up to 20kx magnification to see any surface detail. Some dirt on the surface can make finding that surface much easier. Materials failure analysis is a lot of fun!
Ken Converse owner Quality Images third party SEM service Delta, PA
Dear listserved all, I wish to know the examination dates for the next cycle of MSA certification in Electron Microscopy. I know that this question should have been sent to the business office at MSA, but I hope to have a quicky answer from any of the list members. Cheers and have a good day. Mohammed Yousuf A.Rawoof.
} } 1. Are there ways to unwrap EMSA/MAS data directly into Excel? }
Scott, it's a fairly straightforward process to unwrap the data in MSExcel. I recorded a macro to do it a while ago - although we were on Macs then, not PCs, I think I kept it somewhere... If you e-mail a file to me I could write it again, if you like. It is fairly easy to do yourself - I think it's just a matter of cutting and pasting columns. If you can work out a routine to do it once, you can 'record' what you're doing as a macro. All you have to do for subsequent files is to run the macro again.
} 2. Is there a public domain EELS program that will plot the data and } perhaps do some work with the EMSA/MAS format that will run on a PC or a } Mac? (I would prefer PC because our Macs are old and there is not much } chance of getting new ones here.) } } Thanks in advance. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } PO Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } } } } }
I am alarmed by your statement. It is Evex Analytical's policy to "always" perform an on-site installation of "any" new detector install or detector repair.
Please be more specific on your customer's name, location, detector serial number, date of service. .
Are you absolutely sure the customer you mentioned sent a "Kevex" detector to "Evex Analytical"? Your prompt reply is appreciated.
Thank you Evex Analytical Peter Tarquinio
-----Original Message----- } From: Kenneth Converse {qualityimages-at-netrax.net} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} ; Arnold, Jim {Jim.Arnold-at-alliedsignal.com}
I am working with self-assembled microstructures that are composed of a sterol and a phosphatidylcholine. I need to be able to attach these structures to glass or plastic walls of a chamber they grow in. What would you recommend? I have tried various commercially coated glass slides, super glue, jewel glue, leather glue, and almost any other glue I could think of. The problem is that my structures grow in an aqueous solution, and I was not able to find an adhesive which would not only glue the structures (super glue did the job actually), but also not dissolve in water.
I would greatly appreciate your suggestions and comments.
Thank you in advance -- Yevgeniya Zastavker.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Yevgeniya V. Zastavker Massachusetts Institute of Technology, Biophysics 77 Massachusetts Ave, Room 13-2038 Cambridge, MA 02139 (617) 253-4826
Sorry to bombard you with questions. This could be the wrong list, but I thought to try anyway. I am looking for crystallographic data of various sterols, and in particular (MAJORLY) I need information on the crystal angles of various sterols. Does anybody know of a good source for this information? I would greatly appreciate your advice.
Thank you very much in advance -- Yevgeniya Zastavker
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Yevgeniya V. Zastavker Massachusetts Institute of Technology, Biophysics 77 Massachusetts Ave, Room 13-2038 Cambridge, MA 02139 (617) 253-4826
The Electron Microscopy Laboratory at New Mexico State University has an open position for an Electron Microscopy Specialist. The laboratory provides transmission and scanning electron microscopy and some light microscopy services for the university research community and a few external organizations, in biological, physical and materials sciences fields. Qualifications: bachelor's degree minimum, master's degree desirable, with at least four years of electron microscopy experience. The preferred candidate will have experience with energy-dispersive x-ray analysis. Experience with digital image capture and analysis, fluorescence microscopy, immunocytochemistry (including immunogold labeling), and laser scanning confocal microscopy is desirable. The candidate must be competent with sample preparation techniques, including vacuum evaporation, sputter coating, critical point drying, support film production, low temperature embedding, and photographic film processing and printing. The successful candidate must be able to work well with researchers, staff, and students, and be able to train graduate and undergraduate students for independent work with relevant techniques and equipment. Duties: operations and routine maintenance of transmission and scanning electron microscopes and associated equipment; fixation, embedding, semi-thin and ultrathin sectioning, staining, and coating of samples; supervision of facility users; record keeping, including billings, budgets, maintenance of instrument and research logs, and researching and designing specimen preparation protocols as required. Salary: DOQ Website: www.nmsu.edu/~personal/postings/professional/ Screening of applicants will begin May 1, 1999 and continue until a candidate is chosen. Applications should include a resume, letter of application and three letters of recommendation.
Apply to: Dr. Reed Dasenbrock Associate Dean/Director Arts and Sciences Research Center New Mexico State University MSC RC, Box 30001 Las Cruces, NM 88003-8001 rdasenbr-at-nmsu.edu
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We have an ETEC SEM available if anyone wants it. (Stockton, CA) It is = in working condition. We bought it new in 1973 and it has been under = contract since that time until Feb. 1, 1999. We also have extra ETEC = parts (power supplies, modules, etc.) We must take it out by March 24, 1999. If anyone wants it, please let me = know or we start chopping it on that day. Thanks Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
id {1DCJVQ86} ; Mon, 8 Mar 1999 12:10:10 -0800 Message-ID: {3D55EE50922DD21192CC00A0C9A9D8270120F8-at-cas.csudh.edu}
Dear subscribers,
I want to purchase a microtome that sections both paraffin and plastic embedded tissue. I have not purchased a rotary microtome before and I would like your suggestions as to what companies handle rotary microtomes at the present time. I have an ultramicrotome and I know that both types of media can be cut on it but I need a second microtome and have about $12,000 to spend.
Laura J. Robles
Laura J. Robles, PhD. Associate Dean, Student Academic Advancement Professor of Biology MBRS Program Director College of Arts and Sciences California State University, Dominguez Hills 1000 East Victoria Street, Carson CA 90747 310 243-3389, FAX 310 516-4268 lrobles-at-cas.csudh.edu
I think an inexpensive inkjet is the way to go. However, you must try to set the DPI of the printer to match the resolution setting of your digital images. If you capture a digital image at 1024 X800 for instance, you have about 820K of information. Now lets say you plan to print a 4X5 image similar to a Polaroid, then your printer should be no less than 300dpi {4"x5" x (300)2=1.8Mpixel} to accommodate the amount of pixel information from the capturing rate of the image. Hence, a 2048X1600 resolution setting captures 3.2 Mpixels of info, so a 400DPI setting should be used. The idea being to match the capturing info with the amount of pixels the printer can resolve to minimize interpolation...be it upwards or downwards. Not sure what the human eye can resolve tho.
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We have an ETEC SEM available if anyone wants it. (Stockton, CA) It is = in working condition. We bought it new in 1973 and it has been under = contract since that time until Feb. 1, 1999. We also have extra ETEC parts (power supplies, modules, etc.) We must take it out by March 23, 1999. If anyone wants it, please let me = know or we start chopping it on that day. Thanks Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
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Dear List, Can anyone explain what does "Reflective" means in Laser Terminology .
I read through a article in a Magazine regarding Laser Marker and in the = article the mentioned that Wood and Paper is 100% reflective . I am lost = and confuse
M.E.Lim Sr Regional Support Engineer QES(Asia Pacific) Sdn Bhd Tel : 603-7241188 ext 214 Fax : 603-7244488 Emails : melim-at-qes.po.my
Can anyone give me tips how to prepare micro-injected cells for TEM. Which cell culture support is best for detaching the cells? Is there any compound which we could micro-inject as a marker?
Try gap-filling super glue (crazy glue), available at any hobby store or it should be at a hardware store. No particular brand, they all work--it's the gap-filling that seems to be important. I used this to glue gelatin specimen blocks to the stage of a Vibratome which was then flooded with phosphate buffer. It holds under water if you let it set. This doesn't take long.
The problem may be the gap-filling property--it may cover your microstructures.
Phil
} I am working with self-assembled microstructures that are composed of a } sterol and a phosphatidylcholine. I need to be able to attach these } structures to glass or plastic walls of a chamber they grow in. What } would you recommend? I have tried various commercially coated glass } slides, super glue, jewel glue, leather glue, and almost any other glue I } could think of. The problem is that my structures grow in an aqueous } solution, and I was not able to find an adhesive which would not only glue } the structures (super glue did the job actually), but also not dissolve in } water. } } I would greatly appreciate your suggestions and comments. } } Thank you in advance -- Yevgeniya Zastavker. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Yevgeniya V. Zastavker } Massachusetts Institute of Technology, Biophysics } 77 Massachusetts Ave, Room 13-2038 } Cambridge, MA 02139 } (617) 253-4826
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 Voice: (608) 833-2885 Fax: (608) 836-1969 (please make sure my name is on any fax) oshel-at-terracom.net
May I offer a word of caution about the approach below?
While you're correct in saying you need to optimize printing conditions for your image and printer, there will probably be a little more to it than setting up comparable number of pixels. Your image is probably either a 256-level gray scale or a 16 million color (256 levels of red, green and blue). An inkjet can't print that kind of color depth in each pixel. The concepts of "halftoning" or "dithering" need to be considered. As I'm far from the expert, and we don't need a complete textbook on the listserver anyway, so I'll refer you to chapter 2 of "The Image Processing Handbook" by John Russ. As we would expect from Dr. Russ, the material is excellent.
That does bring up a question for me, though. The new HP inkjets have a "PhotoREt" technology which, I believe is supposed to be able to vary the size of the dots it produces, therefore producing better photo-printing results. Has anyone determined whether this is true, or just hype?
Jim Passmore Sr. Analytical Chemist Cryovac Division Sealed Air Corporation
---------- } From: Harry.Ekstrom } To: Microscopy } Subject: FW: Printers for SEM Images } Date: Monday, March 08, 1999 5:11PM } ------------- } } I think an inexpensive inkjet is the way to go. However, you must try to } set the DPI of the printer to match the resolution setting of your digital } images. If you capture a digital image at 1024 X800 for instance, you have } about 820K of information. Now lets say you plan to print a 4X5 image } similar to a Polaroid, then your printer should be no less than 300dpi } {4"x5" x (300)2=1.8Mpixel} to accommodate the amount of pixel information } from the capturing rate of the image. Hence, a 2048X1600 resolution setting } captures 3.2 Mpixels of info, so a 400DPI setting should be used. The idea } being to match the capturing info with the amount of pixels the printer can } resolve to minimize interpolation...be it upwards or downwards. Not sure } what the human eye can resolve tho. } } Good Luck, } Harry Ekstrom } }
Does anyone have or know of a good reference on EM lab safety? I thought there was a book called "Safety in the EM Laboratory"...but I haven't been able to find it. Probably imagined it.
FEI Company Celebrates 50 Years Philips Electron Microscopy with an Anniversary Image Contest (Calling all microscopists!)
Fifty years ago, we delivered the first Philips electron microscope. Since then, our TEMs and SEMs are used for all kinds of applications.
To celebrate the occasion, we're inviting all Philips electron microscope users all over the world to join our special Anniversary Image Contest.
To enter, simply submit prints of one or two of your best images made with a Philips electron microscope, showing the original data bar. Prints only, please! Closing date: 31 July 1999
WIN 1,000 EURO*!
Our jury panel of experts will select the best ten images, five in Life Science and five in Material Science. All ten winners will each be awarded a prize of 1000 Euro*. The winners will be announced in August 1999 on the FEI website at www.feic.com.
The following details must accompany each entry: * Name and contact address of owner * Category: Life Science or Material Science * Description of subject * Type of instrument used * Electron optic magnification * Magnification of print
Please submit your entry to: FEI Company 50 Years Philips EM Celebration P.O. Box 218 5600 MD EINDHOVEN The Netherlands
Digital images cannot be accepted for practical reasons. All submissions must be free of any legal obligations. All entries remain property of their original owners, but contestants consent to the use of their entries for promotional purposes by FEI Company without further compensation. Prizes are not transferable. Taxes are the sole responsibility of the winners. Contest rules are available on the company's website (www.feic.com) or can be requested by fax to +31 40 276 6587. FEI Company will not enter into any other correspondence regarding this contest.
*Actual prize will be the equivalent value in the winner's local currency
Yev, You might try Cell-Tak from Collaborative Biomedical Products, Two Oak Park, Bedford, Mass. 617-275-0004. This is the isolated mussel adhesion protein (map) the marine mussels and barnacles use to attach themselves to rocks and boats etc. It is commonly used by people perform atomic force microscopy to attach their molecules to a surface.
Images & Info at http://www.molbio.princeton.edu/confocal {http://www.molbio.princeton.edu/confocal}
Joseph Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University jgoodhouse-at-molbio.princeton.edu {mailto:Jgoodhouse-at-molbio.princeton.edu}
609-258-5432
-----Original Message----- From: Yevgeniya Zastavker [mailto:zhenya-at-critical.mit.edu] Sent: Monday, March 08, 1999 10:43 AM To: microscopy-at-Sparc5.Microscopy.Com Cc: Yevgeniya Zastavker Subject: adhesive for lipids
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I am working with self-assembled microstructures that are composed of a sterol and a phosphatidylcholine. I need to be able to attach these structures to glass or plastic walls of a chamber they grow in. What would you recommend? I have tried various commercially coated glass slides, super glue, jewel glue, leather glue, and almost any other glue I could think of. The problem is that my structures grow in an aqueous solution, and I was not able to find an adhesive which would not only glue the structures (super glue did the job actually), but also not dissolve in water.
I would greatly appreciate your suggestions and comments.
Thank you in advance -- Yevgeniya Zastavker.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Yevgeniya V. Zastavker Massachusetts Institute of Technology, Biophysics 77 Massachusetts Ave, Room 13-2038 Cambridge, MA 02139 (617) 253-4826
I think an inexpensive inkjet is the way to go. However, you must try to set the DPI of the printer to match the resolution setting of your digital images. If you capture a digital image at 1024 X800 for instance, you have about 820K of information. Now lets say you plan to print a 4X5 image similar to a Polaroid, then your printer should be no less than 300dpi {4"x5" x (300)2=1.8Mpixel} to accommodate the amount of pixel information from the capturing rate of the image. Hence, a 2048X1600 resolution setting captures 3.2 Mpixels of info, so a 400DPI setting should be used. The idea being to match the capturing info with the amount of pixels the printer can resolve to minimize interpolation...be it upwards or downwards. Not sure what the human eye can resolve tho.
I just scanned through postings about processing tissue cultures and coverslips for TEM, but I didn't see anything on processing cells grown on 8 well Permanox slides. I will be embedding in PolyBed 812, with a transition through propylene oxide. Unfortunately, the wells don't survive the p.o. step. I would appreciate hearing about any experiences with Permanox slides. Thank you very much.
Sandy Perkins
Laboratory for Neurotoxicity Studies Virginia-Maryland Regional College of Veterinary Medicine Virginia Tech
I agree that you must match the print resolution to the image resolution, but take some exception with your math.
At the crux of the issue is how many printer pixels are required to represent a single image pixel with a satisfactory level of gray or color scale resolution. While a dye sub printer conceptually requires only a single pixel to render the whole range of colors or gray scales, an inkjet printer may require multiple pixels based on the technology used. If an ink jet can only be tunred on and off (like a laser printer), then dithering will be required over a number of pixels to give the appearance of shades of color. More pixels will be needed for smoother or finer gradations. Now I think I heard that new inkjet printers can control the amount of ink at each pixel so that they approach the dye-subs in using only one printer pixel per one image pixel. However, I think better results would be had by allowing something like an 6x6 printer pixel pattern for each image pixel.
Using those assumptions, a 1024x800 image would require 6144x4800 pixels in a 5x4 inch space which requires 1200 dpi printer resolution. Doubling the image resolution to 2048 requires doubling the printer resolution to 2400 dpi. However, the limited resolution (both spatial and color) of the human eye may permit decent images with much less printer resolution, but there would be some loss of image detail.
At 03:11 PM 3/8/99 -0700, Ekstrom, Harry wrote: } } I think an inexpensive inkjet is the way to go. However, you must try to } set the DPI of the printer to match the resolution setting of your digital } images. If you capture a digital image at 1024 X800 for instance, you have } about 820K of information. Now lets say you plan to print a 4X5 image } similar to a Polaroid, then your printer should be no less than 300dpi } {4"x5" x (300)2=1.8Mpixel} to accommodate the amount of pixel information } from the capturing rate of the image. Hence, a 2048X1600 resolution setting } captures 3.2 Mpixels of info, so a 400DPI setting should be used. The idea } being to match the capturing info with the amount of pixels the printer can } resolve to minimize interpolation...be it upwards or downwards. Not sure } what the human eye can resolve tho. } } Good Luck, } Harry Ekstrom
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
I have an address for Electron Microscopy Safety Handbook. 2nd Edition. 1994. $15.00 Barber, V.C. and J.A. Mascorro (Eds)
San Francisco Press, Box 428600 San Francisco, CA 94142-6800
Hopefully it is still in print, Sally -- Sally Burns Center for Electron Optics B5 Center for Integrated Plant Systems E. Lansing, MI 48824 (517) 355-5004 burnssal-at-pilot.msu.edu
We are electropolishing a two-phase alloy containing a Laves phase and a bcc solid solution. The bcc phase is mainly Vanadium and the Laves phase is mainly HfV2. We are experiencing preferential polishing of the bcc phase leading to marginal TEM specimen quality.
Reply to: RE: Electropolishing of Hf alloy Try adding about 5% acetic acid, polish at -40 C, 100 volts.
Alternate polish: (worked on V-20Ti) = 5.3 g lithium chloride Temp=3D -50 C 11.1 g magnesium perchlorate voltage=3D 190-200 500 ml methanol current=3D 40-50 mA 100 ml butyl cellosolve
Above done with a South Bay 550-B single jet polisher in 1985. A notation mentions even grain boundaries. =
Bernie Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439 E-mail {kestel-at-anl.gov} 100 ml butyl cellosolve David E. Luzzi wrote: } We are electropolishing a two-phase alloy containing a Laves phase and a = bcc } solid solution. The bcc phase is mainly Vanadium and the Laves phase is } mainly HfV2. We are experiencing preferential polishing of the bcc phase } leading to marginal TEM specimen quality. } } Our solution is H2SO4 / Methanol / HF / butyl cellusolve } } Does anyone have any suggestions? Thanks in advance. } } David E. Luzzi } Department of Materials Science } University of Pennsylvania } 3231 Walnut Street } Philadelphia, PA 19104-6272 } } 215-898-8366 } 215-573-2128 - fax } luzzi-at-lrsm.upenn.edu {mailto:luzzi-at-lrsm.upenn.edu} } } } } RFC822 header } ----------------------------------- } } Received: from dns2.anl.gov (dns2.anl.gov [146.139.254.3]) by = } horus.et.anl.gov (8.6.11/8.6.11) with ESMTP id NAA23584 for {kestel-at-horus.= et.anl.gov} ; Tue, 9 = } Mar 1999 13:42:50 -0600 } Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com = } [206.69.208.10]) by dns2.anl.gov (8.9.1a/8.6.11) with SMTP id NAA21456; = Tue, 9 Mar 1999 = } 13:42:50 -0600 (CST) } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.= 11) = } id NAA12717 for dist-Microscopy; Tue, 9 Mar 1999 13:07:10 -0600 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id NAA12703 for = } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 9 Mar 1999 13:06:39 -= 0600 } Received: from sol1.lrsm.upenn.edu (SOL1.LRSM.UPENN.EDU [130.91.56.35]) = by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id NAA12692 for = } {Microscopy-at-sparc5.microscopy.com} ; Tue, 9 Mar 1999 13:06:24 -0600 } Received: from Luzzi.sol1.lrsm.upenn.edu (LRSM221PC.LRSM.UPENN.EDU = } [130.91.56.249]) } by sol1.lrsm.upenn.edu (8.8.5/8.8.4) with SMTP } id OAA08509 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 9 Mar 1999 = } 14:23:08 -0500 (EST) } From: "David E. Luzzi" {luzzi-at-sol1.lrsm.upenn.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Subject: Electropolictropoliing of Hf alloy } Date: Tue, 9 Mar 1999 14:22:45 -0500 } Message-ID: {001001be6a62$36779b20$f9385b82-at-Luzzi.sol1.lrsm.upenn.edu} } MIME-Version: 1.0 } boundary=3D"----=3D_NextPart_000_0011_01BE6A38.4DAABAE0" } X-Priority: 3 (Normal) } X-MSMail-Priority: Normal } X-Mailer: Microsoft Outlook 8.5, Build 4.71.2232.26 } X-MimeOLE: Produced By Microsoft MimeOLE V4.72.2106.4 } Importance: Normal } X-MS-TNEF-Correlator: 00000000CB8AF9838A20D111B1C200C04FDF38E4A42E5600 } Errors-to: Microscopy-request-at-sparc5.microscopy.com } Content-Type: multipart/mixed; } boundary=3D"----=3D_NextPart_000_0011_01BE6A38.4DAABAE0" } Content-Length: 3968 } Status: = } --====48555149575755524855===1 Content-Type: text/html; charset="US-Ascii" Content-Transfer-Encoding: quoted-printable
{HTML} {HEAD} {/HEAD} {BODY} {PRE = WIDTH=3D"132"} Reply to: RE: Electropolishing of Hf alloy
{/PRE} {FONT FACE=3D"Geneva" SIZE=3D3 COLOR=3D"#000000"} Try = adding about 5% acetic acid, polish at -40 = C, 100 volts. {BR} {BR} Alternate polish: = (worked on V-20Ti) {BR} {BR} 5.3 = g lithium chloride = Temp=3D -50 C {BR} 11.1 g magnesium perchlorate = voltage=3D 190-200 {BR} 500 ml methanol = current=3D 40-50 = mA {BR} 100 ml butyl cellosolve {BR} {BR} = Above done with a South Bay 550-B single = jet polisher in 1985. {BR} A notation mentions = even grain boundaries. {BR} {BR} Bernie = Kestel {BR} Materials Science Division {BR} = Argonne National Laboratory {BR} Argonne, = Il., 60439 E-mail = <kestel-at-anl.gov> {BR} 100 ml = butyl cellosolve {BR} David E. Luzzi wrote: {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >We are electropolishing = a two-phase alloy containing a Laves phase = and a bcc {BR} >solid solution. The bcc = phase is mainly Vanadium and the Laves phase = is {BR} >mainly HfV2. We are experiencing = preferential polishing of the bcc phase {BR} >leading = to marginal TEM specimen quality. {BR} > {BR} >Our = solution is H2SO4 / Methanol / HF / butyl = cellusolve {BR} > {BR} >Does anyone have = any suggestions? Thanks in advance. {BR} > {BR} >David = E. Luzzi {BR} >Department of Materials Science {BR} >University = of Pennsylvania {BR} >3231 Walnut Street {BR} >Philadelphia, = PA 19104-6272 {BR} > {BR} >215-898-8366 {BR} >215-573-2128 = - fax {BR} > {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} luzzi-at-lrsm.= upenn.edu {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} < {/FONT} {FONT FACE=3D"= Geneva" SIZE=3D1 = COLOR=3D"#0000FF"} {U} mailto:luzzi-at-lrsm.upenn.edu {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} > {BR} > {BR} > {BR} > {BR} >RFC822 = header {BR} >----------------------------------- {BR} > {BR} > = Received: from dns2.anl.gov (dns2.anl.gov = [146.139.254.3]) by {BR} >horus.et.anl.gov = (8.6.11/8.6.11) with ESMTP id NAA23584 for = < {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} kestel-at-= horus.et.anl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} >; Tue, 9 {BR} >Mar = 1999 13:42:50 -0600 {BR} > Received: from = Sparc5.Microscopy.Com (sparc5.microscopy.com = {BR} >[206.69.208.10]) by dns2.anl.gov = (8.9.1a/8.6.11) with SMTP id NAA21456; Tue, = 9 Mar 1999 {BR} >13:42:50 -0600 (CST) {BR} > = Received: (from {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = daemon-at-localhost {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} ) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) {BR} >id NAA12717 for dist-Microscopy; = Tue, 9 Mar 1999 13:07:10 -0600 {BR} > Received: = from no_more_spam.com (Sparc5 [206.69.208.10]) = by {BR} >Sparc5.Microscopy.Com (8.6.11/8.6.11) =
Arizona Imaging and Microanalysis Society Annual Spring Meeting Thursday, March 11, 1999 University of Arizona Student Union Sr. Ballroom
There is no registration fee.
8:30 - 9:00 Registration
9:00 - 9:15 Welcome Dr. Clark Lantz, President AIMS
9:15 - 10:15 Microanalysis Society Tour Speaker - Applications of SEM/EDX to forensic cases and research related to food product and pharmaceutical tampering and counterfeiting. Dr. Frank Platek, US Food and Drug Administration
10:15 - 10:30 Break
10:30 - 10:55 Metals as documents: some uses of imaging and microanalysis in African history. Dr. David Killick, Anthropology, University of Arizona
10:55 - 11:20 Visualizing surfactant aggregation with atomic force microscopy Jon Wolgemuth, Physics, University of Arizona
11:20 - 11:45 Multi-parametric analysis of cell function in 2- and 3- dimensions by spectral imaging microscopy Dr. Ron Lynch, Physiology, University of Arizona
11:35 - 1:30 Lunch AIMS Business Meeting
1:30 - 1:55 In situ molecular imaging of stress proteins and oxidative damage Dr. Claire Payne, Microbiology & Immunology University of Arizona
1:55 - 2:20 Determining the functional signficance of cytoskeletal proteins using microinjection and transfection techniques Dr. Carol Gregorio, Cell Biology and Anatomy University of Arizona
2:20 - 2:45 Fiberoptic Confocal Microscope for In Vivo Imaging Dr. Art Gmitro, Radiology, University of Arizona
2:45 - 3:00 Break
3:00 - 5:00 Student Presentations
5:30 - 7:30 Banquet ($12 for dinner, contact Suzanne Kelly {suekelly-at-ag.arizona.edu} to reserve dinner)
Microscopy Society of America Tour Speaker - Digital manipulation of acquired images: What is possible vs what is ethical Dr. Jack Kinnamon, University of Denver
.................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
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Reply to: RE: Processing of micro-injected cells for TEM Dear Raija,
If you are to microinject cells, then the best support is one that gives = access for your needles. I am sure you would prefer to use coverslips. = The material you use will depend on your cells but glass usually works = well. Get the coverslips that have locator grids etched onto them. It is = important to know if you plan to examine the cells for morphology or if = you want to immunolabel them. I will assume you only want to look at = their morphology.
Once you have micro-injected cells and know where they are on the grid (so = don't plate then at too high a density), fix with aldehyde, post fix with = osmium tetroxide, dehydrate and infiltrate with resin (as you would any = piece of tissue for TEM). During the final stages of infiltration (= acetone or propylene oxide) it is wise to transfer the cells, still on the = glass coverslip, to a glass or metal dish. Plastic will dissolve. =
Embed the glass in resin, cells up, at the bottom of an aluminum weigh = boat. Push the glass to the bottom of the dish and pour unpolymerized = resin over. Polymerize by heat and remove from the aluminum dish. Cut = the thin layer of resin away from the back of the coverslip to expose the = glass. Now you can remove the glass by plunging into liquid nitrogen and = rapidly warming few times. It will cause the block to crack but it does = work. Alteratively you can heat the resin, glass-down, on a hot plate and = slide the glass of.. Either way the cells will remain in the resin, as = will the grid locator lines. You should then be able to locate your cells = and cut sections. This is not as easy as I make it seem but it all has = been done before.
If you don't like the idea of embedding at the bottom of a dish, it is = also possible to embed the glass coverslip, cells to the resin, over BEEM = or gelatin capsules, or over flat embedding molds. You will still have to = remove the glass. You could use plastic coverslips and section them too, = but finding your cells will not be easy.
The second part of your question - is there anything you can inject to = identify the cells by EM. Yes, colloidal gold particles can be prepared = that can be microinjected into cells. However, if the gold suspension is = too concentrated it will block the needle. If it is too dilute, it will be= more difficult to detect by EM. =
Immunocytochemistry by request.
Regards,
Paul Webster
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm http://www.hei.org/htm/apw.htm
Raija Sormunen wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
by arl-img-10.compuserve.com (8.8.6/8.8.6/2.18) id DAA22626; Wed, 10 Mar 1999 03:04:50 -0500 (EST)
Hi,
The Royal Microscopical Society (Oxford England) produced a series of safety notes for its members that included - the microscopes, embedding a= nd fixation.
I have had use of some of these recently so I do know that they are still=
available.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Dear all, thanks for the tips preparing liposomes for TEM observations. In summary, most answers dealed with negativ staining of the liposomes. We' ve tried so and got good results for an overview. I added all replies to this meeages for all who're interested in this topic. Bernward 1.Don Gantz wrotes: To all who desired more info on fixing and staining of liposomes-- I apologize for the delay in my response. We had a major snowstorm and I lost a day. The initial query by Bernward Laube was about evaluating homogeneity of liposome suspensions ( {100nm diam) and estimating particle volumes. We have been using a osmium fix/negative stain technique for a number of years to look at size distribution of VLDL, chylomicrons, and synthetic lipid emulsions. The osmium fix firms up these particles nicely so that flattening is greatly reduced. In fact, based upon metal shadowing exps. the slight enlargement of diameter due to flattening after fixation is negated by slight shrinkage of particles during processing. The result is an excellent estimate of diameter and volume.
In regard to lipid vesicles/liposomes, our experience with smaller ones of 50nm or so prepared by sonication and processed with only negative stain is that they are preserved pretty well. We would expect some flattening although we have not shadowed these. However, with larger liposomes of 100nm+ diam, we believe OsO4 will aid in preservation and stability and presumably reduce flattening.
If one requires differentiation of lamellar structure (uni vs multi) among particles within a population, negative staining is unreliable because of variability of stain penetration. For this purpose, we use vitreous ice cryomicroscopy as we are fortunate to have it available.
In general our fixation/negative staining procedure is as follows:
1) Add 1 volume of 2% OsO4 in Cacodylate buffer to 2 volumes of emulsion or liposome prep. Fix for 30 mins.+
2) Place small aliquot on freshly flow-discharged carbon formvar-coated grid for a few seconds.
3) Remove and blot off excess fluid and immediately stain with 1% NaPT.
4) After a few seconds, remove and blot excess fluid. Air dry.
Optimum concentration is trial and error. Fixed suspension can be diluted as needed. Hope this helps. 2.Sheila Garcia wrotes:I've just finished my Doctorate Thesis, and I worked preparing liposome. I used phosphotungstic acid in 2% aq solution, neutralized with KOH 1M (PTK). But, before it, I used bacitracin 0,1mg/ml aq sol., as a wetting agent ( D.W.Gregory and B.J.S. Pirie-Journal of Microscopy, v.99,pt.3, dec. 1973,.261-265). Take a coated grid (carbon, formvar), put a drop of bacitracin sol. for 2 min. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the liposome (a dilueted solution), draw off again and add a drop of PTK for 2 min. I hope it could help you. Profa. Dra. Sheila Garcia
Check out the following paper:
"Sequential treatment by phosphotungstic acid and uranyl acetate enhances the adherence of lipid membranes and membrane proteins to hydrophobic EM grids", A. N. Barnakov, Journal of Microscopy. Vol 175, Pt 2, August 1994, pp. 171-174. 3.Joe Neilly:Don,
Could you elaborate on how you would fix liposomes w/ Osmium tetroxide. We have tried the negative stain approach with some success but have to live with the obvious artifacts such as flattening. There are likely other artifacts caused by ionic or chemical changes of the stain. The chemical fixation sounds promising but I'm not sure how this could be done on a liquid sample. 4.Charles Garber:For your information, we have never been successful in quenching a sample fast enough (the larger sample used for SEM work) in order to keep the vesicles from rupturing. So we do this solely by freeze fracture TEM.
Do you know the name of Richard Banfield in the UK? He pretty much was the first person to really commercialize a cryo-SEM fracture system, when he owned the former Hexland Ltd. company and which was purchased by Oxford Instruments.
As of about three or four years ago, he confirmed to me that he too, had never been able to visualize liposomes via cryo SEM because of the quench rate and rupturing problem.
Should you figure out a way to quench and see the structures by SEM, a lot of us would like to know the secret! 5.Ming Chen:The easiest way to do is by negative staining technique. A 1-2% PTA (phosphotungstic acid) soultion is commonly used. It only take a few minutes to do and you can examine it under TEM to see the distribution of liposomes right away. 6. L.R. Melsen:We have looked at liposome using routine negative staining protocol. The vesicles will flatten upon drying, but simple math can reconstruct the volume of the sphere. 7. Charles Butterick: Try negative staining with a 2% ammonium molybdate (aq). Take a coated grid (carbon, formvar, etc.), put a drop of liposomes on top of the grid for a minute. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the ammonium molybdate. After a minute, draw off drop as before and allow the grid(s) to dry. Take it to the TEM. The above procedure is only a starting point. The concentration, stain, and times can all be varied to achieve optimum results. You might check out some EM texts on negative staining for other ideas. Good luck. 8. Maria Ericsson: When I've prepared liposome suspensions for TEM I've just done a simple negative staining and it's worked fine. I adsorb the suspension to a carbon coated copper grid for about 1 minute, blot off excess liquid with a filterpaper and stain with 1-2% aqueous Uranyl Acetate for 1 minute and blot on a filterpaper again. (If there is phosphate in your buffer, you will need to wash in a drop of water before the Uranyl Acetate staining) 9. Marc Schmutz:To observe liposomes without cryo systems is quit a difficult purpose. Pure lipid systems can not be easily seen in negative staining (try always Uranyl acetate and PTA or other stains). And they generaly undergo severe structural changes during the staining process. So to calculate the volume I will not try to do it and furthermore I will not believe in a volume calculate from negative staining images. Myself I'm observing routinely liposomes pure or with proteins or DNA associated and I always use cryo TEM. It's really a easy approach and also very rapid. (You don't need more than half a day per specimen) As you said you don't have access to such a apparatus but why you don't consider collaborating with a lab equiped in cryo ? If you need some more infos about cryo you can ask me and I will try to help you.
Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit„tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
This is a multi-part message in MIME format. --------------B73E5AA6C975623FC057F918 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
"...reflector may be either a highly polish metal surface or ...coated. The coating consists of a highly brilliant metallic deposit or a dielectric material." Principles & Practice of Laser Technology.
While both wood & paper are dielectric, their fine porosity would preclude 100% reflective. Typical light reflective number for paper would be 70%. Special paper grades can be much higher. In the case of a coherent laser beam, a porous surface or a surface comprised of fine particles would cause significant scattering.
'Laser marker' refers to a family of products that are used to put bar codes and other identifying markings on products. Since they mark by burning/evaporating away the surface, a marking laser would not work on a 100% reflective surface.
J. Roy Nelson Material Testing Laboratory jrnelson-at-nj1.aae.com
"melim-at-qes.po.my"-at-sparc5.microscopy.com wrote: } } Dear List, } Can anyone explain what does "Reflective" means in Laser Terminology . } } I read through a article in a Magazine regarding Laser Marker and in the article the mentioned that Wood and Paper is 100% reflective . I am lost and confuse } } M.E.Lim } Sr Regional Support Engineer } QES(Asia Pacific) Sdn Bhd } Tel : 603-7241188 ext 214 } Fax : 603-7244488 } Emails : melim-at-qes.po.my --------------B73E5AA6C975623FC057F918 Content-Type: text/x-vcard; charset=us-ascii; name="jrnelson.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for jrnelson Content-Disposition: attachment; filename="jrnelson.vcf"
Dear all, thanks for the tips preparing liposomes for TEM observations. In summary, most answers dealed with negativ staining of the liposomes. We' ve tried so and got good results for an overview. I added all replies to this message for all those who're interested in this topic. Bernward 1.Don Gantz wrotes: To all who desired more info on fixing and staining of liposomes-- I apologize for the delay in my response. We had a major snowstorm and I lost a day. The initial query by Bernward Laube was about evaluating homogeneity of liposome suspensions ( {100nm diam) and estimating particle volumes. We have been using a osmium fix/negative stain technique for a number of years to look at size distribution of VLDL, chylomicrons, and synthetic lipid emulsions. The osmium fix firms up these particles nicely so that flattening is greatly reduced. In fact, based upon metal shadowing exps. the slight enlargement of diameter due to flattening after fixation is negated by slight shrinkage of particles during processing. The result is an excellent estimate of diameter and volume.
In regard to lipid vesicles/liposomes, our experience with smaller ones of 50nm or so prepared by sonication and processed with only negative stain is that they are preserved pretty well. We would expect some flattening although we have not shadowed these. However, with larger liposomes of 100nm+ diam, we believe OsO4 will aid in preservation and stability and presumably reduce flattening.
If one requires differentiation of lamellar structure (uni vs multi) among particles within a population, negative staining is unreliable because of variability of stain penetration. For this purpose, we use vitreous ice cryomicroscopy as we are fortunate to have it available.
In general our fixation/negative staining procedure is as follows:
1) Add 1 volume of 2% OsO4 in Cacodylate buffer to 2 volumes of emulsion or liposome prep. Fix for 30 mins.+
2) Place small aliquot on freshly flow-discharged carbon formvar-coated grid for a few seconds.
3) Remove and blot off excess fluid and immediately stain with 1% NaPT.
4) After a few seconds, remove and blot excess fluid. Air dry.
Optimum concentration is trial and error. Fixed suspension can be diluted as needed. Hope this helps. 2.Sheila Garcia wrotes:I've just finished my Doctorate Thesis, and I worked preparing liposome. I used phosphotungstic acid in 2% aq solution, neutralized with KOH 1M (PTK). But, before it, I used bacitracin 0,1mg/ml aq sol., as a wetting agent ( D.W.Gregory and B.J.S. Pirie-Journal of Microscopy, v.99,pt.3, dec. 1973,.261-265). Take a coated grid (carbon, formvar), put a drop of bacitracin sol. for 2 min. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the liposome (a dilueted solution), draw off again and add a drop of PTK for 2 min. I hope it could help you. Profa. Dra. Sheila Garcia
Check out the following paper:
"Sequential treatment by phosphotungstic acid and uranyl acetate enhances the adherence of lipid membranes and membrane proteins to hydrophobic EM grids", A. N. Barnakov, Journal of Microscopy. Vol 175, Pt 2, August 1994, pp. 171-174. 3.Joe Neilly:Don,
Could you elaborate on how you would fix liposomes w/ Osmium tetroxide. We have tried the negative stain approach with some success but have to live with the obvious artifacts such as flattening. There are likely other artifacts caused by ionic or chemical changes of the stain. The chemical fixation sounds promising but I'm not sure how this could be done on a liquid sample. 4.Charles Garber:For your information, we have never been successful in quenching a sample fast enough (the larger sample used for SEM work) in order to keep the vesicles from rupturing. So we do this solely by freeze fracture TEM.
Do you know the name of Richard Banfield in the UK? He pretty much was the first person to really commercialize a cryo-SEM fracture system, when he owned the former Hexland Ltd. company and which was purchased by Oxford Instruments.
As of about three or four years ago, he confirmed to me that he too, had never been able to visualize liposomes via cryo SEM because of the quench rate and rupturing problem.
Should you figure out a way to quench and see the structures by SEM, a lot of us would like to know the secret! 5.Ming Chen:The easiest way to do is by negative staining technique. A 1-2% PTA (phosphotungstic acid) soultion is commonly used. It only take a few minutes to do and you can examine it under TEM to see the distribution of liposomes right away. 6. L.R. Melsen:We have looked at liposome using routine negative staining protocol. The vesicles will flatten upon drying, but simple math can reconstruct the volume of the sphere. 7. Charles Butterick: Try negative staining with a 2% ammonium molybdate (aq). Take a coated grid (carbon, formvar, etc.), put a drop of liposomes on top of the grid for a minute. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the ammonium molybdate. After a minute, draw off drop as before and allow the grid(s) to dry. Take it to the TEM. The above procedure is only a starting point. The concentration, stain, and times can all be varied to achieve optimum results. You might check out some EM texts on negative staining for other ideas. Good luck. 8. Maria Ericsson: When I've prepared liposome suspensions for TEM I've just done a simple negative staining and it's worked fine. I adsorb the suspension to a carbon coated copper grid for about 1 minute, blot off excess liquid with a filterpaper and stain with 1-2% aqueous Uranyl Acetate for 1 minute and blot on a filterpaper again. (If there is phosphate in your buffer, you will need to wash in a drop of water before the Uranyl Acetate staining) 9. Marc Schmutz:To observe liposomes without cryo systems is quit a difficult purpose. Pure lipid systems can not be easily seen in negative staining (try always Uranyl acetate and PTA or other stains). And they generaly undergo severe structural changes during the staining process. So to calculate the volume I will not try to do it and furthermore I will not believe in a volume calculate from negative staining images. Myself I'm observing routinely liposomes pure or with proteins or DNA associated and I always use cryo TEM. It's really a easy approach and also very rapid. (You don't need more than half a day per specimen) As you said you don't have access to such a apparatus but why you don't consider collaborating with a lab equiped in cryo ? If you need some more infos about cryo you can ask me and I will try to help you.
Bernward Laube University of Bielefeld =46aculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=D1tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
We use 4 and 8 well Labtek=A9 Chamber Slides (permanox slides).... substituting Ethanol for the Propylene Oxide. They are my favorite slides for processing cell cultures.
See the following publication for more information:
"Subcellular Localization of SV2 and Other Secretory Vesicle Components in PC12 Cells by an Efficient Method of Preembedding EM Immunocytochemistry for Cell Cultures", Tanner, Ploug and Tao-Cheng, Journal of Histochemistry and Cyrtochemistry, Vol. 44, No. 12, pp. 1481-1488, 1996.
If you have any questions.. feel free to contact me.
Virginia Tanner Crocker
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******************************************************************* Virginia Tanner Crocker Biologist NIH, NINDS EM Facility, Bldg 36, Room 3B24 Bethesda, MD 20892
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Thanks to all who replied. The ETEC has been spoken for. If for some = reason it does not leave as scheduled I will keep the names of those who = responded, just in case, since it MUST GO.
Thanks again, Judy Murphy
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
by mail-ewr-3.pilot.net (Pilot/8.8.8) with ESMTP id MAA19999; Wed, 10 Mar 1999 12:27:37 -0500 (EST) Received: from ridexch1.rid.com ([148.189.116.16]) by mailgw.bi-pharm.com with ESMTP id MAA24037; Wed, 10 Mar 1999 12:28:05 -0500 (EST) Received: by RIDEXCH1 with Internet Mail Service (5.5.2232.9) id {F437PDQP} ; Wed, 10 Mar 1999 12:27:36 -0500 Message-ID: {5063A0AB7328D211BCAA0008C7A4467704758B-at-RIDMSG05}
Sandy:
The p.o. step is not necessary. Dehydration with any of the 812 replacements can be done through EtOH alone. I use at least 3x 100%, then grade thru the EtOH:epoxy at 2:1, 1:1 and 1:2, then into pure resin. The only caveat is that the EtOH and resin must be very carefully mixed--both to ensure complete mixing and to avoid the creation of bubbles in the mix. I also use only freshly prepared resin once I reach the 1:1 stage, but have had no problem using older batches that were stored at -20 C and thawed just prior to use.
Roger Moretz Toxicology & Safety Assessment
} -----Original Message----- } From: Sandy Perkins [SMTP:skperkin-at-vt.edu] } Sent: Tuesday, March 09, 1999 11:01 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM-cells on Permanox slides } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi- } } I just scanned through postings about processing tissue cultures and } coverslips for TEM, but I didn't see anything on processing cells grown on } 8 well Permanox slides. I will be embedding in PolyBed 812, with a } transition through propylene oxide. Unfortunately, the wells don't } survive } the p.o. step. I would appreciate hearing about any experiences with } Permanox slides. Thank you very much. } } Sandy Perkins } } Laboratory for Neurotoxicity Studies } Virginia-Maryland Regional College } of Veterinary Medicine } Virginia Tech } }
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Dear all, thanks for the tips preparing liposomes for TEM observations. I= n summary, most answers dealed with negativ staining of the liposomes. We' ve tried so and got good results for an overview. I added all replies to this message for all those who're interested in this topic. Bernward 1.Don Gantz wrotes: To all who desired more info on fixing and staining of liposomes-= - I apologize for the delay in my response. We had a major snowstorm and I lost a day. The initial query by Bernward Laube was about evaluating homogeneity of liposome suspensions ( {100nm diam) and estimating particle volumes. We have been using a osmium fix/negative stain technique for a number of years to look at size distribution of VLDL, chylomicrons, and syntheti= c lipid emulsions. The osmium fix firms up these particles nicely so that flattening is greatly reduced. In fact, based upon metal shadowing exps. the slight enlargement of diameter due to flattening after fixation is negated by slight shrinkage of particles during processing. The result i= s an excellent estimate of diameter and volume.
In regard to lipid vesicles/liposomes, our experience with smaller ones of 50nm or so prepared by sonication and processed with only negative stain is that they are preserved pretty well. We would expect some flattening although we have not shadowed these. However, wit= h larger liposomes of 100nm+ diam, we believe OsO4 will aid in preservation and stability and presumably reduce flattening.
If one requires differentiation of lamellar structure (uni vs multi) among particles within a population, negative staining is unreliable because of variability of stain penetration. For this purpose, we use vitreous ice cryomicroscopy as we are fortunate to have it available.
In general our fixation/negative staining procedure is as follows:
1) Add 1 volume of 2% OsO4 in Cacodylate buffer to 2 volumes of emulsion or liposome prep. Fix for 30 mins.+
2) Place small aliquot on freshly flow-discharged carbon formvar-coated grid for a few seconds.
3) Remove and blot off excess fluid and immediately stain with 1% NaPT.
4) After a few seconds, remove and blot excess fluid. Air dry.
Optimum concentration is trial and error. Fixed suspension can be diluted as needed. Hope this helps. 2.Sheila Garcia wrotes:I've just finished my Doctorate Thesis, and I worked preparing liposome. I used phosphotungstic acid in 2% aq solution, neutralized with KOH 1M (PTK). But, before it, I used bacitracin 0,1mg/ml aq sol., as a wetting agent ( D.W.Gregory and B.J.S. Pirie-Journal of Microscopy, v.99,pt.3, dec. 1973,.261-265). Take a coated grid (carbon, formvar), put a drop of bacitracin sol. for 2 min. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the liposome (a dilueted solution), draw off again and add a drop of PTK for = 2 min. I hope it could help you. Profa. Dra. Sheila Garcia
Check out the following paper:
"Sequential treatment by phosphotungstic acid and uranyl acetate enhances the adherence of lipid membranes and membrane proteins to hydrophobic EM grids", A. N. Barnakov, Journal of Microscopy. Vol 175, Pt 2, August 1994, pp. 171-174. 3.Joe Neilly:Don,
Could you elaborate on how you would fix liposomes w/ Osmium tetroxide. W= e have tried the negative stain approach with some success but have to live with the obvious artifacts such as flattening. There are likely other artifacts caused by ionic or chemical changes of the stain. The chemical fixation sounds promising but I'm not sure how this could be done on a liquid sample. 4.Charles Garber:For your information, we have never been successful in quenching a sample fast enough (the larger sample used for SEM work) in order to keep the vesicles from rupturing. So we do this solely by freeze fracture TEM.
Do you know the name of Richard Banfield in the UK? He pretty much was the first person to really commercialize a cryo-SEM fracture system, when he owned the former Hexland Ltd. company and which was purchased by Oxfor= d Instruments.
As of about three or four years ago, he confirmed to me that he too, had never been able to visualize liposomes via cryo SEM because of the quench rate and rupturing problem.
Should you figure out a way to quench and see the structures by SEM, a lo= t of us would like to know the secret! 5.Ming Chen:The easiest way to do is by negative staining technique. A 1-2% PTA (phosphotungstic acid) soultio= n is commonly used. It only take a few minutes to do and you can examine it under TEM to see the distribution of liposomes right away. 6. L.R. Melsen:We have looked at liposome using routine negative staining protocol. The vesicles will flatten upon drying, but simple math can reconstruct the volume of the sphere. 7. Charles Butterick: Try negative staining with a 2% ammonium molybdate (aq). Take a coated grid (carbon, formvar, etc.), put a drop of liposomes on top of the grid for a minute. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the ammonium molybdate. After a minute, draw off drop as before and allow the grid(s) to dry. Take it to the TEM. The above procedure is only a starting point. The concentration, stain, and times can all be varied to achieve optimum results. You might check out some EM text= s on negative staining for other ideas. Good luck. 8. Maria Ericsson: When I've prepared liposome suspensions for TEM I've just done a simple negative staining and it's worked fine. I adsorb the suspension to a carbon coated copper grid for about 1 minute, blot off excess liquid with a filterpaper and stain with 1-2% aqueous Uranyl Acetate for 1 minute and blot on a filterpaper again. (If there is phosphate in your buffer, you will need to wash in a drop of water before the Uranyl Acetate staining) 9. Marc Schmutz:To observe liposomes without cryo systems is quit a difficult purpose. Pure lipid systems can not be easily seen in negative staining (try always Uranyl acetate and PTA or other stains). And they generaly undergo severe structural changes during the staining process. So to calculate the volume I will not try to do it and furthermore I will not believe in a volume calculate from negative staining images. Myself I'm observing routinely liposomes pure or with proteins or DNA associated and I always use cryo TEM. It's really a easy approach and also very rapid. (You don't need more than half a day per specimen) As you said you don't have access to such a apparatus but why you don't consider collaborating with a lab equiped in cryo ? If you need some more infos about cryo you can ask me and I will try to help you.
Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=D1tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
Hello, Does anyone have a suggesstion on how to fix/reset the advance mechanism to a reichert e ultramicrotome. The mechanism on the left side (0.5 um to 2 um advance) does not work, I have to get close with the coarse knife advance then use the electronic advance which makes alignment tedious. Thanks
We are thinking of purchasing a negative scanner for use with TEM negatives from our Jeol 2000-FX, and also for SEM negatives. A scanner has been recommended to us: the Agfa Duoscan T2500, which has a resolution (hardware) of 1250 x 2500 dpi and a density of 3.4D. (The scans would be output to a Kodak DS 8650 PS printer)
We need the quality of the scans to match the quality of the standard darkroom enlarger if possible, as we would like to 'go digital' at least for routine work. Does anyone have experience of routine negative scanning for TEM prints, with this or other scanners, and if so, is it realistic to expect such high quality?
Also, what additional image processing software would people recommend we got to go along with this?
Any advice would be appreciated.
Caspar
Caspar McConville, Ph.D. Technical Specialist New York State College of Ceramics Alfred University
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Dear all, thanks for the tips preparing liposomes for TEM observations. I= n summary, most answers dealed with negativ staining of the liposomes. We' ve tried so and got good results for an overview. I added all replies to this message for all those who're interested in this topic. Bernward 1.Don Gantz wrotes: To all who desired more info on fixing and staining of liposomes-= - I apologize for the delay in my response. We had a major snowstorm and I lost a day. The initial query by Bernward Laube was about evaluating homogeneity of liposome suspensions ( {100nm diam) and estimating particle volumes. We have been using a osmium fix/negative stain technique for a number of years to look at size distribution of VLDL, chylomicrons, and syntheti= c lipid emulsions. The osmium fix firms up these particles nicely so that flattening is greatly reduced. In fact, based upon metal shadowing exps. the slight enlargement of diameter due to flattening after fixation is negated by slight shrinkage of particles during processing. The result i= s an excellent estimate of diameter and volume.
In regard to lipid vesicles/liposomes, our experience with smaller ones of 50nm or so prepared by sonication and processed with only negative stain is that they are preserved pretty well. We would expect some flattening although we have not shadowed these. However, wit= h larger liposomes of 100nm+ diam, we believe OsO4 will aid in preservation and stability and presumably reduce flattening.
If one requires differentiation of lamellar structure (uni vs multi) among particles within a population, negative staining is unreliable because of variability of stain penetration. For this purpose, we use vitreous ice cryomicroscopy as we are fortunate to have it available.
In general our fixation/negative staining procedure is as follows:
1) Add 1 volume of 2% OsO4 in Cacodylate buffer to 2 volumes of emulsion or liposome prep. Fix for 30 mins.+
2) Place small aliquot on freshly flow-discharged carbon formvar-coated grid for a few seconds.
3) Remove and blot off excess fluid and immediately stain with 1% NaPT.
4) After a few seconds, remove and blot excess fluid. Air dry.
Optimum concentration is trial and error. Fixed suspension can be diluted as needed. Hope this helps. 2.Sheila Garcia wrotes:I've just finished my Doctorate Thesis, and I worked preparing liposome. I used phosphotungstic acid in 2% aq solution, neutralized with KOH 1M (PTK). But, before it, I used bacitracin 0,1mg/ml aq sol., as a wetting agent ( D.W.Gregory and B.J.S. Pirie-Journal of Microscopy, v.99,pt.3, dec. 1973,.261-265). Take a coated grid (carbon, formvar), put a drop of bacitracin sol. for 2 min. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the liposome (a dilueted solution), draw off again and add a drop of PTK for = 2 min. I hope it could help you. Profa. Dra. Sheila Garcia
Check out the following paper:
"Sequential treatment by phosphotungstic acid and uranyl acetate enhances the adherence of lipid membranes and membrane proteins to hydrophobic EM grids", A. N. Barnakov, Journal of Microscopy. Vol 175, Pt 2, August 1994, pp. 171-174. 3.Joe Neilly:Don,
Could you elaborate on how you would fix liposomes w/ Osmium tetroxide. W= e have tried the negative stain approach with some success but have to live with the obvious artifacts such as flattening. There are likely other artifacts caused by ionic or chemical changes of the stain. The chemical fixation sounds promising but I'm not sure how this could be done on a liquid sample. 4.Charles Garber:For your information, we have never been successful in quenching a sample fast enough (the larger sample used for SEM work) in order to keep the vesicles from rupturing. So we do this solely by freeze fracture TEM.
Do you know the name of Richard Banfield in the UK? He pretty much was the first person to really commercialize a cryo-SEM fracture system, when he owned the former Hexland Ltd. company and which was purchased by Oxfor= d Instruments.
As of about three or four years ago, he confirmed to me that he too, had never been able to visualize liposomes via cryo SEM because of the quench rate and rupturing problem.
Should you figure out a way to quench and see the structures by SEM, a lo= t of us would like to know the secret! 5.Ming Chen:The easiest way to do is by negative staining technique. A 1-2% PTA (phosphotungstic acid) soultio= n is commonly used. It only take a few minutes to do and you can examine it under TEM to see the distribution of liposomes right away. 6. L.R. Melsen:We have looked at liposome using routine negative staining protocol. The vesicles will flatten upon drying, but simple math can reconstruct the volume of the sphere. 7. Charles Butterick: Try negative staining with a 2% ammonium molybdate (aq). Take a coated grid (carbon, formvar, etc.), put a drop of liposomes on top of the grid for a minute. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the ammonium molybdate. After a minute, draw off drop as before and allow the grid(s) to dry. Take it to the TEM. The above procedure is only a starting point. The concentration, stain, and times can all be varied to achieve optimum results. You might check out some EM text= s on negative staining for other ideas. Good luck. 8. Maria Ericsson: When I've prepared liposome suspensions for TEM I've just done a simple negative staining and it's worked fine. I adsorb the suspension to a carbon coated copper grid for about 1 minute, blot off excess liquid with a filterpaper and stain with 1-2% aqueous Uranyl Acetate for 1 minute and blot on a filterpaper again. (If there is phosphate in your buffer, you will need to wash in a drop of water before the Uranyl Acetate staining) 9. Marc Schmutz:To observe liposomes without cryo systems is quit a difficult purpose. Pure lipid systems can not be easily seen in negative staining (try always Uranyl acetate and PTA or other stains). And they generaly undergo severe structural changes during the staining process. So to calculate the volume I will not try to do it and furthermore I will not believe in a volume calculate from negative staining images. Myself I'm observing routinely liposomes pure or with proteins or DNA associated and I always use cryo TEM. It's really a easy approach and also very rapid. (You don't need more than half a day per specimen) As you said you don't have access to such a apparatus but why you don't consider collaborating with a lab equiped in cryo ? If you need some more infos about cryo you can ask me and I will try to help you.
Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=D1tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
I'm trying to cobble together a new probe. So far, I have one WDS spectrometer which used to be on an 840. Can anyone tell me exactly which JEOL SEMs it will fit on to? I am pretty sure it will go onto 6300 and 6400, and maybe also the 733. So far I've had no luck getting this info from JEOL, but if someone within their organisation can help, great. Also, does anyone know whether a two-crystal spectro can be transformed into a 4-crystal type? I've been told that it can't be done in the field, but can the factory do it?
Anybody got any more of these spectros that they'd be willing to sell? And maybe also an 840 or 840A?
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I have been very happy with the results from my Polaroid Sprintscan 45 (~$8500). For a TEM neg, it will digitize at 2000dpi and it will output 12 bits. I can't remember the optical density but I think it is similar to what you are quoting here, but you can look it up on their web site. For a 35 mm slide, it will do 4000 dpi. You have to ask them for a special TEM negative carrier that fits into their 4x5 holder. I use 300 dpi as the standard for what kind of enlargement I can get because my HP 890C (which does a great job on photo deluxe paper) is 300 dpi and the two sub-dye printers that I have access to are both 300 dpi. This gives a usable enlargement factor of 2000/300 = 6.7x printing to these printers. It is also very quick. I think that Polaroid could definitely improve their user control interface a bit, but for the most part it works well. I have had some problems with the computer recognizing the scanner on two systems, (one with a Sprintscan 35 and the other with a Sprintscan 45) that have flatbed scanners attached and that are on. The solution is simply to turn the flatbed power off and reopen the program.
I think that the Duoscan is a flatbed. You have to be careful with putting the negatives on the glass because you can get Newton rings in you images. I have a Umax Powerlook II flatbed that is 600 x 1200 dpi that I sometimes use and you can sometimes see them. You need a mask to lift it off the glass. I have asked the listserver in the past if that defocuses the scanned image, but I did not get a satisfactory answer. I use the flatbed as a contact printer by scanning 6 images at once held in Neg-a-file sheets and digitizing to 150 dpi. This is the minimum value that I can still read the numbers on the JEOL negatives. This is actually better than making contacts prints in the darkroom, because I can select areas to adjust the contrast and brightness independently. This makes BF, DF, and diffraction images come out well even if they are on one sheet.
I highly recommend Adobe Photoshop (version 4 is what I have) coupled with John Russ' Image Processing Toolkit Plug-ins. I Import the images as 12 bits (16bit), adjust the levels to what I want, and then convert the images to 8 bits. You can then use all of Photoshop's features.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Caspar McConville To: Microscopy List Server -----------------------------------------------------------------------.
We are thinking of purchasing a negative scanner for use with TEM negatives from our Jeol 2000-FX, and also for SEM negatives. A scanner has been recommended to us: the Agfa Duoscan T2500, which has a resolution (hardware) of 1250 x 2500 dpi and a density of 3.4D. (The scans would be output to a Kodak DS 8650 PS printer)
We need the quality of the scans to match the quality of the standard darkroom enlarger if possible, as we would like to 'go digital' at least for routine work. Does anyone have experience of routine negative scanning for TEM prints, with this or other scanners, and if so, is it realistic to expect such high quality?
Also, what additional image processing software would people recommend we got to go along with this?
Any advice would be appreciated.
Caspar
Caspar McConville, Ph.D. Technical Specialist New York State College of Ceramics Alfred University
One approach might be to plunge freeze on filmed grids and then clamp these to a standard cryoSEM specimen support. The support could have a hole drilled through it for STEM. Because of the water film thickness, it could be sublimed by freeze drying, although any dissolved salts would be left behind.
A similar approach would be to quench the liposomes on something like aluminium foil, in a size & shape which could then be clamped under a thin ring which is drilled to screw to a standard suppport. These attachments can be done quite easily under a shallow depth of liquid nitrogen in a polystyrene container.
Hoping these ideas are useful to someone
Keith Ryan Marine Biological Association Plymouth, UK
Hello, The Newtonian Ring problem might be corrected the way it was done on anti-Newton glass slides, in days gone by. They used glass which was slightly etched on the side which went next to the film. I am not suggesting you etch the glass face of your scanner but it might be worth experimenting on a spare piece of glass. The etch is ever so slight; maybe just acid fumes would do the trick. This didn't seem to degrade projected slides. Just a suggestion. Good luck.
Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. Number 499, Post Office Box 19400 Austin, Texas 78760 Phone 512/282-5507 Fax 512/280-0702
TEM & SEM Maintenance -----Original Message----- } From: Walck. Scott D. {walck-at-ppg.com} To: Caspar McConville {mcconville-at-olsen.alfred.edu} ; Micro {microscopy-at-Sparc5.Microscopy.Com}
Tamara, The copy I have of "Safety in the EM Lab..." is from San Francisco Press, ISBN 0-911302-56-5, 1985. You should be able to get it ordered with that info. Good luck.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor 09 Mar 1999 4:30 PM CRC-Electron Microscopy Lab Ofc: 704-355-1267 Carolinas Medical Center Lab: 704-355-7220 P.O. Box 32861 Fax: 704-355-7648 Charlotte, NC 28232-2861 USA Eml: WWiggins-at-Carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
} -----Original Message----- } From: Tamara Howard [SMTP:howard-at-cshl.org] } Sent: Tuesday, March 09, 1999 9:49 AM } To: Microscopy Listserver } Subject: EM safety book? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone have or know of a good reference on EM lab safety? I thought } there was a book called "Safety in the EM Laboratory"...but I haven't been } able to find it. Probably imagined it. } } Thanks for any help anyone can give! } } Tamara Howard } CSHL } }
I think you are making an excellent choice with the new Agfa DuoScan T2500. You will appreciate the 3.4 D on this unique flatbed scanner. The glass less negative carriers fit into the lower tray, which slides into a precise position. The negatives will not contact any surface. I have just ordered also a DuoScan T2500. You will learn more about it at:
One of the most versatile imaging software is CorelDraw8 and CorelPHOTO-PAINT8. It offers the highest image format manipulation with Plug-ins, for most applications. Highly recommended.
We are looking for commercially available options for being able to transfer a metallographic type sample from a sample preparation workstation and into an SEM while under vacuum or inert atmosphere. We have a number of make and model SEMs therefore just any general information is fine on this subject.
thanks,
Mark A. Wall
Mr. Mark A. Wall Sr. Scientific Assoc. L-350 Chemistry & Materials Science Directorate Lawrence Livermore National Laboratory Livermore, CA USA 94550
I think you are making an excellent choice with the new Agfa DuoScan T2500. You will appreciate the 3.4 D on this unique flatbed scanner. The glass less
negative carriers fit into the lower tray, which slides into a precise position. The negatives will not contact any surface. I have just ordered also a DuoScan T2500. You will learn more about it at:
One of the most versatile imaging software is CorelDraw8 and CorelPHOTO-PAINT8. It offers the highest image format manipulation with Plug-ins, for most applications. Highly recommended.
Laszlo J. Veto
Caspar McConville wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are thinking of purchasing a negative scanner for use with TEM } negatives from our Jeol 2000-FX, and also for SEM negatives. A } scanner has been recommended to us: the Agfa Duoscan T2500, which } has a resolution (hardware) of 1250 x 2500 dpi and a density of 3.4D. } (The scans would be output to a Kodak DS 8650 PS printer) } } We need the quality of the scans to match the quality of the standard } darkroom enlarger if possible, as we would like to 'go digital' at } least for routine work. Does anyone have experience of routine } negative scanning for TEM prints, with this or other scanners, and if } so, is it realistic to expect such high quality? } } Also, what additional image processing software would people } recommend we got to go along with this? } } Any advice would be appreciated. } } Caspar } } Caspar McConville, Ph.D. } Technical Specialist } New York State College of Ceramics } Alfred University
I just want to say THANK YOU to all who responded to my question regarding the electrolytical thinning of Al. Now I own a collection of serveral different recipes! If anybody else is interested in it, just let me know.
Cheers,
Petra -------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu Visit our WWW site! http://www.crpcu.lu/~wahlbrin
My first posting was placed whilst away from the office, now I am back I can give you the full details of the safety data mentioned.
1. Safety in the Electron Microscope Room - S. K. Chapman, Microscopy &=
Analysis, March 89, 27-29, (only two pages of text)
2. Routine Handling of Resins, RMS E.M. Safety Committee, Single sheet handout
OR from one of the same group
3. Resins: Toxicity, Hazards and Safe Handling - B. E. Causton, Proceedings RMS, Vol 16/4, June 81, 265-269
4. Routine Handling of Fixatives, RMS E.M. Safety Committee, Single shee= t handout.
I am prepared to scan in part of 1, plus 2 and 3 and send as attachments direct to those who ask.
As you can see I was not quite correct they are not all Royal Microscopic= al Society publications. Their address is 37/38 St. Clements, Oxford OX4 1AJ= , England
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
I believe the two scanners are both excellent, although I have a preference for the Polaroid, that I am also going to buy (if can get the money).
As far as being capable of being able to use the scanner instead ot the darkroom, I strongly believe that both scanner technology, and computer technology is not yet capable of completely replacing a darkroom, unfortunately, for all its applications.
Would appreciate very much a feedback on this subject.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
May I offer a word of caution about the approach below?
While you're correct in saying you need to optimize printing conditions for your image and printer, there will probably be a little more to it than setting up comparable number of pixels. Your image is probably either a 256-level gray scale or a 16 million color (256 levels of red, green and blue). An inkjet can't print that kind of color depth in each pixel. The concepts of "halftoning" or "dithering" need to be considered. As I'm far from the expert, and we don't need a complete textbook on the listserver anyway, so I'll refer you to chapter 2 of "The Image Processing Handbook" by John Russ. As we would expect from Dr. Russ, the material is excellent.
That does bring up a question for me, though. The new HP inkjets have a "PhotoREt" technology which, I believe is supposed to be able to vary the size of the dots it produces, therefore producing better photo-printing results. Has anyone determined whether this is true, or just hype?
Jim Passmore Sr. Analytical Chemist Cryovac Division Sealed Air Corporation
---------- } From: Harry.Ekstrom } To: Microscopy } Subject: FW: Printers for SEM Images } Date: Monday, March 08, 1999 5:11PM } ------------- } } I think an inexpensive inkjet is the way to go. However, you must try to } set the DPI of the printer to match the resolution setting of your digital } images. If you capture a digital image at 1024 X800 for instance, you have } about 820K of information. Now lets say you plan to print a 4X5 image } similar to a Polaroid, then your printer should be no less than 300dpi } {4"x5" x (300)2=1.8Mpixel} to accommodate the amount of pixel information } from the capturing rate of the image. Hence, a 2048X1600 resolution setting } captures 3.2 Mpixels of info, so a 400DPI setting should be used. The idea } being to match the capturing info with the amount of pixels the printer can } resolve to minimize interpolation...be it upwards or downwards. Not sure } what the human eye can resolve tho. } } Good Luck, } Harry Ekstrom } }
The M & M '99 Golf tournament will be Sunday, Aug. 1, 1999 just outside Portland, OR. Hole sponsorships are available for $65.00 each on a first come, first serve basis. Prizes for longest drive, longest putt etc. can also be donated. In addition, Logo gifts to be distributed to all participants will also be most welcome. To reserve your holes or to supply gifts/prizes etc. please notify me as soon as possible.
Thank you,
John Arnott Chairman
-- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
We have an aging BioRad Confocal (1991!)with the associated computer and software (COMOS ver.7.0a). Has anyone replaced/upgraded their computer on this system other than just purchasing an upgrade from BioRad? What problems were encountered and what were some solutions? Thanks in advance for any info.
History of unit is below: Like an old car, different parts of the computer are beginning to need replacement, but I understand that several of the boards are specialized propietary boards (control of scan head and frame grabber) and cannot be replaced with conventional boards. We want to "upgrade" the computer, ie new mother board, more memory, etc... The existing mother board is not compatible with any device drivers other than SCSIs.
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu ********************************************
I need to know the wavelength of the main ~ 441.6nm emission line from our He-Cd laser source to at least 5 significant figures, if possible (wavelength in air atmosphere). Any references containing other related data (emission lines) would be useful too.
-- ***************************************** Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
I have recently taken over an SEM with an AN 10000 analyser and I was wondering if anyone out there kows if it is possible to get the results files out to a PC ?
Any help greatly appreciated.
******************************************* Robert McDonald EPMA & SEM Laboratories Dept Geography - Earth Sciences Division Gregory Building LilyBank Gardens University of Glasgow Glasgow G12 8QQ Scotland, UK email: robert-at-earthsci.gla.ac.uk Tel:- +44 (0)141 330 5505/5442 FAX:- +44 (0)141 330 4817 ********************************************
Reply to: RE: SEM vacuum transfer = For many years I used a simple permanent magnet to lift and transfer = samples mounted on metallographic mounts with a steel washer epoxied onto = the back surface. A hole in the circular magnet allowed a push rod to = release the mount from the magnet. Possibly a small electromagnet could = be made that would be easier to operate in your system-or even a vacuum = powered suction cup to eliminate magnetic fields. Most likely you will = need to make one. = Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439
Phone: (630) 252-4945 E-mail {kestel-at-anl.gov} Mark Wall wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
{HTML} {HEAD} {/HEAD} {BODY} {PRE = WIDTH=3D"132"} Reply to: RE: SEM vacuum transfer =
{/PRE} {FONT FACE=3D"Geneva" SIZE=3D3 COLOR=3D"#000000"} For = many years I used a simple permanent magnet = to lift and transfer samples mounted on = metallographic mounts with a steel washer = epoxied onto the back surface. A hole in = the circular magnet allowed a push rod to = release the mount from the magnet. Possibly = a small electromagnet could be made that = would be easier to operate in your system-or = even a vacuum powered suction cup to eliminate = magnetic fields. Most likely you will need = to make one. {BR} {BR} Bernard Kestel {BR} = Materials Science Division {BR} Argonne = National Laboratory {BR} Argonne, Il., = 60439 {BR} {BR} Phone: (630) 252-4945 = E-mail <kestel-at-anl.gov> {BR} Mark = Wall wrote: {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >-----------------------------------------------------------------------= - {BR} >The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America {BR}
Peggy, Can you please contact me with your address, at your convenience? My computer crashed and I lost your e-mail, and snail mail addresses. I'll send the nerve staining info out when you reply. Best wishes Ronnie Houston Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children 2222 Welborn Street Dallas, TX 75219
I am looking for any references that can give the emission wavelength of a HeCd laser to five significant figures (in air/vacuum). The line inparticular is the 441.6 nm line, but a table of up to date measurements would be useful too.
-- ***************************************** Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
} We are thinking of purchasing a negative scanner...
Check out the Imacon FlexTight II. This is an affordable drum scanner with magnetic carriers for various size media (2x2 skides up to at least 8x10 sheeets) that make it as easy to use as a flatbed scanner. The unit has 5,760 dpi optical resolution (although this may drop to 4,800 dpi for something the size of an EM negative), 12-bit grayscale, 24, 32 and 48-bit color, and 14 bits (4.1 OD) of dynamic range, and it is fast. For image editing, Photoshop is the best. Period.
} That does bring up a question for me, though. The new HP } inkjets have a "PhotoREt" technology which, I believe is } supposed to be able to vary the size of the dots it produces, } therefore producing better photo-printing results. Has anyone } determined whether this is true, or just hype?
HP's statement, in itself, is explicid, so I doubt they could say it if it weren't true. I own one of these printers (720C) and looking closely it is hard to tell just how small the dots are, or when and where there being used. Appearance is very close to "random dithering". A couple of clues which would lead you to believe the technology is not just hype: (1) the suggested DPI for the printer is 300dpi which is beyond what would be calculated for a normal 600dpi dithering printer, and (2) dithering can not be seen at all for the primary colors reb, green & blue as created with cyan, magenta and yellow.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
A researcher would like to be able to tell the difference between apototic cells and necrotic and/or other degenerating cells in an invertebrate nervous system, using TEM. So far the only general statement I've come across is that apototic cells will undergo autophagy within their plasma membranes whereas necrotic cells tend to spill their contents and get cleaned up by other cells.
Any additional tips will be appreciated!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I've just found a neat screen capture (and more) utility called HyperSnap-DX, a free trial version can be had from www.hyperionics.com, and registration is only $25!
cheers
rtch
} Have a couple of avi movies here and we need some stills printed from them. } Anyone have any ideas/shareware/freeware? } } } Scott D. Whittaker 218 Carr Hall } EM Technician Gainesville, FL 32610 } University Of Florida ph 352-392-1184 } ICBR EM Core Lab fax 352-846-0251 } sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ } The home of " Tips & Tricks " }
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I'm not a microscopy expert, but a retired computer systems engineer.
An inkjet can produce high color if it uses CMYK inks (vs RGB). Photo-realistic inkjet printers don't dither (screen) or halftone such as with b&w printing. They attempt to produce a 1-1 relationship with the input color data. } } That does bring up a question for me, though. The new HP } inkjets have a "PhotoREt" technology which,
H-P's RET technology has been around for quite some time. I find it introduces distracting patterns into the hardcopy. H-P sat on its laurels for quite some time, and most experts say that they have fallen behind. Unless they've changed, H-Ps use RGB ink.
I find that the Epson and newer Cannon printers do a credible job of photo-color, both are CMYK.
However, you will find that the proper software will make a bigger difference in the results than the printer. If you want high color fidelity, then I recommend Adobe Photoshop, (even if you only use it for printing), calibrated for the printers ink that you use. (Epson and Cannon ink files are provided for Photoshop.) If you expect to manipulate color on the computer screen and then print the same colors on the printer, your computer has to have a "color system" such as Kodak ColorSync (adjusts for your scanner, screen and printer). Color Systems are pretty much free on the Macintosh, but you are may be out of luck if you have a Windows box (I've not found one that works very well).
Your Color System has to "know" your hardware unless you have a means of calibrating. (UMAX scanners provide a means to calibrate from a Kodak target.)
The confusion arises out of the difference between the well established printing industry and the newer technology of photorealistic inkjet printing.
In short: make sure you can calibrate your scanner for color and that it comes with a color system,
that ink files are available for your printer's ink for use by photoshop, (even if you don't use Photoshop, other programs use the PS as a standard),
I will agree wholehearted that Adobe PhotoShop is best, but I'll bet that most people never learn it properly to get their $600 worth. A very impressive PhotoShop clone is Ulead Photo Impact ( http://www.ulead.com {http://www.ulead.com} ) for ~$90.00 which does what most people use PhotoShop for (TWAIN compliant, Image resizing and level adjustments), plus Web-based image production is built in, not an add-on.) Try their 15-day fully functional demo. Enough said on image editors.
Just passing along some unbiased information.
Walt Bobrowski Subcellular Pathology Parke-Davis Research 2800 Plymouth Road Ann Arbor, MI 48105
Over the years our workforce has dwindled to the point that I find myself = working alone in the lab (metalographic sample prep/optical microscopy + = image analysis) almost all the time. The lab doesn't have windows to the = hallway, so it is difficult for people walking by to see if I am in the = lab, and if I am OK. My manager is concerned about my safety and is asking = me for suggestions. What do other labs do to make working alone safe? Everett Ramer Federal Energy Technology Center
I have replaced the original Compaq 386 with a Dell 486. I had no problems with the BioRad MRC 600 system and comos software. BioRad people have told of problems with some computers but I haven't seen any.
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
This announcement is for colleagues of Don W. Fawcett, M.D. ("A textbook = of histology", Bloom & Fawcett and "The Cell"), Professor Emeritus, = Harvard University. =
This coming Sunday (3/14/99) he will be celebrating his 82nd birthday. A = few e-mail greetings might surprize and please him. His e-mail address is = {DFawc20586-at-aol.com} . =
I am sure he would appreciate good wishes from anyone inspired by his = books and papers too.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm --====48525552525549495651===1 Content-Type: text/html; charset="US-Ascii" Content-Transfer-Encoding: quoted-printable
{HTML} {HEAD} {/HEAD} {BODY} {FONT FACE=3D"Monaco" = SIZE=3D1 COLOR=3D"#000000"} Dear Microscopists and Histologists, {BR} {BR} This = announcement is for colleagues of Don W. = Fawcett, M.D. ("A textbook of histology", = Bloom & Fawcett and "The Cell"), = Professor Emeritus, Harvard University. = {BR} {BR} This coming Sunday (3/14/99) he will = be celebrating his 82nd birthday. A few = e-mail greetings might surprize and please = him. His e-mail address is < {/FONT} {FONT FACE=3D"Monaco" = SIZE=3D1 COLOR=3D"#0000FF"} {U} DFawc20586-at-aol.com {/U} {/FONT} {FONT FACE=3D"= Monaco" = SIZE=3D1 COLOR=3D"#000000"} >. {BR} {BR} I am sure he would = appreciate good wishes from anyone inspired = by his books and papers too. {BR} {BR} Paul Webster, = Ph.D {BR} House Ear Institute {BR} 2100 West Third = Street {BR} Los Angeles, CA 90057 {BR} phone:213 = 273 8026 {BR} fax: 213 413 6739 {BR} e-mail: {/FONT} {FONT = FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#0000FF"} {U} pwebster-at-hei.org {/U} {/FONT} {= FONT = FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#000000"} {BR} {/FONT} {FONT FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#0000FF"} {U} http://www.hei.= org/htm/aemi.htm {/U} {/FONT} {/BODY} {/HTML} --====48525552525549495651===1 Content-Type: application/quickmail Content-Transfer-Encoding: base64
by newton.wadsworth.org (8.8.8/8.8.8) with SMTP id QAA21360 for {microscopy-at-msa.microscopy.com} ; Thu, 11 Mar 1999 16:59:34 -0500 (EST) Sender: tivol-at-wadsworth.org Message-ID: {36E839DF.41C6-at-wadsworth.org}
Jonathan Barnard wrote: Dear Johnathan,
} I need to know the wavelength of the main ~ 441.6nm emission } line from our He-Cd laser source to at least 5 significant figures, } if possible (wavelength in air atmosphere). Any references } containing other related data (emission lines) would be useful too. } Do such variables as pressure and composition (e.g., humidity) of the air allow the wavelength to be determined to 5 sig figs? The refractive index of air is related to pressure, and I think the cor- rect expression is n-1 = kP. Since P can easily vary by ~3% at sea level, depending on k, n could vary by more than 10^-5. The humidity might be even more important (especially for any other emission lines where water has n very different from that of air). Yours, Bill Tivol
We're trying to visualize meiotic spindle microtubules using LM and TEM. = Our methods for fixing and embedding insect testes are fairly = standard, however we are using 8% tannic acid in our fixative. This = appears to be what others have used, but we were wondering if anyone has = any warnings, hints or suggestions regarding microtubule preservation = and TEM.
We are also experimenting with different methods of preparing insect = testes for immunocytochemistry and LM. We are using various antibodies = to microtubules and nucleoproteins. We would like to optimize our = methods for the preservation of these structures. Is "live" tissue best = or will fixed tissue suffice? What is the best way to get the cells = spread onto slides? We've tried thumb "squashing" and cytospinning, but = are not satisfied with the results. Any suggestions would be greatly = appreciated!
Thanks,
Laura K. Garvey Dept. of Molecular and Cell Biology University of Connecticut
} } I have recently taken over an SEM with an AN 10000 analyser and I was } wondering if anyone out there kows if it is possible to get the results } files out to a PC ? } Bob-
Several years ago I wrote a nunber of utilities that would allow you to do just what you describe. You can write the files to an AN 10000 3.5" floppy (I hope yours *is* a 3.5" system? - if not, my programs won't help), which my utility will then read on a PC, copying the files to the PC in a byte-for-byte format. Then I wrote other utilities that would convert spectra and linescan files into tab-separated text files, and would extract images from studies and convert them, or individual image files, to baseline TIFF 6.0 files. I still use these utilities on a daily basis.
For a long while these were available on an FTP server here, but after aeons of no hits, and in the general progression of computer hardware, this went by the wayside. It would be the work of a few minutes to re-establish this, if there is interest. In the meantime, Bob, you may try contacting Pat Nicholson in the Dept. of Physics and Astronomy at Glasgow. I'm not sure, but I may well have given him copies of these files. In any case, I'll post the URL when I have re-established it.
If you have Windows, you should have everything you need already.
In DOS, the PrintScrn key used to dump a copy of the screen on the dot-matrix printer. In Windows nothing apparently happens. However, if you check the clipboard, PrintScrn snaps a copy of the desktop and stores it for pasting. Try pressing PrintScrn and then try paste into Word, or elsewhere. You will get a bitmap of the screen. Perhaps you don't want the whole screen - then Alt-PrintScrn copies just the active application window to the clipboard.
Using this process you end up with a lot or a little extra window junk around the sides. Then I use MS Imager that came with Office 6.0 (and was available on the MS website) to crop the image down to what I want.
I tried this using the AVI player from MS. I stopped the video, pulled the slider to the frame I wanted, and pressed Alt-PrintScrn. I opened up MS Imager, selected the File, New, Clipboard option and up came my AVI viewer window as a bitmap. I saved copies of it before and after cropping. I will send those to you directly. They are from the PICTURE.AVI movie that came on the Windows 98 disk.
You can use your imagination to apply this technique for other things, like producing your own instructions with snapshots showing what your EDS screen looks like at various stages.
Hope this Tip and Trick helps.
Warren
At 01:03 PM 3/11/99 +0000, you wrote: } } Have a couple of avi movies here and we need some stills printed from them. } Anyone have any ideas/shareware/freeware? } } } } } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { } GO GATORS } Scott D. Whittaker 218 Carr Hall } EM Technician Gainesville, FL 32610 } University Of Florida ph 352-392-1184 } ICBR EM Core Lab fax 352-846-0251 } sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ } The home of " Tips & Tricks "
A Mac application that delivers the most commonly used tools in Photoshop (e.g. adjusting levels, assembling RGB images and montages) is Color-It!, from MicroFrontiers. It retails for about $50.
Glen
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
On Thu, 11 Mar 1999, Bobrowski, Walter wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I will agree wholehearted that Adobe PhotoShop is best, but I'll bet that } most people never learn it properly to get their $600 worth. A very } impressive PhotoShop clone is Ulead Photo Impact ( http://www.ulead.com } {http://www.ulead.com} ) for ~$90.00 which does what most people use } PhotoShop for (TWAIN compliant, Image resizing and level adjustments), plus } Web-based image production is built in, not an add-on.) Try their 15-day } fully functional demo. Enough said on image editors. } } Just passing along some unbiased information. } } Walt Bobrowski } Subcellular Pathology } Parke-Davis Research } 2800 Plymouth Road } Ann Arbor, MI 48105 } } TEL: (734) 622-7814 } FAX: (734) 622-3478 } Mailto:Walter.Bobrowski-at-WL.COM {mailto:Walter.Bobrowski-at-WL.COM} } } } }
The nice thing about HyperSnap is that you can copy just any selected rectangular portion from your screen, and then either print it, fool around with it, or save it in an astonishing number of formats.
Ritchie
} } If you have Windows, you should have everything you need already. } } In DOS, the PrintScrn key used to dump a copy of the screen on the } dot-matrix printer. In Windows nothing apparently happens. However, if you } check the clipboard, PrintScrn snaps a copy of the desktop and stores it } for pasting. Try pressing PrintScrn and then try paste into Word, or } elsewhere. You will get a bitmap of the screen. Perhaps you don't want the } whole screen - then Alt-PrintScrn copies just the active application window } to the clipboard. } } Using this process you end up with a lot or a little extra window junk } around the sides. Then I use MS Imager that came with Office 6.0 (and was } available on the MS website) to crop the image down to what I want. } } I tried this using the AVI player from MS. I stopped the video, pulled the } slider to the frame I wanted, and pressed Alt-PrintScrn. I opened up MS } Imager, selected the File, New, Clipboard option and up came my AVI viewer } window as a bitmap. I saved copies of it before and after cropping. I will } send those to you directly. They are from the PICTURE.AVI movie that came } on the Windows 98 disk. } } You can use your imagination to apply this technique for other things, like } producing your own instructions with snapshots showing what your EDS screen } looks like at various stages.
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Everett, if the door to your lab is of a "simple" configuration, it would be easy to replace it with a "windowed" door as long as your boss is willing to accept the cost as a price towards improved safety. Obviously, not a complete answer to the problem but a first step of common sense.
Mike Bucker Feed Microscopy Consolidated Labs of Va
} } } "EVERETT RAMER" {Everett.Ramer-at-fetc.doe.gov} 03/11 4:23 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Over the years our workforce has dwindled to the point that I find myself working alone in the lab (metalographic sample prep/optical microscopy + image analysis) almost all the time. The lab doesn't have windows to the hallway, so it is difficult for people walking by to see if I am in the lab, and if I am OK. My manager is concerned about my safety and is asking me for suggestions. What do other labs do to make working alone safe? Everett Ramer Federal Energy Technology Center
Everett and all: I work at Dow Chemical and we have a fairly rigorous "lone operator" system. If we are working in an isolated area or at a time when there are few people in the building (evenings, weekends, etc.) we carry little alert transmitters which call out to plant security (as well as within the building). The building receivers are location sensitive and we have a "lone-operator" login book at the building entrance with a building map. The lone operator marks the map as s/he signs in. Between the map and the alert location, they can find us pretty fast.
This is pretty elaborate, but maybe a mini version using something like the "First Alert" products would work ("Help, I've fallen and I can't get up") - have it tied into a siren or flashing light outside your lab so that anyone along the corridor would know there was a problem. Maybe even carry a cordless phone with an autodial button to your site security - caller ID would get them to you pretty quick.
This is definitely NOT a trivial matter. It is always disconcerting to me when I am working in an area with nobody else around - I hope you get an effective solution soon.
there's lots of stuff on t.e,m, of apoptosis in vertebrate cells (it's very popular in HIV, cancer, inflammation response etc) and much of it indicates visible nuclear changes but relative stability of cytoplasm compared with necrosis. There was a review article (as a good starting point): Microscopical Study of Cell Death via Apoptosis by S. Verhaegen in MIcroscopy and Analysis, January 1998 pp5-7
but you could do a reference or citation 'trawl' on the authors: Kerr, J.F.; Wyllie, A.H. or Currie
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: Tina Carvalho To: Microscopy Listserver
Hi, All-
A researcher would like to be able to tell the difference between apototic cells and necrotic and/or other degenerating cells in an invertebrate nervous system, using TEM. So far the only general statement I've come across is that apototic cells will undergo autophagy within their plasma membranes whereas necrotic cells tend to spill their contents and get cleaned up by other cells.
Any additional tips will be appreciated!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
by ahmler1.mail.eds.com (8.9.1/8.9.1) with ESMTP id JAA30032; Fri, 12 Mar 1999 09:51:43 -0500 (EST) Received: from SYS1.BSCO.COM (sys1.bsco.com [198.132.68.84]) by nnsa.eds.com (8.9.1/8.9.1) with SMTP id JAA27069; Fri, 12 Mar 1999 09:51:12 -0500 (EST) Received: from rfurdanowiczw [10.224.0.46] by SYS1.BSCO.COM (IBM VM SMTP Level 310) via TCP with SMTP ; Fri, 12 Mar 1999 09:51:21 EST Reply-To: {rwafu-at-bsco.com} {microscopy-at-Sparc5.Microscopy.Com}
Yes, on my AN10000 there is a program called MSDOSCV.SV which converts files between the Link (now Oxford) and PC DOS operating systems. It writes files to 720kB 3.5" floppies that previously have been formatted to that density on a PC.
An alternative that I sometimes resort to is to capture on a PC output meant to go to the Facit printer over the serial connection.
If you need more details, contact me directly or seek support from Oxford (they are very helpful), for example Ruth Murray ( ruth-at-oxford.usa.com ).
Valdemar Furdanowicz Research Labs Bethlehem Steel Co. valdemar-at-fast.net or rwafu-at-bsco.com
-----Original Message----- } From: Robert McDonald [mailto:R.McDonald-at-geology.gla.ac.uk] Sent: Thursday, March 11, 1999 10:08 AM To: microscopy-at-sparc5.microscopy.com
Hi All:
I have recently taken over an SEM with an AN 10000 analyser and I was wondering if anyone out there kows if it is possible to get the results files out to a PC ?
Any help greatly appreciated.
******************************************* Robert McDonald EPMA & SEM Laboratories Dept Geography - Earth Sciences Division Gregory Building LilyBank Gardens University of Glasgow Glasgow G12 8QQ Scotland, UK email: robert-at-earthsci.gla.ac.uk Tel:- +44 (0)141 330 5505/5442 FAX:- +44 (0)141 330 4817 ********************************************
Differentiating apoptosis and necrosis morphologically is based primari= ly upon nuclear changes, although there are characteristic cytoplasmic cha= nges as well. In general (note the wiggle words), necrotic cells swell and = lyse, whereas apoptotic cells shrink and fragment. Chromatin in apoptotic ce= lls forms electron-dense crescents at the nuclear envelope, then breaks up.=
Apoptotic cells fragment into "apoptotic bodies" that may contain bits = of chromatin. A good place to see characteristic ultrastructural changes = of apoptosis is in lymphoid tissues, where lymphocytes die via apoptosis a= nd then are phagocytosed by resident macrophages (the ones that are someti= mes called "tingible body macrophages" because of the staining properties o= f the apoptotic cell remnants in them.)
Differenting apoptosis from necrosis is a tricky deal. Apoptotic cells= may undergo secondary necrosis, during which they swell and lyse. So, just=
because you see necrotic cells doesn't mean that they didn't die apoptotically. Like everything else, it's complicated; there is a cont= inuum of change with apoptosis and necrosis at opposite poles and a lot of st= uff in between!
There are lots of good reviews on this topic. The Aug 28, 1998 issue o= f Science had a special section on apoptosis, and on page 1302, there is = a series of three electron micrographs of neurons undergoing apoptosis. = One of the first reviews of the subject contains the best collection of micrographs I've found - "Cell death: the significance of apoptosis" in= the International Review of Cytology 68:251-306, 1980. Another good revie= w was in Amer J of Pathol 146:3-15, 1995. The title is "Apoptosis, oncosis, = and necrosis: an overview of cell death."
Hope this helps!
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064-6202 =
In the past I have been on jobs where a great deal of the time was spent isolated. Now days I think it is a huge safety liability. One time I had appendicitis and had to drive myself to 50 miles on back roads to a clinic with my knees up on the stearing wheel. The point here is, even though you work in a laboratory complex, if something happened you probably wouldn't get help until the cleaning crew found you. Bad news! the other issue is: Life is short and work is long, and working alone sucks. It's not emotionally healthy. Make a change. Consolidate in with other workers.
Bob Derm Imaging Center Microscopist Ex-wood cutter
On Thu, 11 Mar 1999, EVERETT RAMER wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Over the years our workforce has dwindled to the point that I find myself working alone in the lab (metalographic sample prep/optical microscopy + image analysis) almost all the time. The lab doesn't have windows to the hallway, so it is difficult for people walking by to see if I am in the lab, and if I am OK. My manager is concerned about my safety and is asking me for suggestions. What do other labs do to make working alone safe? } Everett Ramer } Federal Energy Technology Center } }
The New York State Center for Advanced Thin Film Technology University at Albany - State University at New York announces the following positions:
Senior Analytical Specialist (Two Positions Available) The New York State Center for Advanced Thin Film Technology at the University at Albany - SUNY is a fast growing, high technology research and development program with a mission of supporting industry and creating new jobs. This position will serve as a primary, materials characterization, support person for the Center's scientific and technical staff in our advanced surface science facilities.
Job responsibilities include: performing day to day operation and support of the advanced surface science laboratories; assisting in data acquisition and analysis; participating in selected research projects to ensure contractual deliverables are met; providing materials characterization of microelectronic, optoelectronic and photonic samples for faculty and staff; providing instrumentation training to staff and student; and working with students on the advanced analytical tools.
The position requires: a Ph.D. in materials science or related field such as physics or chemistry and a minimum of three years experience in the characterization of microelectronic, optoelectronic or photonic materials or a bachelors degree with 10 years of relevant work experience; demonstrated ability to work in a high energy, team oriented environment; and excellent communication and analytical skills. Preference will be given to those candidates with the required work experience with Auger Electron Spectroscopy, X-Ray Photoelectron Spectroscopy, Scanning Electron Microscopy, X-Ray diffractometry, Atomic and Scanning Tunneling Microscopy, or Transmission Electron Microscopy. Salary and Benefits are highly competitive and dependent upon experience.
Please submit a resume and cover letter to:
Jacqueline DiStefano NYS Center for Advanced Thin Film Technology CESTM B110 251 Fuller Road Albany, NY 12203
The SUNY/Research Foundation is an Equal Opportunity/Affirmative Action Employer.
Well, the best method for presevering wonderfully empheral / delicate structures is cryo-preservation and freeze subsitution. I see you are a little far from us to come and try some freezing, but maybe you have some ready access to some rapid freezing.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
I'll second the opinion about getting your money's worth out of Adobe Photoshop. I've used PhotoImpact (we have v 3 here at work; I've not used v 4 which is out already) and agree it's good. My favorite, though, is Paintshop Pro (Jasc, Inc.) which I bought for home after trying a download version. I find it is a little more intuitive than PhotoImpact (at least for me!). It handles layered images like Photoshop, and runs most Photoshop plugins. I even use it with the Image Processing Toolkit (Dr. John Russ); most of the plugins run without problem, although a few tend to crash. PhotoImpact also handles layers, I believe, but with an object-oriented approach. I haven't tried the IP Toolkit plugins in PhotoImpact.
Paintshop Pro can probably be had for a little less than Photoimpact. List price is probably $90 or $100, but I've seen it on some of the on-line computer stores for much less. I think one may have even had it for something like $58 (?). Check out info & demo at http://www.jasc.com
Disclaimer: I have no ties to any of these software packages!
Jim Passmore Cryovac Division Sealed Air Corp.
---------- } From: Walter.Bobrowski } To: Microscopy } Subject: RE: Photo Editors } Date: Thursday, March 11, 1999 3:52PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think you are making an excellent choice with the new Agfa DuoScan T2500. You will appreciate the 3.4 D on this unique flatbed scanner. The glass less
negative carriers fit into the lower tray, which slides into a precise position. The negatives will not contact any surface. I have just ordered also a DuoScan T2500. You will learn more about it at:
One of the most versatile imaging software is CorelDraw8 and CorelPHOTO-PAINT8. It offers the highest image format manipulation with Plug-ins, for most applications. Highly recommended.
Laszlo J. Veto
Caspar McConville wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are thinking of purchasing a negative scanner for use with TEM } negatives from our Jeol 2000-FX, and also for SEM negatives. A } scanner has been recommended to us: the Agfa Duoscan T2500, which } has a resolution (hardware) of 1250 x 2500 dpi and a density of 3.4D. } (The scans would be output to a Kodak DS 8650 PS printer) } } We need the quality of the scans to match the quality of the standard } darkroom enlarger if possible, as we would like to 'go digital' at } least for routine work. Does anyone have experience of routine } negative scanning for TEM prints, with this or other scanners, and if } so, is it realistic to expect such high quality? } } Also, what additional image processing software would people } recommend we got to go along with this? } } Any advice would be appreciated. } } Caspar } } Caspar McConville, Ph.D. } Technical Specialist } New York State College of Ceramics } Alfred University
I am pseudocolorizing some data we have of cellular responses from striatum of rats learning a T-maze from Ann Graybiel's tetrode recording project. What I am looking for is any "standard" pseudocolor tables used by imagers to colorize 0-255 grey levels into RGB values.
I can apply a standard spectrum from 0 to 255 running from magenta (255 0 255 RGB) through blue, cyan, green, yellow to red (255 0 0) or start with blue and go to red in a linear way. What this produces is colors that are mostly in the middle. What I think imagers must be using are tables weighted toward the red and blue, so transitions show up better. I can play with my tables in Excel to boost the red and blue ends and then apply them using Paint Shop Pro - what I was wondering was if you have or know where I can find any standard tables, preferably of numerical RGB values used by the pros in the field.
Hi, A student in the lab is looking at museum samples of dinosaur bones and teeth on the SEM. He could get access to more samples if he could restore them to their original condition ( ie, remove the gold). Is there a good nondestructive way to do this?
Kim DeRuyter Histology and Electron Microscopy Labs University of Alaska Fairbanks
In a message dated 3/12/99 9:55:46 AM Hawaiian Standard Time, underwoo-at-u.washington.edu writes:
{ { Bad news! the other issue is: Life is short and work is long, and working alone sucks. It's not emotionally healthy. } }
Perhaps it is true for some that working alone is not emotionally healthy. However, if you ARE emotionally healthy, then working alone can be emotionally healthy and spiritually healthy also. It can be a time for focussed energy, contemplative thought, creative thought - all without interruption. Personally I love working alone and after the work I enjoy the company of my family and friends.
} From: Robert Underwood {underwoo-at-u.washington.edu}
} } In the past I have been on jobs where a great deal of the time was spent } isolated. Now days I think it is a huge safety liability. One time I had } appendicitis and had to drive myself to 50 miles on back roads to a clinic } with my knees up on the stearing wheel. The point here is, even though } you work in a laboratory complex, if something happened you probably } wouldn't get help until the cleaning crew found you. Bad news! the other } issue is: Life is short and work is long, and working alone sucks. It's } not emotionally healthy. Make a change. Consolidate in with other workers.
I spent most of my life farming and ranching. Both rather dangerous occupations. You spend almost all your time alone and in the busy season I might be alone 48 hours at a time so no one would even start looking for me for a couple of days.
Not many people are killed because of the isolation. Your best protection' is to think before you do something dangerous.
Gordon
Gordon Couger gcouger-at-couger.com Owner PRAG-L PRactical AGriculture List www.couger.com/prag-l Stillwater, OK 405 624-2855 GMT -6:00
Can anyone help me acquire an instruction manual for a Reichert-Jung FC4E cyroultramicrotome attachment for a Reichert-Jung Ultracut E ultramicrotome? I would be able to pay copy and mailing expenses.
Thanks
Damian Neuberger Research Scientist damian_neuberger-at-baxter.com
There are some two way pagers that have a man down feature. If the pager is in a horozonal postion for 10 seconds it beeps. If the wearer doesn't acknoledge the beep it sends an alarm to a central station. These pagers have a panic button as well and will serve as regular alpha numeric pagers that allow yes/no acknoledgement from the wearer.
Disclaimer: I own a substantial share of Datalink System that manufactures and selling these. See www.rfdata.net. Gordon
Gordon Couger gcouger-at-rfdata.net Datalink Systems www.rfdata.net Stillwater, OK 405 624-2855 GMT -6:00 =================================
Everett Ramer's question regarding working-alone has ellicited thoughtful and poignant reply. Useful reply as well, by Jerry Heeschen.
To level this solemn feeling, here is mine originally just shared with Everett.
Nathan Haese - at work, alone in a garage, on a new microscope, perhaps too alone.
Everett,
At a large chemical company that I once worked for, we had a badge with a radio alert button that we wore when we worked alone on weekends. Pressing the button would alert the gate guard to come find us in the lab=
Does any one have an instruction/operations manual for an American Optical microtome knife sharpener? Or if it's basic enough to enlighten me just put it in an email? Thanks in advance Steve D'Angelo
Let's join and pay our tribute to Drs Richard Henderson & Nigel Unwin for their Aminoff Award.. Cheers!
The Royal Swedish Academy of Sciences has decided to award the Gregori Aminoff prize in crystallography for 1999 to dr. Richard Henderson and dr. Nigel Unwin, MRC Laboratory of Molecular Biology, Cambridge, England, for their development of methods for structure determination of biological macromolecules using electron diffraction. The prize is presented at the Annual Meeting of the Academy 31. March 1999. The Aminoff symposium 29 - 30. March is organised to the honour of the prize-winners.
The symposium is supported by the Academy through its Nobel Institute for Chemistry.
Aminoff Symposium Structure Determination of Macromolecules with Electron Diffraction 29 - 30. March 1999
Monday 29. March
13.00 - 13.15 Opening of the symposium: Erling Norrby Introduction: Ivar Olovsson
General and non-biological Systems
13.15 - 14.15 Atomic Resolution Electron Microscopy in Biology Richard Henderson
14.15 - 15.00 Imaging Individual Atoms by Electron Microscopy Sven Hovmller, Stockholm
15.00 - 15.30 Coffee/Tea
Diffraction Studies of Membrane Proteins
15.30 - 16.30 Different methods for the study of membrane structures. Matti Saraste, EMBL, Heidelberg
16.30 - 17.15 Electron Crystallography of Membrane-bound Enzymes Hans Hebert, Stockholm
17.15 - 18.00 Structural Studies on the Cytochrome bc1 Complex So Iwata, Uppsala
18.00 - 18.30 General discussion
18.30 Dinner in the Club House of the Academy
Tuesday 30. March
Studies of Virus Structures
09.00 - 10.00 Combination of Different Methods in Virus Studies Michael Rossmann, Purdue
10.00- 10.45 Virus Studies, Essence of Supermolecular Symmetry Holland Cheng, Stockholm
10.45 - 11.15 Coffee/Tea
Non-crystalline Materials
11.15 - 12.00 Visualization of Single Protein Molecules by Electron tomography. Ulf Skoglund, Stockholm
12.00 - 13.00 0 - dimensional Crystallography Marin van Heel, Imperial College
13.00 - 14.30 Lunch in the Club House
Concluding talks
14.30 - 15.30 Making Light Work: The Membrane Proteins of Plant Photosynthesis Werner Khlbrandt, Frankfurt
Getting gold off museum fossil pieces is nay impossible. Al would be easier but basically its the same problem. C coating is good enough for low powers, but again, Curators do not like that dark coating, To view these "hard, dry and non-conducting" specimens uncoated, the best solution is a poor vacuum SEM; a fully fledged Environmental SEM would also do well, but its a more expensive instrument for that job. Poor vacuum SEM's (I believe at least a couple of the major manufacturers make instruments with that facility) use only mechanical pump vacuum in the specimen chamber and because of a vacuum limiting aperture retain high vacuum in the gun chamber. Secondary mode is impossible, but a Robinson detector gives excellent images for this type of work. Magnifications under these conditions are limited to about 2000x, but details in fossils do not warrant higher magnifications; its the SEM's superior depths of field that wins out over light microscopy. Kim - all you require now is one of those scopes! Years ago I modified an Etec Autoscan to function reversibly as a poor vacuum instruments. It worked well but it was a fair bit of trouble to accomplish the required modifications. Our online contain a link to an archive collated by Scott Wight. This contains listserver contributions concerned with Environmental and Poor Vacuum SEM. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, March 13, 1999 6:55 AM, Kim DeRuyter [SMTP:fnksd1-at-uaf.edu] wrote: } } Hi, } A student in the lab is looking at museum samples of } dinosaur bones and } teeth on the SEM. He could get access to more samples if } he could restore } them to their original condition ( ie, remove the gold). } Is there a good } nondestructive way to do this? } } Kim DeRuyter } Histology and Electron Microscopy Labs } University of Alaska Fairbanks }
Due to the response (and fun) of our contest at last years MSA/MAS Conference, we will repeat the contest this year in Portland. The concept of the contest is based on composite micrographs, each made up from two or more images - one of which must be microscopical in nature. Prizes of value will be awarded and one will not have to be present to win. If of interest, kindly advise by return email and I will see that you receive full contest detail. Don Grimes, Microscopy Today
Many years ago I worked in a hospital basement EM lab. One day my boss and I came out of the inner TEM room and wondered why the hallways were deserted. Then a fire marshall came by demanding to know why we hadn't vacated the building during the fire drill!
The following email notice arrived from our university safety office: ------------
Kim
I seem to remember a story - a long time ago - about about dipping the sample in liquid mercury - the gold is taken into the liquid as an amalgam and leaves the specimen clean. I have never tried it. I do not know what any Safety person would say about that these days!
Keith Ryan Marine Biological Association of the UK Citadel Hill Plymouth Devon PL1 2PB England
In a message dated 3/12/99 9:45:34 PM Mid-Atlantic Standard Time, fnksd1-at-uaf.edu writes:
{ { Hi, A student in the lab is looking at museum samples of dinosaur bones and teeth on the SEM. He could get access to more samples if he could restore them to their original condition ( ie, remove the gold). Is there a good nondestructive way to do this?
Kim DeRuyter Histology and Electron Microscopy Labs University of Alaska Fairbanks } } Good Morning! A less destructive way to analyze these samples would be to coat them with carbon instead of gold, and then ashing the carbon off with O2. Gold is tricky to remove on most materials (I work mostly with semiconductors), but would be very difficult to remove on dinosaur bones. (This is assuming your samples are small enough to fit into an available asher or RIE tool). There are several labs that could perform both the coating and ashing - let me know if you have trouble locating one close to your area. Lisa Montanaro Consultant, MME
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im, A good way to handle specimens of this sort is to make casts of the surface and examine the casts in the SEM. This way there is no damage to the original artifact. I had an anthropology grad student do this with human teeth for his thesis research. He worked out a very inexpensive, low tech, but reliable method to do the casting. He can be reached at the following for full details of his method: Dr. Chris Schmidt Indianapolis University cschmidt-at-indy.edu
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
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Hi, A student in the lab is looking at museum samples of dinosaur bones and teeth on the SEM. He could get access to more samples if he could restore them to their original condition ( ie, remove the gold). Is there a good nondestructive way to do this?
Kim DeRuyter Histology and Electron Microscopy Labs University of Alaska Fairbanks
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by dj.stud.ntnu.no (8.9.1/8.9.3) with ESMTP id QAA03224; Mon, 15 Mar 1999 16:01:30 +0100 (MET)
I have the same problem with the Microm-Heidelberg, Heavy duty Microtome HM 350. We would also be able to pay copy and mailing expenses.
Gary. NTNU Norway. } } } Can anyone help me acquire an instruction manual for a Reichert-Jung FC4E } cyroultramicrotome attachment for a Reichert-Jung Ultracut E } ultramicrotome? I would be able to pay copy and mailing expenses. } } Thanks } } Damian Neuberger } Research Scientist } damian_neuberger-at-baxter.com } } }
Two postdoctoral fellowships are available immediately in the Supramolecular Structure and Function group in the Bioengineering & Physical Science Program at the NIH. Our laboratory is looking for a physical scientist (e.g., physics or materials) and biological scientist (e.g., biophysics) to help develop and apply new methods based on electron microscopy and spectroscopy. There is considerable flexibility in the scope of research which includes the following:
(i) development of EELS spectrum-imaging in the STEM to map phosphorus, calcium and other elements in macromolecular assemblies and cells.
(ii) development of energy-filtered TEM to establish elemental detection limits and to map elemental distributions.
(iii) development of x-ray microanalysis and cryo-preparation techniques for studies in cell biology.
(iv) image processing techniques to determine the structures of large macromolecular assemblies using cryo-EM and STEM.
(v) applications of any of the above methods to biomedical research in collaboration with investigators in other NIH laboratories.
Our laboratory is equipped with field-emission scanning transmission electron microscopy (STEM), electron energy loss spectroscopy (EELS), energy-filtered electron microscopy (EFTEM), x-ray microanalysis (EDXS), cryo-electron microscopy, and UNIX-based image processing.
Preference will be given to candidates with less than five years of relevant postdoctoral experience. Candidates from the United States or from overseas are welcome to apply.
For additional information see: http://www.nih.gov/od/ors/beps/ssfr/
Please send curriculum vitae and bibliography to:
Dr. Richard Leapman
Biomedical Engineering & Physical Sciences Program
- Eye on Imaging - MSC/SMC Conference May 26-28, 1999
Sponsored by the Microscopical Society of Canada
We are pleased to annouce the 26th annual meeting of the Microscopical Society of Canada. This spring meeting and exhibition will be taking place for three days, May 26-28, 1999, on the campus of the University of Guelph in Guelph, Ontario. Many interesting speakers have agreed to participate including Dr. John Russ (author of the Image Processing Handbook, Dr. P.C. Cheng (multi-photon microscopy and microscope construction) , Dr. Chris Yip (AFM), Dr. Nestor Zaluzec (Tele-Presence microscopy), Dr. Nick White (3-D quantitative analysis and multi-photon microscopy) and Dr. Brian Kaye (Fractal analysis) amongst others. We are also offering a variety of afternoon workshops as well as a commercial exhibition offering a full range of Microscopy and Imaging equipment and supplies. Please visit our web site for information and registration packages:
http://www.uoguelph.ca/botany/rootlab/msc99.htm
Please pass this link along to anybody that may be interested. Deadline for submission of abstracts and pre-registration is April 6, 1999. Hope you can make it,
George Harauz Microscopical Society of Canada Chairman, Local Organizing Committee University of Guelph Guelph, Ontario gharauz-at-uoguelph.ca
You might look into utilities that allow you to adjust the color LUT and data separately of each other. Then you should be able to stretch the colors to fit like you want. If that is not easily possible with your software, then you might try playing with the gamma, contrast and brightness on your image before applying the pseudo-color and you should be able to get your weighting as you like it.
If you absolutely need help, I might be able to fabricate a color table here if you can send me a typical image and your color table.
At 03:46 PM 3/12/99 -0500, you wrote: } Hi computer imagers,, } } I am pseudocolorizing some data we have of cellular responses from striatum of } rats learning a T-maze from Ann Graybiel's tetrode recording project. What I } am looking for is any "standard" pseudocolor tables used by imagers to colorize } 0-255 grey levels into RGB values. } } I can apply a standard spectrum from 0 to 255 } running from magenta (255 0 255 RGB) through blue, cyan, green, yellow to red } (255 0 0) or start with blue and go to red in a linear way. What this produces } is colors that are mostly in the middle. What I think imagers must be using } are tables weighted toward the red and blue, so transitions show up better. I } can play with my tables in Excel to boost the red and blue ends and then apply } them using Paint Shop Pro - what I was wondering was if you have or know where } I can find any standard tables, preferably of numerical RGB values used by the } pros in the field. } } } ------------------------------------------------------------------ } |Glenn Holm {mailto:karuzis-at-wccf.mit.edu} | } |Graybiel Lab (617)253-5780;fax (617)253-1599 | } |M.I.T Dept. of Brain + Cog. Sci. Cambridge, MA 02139 | } ------------------------------------------------------------------
Kim DeRuyter wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } A student in the lab is looking at museum samples of dinosaur bones and } teeth on the SEM. He could get access to more samples if he could restore } them to their original condition ( ie, remove the gold). Is there a good } nondestructive way to do this? } } Kim DeRuyter } Histology and Electron Microscopy Labs } University of Alaska Fairbanks Kim, One thing that no one has mentioned, yet, is low kV operation. If your instrument will operate at 5kV or lower (perferably around 1kV) and you limit your beam current, you should be able to view uncoated specimens at at least a couple of kX. Some intruments will go much higher at that voltage range. Grains of quartz may still present a problem because SiO2 is such a good insulator, but many other minerals will work fine under those conditions.
Ken Converse owner Quality Images third party SEM service Delta, PA
I have set up an FTP site at prism.mit.edu, port 2101, with the files I mentioned that will convert LINK/Oxford AN10/eX/L files. Apologies if the documentation is sparse!
To access a non-standard port in FTP you will need to know how your FTP program works. In Netscape, use the following URL:
ftp://prism.mit.edu:2101
In WS_FTP you have to change the port number in the Advanced tab. From a command line ftp program (e.g. Unix, or DOS from Win 95/98/NT), first invoke the program without a server name, i.e. just enter
ftp
The program responds with the ftp prompt ftp} . Then you enter:
open prism.mit.edu 2101
The user is anonymous, and the password is unimportant. I don't know how you would do it with other FTP programs. Sorry about the difficulty, but I am already using the standard FTP port for another, non-public, purpose!
I think you are making an excellent choice with the new Agfa DuoScan T2500. You will appreciate the 3.4 D on this unique flatbed scanner. The glass less
negative carriers fit into the lower tray, which slides into a precise position. The negatives will not contact any surface. I have just ordered also a DuoScan T2500. You will learn more about it at:
One of the most versatile imaging software is CorelDraw8 and CorelPHOTO-PAINT8. It offers the highest image format manipulation with Plug-ins, for most applications. Highly recommended.
Laszlo J. Veto
Caspar McConville wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are thinking of purchasing a negative scanner for use with TEM } negatives from our Jeol 2000-FX, and also for SEM negatives. A } scanner has been recommended to us: the Agfa Duoscan T2500, which } has a resolution (hardware) of 1250 x 2500 dpi and a density of 3.4D. } (The scans would be output to a Kodak DS 8650 PS printer) } } We need the quality of the scans to match the quality of the standard } darkroom enlarger if possible, as we would like to 'go digital' at } least for routine work. Does anyone have experience of routine } negative scanning for TEM prints, with this or other scanners, and if } so, is it realistic to expect such high quality? } } Also, what additional image processing software would people } recommend we got to go along with this? } } Any advice would be appreciated. } } Caspar } } Caspar McConville, Ph.D. } Technical Specialist } New York State College of Ceramics } Alfred University
} From: Self {miller.TEX.TAFA} To: Microscopy-at-MSA.Microscopy.Com
Howdy all, There is a graduate student here who is trying to look at a possible stem cell line under TEM. The problem is that they are quite small, don't seem to pellet very well, and he has only been able to get me a few thousand cells at a time. We have tried to embed the cells in 2% Agar after fixation but this hasn't worked. Is it possible to filter the media and cells, and then process the filter with the cells stuck onto it? Maybe use a cytospin to spin the cells into a filter? Is there anyone with experience with this? Any suggestions or ideas would be greatly appreciated.
Thanks in advance Frank Herbert Technician Integrated Microscopy Core Department of Cell Biology Baylor College of Medicine
Somewhere in the deep receses of my mind I recall a cytochemical test for arsenic. I think it was for EM but I am not sure. Does anyone out there know it??
Or was it all just a bad dream?
Greg Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
We are trying to equip our lab with a reflected light BF/DF microscope (+ trinocular head, B&W video system, 4X5" Polaroid system), for two basic needs: 1. viewing and photographing our fine metal, ceramic, and carbide powders for QA purposes (e.g. visual standards), and 2. viewing and photographing mounted/polished thermal spray powder and coating samples prepared at our parent company's facility.
Our budget allows for a used (reconditioned or demo) top-name microscope (e.g. Nikon, Olympus, Leitz, Zeiss) or a new lesser- name scope (e.g. Meiji...) with the same basic features. Not being professional microscopists ourselves, we ask you to comment on aspects of the above tradeoff, based on your experience. Our main concern is flatness of field and sharpness of image at magnifications up to 500X. Thanks for your help!
Sincerely,
Robert A. Miller TAFA Material Technologies, Inc. 1702 Mykawa Road Pearland TX 77581 USA PHONE: 281-485-7765 FAX: 281-485-0211 EMAIL: miller-at-tafa.com
by dogwood.botany.uga.edu (8.9.1/8.9.1) with SMTP id RAA17081 for {Microscopy-at-Sparc5.Microscopy.com} ; Mon, 15 Mar 1999 17:56:43 -0500 (EST) Message-Id: {199903152256.RAA17081-at-dogwood.botany.uga.edu} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Hi, I'm trying to find out if the elemental imaging system on my scope (a Zeiss 902A TEM) is working...or if it is a problem with the sample. Does anyone have a sample they could spare for elemental imaging?
In October, I had a great sample grid (30nm liver sections with no stain). We were able to get nice images of iron. The scope had its annual service in November and the elemental imaging system has not worked correctly since that time. The serviceman is suggesting that the sample is fried (no pun intended). I think the scope is whacked. So I'm trying to get another liver sample (my source for the first sample retired) or I'm willing to try something else. I need a "known" sample, cut 30 nm thick, no support film (600 mesh grids help) or staining.
Any help or suggestions would be GREATLY appreciated!
Sincerely, Beth Richardson
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm interested. Elaine
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
hello members. here is a probable dumb question. Maybe an impossible situation.
Suppose that I have a prepared microscope slide-- 1"x3" glass slide with specimen under cover slip. Ordinary LM analysis works fine. Is there some other analysis method besides confocal that would offer better resolution and increased depth of field? The idea is to not have to prepare TEM specimens.
Is this possible? Gary Gaugler, Ph.D. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Thanks to everyone for your fantastic responses to my request for a Reichert-Jung FC4E manual. The folks from Leica have been especially helpful and quick to respond and I'm most grateful for the help.
As Valdemar Furdanowicz wrote you, you can transfer LINK files to the PC = with their MSDOSCV program. However, you can also use the serial port of the = LINK=20 computer and it can be a little bit faster. What I don't know, however, = is whether you can connect the the two machines directly. When I worked in the electron = beam lab of Hungalu Engineering, we had a little box between the serial ports=20 of the machines, made by Oxford. You can try any terminal program on the PC if it has an option for the = well-known XMODEM protocol. Once transferred, the files have to be converted from one format to = another , because the numbers are formatted differently on the LINK and the PC. With fixed point numbers the only difference is the order of the bytes, = but with floating point numbers the problem is more serious.=20 Your opportunities for file conversion are very limited with the MSDOSCV = program.=20 Of course you have to know the structure of the files, if you want to = convert them.=20 You can find this information in the Oxford manuals.
I wrote some programs to solve the above mentioned problems: 1. File transfer: with our XMODEM program we can transfer multiple files = using wildcards in=20 the file specifications. For this purpose the PC program prepares the = necessary LINK macros, which are sent as a first step. 2. File inspection: with a little utility program one can look at the = files obtained from the LINK computer and compare their content with the documentation.=20 3. File conversion: I have worked out simple conversion programs for = image files, DIGISCAN image anal. result files and X-ray spectra.
With a private letter I can send you anything from these programs, but = there are still some minor=20 problems with that. When I worked for Hungalu, I was the only user of my = programs, and I didn't have enough time for a better user interface. They are written = for the DOS platform, and they send Hungarian messages to the screen. Now I work as a computer programmer in a very different field, and =
in our group we are using formvar-coated grids to stabilize ultrathin sections for immunogold-labelling. Sometimes the formvar is *heavily* labelled by gold particles. This is no= t due to binding of the gold grains, but depends only on the primary antibo= dy that had been used. It can happen both with Protein A-gold or goat anti rabbit-gold, but using other primary antibodies the formvar is clean. What is the reason for this binding of antibodies to formvar? Is there any blocking reagent that would provide this? Thanks in advance
Birgit
-------------------------------------------------------------------------= ----- Dr. Birgit Neubohn Institut fuer Pflanzengenetik und Kulturpflanzenforschung (IPK) (Institute of Plant Genetics and Crop Plant Research) Corrensstr. 3 D-06466 Gatersleben, Germany
there might be a number of other sources for your problem beside the microscope itself. You could have trouble with your camera and/or image processing system. Please give more details what equipment you are using to acquire an elemental map (SIT or CCD camera, what software, etc. ...) Due to the principle of the energy-filter of your microscope (Castaing-Henry electrostatic mirror type) you would probably not be able to see an image at all if it would malfunction. But to make things clear you could insert a fairly thick biological sample and check: 1. If you the a normal spectrum on the final screen if you are switching from image mode to spectrum mode and 2. if you are able to see the contrast change if you acquire filtered images before and above the carbon edge. If both works fine, you should search the problem elsewhere. Are the intensities of the filtered images before and on the iron edge high enough to allow the system to calculate a useful result? Did you change any other important parameters since your last successful try (slit width, energy losses, acquisition time, background correction, ...)? You can contact me directly if you like to discuss this a little more in detail.
Cheers,
Petra
beth-at-dogwood.botany.uga.edu wrote:
} Hi, } I'm trying to find out if the elemental imaging system on my scope (a Zeiss } 902A TEM) is working...or if it is a problem with the sample. } Does anyone have a sample they could spare for elemental imaging? } } In October, I had a great sample grid (30nm liver sections with no stain). } We were able to get nice images of iron. The scope had its annual service } in November and the elemental imaging system has not worked correctly since } that time. The serviceman is suggesting that the sample is fried (no pun } intended). I think the scope is whacked. So I'm trying to get another liver } sample (my source for the first sample retired) or I'm willing to try } something else. I need a "known" sample, cut 30 nm thick, no support film } (600 mesh grids help) or staining. } } Any help or suggestions would be GREATLY appreciated! } } Sincerely, } Beth Richardson -------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu Visit our WWW site! http://www.crpcu.lu/~wahlbrin
Perhaps a second-hand Reichert Zetopan equipped for reflected light could fit the bill.
These are available from time to time at +/- decent prices in the USA (about $ 1000 - 2000, depending on condition and accessories mounted on the stand). (Some) spare parts are available in Germany or Austria.
Reichert had objectives for incident light for Zetopan 5.5x/0.15; 11x/0.25; 32x/0.65, 45x/0.95. I don't know about higher magnifications... The objectives mentioned are still available trough at least one German dealer.
I don't know about their quality regarding flatness of field and sharpness: I use a Zetopan equipped for biological work/transmitted light. Contact me offline if you are interested (I have no financial interests in this).
Yvan Lindekens. ---------- } Van: Bob Miller {miller-at-tafa.com} } Aan: microscopy-at-sparc5.microscopy.com } Onderwerp: Instrument Quality vs. Budget } Datum: maandag 15 maart 1999 23:32 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We are trying to equip our lab with a reflected light BF/DF } microscope (+ trinocular head, B&W video system, 4X5" Polaroid } system), for two basic needs:
Dear Kim, an old method which I used a long while ago and now and then recently to remove gold from calcareous surfaces of REM-samples is to leave the sample in a solution of NaCN and NaOH and let air bubble through by means of a small glass pipette. The amount of NaCN is about 1-2% and NaOH should be 1 N or more. You may try it on a fresh part of bone perhaps it works. Dietmar Keyser PLEASE NOTE THE CHANGE IN TELEFON AND FAX Dr.Dietmar Keyser Zoologisches Institut und Museum Martin-Luther-King Platz 3 D-20146 Hamburg Germany Tel. +49 40/428 38 4232 Fax: +49 40/428 38 3937 E-mail: Keyser-at-zoologie.uni-hamburg.de
EUREM XII 12th European Congress on Electron Microscopy Brno, Czech Republic, July 9-14, 2000 http://www.eurem2000.isibrno.cz/ -------------------------------------------------------- Second Circular and Call for papers will be distributed in May 1999 to pre-registered scientists. DO NOT FORGET TO PRE-REGISTER YOURSELF AS SOON AS POSSIBLE! PRE-REGISTRATION FORM CAN BE COMPLETED DIRECTLY ON THE WEB SITE:
http://www.eurem2000.isibrno.cz/regform.html
Petr Schauer
+---------------------------------------------------------------------+ | Dr. Petr Schauer, Vicechairman of the Or- | tel.: (+420 5) 41514313 | | ganization Committee of the 12th EUROPEAN | fax : (+420 5) 41514404 | | CONGRESS ON ELECTRON MICROSCOPY | (+420 5) 41514337 | | (Brno, Czech Republic, July 9 - 14, 2000) | e-mail: petr-at-isibrno.cz | | Kralovopolska 147, CZ-612 64 Brno | eurem2000-at-isibrno.cz | | Czech Republic |www.eurem2000.isibrno.cz | +---------------------------------------------------------------------+
Greetings Friends, Does anyone have tips for critical point drying and SEM viewing of fresh fish samples including skin/scale layers? They seem to be VERY oily, and I am wondering if this will hamper the CPD process or muck up the vacuum in the SEM. Any food or fish scientists out there? Thanks, Linda lfox1-at-wpo.it.luc.edu
{bold} "Analyzing Materials Interfaces at Atomic Resolution" {/bold}
There will be a Materials Science symposium at Scanning 99 entitled "Analyzing Materials Interfaces at Atomic Resolution". Scanning 99 is being held at the Hyatt Regency O'Hare in Rosemont, IL from April 11-14 1999. The "Analyzing Materials at Atomic Resolution" symposium is scheduled for Tuesday April 13th. The details of the conference can be found at http://www.scanning.org or can be requested from Mary Sullivan (e-mail: scanning-at-fams.org, tel:201-818-1010). Registration for members of the Midwest Microscopy and Microanalysis Society is at the reduced rates of $150 (regular $ 235) for the whole conference or $50 (regular $ 95) for a single day (all attendees of the MMMS symposium held at UIC last May are members of MMMS).
{bold} Speakers {/bold}
9.00 {bold} M. Haider-CEOS GmbH, Germany {/bold}
"Towards sub-Angstrom resolution by correction of spherical aberration"
9.30 {bold} O. Krivanek-University of Washington {/bold}
"Towards sub-Angstrom electron probes by Cs-corrected STEM."
10.00 Break
10.30 {bold} E. M. James-University of Illinois at Chicago {/bold}
"Atomic resolution scanning transmission electron microscopy on the 200kV FEGTEM"
11.00 {bold} S. J. Pennycook-Oak Ridge National Lab {/bold}
"Probing the Origin of Interfacial Properties by STEM"
11.30 {bold} D. A. Muller-Lucent Technologies {/bold}
"The End of the Roadmap for Silicon Dioxide: The Electronic Structure of Hyper-Thin Gate
Oxides at the Atomic Scale"
12.00 {bold} D. B. Williams-Lehigh University {/bold}
"Atomic-Resolution X-ray Microanalysis in the AEM"
12.30 Lunch
2.00 {bold} L. D. Marks-Northwestern University {/bold}
"Picometer structure determination using Electron Diffraction"
2.30 {bold} J. M. Gibson -University of Illinois at Urbana-Champaign {/bold}
"Statistical Measurement of Electron Scattering Fluctuations in Amorphous
Materials - A new Structural Tool"
3.00 Break
3.30 {bold} M. Gajdardziska-Josifovska-University of Wisconsin at Milwaukee {/bold}
"Quantitative surface microscopy and diffraction over the length scales: Morphology and
crystallography of polar oxide surfaces. "
4.00 {bold} M. Tanaka-NRIM, Tsukuba, Japan {/bold}
"Nano-behavior of Small Metal Particles in the Electron Beam"
I would like to establish some contacts with the TEM comunity in Brazil and I thought that perhaps you could give me some information concerning TEM-oriented Materials Science groups in Brazil.
Regards,
LF Giles
***************************************** Luis Felipe Giles, PhD
MPI of Microstructure Physics Weinberg 2, D-06120, Halle (Saale) Germany
} Kim DeRuyter wrote: } ... } } Hi, } A student in the lab is looking at museum samples of dinosaur bones and } teeth on the SEM. He could get access to more samples if he could restore } them to their original condition ( ie, remove the gold). Is there a good } nondestructive way to do this? } } ...
Of all the suggestions, I believe you'll want to develop the casting procedure for your facility. I can give you the e-mail address for Dr Robert Pastor who can let you know which resin materials he used, but he used two ... one for the field while casting the pliable negative mold of ancient Pakastani teeth, and another casting resin for the durable positive mold in the lab. You'd then be able to coat with gold and archive these specimens for all types of future study ... and I was quite impressed with the detail of the micr-wear and lack of artifacts.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
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I sent the following message out yesterday with the incorrect Email. I talked to Chris who has gone from human teeth to mastodon teeth (a bit large to cast for SEM!!) and he would be happy to provide details and answer questions about his casting method. The correct Email is: cschmidt-at-uindy.edu
Sorry for the mistake. Debby -------------
Kim, A good way to handle specimens of this sort is to make casts of the surface and examine the casts in the SEM. This way there is no damage to the original artifact. I had an anthropology grad student do this with human teeth for his thesis research. He worked out a very inexpensive, low tech, but reliable method to do the casting. He can be reached at the following for full details of his method: Dr. Chris Schmidt Indianapolis University cschmidt-at-indy.edu
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
(SMTPD32-4.06) id A372AEA00BC; Tue, 16 Mar 1999 12:22:58 EST Message-Id: {3.0.3.32.19990316123207.00ea5d10-at-mail.map.com} X-Sender: mme-at-mail.map.com X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.3 (32)
Dear Linda,
The world class expert on fish scales is Dr. Gilbert Hartley in the UK. You can probably get his contact information by contacting the Royal Microscopical Society, 37/38 St. Clements Street, Oxford, England.
Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 07:44 AM 3/16/99 -0600, Linda Fox wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If you are growing stem cells in a culture well plate and they're fairly confluent you can do all the processing in the dish, including embedding. This saves alot of time as you dont have to spin between changes, etc. If the plates are Permanox, any resin will work as will PO. However if they're in something like Falcon or Corning you need to use Eponate 12 and skip the PO. I've tried lots of resins and most will partially dissolve the plastic. After polymerization you simply pop the capsules off the dish and section. This works very well if you have a fairly large population of cells. Otherwise you just need to cut several blocks. I can send you our protocol if you'd like. Good luck!
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
I believe that the 1 N NaOH is unecessarily strong to accomplish the purpose and would result in damage to some specimens. A very brief exposure to NaCN with a stronger oxidant such as ammonium persulfate or hydrogen peroxide would probably accomplish the result without prolonged exposure to the reagents which would lead to their becoming more deeply absorbed into capillaries that may exist.
John Twilley Conservation Scientist
KEYSER.DIETMAR wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Kim, } an old method which I used a long while ago and now and then recently } to remove gold from calcareous surfaces of REM-samples is to leave } the sample in a solution of NaCN and NaOH and let air bubble through } by means of a small glass pipette. The amount of NaCN is about 1-2% } and NaOH should be 1 N or more. You may try it on a fresh part of } bone perhaps it works. } Dietmar Keyser } PLEASE NOTE THE CHANGE IN TELEFON AND FAX } Dr.Dietmar Keyser } Zoologisches Institut und Museum } Martin-Luther-King Platz 3 } D-20146 Hamburg } Germany } Tel. +49 40/428 38 4232 } Fax: +49 40/428 38 3937 } E-mail: Keyser-at-zoologie.uni-hamburg.de
We are in the final phases of deciding which confocal microscope to purchase. We are down to the following two which are comparably equipped and priced. We are having difficulty resolving this decision and would like some input from those who may have some experience with these instruments, the software and support service.
Here are our two choices: ******* Nikon PCM - 2000 MultiLine 3 Confocal w/Compix Software
8 bit capture 2 monitors 3 PMT's Fiber Optic Image capture ******* Olympus Flouview 12 bit capture 1 monitor 2 PMT's Direct optical light *******
We are experienced with standard light microscopes, SEM and TEM but are novices with the subtleties of making a good choice in the purchase of a confocal microscope. Please ask me about any additional information I haven't listed that is pertinent to a clear comparison.
Any suggestions about these instruments and the issues we need to consider would be greatly appreciated.
Thanks in advance for your help, Gerald Harrison Biochemistry/Dental Univ. of Penn.
I am forwarding this message for a Ph. D student in our department.
"Does anyone have references for or a working procedure for differentiating between macrophages and neutrophils in tissue sections processed for TEM. I am currently trying to determine the lineage of inflammatory cells present in dog muscle. These cells harbor a parasite, H. americanum. Preliminary TEM data suggests that these cells are modified macrophages but I need to positively confirm the lineage. I would be grateful for any information or ideas as to how to positively determine that these cells are really macrophages or of a monocyte lineage".
Thanks.
Phoebe J. Doss Manager, Electron Microscope Lab Oklahoma State University
I've read with interest the comments regarding negative scanners for TEM because I'm currently starting to look for scanners for X-ray film. Questions:
1) The resolution info. I've gotten for x-ray film scanners is that they can scan to 50 micron resolution. How do I convert between the dpi resolution quoted on the microscopy listserver to this type of number? My back of the envelope calculations seems wacky when I convert.
2) The x-ray film is much thicker and darker than TEM film. Do the TEM scanner folk talk about what the scanners are capable of blasting through?
At 15:44 16/03/99 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
U.S.A. Contact Jim Hilton Advanced Database Systems 7931 S. Broadway #322 Littleton CO 80122 U.S.A. Tel: + 1 303 761-5635 Fax: + 1 303 761-592
} ***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
We are thinking about revising the user fees (including tech support) for our TEM, SEM and all related equipments. However, before doing so, we would very much appreciate some feedback about user fee structures in place and current rates. This is a centralized, university EM facility serving both biological and material people.
Secondly, the university acquired a new confocal microscope and it is going to be a part of the EM facility. I would also like to get some user fee structures for this instrument.
Thanks in advance,
Soumitra Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
} Date: Mon, 15 Mar 1999 15:01:29 -0600 } From: Frank herbert {fherbert-at-bcm.tmc.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM of Lymphocytes } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Howdy all, } There is a graduate student here who is trying to look at a possible stem } cell line under TEM. The problem is that they are quite small, don't seem } to pellet very well, and he has only been able to get me a few thousand } cells at a time. } We have tried to embed the cells in 2% Agar after fixation but this hasn't } worked. } Is it possible to filter the media and cells, and then process the filter } with the cells stuck onto it? Maybe use a cytospin to spin the cells into a } filter? } Is there anyone with experience with this? Any suggestions or ideas would } be greatly appreciated. } } } Thanks in advance } Frank Herbert } Technician } Integrated Microscopy Core } Department of Cell Biology } Baylor College of Medicine } I presume, from your description, the cells are non-adherent?
Pellet the cells. Remove media with pipet. Add small amount of 2-4% glut (depends on size of tube--?maybe 1/2 ml). Resuspend cells, place into very tiny tube with pointed end, and IMMEDIATELY pellet. Let sit in fix. Do not resuspend pellet. After an hour or so, remove most of the fix, and cut off the tip of the tube just below cells, then slice the tube above the pellet. You end up with cells in the center of a log of plastic. Gently dislodge cells with paper clip. Drain with wedge of filter paper, but don't allow to dry out--should be the consistency of thick oat meal. Cut into 1 mm cubed pieces if necessary. Coat each pile of cells with 1% molten, cooled agar (Do not fix the agar in glut). Wash in buffer. Fix in Os. Proceed as with tissue. This method was printed in Microscopy Today sometime back. Also try:
Miller SE. 1983. Electron microscopy of tissue culture. In Jones BR, Hayes RL (eds.) Techniques in Electron Microscopy: A Laboratory Workbook. Burgess Publishing Co., Minneapolis, MN. pp. 478-512.
OR reprinted as
Miller SE. 1985. Electron microscopy of tissue culture. In Jones BR (ed.) Electron Microscopy: 41 Exercises by 17 Scientists. Library Research Associates, Monroe, NY. pp. 293-315.
Good luck, Sm
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I got 12.7 um for 2000 dpi (1/2000 in x 25400 um/in) and half that if it is 4000 dpi. You should know that the Fuji plate system was originally developed for X-ray plates and was at 50 um. They had to get that down to at least 25 um for TEM usage. They key to blasting is optical density of the scanner (assuming that you have enough light and exposure control). It should be at least 3.4 or higher but costs as this goes up. See the book, Real World Photoshop (3,4,or 5). It has a very good section on scanners and buying considerations. You should also get something that outputs with a depth of 12 bits per channel or higher. It really helps in getting contrast and brightness correct in the final TEM image. I would avoid scanner software that automatically converts it to 8 bits -you can do better. You do need a software platform (NIH Image, Photoshop, Digital Micrograph, etc.) that will handle 16 bit images so that you can convert them to 8 bits.
-Scott ---------- } From: "rgriffin-at-eng.uab.edu"-at-Sparc5.Microscopy.Com To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
I've read with interest the comments regarding negative scanners for TEM because I'm currently starting to look for scanners for X-ray film. Questions:
1) The resolution info. I've gotten for x-ray film scanners is that they can scan to 50 micron resolution. How do I convert between the dpi resolution quoted on the microscopy listserver to this type of number? My back of the envelope calculations seems wacky when I convert.
2) The x-ray film is much thicker and darker than TEM film. Do the TEM scanner folk talk about what the scanners are capable of blasting through?
} Date: Tue, 16 Mar 1999 08:04:52 +0100 } From: Birgit Neubohn {neubohn-at-ipk-gatersleben.de} } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM: antibodies bind to formvar } =20 } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Hello, } =20 } in our group we are using formvar-coated grids to stabilize ultrathin } sections for immunogold-labelling. } Sometimes the formvar is *heavily* labelled by gold particles. This is no= t } due to binding of the gold grains, but depends only on the primary antibo= dy } that had been used. It can happen both with Protein A-gold or goat anti } rabbit-gold, but using other primary antibodies the formvar is clean. } What is the reason for this binding of antibodies to formvar? } Is there any blocking reagent that would provide this? } Thanks in advance } =20 } Birgit } =20 } =20 } -------------------------------------------------------------------------= ----- } Dr. Birgit Neubohn } Institut fuer Pflanzengenetik und Kulturpflanzenforschung (IPK) } (Institute of Plant Genetics and Crop Plant Research) } Corrensstr. 3 } D-06466 Gatersleben, Germany } =20 } Phone.: (+49) 039482 5447 } Fax: (+49) 039482 5139 } e-mail: neubohn-at-ipk-gatersleben.de } -------------------------------------------------------------------------= -----=9D } =20 We use Formvar routinely for ultrathin cryosections. There should be no=20 gold on the plastic. You should block your sections with a protein=20 before the primary, and use protein in the antibody solutions as well. =20 Bovine serum albumen (BSA) and fetal calf serum (FCA) are commonly used. = =20 We wash grids in 5% FCS in PBS 10 min, PBS with 100 mM ammonium=20 chloride--quenches aldehydes in cryo sections (you don't say what kind=20 of sections), PBS, then primary, then 6 washes in FCS/PBS, then=20 secondary in PBS with FCS, 6 more washes; wash off protein with water=20 before proceeding. Next step will depend on what kind of sections you=20 have. Results: no background.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710=20 Ph: 919 684-3452 FAX: 919 684-8735
I am looking for configuration details for a PCVisionplus framegrabber - = specifically, the board jumper settings. These are all listed in the = hardware manual (which we have mislaid). If anyone has this info, could = you please email or fax me, as below. The PCVisionplus card was very popular a few years ago, as one of the = preferred framegrabbers for a variety of image analysis packages (JAVA, = Optimas, etc).
Cheers,
David Vowles Senior Technical Officer Electron Microscopy Unit Australian National University PO Box 475 Canberra ACT 2601 Australia Tel: +61 2 6249 3543 Fax: +61 2 6279 8525 Email: vowles-at-rsbs.anu.edu.au
rgriffen asks ... } } ... } } 1) The resolution info. I've gotten for x-ray film scanners is that they } can scan to 50 micron resolution. How do I convert between the dpi } resolution quoted on the microscopy listserver to this type of number? My } back of the envelope calculations seems wacky when I convert.
My calculations at this late hour would imply 50 microns converts to 500 "per inch", but since "resolution" is the distance between two resolvable points (with something between), a 50u resolution scanner would need scan "optically" at 1000 pixels/inch (the distance between the centers of 2 black pixels with a white pixel between).
} } 2) The x-ray film is much thicker and darker than TEM film. Do the TEM } scanner folk talk about what the scanners are capable of blasting through?
This is a good question ... most transparency and film scanners don't do very well for dense areas. Presumably if the manufacturer advertises a relatively high "dynamic range" it will do better ... but do evaluate for detail and noise in those areas.
} I am wonder if Dietmar method could be used for beetles... } } Regards } } Ricardo } www.coleoptera.org
Hello Ricardo! Especially for insects have a look at:
C. Arno': Removal of gold coatings from biological SEM specimens, European Microscopy and Analysis, July 1998, p.13
In this suggestion bromine vapors are used. If you have problems to get this article, let me know your fax number. I then will send a fax copy -- Gerhard Frank UNIVERSITAET ERLANGEN-NUERNBERG Phone: +49 9131 85 28606 Institut fuer Werkstoffwissenschaften VII Fax: +49 9131 85 28602 Cauerstr. 6, D-91058 Erlangen, Germany frank-at-ww.uni.erlangen.de
Dear Dr. Neubohn, In our experience there is a huge variety in stickiness of primary rabbit antibodies. We have the impression that this originates from different degrees of hydrophobicity, the more hydrophobic the protein, the easier it sticks to a hydrophobic surface (formvar, resins and so on). If this problem shows itself it is almost always with rabbit species. Possible solutions are: 1. make the primary less hydrophobic. Some have done this by pre-incubating the primary antibody with free protein A, but this would make a detection with protein A/gold complexes impossible. If you need a reference I can look it up. An alternative is sometimes, but not always, to use Fab or Fab2 fragments of the primary, but then you would have to use a (goat)-anti-rabbit conjugate. 2. make the substratum less hydrophobic. In general parlodion films seem to be less sticky, but with formvar films possibilities are to use glow discharged films, and a carbon coating. An appropriate blocking step will deal efficiently with hydrophobic binding sites on the substratum. Albumin is suited and a relatively long blocking time helps, as more albumin molecules tend to share larger surface areas with the substratum, resulting in a more efficient blocking with time. If you work with resin sections, you might consider to use Tween-20 (0.1%, not less!) in PBS for an incubation buffer. At concentrations above the critical micel concentration the detergent will prevent hydrophobic interactions like the one you describe. This approach may not be suited for cryo sections, since they are much easier affected by detergents. Good luck, Jan =========================== Jan Leunissen AURION http://www.aurion.nl Costerweg 5 The Netherlands phone: 31-317-497676 fax: 31-317-415955 You will find more technical info on our web site
rgriffin, I've recently had a few attempts at using my Linotype-Hell Saphir Ultra scanner to digitise X-ray radiographs and topographs. I usually use the scanner for TEM negs, and it does a fair job for those as long as they're not too full of contrast (haven't gone in the darkroom since I bought it). I have found it woefully inadequate for X-ray images recorded on nuclear emulsion (grain size 5 or 10 um). The 'real' (supposedly non-interpolated) resolution of the scanner is 1200 dpi, even though it will interpolate up to 5000 dpi. (2.54 cm/1200 = 21 um pixel size). Nearly all of the image detail is lost in a scanned image, even at the maximum resolution setting. And yes, you have to have a fairly weak-looking negative to cope with the contrast produced by a gold wire in a radiograph. My current way of dealing with these images is to use a digital camera on a microscope, which is fine for detail of wire bonds, etc. However, I know I'll have to go into the darkroom today since I have some topographs which are several inches square. I'll scan the prints. In a couple of weeks I'll have a macro lens for the camera and may be able to take pictures from a light box. Unless scanners have improved massively since last year, I wouldn't recommend using one as your only way of getting an image. (I believe 2000 dpi is standard now, and maybe 15% better range in optical density.)
Tel. +44 1327 356363 Fax. +44 1327 356389 } ---------------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------. } } } I've read with interest the comments regarding negative scanners for TEM } because I'm currently starting to look for scanners for X-ray film. } Questions: } } 1) The resolution info. I've gotten for x-ray film scanners is that they } can scan to 50 micron resolution. How do I convert between the dpi } resolution quoted on the microscopy ListServer to this type of number? My } back of the envelope calculations seems wacky when I convert. } } 2) The x-ray film is much thicker and darker than TEM film. Do the TEM } scanner folk talk about what the scanners are capable of blasting through? } }
I have been using a system by Orion with my JEOL 840 SEM. I have been using the DOS version and am in the process of upgrading to the Windows NT version. You can visit their web site at: http://www.OrionMicroscopy.com/
Louie Kerr
At 3:44 PM -0600 3/16/99, "rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX) 508-292-0289 (Cell phone)
Hi Linda, Although I haven't tried this with oily fish scales, I've managed to defat raw food samples (at different stages of refining and production). In preparation for CPD, samples can be dehydrated through a gradient series of ethanol or acetone. If the sample still contains oil, you might try hexane followed by acetone before going to CPD. A common alternative to CPD is to use hexamethyldisilizane (HMDS) x3 following the final 100% ethanol wash. Your samples can then be air dried. Do some trial runs on samples on test samples. The HMDS is available from EMS or other EM supply companies. Let me know how this project works out. Rosemary
We have prepared fish parts including scales for SEM and TEM without problems. The standard preparation protocols of fixation, washes, dehydration and CPD should result in clean dry samples. One problem with scales is that they sometimes tend to curl up as they dry. Usually we just work around that problem but you can try to process them more slowly or try to hold them flat through the processing.
Louie
At 7:44 AM -0600 3/16/99, Linda Fox wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX) 508-292-0289 (Cell phone)
On the general subject of removing sputtered of evaporated metal from SEM samples. I am told that removing silver can be a relatively simple. The following paper describes a method using "Farmer's" reducer, a dilute aqueous solution of potassium ferricyanide and sodium thiosulphate.
"Silver as a removable conductive coating for scanning electron microscopy" by A.A. Mills Dept. Geology, University of Leicester. UK
Best regards Mike Wombwell Polaron range Business Manager V G Microtech The Birches Industrial Estate Imberhorne Lane East Grinstead West Sussex RH19 1UB UK Direct line: +44 (0)1342 310296 Switchboard: +44 (0)1342 327211 Fax: +44 (0)1342 315074 http://www.vgmicrotech.com/polaron-range E&OE
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At 03:44 PM 3/16/99 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Our company, E.L.I. Belgium has designed a very powerful image grabbing s= ystem for SEMs: Orion 5 for Windows.
This system connects to ANY SEM. Its resolution goes up to 8k x 8k (only depending on the SEM resolution). A lot of useful functions are easily available: zoom, distance measuremen= ts, image extraction, text inclusions,...=20 Orion is the only frame grabber with FULL electrical isolation between th= e SEM and the PC. Orion is fully programmable with macro commands. The Orion system is fully configurable: up to 64 image sizes stored in a driver file. Orion runs under Windows 3.1x, Windows 95 and Windows NT. Continuous software updates guarantee the evolution of the Orion system.=20 It is a fully passive device that tracks the SEM scanning signals - easy connection Orion is open to the outside world: all =93 popular =94 image file format= s (TIFF, BMP, TGA, JPG...) can be used.
Please visit our web site (orionmicroscopy.com) to get more info and also= to download a demo program to evaluate the different functions without any cnnection to an SEM or contact us directly to know our nearest dealer.
Best regards,
Paul Vanderlinden. Sales Manager.
********************************************************************* See our web site: http://www.orionmicroscopy.com
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{html} {div} At 03:44 PM 3/16/99 -0600, you wrote: {/div} {div} >----------------------------------------------------------------= -------- {/div} {div} >The Microscopy ListServer -- Sponsor: The Microscopy Society of America {/div}
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At 03:44 PM 3/16/99 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Our company, E.L.I. Belgium has designed a very powerful image grabbing s= ystem for SEMs: Orion 5 for Windows.
This system connects to ANY SEM. Its resolution goes up to 8k x 8k (only depending on the SEM resolution). A lot of useful functions are easily available: zoom, distance measuremen= ts, image extraction, text inclusions,...=20 Orion is the only frame grabber with FULL electrical isolation between th= e SEM and the PC. Orion is fully programmable with macro commands. The Orion system is fully configurable: up to 64 image sizes stored in a driver file. Orion runs under Windows 3.1x, Windows 95 and Windows NT. Continuous software updates guarantee the evolution of the Orion system.=20 It is a fully passive device that tracks the SEM scanning signals - easy connection Orion is open to the outside world: all =93 popular =94 image file format= s (TIFF, BMP, TGA, JPG...) can be used.
Please visit our web site (orionmicroscopy.com) to get more info and also= to download a demo program to evaluate the different functions without any cnnection to an SEM or contact us directly to know our nearest dealer.
Best regards,
Paul Vanderlinden. Sales Manager.
********************************************************************* See our web site: http://www.orionmicroscopy.com
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{html} {div} At 03:44 PM 3/16/99 -0600, you wrote: {/div} {div} >----------------------------------------------------------------= -------- {/div} {div} >The Microscopy ListServer -- Sponsor: The Microscopy Society of America {/div}
Several people in the recent past have asked about the discontinued Leafscan 45 film scanner. Leaf transferred the technology to Bremson, Inc. of Kansas. Their web page can be found at www.bremson.com . The page detailing the specs for the scanner can be found at www.bremson.com/products/45hsprod.htm .
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798 cook-at-aaem.amc.anl.gov
The biggest difference between these cell types is in the nucleus. Neutrophils have a multilobed nucleus, and in thin sections this will a= ppear as multiple small, unconnected profiles. The nucleus of macrophages ma= y be deeply indented, but it will usually not be seen as multiple, unconnect= ed profiles. The cytoplasm of neutrophils is generally more electron dens= e than macrophages, and there will be more granules in the neutrophil cytoplas= m. Macrophages will have a distinct Golgi region with lots of vesicles and=
secretory granules of many sizes.
The best way to learn to differentiate the cell types is to get a buffy= coat sample and fix it without disturbing the cell layers. Neutrophils and eosinophils will layer nearest to the erythrocyte layer, lymphocytes an= d macrophages will be next, and platelets will be at the top. I've got a= n easy procedure for processing buffy coats for TEM, if you'd like to have it.=
Jane A. Fagerland, Ph.D. Abbott Laboratories Abbott Park IL 60064-6202 =
We're new to making density measurements on digital images. I'm looking for basic papers/books on the topic. I already have John Russ's "The Image Processing Handbook" and "Computer Assisted Microscopy". Is there any other good stuff out there?
Dear Fellow Microscopists, What is the best way of determining Carbon in solution in Ni-base superalloys? Chemical analysis strikes to me as one of the best ways, but I don't have enough material. Thanks. Anita
} Date: Wed, 17 Mar 1999 15:01:15 -0600 } To: Histonet-at-Pathology.swmed.edu } From: Sue Danielson {sdaniels-at-post.its.mcw.edu} } Subject: Laboratory Technologist position } } The Department of Neurology at the Medical College of Wisconsin has an immediate opening in their Neuromuscular Diagnostics Laboratory for a Clinical Laboratory Technologist. Our fast-paced, growing laboratory receives muscle & nerve biopsies from hospitals & clinics throughout Wisconsin and Northern Illinois. } } Applicants should possess a Bachelors' degree in the Biological or related Sciences or an MT/HT certification. Laboratory experience in histology (cryosectioning and muscle enzyme histochemistry) and transmission electron microscopy techniques a plus. Excellent organizational skills and ability to work independently as well as with physicians and medical students is essential. } } Excellent salary and benefits package offered including health and dental insurance, 403B retirement plan and tuition reimbursement. This is a full-time, 40hr/wk salaried position (no weekend or evening hrs required) available immediately. } } Applicants may contact/submit resume to: } } Susan K. Danielson, MS } Neuromuscular Laboratory Coordinator } Froedtert Memorial Lutheran Hospital - West } Muscle/Nerve Lab, Rm 1132 } 9200 W. Wisconsin Ave. } Milwaukee, WI 53226 } } Ph: (414) 259-3836 } Fax: (414) 454-7905 } email: sdaniels-at-mcw.edu }
Gerald- I have worked with the Fluoview system from Olympus, and the BioRad MRC600, but not the Nikon system. I prefer the BioRad over Olympus. My advice is to sit down and work with the software, with each of the systems you are considering. Time at the scope is the only way to understand the quirks of each. If you don't have the time, assign someone you trust to do the comparison. Get both scopes set up as DEMOS for at least 2-4 weeks. Run them trough their functions, collect images, edit images, transport to various other SW programs, print to various printers, etc. All systems have bugs, most are in the software, file formats, the ability to transfer images from one platform to another is essential... find the bugs, decide if you can live with them.
Good luck -Mike
disclaimer- I have no financial interest in any of the above mentioned companies.
On Tue, 16 Mar 1999, Gerald Harrison wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear fellow microscopists, } } We are in the final phases of deciding which confocal microscope to } purchase. We are down to the following two which are comparably equipped } and priced. We are having difficulty resolving this decision and would like } some input from those who may have some experience with these instruments, } the software and support service. } } Here are our two choices: } ******* } Nikon PCM - 2000 MultiLine 3 Confocal w/Compix Software } } 8 bit capture } 2 monitors } 3 PMT's } Fiber Optic Image capture } ******* } Olympus Flouview } 12 bit capture } 1 monitor } 2 PMT's } Direct optical light } ******* } } We are experienced with standard light microscopes, SEM and TEM but } are novices with the subtleties of making a good choice in the purchase of a } confocal microscope. Please ask me about any additional information I } haven't listed that is pertinent to a clear comparison. } } Any suggestions about these instruments and the issues we need to } consider would be greatly appreciated. } } Thanks in advance for your help, Gerald Harrison } Biochemistry/Dental } Univ. of Penn. } } }
Thanks to the several contributors who posted very helpful information to me regarding our choice between a Nikon and Olympus confocal. We were restricted to these two choices by budgetary constraints.
I have just emerged from what is the last meeting for making this decision and we went over the many good questions and comments I've received. They were a significant aid in our being able to focus on the kind of unique considerations facing us.
We believed, from on-site demonstrations of both instruments and comments I received from the List, that image quality could be considered equivalent. Olympus was also including an additional remote workstation to compensate for the advantage of the Nikon's Compix compatibility with Windows NT. We were swayed by the apparently more powerful software for the Nikon, but because we will be a multi-user facility primarily doing routine visible fluorescence imaging, and the person who will work with users is already familiar with the Olympus package and believes it is easier for a novice to learn basic image acquirement, we will go with the Olympus.
Again, the department personnel involved in making this choice are very grateful for all the helpful input.
Gerald Harrison Biochemistry/Dental Univ. of Penn.
When I posted this letter for the first time, something went wrong, and = the last 10 percent were lost.
As Valdemar Furdanowicz wrote you, you can transfer LINK files to the PC = with their MSDOSCV program. However, you can also use the serial port of the = LINK=20 computer and it can be a little bit faster. What I don't know, however, = is whether you can connect the the two machines directly. When I worked in the electron = beam lab of Hungalu Engineering, we had a little box between the serial ports=20 of the machines, made by Oxford. You can try any terminal program on the PC if it has an option for the = well-known XMODEM protocol. Once transferred, the files have to be converted from one format to = another , because the numbers are formatted differently on the LINK and the PC. With fixed point numbers the only difference is the order of the bytes, = but with floating point numbers the problem is more serious.=20 Your opportunities for file conversion are very limited with the MSDOSCV = program.=20 Of course you have to know the structure of the files, if you want to = convert them.=20 You can find this information in the Oxford manuals.
I wrote some programs to solve the above mentioned problems: 1. File transfer: with our XMODEM program we can transfer multiple files = using wildcards in=20 the file specifications. For this purpose the PC program prepares the = necessary LINK macros, which are sent as a first step. 2. File inspection: with a little utility program one can look at the = files obtained from the LINK computer and compare their content with the documentation.=20 3. File conversion: I have worked out simple conversion programs for = image files, DIGISCAN image anal. result files and X-ray spectra.
With a private letter I can send you anything from these programs, but = there are still some minor=20 problems with that. When I worked for Hungalu, I was the only user of my = programs, and I didn't have enough time for a better user interface. They are written = for the DOS platform, and they send Hungarian messages to the screen. Now I work as a computer programmer in a very different field, and = microscopy remained only a hobby for me. Well, I would like to port my programs to Windows = for my previous colleagues,=20 but I can work on them only in my spare time. =20 (And maybe, I would like to earn a little money if they can afford to = pay.) Anyhow, you can try to convert the image files with Photoshop. They can = be read by Photoshop as raw bitmaps. Here you don't have byte order problems, because the = LINK machine usually stores=20 every pixel on one byte. Even black and white pictures use eight bits = for every single pixel.=20 (Such files can be packed with PKZIP on the PC with 50 to 100:1 ratios.) There is only one thing you have to do: remove the header from the beginning of the file. As far as I can remember, it is 256 bytes long.
Good luck!
******************************************* Mr. L=E1szl=F3 Varga M=C1V Informatika Ltd. H-1012 Budapest, Krisztina Krt. 37/a Hungary Tel: (36-1) 457-9387 email: varguc-at-freemail.c3.hu ********************************************=09 -------------------------------------------------------------------------= ----------- Robert McDonald wrote: Hi All:
I have recently taken over an SEM with an AN 10000 analyser and I was wondering if anyone out there kows if it is possible to get the results files out to a PC ?=20
I'm looking for spares for a Durst M800 Enlarger and a Fujimoto G70 Dichro = Enlarger. Some bright spark has dropped the negative carrier in both = cases and broken the glass plates. If I can get a replacement negative = carrier or just replacement glass plates that would be great. I'd also = like to get some specs on the plates, so if need be I can get replacement = plates made up. =20 If anyone has a complete unit that they want to get rid of rather then the = parts, let me know, we might be in a position to buy for a reasonable = price. =20
Thanks=20
George
If at first you do succeed, try not to look surprised!
George Theodossiou Dept Applied Physics RMIT GPO Box 2476V Melbourne 3001 Victoria Australia Ph: +61 3 9925 3394 Fax: +61 3 9925 5290 Email: george.theodossiou-at-rmit.edu.au
Dear All, We are in the process of mulling over the question of how much we should charge for various services on offer within the department to in-house users, external users and industry. The aim is to revise the charges at some practical level, neither frighteningly expensive nor laughably cheap. However, in order to come up with realistic values, I thought that some input on what the wider world charges would be of use. Of course, I could just sit down and calculate a figure for each of the things that we can do, but this is complicated as some of the variables are unknown - we don't get an electricity or water bill!! Also, there will be some discrepancies between different countries due to currency and the cost of living, but nevertheless, to know would be of undoubted help. So, please let me know how you price microprobe usage, XRF usage and sample preparation, and thin-section preparation. Yours gratefully, Malc. Dr MP Roberts Department of Geology Rhodes University Grahamstown 6140 South Africa Tel: +27 46 6038316 Fax: +27 46 6229715 ******************************* "A sleep is as good as a walk to a legless insomniac" Anon.
} ---------- } From: John Best[SMTP:jbest-at-elmdas.com] } Sent: Wednesday, March 17, 1999 1:53 PM } To: rgriffin-at-eng.uab.edu } Cc: Best, Christine (BEST); jbest-at-elmdas.com } Subject: Digitizing Philips 515 } } Dear Dr. Griffin, } } Your request for information regarding updating a Philips 515 with } digital imaging was recently forwarded to me. } } We manufacture just such a system, and have installed it on Philips 500 } series SEM's owned and operated at several Philips manufacturing or } research facilities. } } It has active control of the SEM probe, and will therfore add features } like frame averaging to your SEM. Perhaps the best approach is to check } out our website at http:\\www.elmdas.com. } } I look forward to hearing from you. } } Regards, } John Best } } } ELMDAS Co. ELectron } Microscopy } Data } Acquisition } Systems } } Mailing & Shipping: RD1 Box 62A, } Alexandria, PA 16611 } Phone: 814-669-4474 } Email: jbest-at-elmdas.com } Our Website: http://www.elmdas.com }
I'm looking for feedback from labs having experience with insurance companies that provide programs replacing traditional manufacturer's service contracts on scopes and other lab equipment. According to the companies' claims these programs provide substantial savings over regular contracts, with service continuing to be from the provider of choice. In other words, they seem to be saying that we can save huge amounts of money with no decline in service. This is a very attractive proposition for the cost-cutters, but can it be real?
Feel free to reply directly to me, if you like. (And thanks to those of you who have already spoken with me by phone!)
Thanks. Randy
Randy Tindall Electron Microscope Core College of Veterinary Medicine University of Missouri - Columbia Phone: 573-882-8304
I've received several requests for this protocol. Phil Oshel asked tha= t I write it up for Microscopy Today, so it will be available to subscriber= s there, as well.
PREPARATION OF BLOOD SAMPLES FOR ELECTRON MICROSCOPY
1. Collect blood in standard EDTA tube.
2. Spin blood sample in clinical centrifuge to obtain visible buffy co= at (about 3500 rpm for 20 minutes).
3. Carefully remove straw-colored plasma layer with pipette, without disturbing the buffy coat layer. It won't hurt to leave some plasma ab= ove the buffy coat.
4. Gently add buffered glutaraldehyde by running it dropwise down the = sides of the tube, again without disturbing the buffy coat layer. 3% glut in= 0.1M cacodylate or Sorensen's phosphate buffer, pH 7.2- 7.4 works fine.
5. After about 30 minutes, test the plug to see if it's fixed - it sho= uld feel rubbery with gentle prodding. If it is not firm, replace the fixa= tive with fresh and allow it to fix for another 15 - 30 minutes.
6. Remove the fixed buffy coat plug from the tube. A small spatula or=
sharpened wooden stick works well. This step can get a little messy, b= ut the idea is to get the plug out as a coin-shaped disk with only a small amo= unt of adherent erythrocytes on the underside. Erythrocytes can be washed off= with buffer, if needed.
7. Place the plug with the erythrocyte-side down on dental wax and cut= out a long transverse slice. Take the slice and slice it again crossways int= o tissue-sized blocks (1-2 mm thick). Each block will contain all cell t= ypes with erythrocytes on the bottom, then granulocytes (neutrophils, eosino= phils, and basophils), then macrophages (often mixed with large lymphocytes), = small lymphocytes, and finally platelets at the top.
8. Place the blocks into a processing vial, and fix for an additional = hour in glutaraldehyde.
9. Wash in buffer, 2 x 10 minutes each.
10. Post-fix in 1% osmium tetroxide in 0.1 M buffer (same type as used = in glut fixative).
11. Rinse several times in distilled water.
12. Dehydrate in ethanol: 50, 70, and 95%, 15 minutes in each.
13. Absolute ethanol: 2 x 15 minutes each.
14. Propylene oxide: 2 x 15 minutes each.
15. Propylene oxide: epoxy resin (1:1) - 1 hour.
16. 100% resin - 2 x 8 hours
17. Orient for embedding so that the largest face of the block will be sectioned (to get all the cell layers in the section).
18. Polymerize blocks as usual.
The dehydration and infiltration schedule is what I've used and know wo= rks. Other methods will most likely work (e.g., acetone dehydration, differe= nt infiltration schedules).
If there are any further details needed, please contact me.
Good luck!
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064-6202 =
Does the MSA have a web site or something that lists EM & LM job postings? Someone told me that it exists but I don't know where to look. So please drop me a line and I'll fish around in the listings.
With things going swimmingly well,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Dear Robin, I have had the Quartz PCI digital imaging system for several years and I am very happy with it. The software runs on Windows 95, NT and 98, is very robust (even my students can't crash it) and it can attach to any SEM. It really transforms older SEMs. You can contact them at: http://www.quartzimaging.com/ You wrote: } } I need vendors that can sell me the set-up for getting digital images from } our Phillips 515 SEM. Any suggestions? } } Robin Griffin } rgriffin-at-eng.uab.edu } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Regarding the question about charges in a centralized EM facility--We have the same situation but without sem. The federally mandated cost accounting standard has made things difficult but since all universities must now comply, we have no choice. Because of this, your rates should vary based on cost of equipment (and therefore depreciation), cost of personnel and the number of hours the facility is used by paying customers. Our facility charges $25.00 per hour for a Hitachi H7000 TEM and will be charging $35.00 for a new TEM with digital capabilities. Technical help is 35.00 per hour, an inverted multi-photon confocal is $25 per hour and a 3 laser upright confocal with nomarski is $15 per hour. We offer two hours of training at no charge. Use of the darkroom which covers chemicals and the service on our processor is $10 per hour. We charge cost plus shipping on any supplies or chemicals people use. After hours rates are the same. These rates vary yearly depending upon usage. Unfortunately we can't charge what it really cost to operate the facility or we'd have no business--so the university subsidizes us. Hope this helps,
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
It's me again. I've just been asked to speak to a group of high school kids about EM, and how I got to where I am today (I'm not gonna tell them about all the little people I squashed along the way). Actually what I wanted to know is if there are any folks out there doing Forensic EM. You know analysis of blood, guts, bullets, things like that. You know that if I gross the kids out they'll think that EM is coolest thing since ice cubes. So if anybody does that sort of stuff or knows of anybody I can talk to about it, let me know. Cuz y'all know there's nothing better than gettin' a great big EEEEWWWW out of the young'uns.
Gotta go, I hear my microtome calling me,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Dear Robin, We (Soft Imaging System SIS)have a few options that you may want to look at for acquiring digital images from your SEM. This is where you can find a listing of our products on the web: http://www.soft-imaging.de/products/p_one.htm
The first solution is our ADDA II, slow-scan interface for active or passive digital image acquisition from SEM/STEM. Technical data: Resolution: 4096 x 4096 pixel, 4096 gray values. 8 analog (ADC) inputs. 16 logical input and output channels. http://www.soft-imaging.de/products/hardware/h_add.htm
The second solution is our framegrabber (the grabBit) and our software (analySIS) for acquisition of standard video images generated on your microscope. The image quality, S/N ratio, is much worse for video images than it is for the slow-scan interface.
Our software is a state-of-the-art image acquisition, processing, analysis, and archiving package. We have application specific modules, like STEREO, for generating and viewing height mapped images from a stereo pair.
Please contact me if you have any further questions about either of these solution. I would be glad to send out brochures describing our products, or discuss your specific application. By telephone, (888) FIND SIS, or e-mail ac-at-soft-imaging.com
-Sincerely,
Andrew Cahill Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 web: www.soft-imaging.com, www.soft-imaging.de email: ac-at-soft-imaging.com
At 03:44 PM 3/16/99 -0600, you wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
We bought several large Plexiglas developing tanks from Energy Beam Sciences for developing TEM negatives. Several of them split their seams after a few months and we sent them out for repair. After a few months, they split again. Has anyone else had problems with these tanks? They are model DT-111 and are 1-1/2 gallon size (approx. 8-3/4" x 12" x 8").
Please contact me off-line.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Dear Listers, We are planning to purchase a cold stage for a Zeiss Axioplan and would welcome your experience with vendors in this field. It will primarily be used for imaging food products at temps from -20C to -120C. Thanks in advance! Rosemary
Randy- from my experience these Insurance policies are valid, however the biggest variable is timeliness of service. You will not be at the top of the priority list once you leave the manufacturer and use a middle man to schedule your service needs. -Mike
On Thu, 18 Mar 1999, Tindall, Randy D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } I'm looking for feedback from labs having experience with insurance } companies that provide programs replacing traditional manufacturer's service } contracts on scopes and other lab equipment. According to the companies' } claims these programs provide substantial savings over regular contracts, } with service continuing to be from the provider of choice. In other words, } they seem to be saying that we can save huge amounts of money with no } decline in service. This is a very attractive proposition for the } cost-cutters, but can it be real? } } Feel free to reply directly to me, if you like. (And thanks to those of you } who have already spoken with me by phone!) } } Thanks. } Randy } } } Randy Tindall } Electron Microscope Core } College of Veterinary Medicine } University of Missouri - Columbia } Phone: 573-882-8304 } } }
We encircle our tissue samples using a PAP pen to keep the solutions in place during immunocytochemical procedures. We just received a new PAP pen and we think they changed the formula!!
The PAP material lifts off the slides during incubation whereas with the old pens the material stayed put until we removed it.
Do you all have any suggestions regarding something else we could use in place of the PAP pen??
MSA's Placement Office is the location to look for this information. It is located on the MSA WWW Site. Look under General Society Information. at http://www.msa.microscopy.com
} } } Hidee-ho Boarders, } } Does the MSA have a web site or something that lists EM & LM job } postings? Someone told me that it exists but I don't know where to look. } So please drop me a line and I'll fish around in the listings. } } With things going swimmingly well, } } } Paula :-) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } phone: 510-642-2085 } fax: 510-643-6207 } http://biology.berkeley.edu/EML
Three years ago almost to the day we had a discussion on ENTOMO-L about confocal laser microscopes, which several contributors believed would eventually largely replace the use of electron microscopes in insect taxonomy and morphology. One huge advantage is that images are "captured" directly into a computer, where they can be stored and manipulated later. As Richard L. Brown wrote, with regard to insect genitalia, "The confocal can take images every two microns (or more) from the top to bottom and then reconstruct the photo-sections to provide a flat or 3-D image (with resolution comparable to SEM at lower magnifications). The stored data from all the sections can then be compiled and used to provide an image at any level of the structure or a movie or photo of the whole genitalia as it rotates, allowing one to see areas of the genitalia that are obscured by overlaying structures."
Our University has now purchased such a microscope system, and it has recently been installed. Can a confocal laser microscope be used to view entire objects, or just slide mounts?? I called the technician who will be training us in its use, and he had only heard of using it with microscope slides. I went through the email discussion we had on the list, and nowhere is this limitation stated. My desire is to use it with very small beetles (0.6 mm up to whatever the limit is, max. for my taxa about 12 mm) which cannot be dissected or slide mounted because they are type material or otherwise special specimens.
Perhaps we could hear generally, again, the potential uses for these instruments in entomological research. Surely much has happened since Mar. 1996?
Lawrence ..................................................................... Lawrence R. Kirkendall lawrence.kirkendall-at-zoo.uib.no Associate Professor FAX: +47 55 58 96 73 Univ. Bergen Dept. of Zoology TELEPHONE:+47 55 58 23 42 Allegaten 41, N-5007 BERGEN Norway time = GMT + 1 hour Home ph. (if you can't beat the time zone differences):+47 55 95 00 34
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptiste (surname) of Bejsak (Bayshark)-Collorado-Mansfeld
Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., P.O.Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
http://www.coleoptera.org
Home address: 32 Girrawheen Ave. Kiama NSW 2533 Australia
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
We are having problems with our Kevex 770 keyboard. They seem to = print out random letters and numbers when you press certain keys. Does = anyone have a spare that they are willing to part with. thanks.
______________________ Roberto Garcia Senior Analyst, Metallography North Carolina State University Analytical Instrumentation Facility Box 7531, Room 303 EGRC Raleigh, NC 27695-7531 rgarcia-at-unity.ncsu.com http://spm.aif.ncsu.edu/aif
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD} {META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {STYLE} {/STYLE}
{META content=3D'"MSHTML 5.00.0910.1309"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT size=3D3} Hi, {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D2} We are having problems with our = Kevex 770=20 keyboard. They seem to print out random letters and numbers when you = press=20 certain keys. Does anyone have a spare that they are willing to part = with.=20 thanks. {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D3} ______________________ {BR} Roberto Garcia {BR} Senior = Analyst,=20 Metallography {BR} North Carolina State University {BR} Analytical = Instrumentation=20 Facility {BR} Box 7531, Room 303 EGRC {BR} Raleigh, NC=20 27695-7531 {BR} rgarcia-at-unity.ncsu.com {BR} http://spm.aif.ncsu.edu/aif {/FONT= } {/DIV} {/BODY} {/HTML}
I've just been asked to speak to a group of high school kids about EM, and how I got to where I am today (I'm not gonna tell them about all the little people I squashed along the way). Actually what I wanted to know is if there are any folks out there doing Forensic EM. You know analysis of blood, guts, bullets, things like that. You know that if I gross the kids out they'll think that EM is coolest thing since ice cubes. So if anybody does that sort of stuff or knows of anybody I can talk to about it, let me know. Cuz y'all know there's nothing better than gettin' a great big EEEEWWWW out of the young'uns.
I would be glad to talk to you about forensic applications of scanning electron microscopy as that is my job, however......you should understand "forensic EM" goes far beyond the common opinion of " analysis of blood, guts, bullets, things like that". An electron microscopist performing airborne particle analysis for possible occupational/environmental exposure to harmful particulates or the pathologist using electron microscopy to identify a pathogen/toxin induced lesion or foreign object are also performing forensic EM analyses. Forensic means "Of or used in legal proceedings or in public debate" (The American Heritage Dictionary, 2nd Edition). The investigative accountant analyzing data in the investigation of corporate illegal dealings for prosecution and the forensic scientist in the laboratory matching bloody fibers or a bullet found at a crime scene are both performing forensic analyses.
Is the point of your giving this talk to "gross them out" and generate " a great big EEEEWWWW out of the young'uns" or are you teaching to really stimulate some young minds and show them other areas of science that use microscopy for possible careers? High school students are not "young'uns" and you may have a profound impact on a few futures. You can either encourage and stimulate OR you can deter further interest altogether. Many of us were originally guided to our profession by a good speaker or teacher.
As one who both instructs and lectures extensively on forensic applications of electron microscopy, I have found the audiences (high schools, colleges, forensic science and microscopist societies) all enjoy the case history (not gore) from an investigation. The myriad of television shows on the subject speak to the popularity of this current "hot" topic. I'm sure the international emphasis on the "Crime of the Century" has contributed greatly to the recent popularity in forensic science and scientific evidence. You propose to SHOCK your audience with raw crime scene photos when the students are not used to seeing "the real thing". This is counter productive as they will be discussing the shock image(s) the whole time you are trying to explain the science/EM in the investigation. The "gore" is primarily what they will remember and take away with them.
If you really want to TEACH these high school students, show them how to identify objects from two places (exa. - particles such as soil samples from the "crime scene" with particles from a suspect's shoes - SEM/TEM/EDX) or how to find specific particles that confirm a scenario (exa. - specific lake diatoms in "lung fluids" indicating respiration of water from a lake in a possible drowning victim). You can make up your own "scenarios" and supply the "evidence" from the crime scene. Make up a class exercise "crime" and lead them through an investigation of the evidence by EM. Make your SEM/TEM/EDX lecture slides in advance and discuss possible conflicts/artifacts, etc.. You won't need "blood, guts, bullets, things like that" to impress and instruct the students. Leave that for "Hollywood" and the criminal investigator in the proper arena (the court room or instructing other forensic scientists).
You may want to attend the SCANNING 99 meeting in Chicago (April 11-14, 1999) where an all day short course (FORENSIC SCIENCE AND SCANNING MICROSCOPY) will be offered followed by a second day symposium (SCANNING MICROSCOPY APPLICATIONS IN FORENSIC SCIENCE). For information on that International Meeting, call Mary K. Sullivan (201) 818-1010 , web site { http://www.scanning-fams.org } .
Again, I offer my assistance with some other ideas if you are interested. Feel free to contact me off line or by DIRECT E-mail.
Sincerely,
S. Frank Platek Research Biologist/Electron Microscopist Forensic Chemistry Center US Food and Drug Administration 6751 Steger Drive, Cincinnati, OH 45237-3097 (513) 679-2700 , [FAX] (513) 679-2761 E-mail: fplatek-at-ora.fda.gov
DISCLAIMER: THE OPINIONS EXPRESSED ARE SOLELY MY OWN AND DO NOT REPRESENT THOSE OF THE U.S. FOOD AND DRUG ADMINISTRATION OR ANY OTHER AGENCY OF THE U.S. FEDERAL GOVERNMENT OR THE FOUNDATION FOR ADVANCES IN MEDICINE AND SCIENCE (FAMS, INC.).
We purchased two of those tanks this past January. One of them had a leak, but it was suggested to use a silicon adhesive to repair the split (instead of returning the tank). So far the tank is fine. In retrospect, I should have stuck with the stainless steel developing tanks. } } } Bruce
Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124-4305
} ---------- } From: Walck. Scott D.[SMTP:walck-at-ppg.com] } Sent: Thursday, March 18, 1999 10:17 AM } To: Micro } Subject: Plexiglas developing tank -problems } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We bought several large Plexiglas developing tanks from Energy Beam } Sciences } for developing TEM negatives. Several of them split their seams after a } few } months and we sent them out for repair. After a few months, they split } again. Has anyone else had problems with these tanks? They are model } DT-111 and are 1-1/2 gallon size (approx. 8-3/4" x 12" x 8"). } } Please contact me off-line. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P.O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } }
We have been experimenting with telepresence for this purpose. One can get high quality zoom-pan cameras with over-the-web control for less than about $1000 or a simple static camera for about $100. Creating a web page with this view is not very difficult and allows coworkers to monitor each others safety from any place that has network access.
Eric B. Steel, Leader e-mail: eric.steel-at-nist.gov Microanalysis Research Group Office: 301-975-3902 N.I.S.T. FAX: 301-417-1321 100 Bureau Drive, Stop 8371 Gaithersburg, MD 20899-8371
} Over the years our workforce has dwindled to the point that I find myself working alone in the lab (metalographic sample prep/optical microscopy + image analysis) almost all the time. The lab doesn't have windows to the hallway, so it is difficult for people walking by to see if I am in the lab, and if I am OK. My manager is concerned about my safety and is asking me for suggestions. What do other labs do to make working alone safe? } Everett Ramer } Federal Energy Technology Center
When I worked at Sarastro a decade ago, applications specialists in our parent company in Sweden had taken pictures of aphids, orchid seeds, and various other 3D objects very successfully with the confocal. As long as the structures of interest are either autofluorescenct, reflective, or can be stained with a fluorescent tag so that they can be imaged, the confocal is a great instrument for this type of work.
The only caveats: (1) make sure to match the numerical aperture of the objective (which governs the depth of field) to the vertical step height so that you get continiguous information (no gaps in the image) (2) be aware of the size of the image file so that you will have enough room on your computer/tape to handle the file (3) check out various types of reconstruction to see which ones best reproduce the information you are seeking. I seem to remember that "look through projections" worked really well but this is an area which has grown tremendously in the recent past. See what is available on your particular system.
Best of luck, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 10:00 PM 3/19/99 +1100, ricardo wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Three years ago almost to the day we had a discussion on ENTOMO-L about } } confocal laser microscopes, which several contributors believed would } } eventually largely replace the use of electron microscopes in insect } } taxonomy and morphology. One huge advantage is that images are "captured" } } directly into a computer, where they can be stored and manipulated later.
Just an aside here, many SEMs can also capture images digitally. Also, (although it involves an additional step) photographs can be scanned at very high resolutions and bit depths, and subsequently digitally manipulated and stored.
} } Our University has now purchased such a microscope system, and it has } } recently been installed. Can a confocal laser microscope be used to view } } entire objects, or just slide mounts??
My $0.02, based on my previous experience...
Several years ago I worked with Prof. Dan Young of our entomology Dept. on doing some coleopteran cranial morphology studies. The samples in question were, in some cases, one-of-a-kind museum pieces; the only known examples of their species. For this reason dissection and platinum coating of the beetle heads was not a possiblility.
We had some good luck using autofluorescence of the exoskeleton to capture striking tilt-stereo images of the beetle heads, which could then be displayed as 3D stereo-pairs. The dense, opaque, and reflective nature of the exoskeletal material was such that the laser didn't penetrate it readily, so traditional confocal z-series and reconstruction was not possible. We arrived at a protocol that worked, however, in order to capture these pictures, most of the advantages of the confocal had to be circumvented. We used a 3.5x lens with the pinhole open as far as it would go so that we could get a large area and depth of field to capture the large (for microscopy; about 3 x 2 mm) specimens.
It is my opinion that we would have been much better off using a good quality dissecting scope with a and a color video camera to obtain our tilt-stereo images. For smaller and less opaque organelles, there is the possibility that the confocal would work quite well. But for 3D tilt-stereo capture of the larger external features, I would recommend that people try out a good quality, video-equipped dissecting scope.
-- Charles Thomas 4D Software Engineer 4D Imaging Coordinator Integrated Microscopy Resource Univ. of Wisconsin-Madison 1675 Observatory Dr. Madison, WI 53706 608-263-6288 Office 608-265-4076 Fax
cthomas-at-facstaff.wisc.edu
VISIT OUR WEB SITE: http://www.bocklabs.wisc.edu/imr/facility/4D/4d.htm
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A colleague is looking for the following: Leitz Accessory 10-419-613 4x5 cone for the Photomacroscopes (connection point is circular; needs to fit into an MPS 11 manual shutter)
If your lab has one and is interested in selling it, please contact Dennis O'Leary of MicroOptical Methods: 518-482-8200
Thanks Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
I am wondering how many microscopists are suffering from RSI (repetitve stress injuries). I have recently developed DeQuervain's tendonitis in my thumb and I am wondering if it is from moving the specimen drive control knob on the E.M. with my thumb for many years. Not to mention the knobs on the microtome. We work with alot of knobs, some easy to turn and some not Here at Stanford, 3 out of 5 electron microscopists ( 2 are no longer working here but not because of RSI), suffer from hand problems. I thought I would take a survey to find out the prevalence of this condition among microscopists.
Olympus has discontinued a dual output module for their AX and BX line of optical microscopes. This accessory allows two lamps (e.g., halogen and mercury) to be mounted to a vertical illuminator; either source can be used by flipping a mirror. I am told the decision to discontinue the accessory was due to liability concerns that could arise if an operator switched from a halogen to UV lamp source without engaging an appropriate fluorescence cube (with barrier filter).
Several microscope vendors have told me that third-party accessories are available, but can or will not provide any documentation. One company mentioned is 20-20 Technology.
I am seeking referrals to a company providing such an accessory, or a microscopist who wishes to part with one.
Thank you.
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center http://members.tripod.com/~James_Martin/dasrhome.htm
Research Scientist in Chemistry Williams College www.williams.edu http://members.tripod.com/~James_Martin
Does anyone know of an authoritative source explaining the deduction of quantitative information (such as void volume, etc.) from stereomicroscopy of membrane microstructures? Thanks, Sai
******************************************************************** Sai S. Prakash, M.S. Doctoral Student / Graduate Research and Teaching Assistant (Research Area: Cryo-SEM of Coating Microstructures) Chemical Engineering and Materials Science Department University of Minnesota 421 Washington Avenue SE, Box 137 Minneapolis, MN 55455. USA. Tel: 612 625 4809 Fax: 612 626 7246 Email: sprakash-at-cems.umn.edu ********************************************************************
Is anyone familiar with reasonably priced software programs that can manipulate 16-bit gray scale images? While Adobe Photoshop can read 16-bit images, you have to change the images to 8-bit before you can run filters etc. Gatan's Digital Micrograph can work with 16-bit images, but its price is a little steep for equipping several computers. What are the capabilities of other packages? I would like to use the software to work with diffraction patterns as well as images.
TIA, Henk
Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility (614) 292-0674 "An optimist believes that we live in the best of all possible worlds. A pessimist fears that this is true."
WOW!! Thanks for all the advice and info!! You folks are really wonderful!
I'm sorry to take so long to get back to you; I have been "down and out" with a killer cold. Then trying to get caught up on all the "urgent" jobs that always seem to appear just when all you really feel good for is a doorstop.
I'll try to answer everyone at once, to not clutter the list with a bunch of messages. Yes, I think it must be a Zetopan. That name rings a faint bell in the old belfrey. But it is an older model than the one in the image Yvan kindly sent. No, I don't have an image of mine; but I was able to dig up the original inventory entry for it. It was purchased in 1955! And retired to storage in 1981, apparently because it was impossible to get a bulb that would fit it. Yvan, that is interesting about the "Reichert adapter" for the bulb you mentioned. We have quite a few bulbs (apparently bought for it), but they are not long enough to reach the contact at the base of the socket. This was no doubt the problem way back then - our "Mr. Fix-it" (who keeps this place running) has now solved that problem by soldering a tiny washer onto the contact at the base of the socket so that when the bulb is screwed in all the way, it makes contact with the washer. And we have light! The bulbs we have are Osram BPA 554, 6V 5A. There is also a burned-out Osram 8100 with it (was in the socket) but it is even shorter than the BPAs., so didn't meet the contact either. Maybe someone threw out the adapter with the last bulb? Both types of bulbs are the same diameter so have the same tight fit into the bulb housing. There is no identification on the housing as to illuminator type; and the rheostat/transformer has been replaced with one from another make of microscope.
The bulb housing at the back looks like the one in Laslo's image, not Yvan's. There is no "Epi" attachment though; and no "flip-in" phase telescope. Between "6" and "7" in Yvan's image (objective lens turret and eyepiece attachment) my scope has two filter holder slides (empty of course; one with a hole about dime-sized, the other about nickle-sized) and the lever to send the light up the photo tube.
Many, many thanks to Barbara for taking the time to send such detailed instructions re phase set-up! Unfortunately... I can't get past the first step! My scope doesn't have a turret condenser with a brightfield option. I said "dedicated phase" and I should have said "phase only" to be clearer. Sorry. This condenser is strictly one-size-fits-all! It doesn't have an "aperture iris" either. But, it does have a knurled ring around its bottom and a measuring scale (like a ruler) stamped around its front. Turning the knurled ring screws the innards of the condenser up and down through a total of about 15 divisions (I don't know what units) on the scale. It also has the following stamped on its front above the scale: "Objektragerdicke 1,0 mm". I'm sure that means something important! So one can focus the condenser by the usual control that moves the entire gizmo in its fork mount, as well as this "internal" focus. I have, eons ago, set up phase on another scope with a turret condenser, so the steps you detailed are familiar. I don't recall ever using a condenser like this one. But many thanks for the procedure!
Wayne; great! I'd like to have a manual if you have one that might apply to this old microscope. Julian and Susan; I would like bits and pieces ( like a condenser with brightfield option! and brightfield objectives) if you wouldn't have a problem sending them to Canada. I'm appending my address below. Unfortunately we have no money (as usual) for servicing, or new parts. I've just been told to have the manual etc. sent COD, and we'll deal with it when it arrives. I guess it had better not exceed what's in petty cash that day!
Again, many thanks.
Sharon Godkin Canadian Food Inspection Agency Centre for Plant Health 8801 East Saanich Road Sidney, B.C. Canada V8L 1H3
I agree with Jim that the best way to deal with gold on museum specimens, = artwork, taxonomic type specimens, fossils etc is to leave it off in the = first place where possible. The material can sometimes be viewed in a = normal SEM by keeping kV and probe current low, or generally more = conveniently by using one of several types of variable pressure SEM. = Until recently the cheaper versions, working up to about 2 torr, were = restricted to non-SE detection modes (BSE, CL, absorbed current etc) but = several SE detector systems working in this range have become commercially = available. =20 We have just installed a new Robinson detector which allows convenient= BSE and/or SE detection to low kV, in high vacuum or variable pressure = conditions, on a small Hitachi NSEM. (1992) and find the combination mode = in particular gives very good results for topographic imaging. =09 beats explaining a caustic cyanide stew to your friendly curator, I should = think.
cheers
Sally
Dr Sally Stowe Facility Coordinator Australian National University EM Unit Research School of Biological Sciences Box 475, ACT 2601, Canberra, Australia FAX 06 (0)2 6279 8525
James , I don't understand how the light from the mercury illuminator could get to the observers eyes without a filter in place. If a filter position is blank and in place the light from the illuminator has no beam splitter or prisim to direct light to the eyepieces. There fore the light remains enclsoed within the illuminator module... Also there should be a stop on the illuminator to block any light coming through. Leica has the same option built into the stand of their DMR series microscope and does not seem to have a problem with light reaching the observer unfiltered.. This is probably another argument why the Leica is preferred over tthe competition.. Another is being the Lecia optics tend to absorb the harmful UV more so then its inferior Japanese competitors. Did you ever notice the Olympus has the filter shield in place so you don't see the uv light coming out of the objectives at the sample. Thats there for a reason....Harmful UV........ In answer to your question, I do not know of anyone who has one of these devices but would check with some of the larger Olympus dealers. Since it was not such a good idea maybe some of them still have units unsold on the shelf waiting to unload... Good Luck C YA CIP -----Original Message----- } From: James Martin {James.S.Martin-at-williams.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
That Olympus reasoning is absurd! We (Nikon), Zeiss and Leica all still manufacture this part! In any event, here are a few third-party sources where you can get such an item. Try Opti-Quip, Inc., Highland Mills, NY (914) 928-2254 or MicroTec, Inc. Milford, NJ (800) 724-5508. I have used the MicroTec one (product #MIT-001), just specify Olympus adapter fittings.
Good Luck,
Lawrence Kordon Nikon, Inc. Columbia, Maryland nikon-at-jagunet.com
Cono Passione wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } James , } I don't understand how the light from the mercury illuminator could get to } the observers eyes } without a filter in place. If a filter position is blank and in place the } light from the illuminator has } no beam splitter or prisim to direct light to the eyepieces. There fore the } light remains enclsoed } within the illuminator module... Also there should be a stop on the } illuminator to block any light } coming through. } Leica has the same option built into the stand of their DMR series } microscope and does not } seem to have a problem with light reaching the observer unfiltered.. This } is probably another } argument why the Leica is preferred over tthe competition.. Another is } being the Lecia optics } tend to absorb the harmful UV more so then its inferior Japanese } competitors. Did you ever } notice the Olympus has the filter shield in place so you don't see the uv } light coming out of the } objectives at the sample. Thats there for a reason....Harmful UV........ } In answer to your question, I do not know of anyone who has one of these } devices but would check } with some of the larger Olympus dealers. Since it was not such a good idea } maybe some of them } still have units unsold on the shelf waiting to unload... } Good Luck } C YA CIP } -----Original Message----- } } From: James Martin {James.S.Martin-at-williams.edu} } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } Date: Friday, March 19, 1999 10:40 PM } Subject: light microscopy: dual output monitor } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Olympus has discontinued a dual output module for their AX and BX line of } } optical microscopes. This accessory allows two lamps (e.g., halogen and } } mercury) to be mounted to a vertical illuminator; either source can be } } used by flipping a mirror. I am told the decision to discontinue the } } accessory was due to liability concerns that could arise if an operator } } switched from a halogen to UV lamp source without engaging an appropriate } } fluorescence cube (with barrier filter). } } } } Several microscope vendors have told me that third-party accessories are } } available, but can or will not provide any documentation. One company } } mentioned is 20-20 Technology. } } } } I am seeking referrals to a company providing such an accessory, or a } } microscopist who wishes to part with one. } } } } Thank you. } } } } James Martin } } Director of Analytical Services and Research } } Williamstown Art Conservation Center } } http://members.tripod.com/~James_Martin/dasrhome.htm } } } } } } Research Scientist in Chemistry } } Williams College } } www.williams.edu } } http://members.tripod.com/~James_Martin } } } } } }
A point that Cono seems to miss in the criticism of the Olympus scope (with the U.V. filter flange over the objective nosepiece) is that scattered U.V. from the sample is not a function of the quality of the optics or the manufacturer but rather a function of the type of excitation in use.
Some users, particularly biologists, use a mercury or xenon lamp but are not relying upon the U.V. output per se. In some filter combinations a high level of excitation in a certain short wavelength region of the visible spectrum is the most efficient for stimulating emission from a fluorochrome which emits in the longer wavelength region of the visible. Sometimes the cubes which are optimized for this effect limit the amount of short-wave U.V. which reaches the sample. Therefore, scattered light from the sample surface is lower in proportion of U.V. than the proportion emitted from the lamp.
However, people who make use of intrinsic U.V. fluorescence in unlabeled specimens, or cascade effects, often use cubes which pass a significant quantity of U.V. directly to the sample. The scatter or reflected light may be correspondingly high in it's proportion of U.V. Those in forensics and art conservation who rely on this mechanism for all or part of their work are dealing with a different situation by intention rather than accident and must take precautions regardless of the manufacturer.
John Twilley Art Conservation Scientist P.O. Box 2324 Sag Harbor, NY 11963-0113
Cono Passione wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } James , } I don't understand how the light from the mercury illuminator could get to } the observers eyes } without a filter in place. If a filter position is blank and in place the } light from the illuminator has } no beam splitter or prisim to direct light to the eyepieces. There fore the } light remains enclsoed } within the illuminator module... Also there should be a stop on the } illuminator to block any light } coming through. } Leica has the same option built into the stand of their DMR series } microscope and does not } seem to have a problem with light reaching the observer unfiltered.. This } is probably another } argument why the Leica is preferred over tthe competition.. Another is } being the Lecia optics } tend to absorb the harmful UV more so then its inferior Japanese } competitors. Did you ever } notice the Olympus has the filter shield in place so you don't see the uv } light coming out of the } objectives at the sample. Thats there for a reason....Harmful UV........ } In answer to your question, I do not know of anyone who has one of these } devices but would check } with some of the larger Olympus dealers. Since it was not such a good idea } maybe some of them } still have units unsold on the shelf waiting to unload... } Good Luck } C YA CIP } -----Original Message----- } } From: James Martin {James.S.Martin-at-williams.edu} } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } Date: Friday, March 19, 1999 10:40 PM } Subject: light microscopy: dual output monitor } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Olympus has discontinued a dual output module for their AX and BX line of } } optical microscopes. This accessory allows two lamps (e.g., halogen and } } mercury) to be mounted to a vertical illuminator; either source can be } } used by flipping a mirror. I am told the decision to discontinue the } } accessory was due to liability concerns that could arise if an operator } } switched from a halogen to UV lamp source without engaging an appropriate } } fluorescence cube (with barrier filter). } } } } Several microscope vendors have told me that third-party accessories are } } available, but can or will not provide any documentation. One company } } mentioned is 20-20 Technology. } } } } I am seeking referrals to a company providing such an accessory, or a } } microscopist who wishes to part with one. } } } } Thank you. } } } } James Martin } } Director of Analytical Services and Research } } Williamstown Art Conservation Center } } http://members.tripod.com/~James_Martin/dasrhome.htm } } } } } } Research Scientist in Chemistry } } Williams College } } www.williams.edu } } http://members.tripod.com/~James_Martin } } } } } }
Depends on what you mean by reasonably priced. I am a firm believer that Research Systems (RSI) IDL software is the most complete quantitative image analysis software available. It runs on UNIX, VMS, Windows, and Mac OS. I have used all versions except Windows, but I expect it to work like the others. It is not cheap; $1500 single user licence. Has been as low as $500 for .EDU institutions. The software comes with an enormous number of canned routines and its own procedural programming language. It will also interface with external routines and has been used for many years to run the Scanning X-ray Microscope (STXM) at the National Synchrotron Light Source, Brookhaven. It handles any data format you want 8, 16, 24 bit images, 3-D data sets, etc. You do have to spend a week or two learning the basics but I don't consider it difficult. Frequently we ended up hiring work study students to do any special programming. These were biology students not programmers. So if you want something that is complete, extensible, more color tables than you'd ever want, and handles any data format, get IDL. I know this sounds like a commercial but I have no financial interest in RSI or anyother imaging software company.
As an alternative I recommend NIH-Image for the Mac and its equivalent Scion-Image for PCs obtainable from the site shown below. Just remember to check the "calibrate box" and use the import file option. It cannot export 16-bit tiff but with calibrate turned on it will do calculations using 16 bit for projecting 3-D data, numerical calculations,etc. These programs will import image stacks only limited by the amount of available computer ram. For raw data, the import dialog under "custom" requires you to tell how many frames to import. Best of all these programs are free. However, they are not suitable for diffraction analysis. IDL is. Good luck.
http://www.scioncorp.com
} } Is anyone familiar with reasonably priced software programs that can } manipulate 16-bit gray scale images? While Adobe Photoshop can read 16-bit } images, you have to change the images to 8-bit before you can run filters } etc. Gatan's Digital Micrograph can work with 16-bit images, but its price } is a little steep for equipping several computers. What are the } capabilities of other packages? I would like to use the software to work } with diffraction patterns as well as images. } } TIA, } Henk } } Hendrik O. Colijn colijn.1-at-osu.edu } OSU Campus Electron Optics Facility (614) 292-0674 } "An optimist believes that we live in the best of all possible worlds. } A pessimist fears that this is true."
Mario M. Moronne, Ph.D. Material and Life Science Div. M/S 6-2100 University of California Berkeley Lab 1 Cyclotron Rd. Berkeley, CA 94720
ph (510) 486-4236 FAX (510) 528-8076 mariomm-at-ux5.lbl.gov or biostar-at-aldecoa.com
Am headed out of town and don't have time to track down the Reichert manuals, but the phase system you describe sounds amazingly like the "anopteral phase" which I have on my system. Will be back in about a week and will check back with you then.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 08:48 PM 3/19/99 -0500, Sharon Godkin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Check with Media Cybernetics. I am pretty sure that their Image Pro is at least 16 bit now. They are in Silver Spring MD and on the web.
Best of luck Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 08:16 PM 3/19/99 -0500, Hendrik O. Colijn wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The dual output accessory couples two lamp housings to the back end of the vertical illuminator. A mirror is used to reflect light from either lamp housing down the vertical illuminator to a rotating turret, situated over the nosepiece, that contains filter cubes. A variety of excitation/emission cubes would be used for fluorescence examination (UV or visible excitation) using a mercury or xenon source. A brightfield cube would be used for visible light examination using a halogen source. If a mercury or xenon source was used with a normal brightfield cube (not designed for use with a UV source), an observer would not be protected from UV wavelengths reflected or fluoresced from a sample. Of course, this sort of mistake, followed by examination, ought not to occur in practice. However, the possibility does exist with such multipurpose illumination systems. I hope this helps to answer your most reasonable question.
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center http://members.tripod.com/~James_Martin/dasrhome.htm
Research Scientist in Chemistry Williams College www.williams.edu http://members.tripod.com/~James_Martin
On Sat, 20 Mar 1999, John Twilley wrote:
} Cono Passione and James Martin, } } A point that Cono seems to miss in the criticism of the Olympus scope (with the } U.V. filter flange over the objective nosepiece) is that scattered U.V. from } the sample is not a function of the quality of the optics or the manufacturer } but rather a function of the type of excitation in use. } } Some users, particularly biologists, use a mercury or xenon lamp but are not } relying upon the U.V. output per se. In some filter combinations a high level } of excitation in a certain short wavelength region of the visible spectrum is } the most efficient for stimulating emission from a fluorochrome which emits in } the longer wavelength region of the visible. Sometimes the cubes which are } optimized for this effect limit the amount of short-wave U.V. which reaches the } sample. Therefore, scattered light from the sample surface is lower in } proportion of U.V. than the proportion emitted from the lamp. } } However, people who make use of intrinsic U.V. fluorescence in unlabeled } specimens, or cascade effects, often use cubes which pass a significant } quantity of U.V. directly to the sample. The scatter or reflected light may be } correspondingly high in it's proportion of U.V. Those in forensics and art } conservation who rely on this mechanism for all or part of their work are } dealing with a different situation by intention rather than accident and must } take precautions regardless of the manufacturer. } } John Twilley } Art Conservation Scientist } P.O. Box 2324 } Sag Harbor, NY 11963-0113 } } Cono Passione wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } James , } } I don't understand how the light from the mercury illuminator could get to } } the observers eyes } } without a filter in place. If a filter position is blank and in place the } } light from the illuminator has } } no beam splitter or prisim to direct light to the eyepieces. There fore the } } light remains enclsoed } } within the illuminator module... Also there should be a stop on the } } illuminator to block any light } } coming through. } } Leica has the same option built into the stand of their DMR series } } microscope and does not } } seem to have a problem with light reaching the observer unfiltered.. This } } is probably another } } argument why the Leica is preferred over tthe competition.. Another is } } being the Lecia optics } } tend to absorb the harmful UV more so then its inferior Japanese } } competitors. Did you ever } } notice the Olympus has the filter shield in place so you don't see the uv } } light coming out of the } } objectives at the sample. Thats there for a reason....Harmful UV........ } } In answer to your question, I do not know of anyone who has one of these } } devices but would check } } with some of the larger Olympus dealers. Since it was not such a good idea } } maybe some of them } } still have units unsold on the shelf waiting to unload... } } Good Luck } } C YA CIP } } -----Original Message----- } } } From: James Martin {James.S.Martin-at-williams.edu} } } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } } Date: Friday, March 19, 1999 10:40 PM } } Subject: light microscopy: dual output monitor } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Olympus has discontinued a dual output module for their AX and BX line of } } } optical microscopes. This accessory allows two lamps (e.g., halogen and } } } mercury) to be mounted to a vertical illuminator; either source can be } } } used by flipping a mirror. I am told the decision to discontinue the } } } accessory was due to liability concerns that could arise if an operator } } } switched from a halogen to UV lamp source without engaging an appropriate } } } fluorescence cube (with barrier filter). } } } } } } Several microscope vendors have told me that third-party accessories are } } } available, but can or will not provide any documentation. One company } } } mentioned is 20-20 Technology. } } } } } } I am seeking referrals to a company providing such an accessory, or a } } } microscopist who wishes to part with one. } } } } } } Thank you. } } } } } } James Martin } } } Director of Analytical Services and Research } } } Williamstown Art Conservation Center } } } http://members.tripod.com/~James_Martin/dasrhome.htm } } } } } } } } } Research Scientist in Chemistry } } } Williams College } } } www.williams.edu } } } http://members.tripod.com/~James_Martin } } } } } } } } } } } } }
I've been looking at purchasing a dry 60x objective for use on a Nikon E600 microscope (application is temporary and permanent mounts of soil invertebrates, particularly soil mites, nematodes, etc.). The particular lens I'm trying right now is the CFI PLAN APO Phase 60x Dry DM objective. According to the sales rep, a DIC slider is not made specifically for this objective, and Nikon suggests that users try different combinations and choose what they like best. We've mixed and matched what the rep had available, and the best image was with CD DIC slider for the Plan Fluor ELWD (extra long working distance) DLL 60x C Phase objective. This objective is for an inverted microscope and is meant to look through a lot more material than one coverslip. I'd like to know if anyone on the list uses this Plan Apo objective for DIC, and what slider they use. Does this seem right to have to mix and match DIC sliders and objectives? ______________________________________________________________________ Edmond R. Zaborski phone (217) 265-0330 Soil Invertebrate Ecology Center for Economic Entomology FAX (217) 333-8076 Illinois Natural History Survey zaborski-at-uiuc.edu 607 E. Peabody Dr. Champaign, Illinois 61820 U.S.A. http://www.inhs.uiuc.edu/ ______________________________________________________________________
I am a neophyte to the microscopy world and am curious about EBIC and Voltage Contrast. I am wondering if there are good resources on the general mechanics of setting up the system. I also am having trouble finding basic information/ uses for EBIC. The Goldstein et al book does not delve too much into the topic of EBIC and am not sure of any other good SEM resource books.
Sharon Godkin says: } I'll try to answer everyone at once, to not clutter the list with a bunch } of messages. Yes, I think it must be a Zetopan. That name rings a faint } bell in the old belfrey. But it is an older model than the one in the } image Yvan kindly sent.
The *shape* should be roughly the same. The *color* would be black in your case (the old model).
} No, I don't have an image of mine; but I was able to dig up the original } inventory entry for it. It was purchased in 1955!
(:-) [but doesn't the recond say what microscope it is?]
} And retired to storage in 1981, apparently because it was impossible to } get a bulb that would fit it.
Osram 8100 should fit. See below.
} Yvan, that is interesting about the "Reichert } adapter" for the bulb you mentioned. We have quite a few bulbs } (apparently bought for it), but they are not long enough to reach the } contact at the base of the socket. This was no doubt the problem way } back then - our "Mr. Fix-it" (who keeps this place running) has now } solved that problem by soldering a tiny washer onto the contact at the } base of the socket so that when the bulb is screwed in all the way, it } makes contact with the washer. And we have light! The bulbs we have } are Osram BPA 554, 6V 5A. There is also a burned-out Osram 8100 } with it (was in the socket) but it is even shorter than the BPAs., so didn't } meet the contact either. Maybe someone threw out the adapter with } the last bulb?
I think this is EXACTLY the case: somebody "discarded" the bulb adapter (probably because another idiot soldered the bulb TO the adapter instead of just screwing it in - Yvan told me about one such incident).
} Both types of bulbs are the same diameter so have the same tight fit } into the bulb housing.
Hmm... This is to the experts to comment on.
} There is no identification on the } housing as to illuminator type; and the rheostat/transformer has been } replaced with one from another make of microscope.
No worries - the one I got also came with a "foreign" transformer. Who cares (unless you are a collector :-).
} The bulb housing at the back looks like the one in Laslo's image, not } Yvan's. There is no "Epi" attachment though; and no "flip-in" phase } telescope.
No Epi - because it was configured for transmitted light. No flip-in Bertrand lens ("phase telescope") - because yours is an older model (mine is "younger" than yours but still doesn't have Bertrand lens).
} Between "6" and "7" in Yvan's image (objective lens turret } and eyepiece attachment) my scope has two filter holder slides (empty } of course; one with a hole about dime-sized, the other about } nickle-sized) and the lever to send the light up the photo tube.
One slot (the lower narrow one, going 45 degrees) is for a compensator (don't know the details). The wider one is of course for the filter slider. Your swivelling head is identical to mine.
} It also has the following stamped on its front above the scale: } "Objektragerdicke 1,0 mm". I'm sure that means something important!
I suspect it says "object-vertical-thickness 1mm". Might it apply to the specimen slide?
As for bits and pieces - hey, me too! Objectives, condensers, TL DIC, etc.! Please e-mail with whatever Zetopan parts you're willing to part, and how much you'd like for it. -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
Hello Frank and everybody, I hope this will be of some help:
Dr. Pickett-Heaps recently published (Journal of Phycology 34:1088) a solution that, I remember, just mesmerized me with its elegance. "A rapid, highly efficient method... for fixation, dehydration, and embedding of small (e.g. planktonic) cells dispersed in large volumes of culture medium... based on continuous filtration..." -- so that you never really pellet the cells, just gently concentrate them over a cellulose filter in a standard Millipore apparatus while gradually adding the fixatives-washes-dehydrant-resin from the top. Seems worth trying , doesn't it? And yet the setup is VERY simple.
Just contact me directly if you can't get the journal. And if you do try it, I would ask you to please comment about the outcome. I have not yet made the time to try that for myself and will be MOST INTERESTED to hear how it worked for, say, lymphocytes. Or maybe somebody else uses something similar routinely?
Sincerely yours, Vlad.
} } Howdy all, } There is a graduate student here who is trying to look at a possible stem } cell line under TEM. The problem is that they are quite small, don't seem } to pellet very well, and he has only been able to get me a few thousand } cells at a time. } We have tried to embed the cells in 2% Agar after fixation but this hasn't } worked. } Is it possible to filter the media and cells, and then process the filter } with the cells stuck onto it? Maybe use a cytospin to spin the cells into a } filter? } Is there anyone with experience with this? Any suggestions or ideas would } be greatly appreciated. } } } Thanks in advance } Frank Herbert } Technician } Integrated Microscopy Core } Department of Cell Biology } Baylor College of Medicine } } } Vladislav V. Speransky School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Phone: 207 581 2998 Fax: 207 581 2969 Email: vladis-at-maine.maine.edu
The description provided by Sharon and some educated guesses:
..."... The bulbs we have are Osram BPA 554, 6V 5A..."
I didn't found that one in older Osram catalogues, but these doesn't go back to the 50's. I can send some pages from Osram with descriptions of bulbs that could fit and their most important parameters (voltage and current, foot type, dimensions of lamp and filament, relative position of the filament....).
"Longer" bulbs are:
* 8001 (6V, 4.35A, E14 foot, filament (4 x 0.9 mm) at 57mm from the middle contact of the foot, overall dimensions the same as 8100)
* 8025 (6V, 5A, BA20d foot, filament (2.2 x 2 mm) at 54 mm from the "bottom" of the foot, overall dimensions about the same as 8100, length is 60mm instead of 65 in the 8100)
* ...
..."... My scope doesn't have a turret condenser with a brightfield option. I said "dedicated phase" and I should have said "phase only" to be clearer. Sorry. This condenser is strictly one-size-fits-all! It doesn't have an "aperture iris" either. But, it does have a knurled ring around its bottom and a measuring scale (like a ruler) stamped around its front. Turning the knurled ring screws the innards of the condenser up and down through a total of about 15 divisions (I don't know what units) on the scale. It also has the following stamped on its front above the scale: "Objektragerdicke 1,0 mm". ..."...
"Objektragerdicke 1,0 mm" means: thickness of slides (to be used with this condenser): 1mm
As Barbara suggested (unless I misunderstand that) the condenser could be a "MS1.40" condenser, usable with phase and/or Anoptral inserts (the "UV-inserts"), but the MS1.40 is a brightfield condenser usable too for fluorescence and it doesn't has a ".... knurled ring around its bottom and a measuring scale (like a ruler) stamped around its front. Turning the knurled ring screws the innards of the condenser up and down through a total of about 15 divisions...". Could the curled ring device be an insert in the condenser that can be removed? (The MS 1.40 uses magnetic inserts that can easily be removed, you just have to pull these out of the condenser...). If it's the same condenser as the one I have, it should read on the top lens "UV". Sure it's not an adjustable DF-condenser?
Can send you some pictures of different condensers offline if you like... Can't send pictures to the list as it's considered poor netiquette (I had some reactions regarding that after sending the Zetopan picture...).
There seems to be some misunderstanding regarding the 6V, 5A bulb for the "LUX FNI" illuminator for Zetopan:
Uri: "... } I think this is EXACTLY the case: somebody "discarded" the bulb adapter } (probably because another idiot soldered the bulb TO the adapter instead } of just screwing it in - Yvan told me about one such incident)....".
Ussualy the bulb + the adapter were discarded. reichert sold these bulbs soldered in their individual adapter, thus providing a fully precentered and preadjusted bulb for the Zetopan. desoldering the bulb and replace it for an osram 8100 in the original adapter is just a way of fixing the problem that the bulbs are rather rare now...
Sharon: I'm preparing a multipage tif-file containing information and pictures on bulbs, lamphouses, filterholders and condensers for Zetopan. Interested?
Chris, For a number of years I performed what we call "electrical failure analysis" using electron beam techniques on semiconductor devices and integrated circuits. My favorite was Electron Beam Induced Current measurements. I found no definitive text or reference for that or the other measurements, Voltage Contrast, Specimen Absorbed Current, Electron Bean Induced Voltage, Charge Contrast, although I do have a few publications which discuss them. I have included in-depth discussions of these techniques in classes, and have assembled a number of example analyses which help to explain them. Send me a personal note if you have an interest in any of this. Or respond with a particular question that might be of interest to the e-beam community on the list. Jerry
_______________________ Jerome D. Schick, Ph.D. Semiconductor Devices and Electron Microscopy 26 Kuchler Drive LaGrangeville, NY 12540 Bus (914)223-7393 FAX (914)227-2743 jdschick-at-worldnet.att.net
Chris Parks wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello, } } I am a neophyte to the microscopy world and am curious about EBIC and } Voltage Contrast. I am wondering if there are good resources on the } general mechanics of setting up the system. I also am having trouble } finding basic information/ uses for EBIC. The Goldstein et al book does } not delve too much into the topic of EBIC and am not sure of any other } good SEM resource books. } } Thanks } CP
JoAnn, About 5 years ago I developed a bad case of tendonitis which our occupational health adviser attributed to the magnification knob in our SEM. She advised on a modification which our worksops made up. This effected a rapid improvement in my condition, although I still get some discomfort if I do a lot of typing.
Eric
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
Jerome D. Schick, Ph.D. Semiconductor Devices and Electron Microscopy 26 Kuchler Drive LaGrangeville, NY 12540 Bus (914)223-7393 FAX (914)227-2743 jdschick-at-worldnet.att.net
March 22, 1999
We are currently doing a very small R&D project for an electrical contector company which involves SEM EBIC. I would love to read any articles or notes that you may have on the subject.
J. Roy Nelson, Ph.D Material Testing Laboratory Pennington, NJ 08534 (609) 730-0575 FAX 737-7119 jrnelson-at-nj1.aae.com
Hi, Several people have asked for a summary of the responses to this question, = and there were many. Thanks to all for some good suggestions. Everett Ramer Federal Energy Technology Center
The solutions suggested were: 1. call a coworker/secretary/boss/security every hour 2. a pendant call button worn around neck---some have location capabilities= to guide rescuers to wearer's location. 3. a pendant worn around neck that automatically calls for help when the = wearer is in the horizontal position. 4. a pager with built-in motion detector that beeps after several minutes = of inactivity. If wearer does not shut off the beeping, the pager = automatically calls security. 5. a zoom-pan camera with over-the-web control with a web page that allows = coworkers to monitor each other safety from any place that has network = access.
Question: I recently purchased an older B&L microscope w/ 3 objectives and a prism inclined eyepiece. I think this model of microscope is from the 50's or 60's. That is all that I know of the model as there are no other markings that I can find to this regard. I was wanting to get a 4X & 20X objective for it and was wondering if AO Spencer objectives would fit. I have an opportunity to get these over the Web, but cannot try them out prior to purchase. We are planning to use the microscope to augment our childrens' studies and perhaps allow my oldest who is interested in cattle use it to examine parasites.
Replies to my inquiry about vendors of dual illumination modules are summarized below:
Opti-Quip, Inc. Highland Mills, NY (914) 928-2254 Model 1030 Universal Mirror housing will adapt to just about any microscope. The price is $998.00. For the Olympus there are 3 models: Olympus IMT2 (Part number: 1030E), Olympus BH (Part number: 1030-F) and Olympus B MAX (Part number: 1030-G)
MicroTec, Inc. Milford, NJ (800) 724-5508 #MIT-001
Thank you for your assistance.
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center http://members.tripod.com/~James_Martin/dasrhome.htm
Research Scientist in Chemistry Williams College www.williams.edu http://members.tripod.com/~James_Martin
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Hi, I need to get information (specs and quotations) rather quickly on = vibration tables for ultramicrotomes, light microscopes, and an atomic = force microscope. If any manufacturers of such equipment read the list, would you = please contact my e mail directly with information. This is for our new building and we need to get updated info into the = state in a very short time.
Thanks in advance, Judy M.
Judy Murphy, PhD Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
with SMTP (Eudora Internet Mail Server 1.2); Sun, 21 Mar 1999 16:47:32 -0800 Message-ID: {n1290087840.86882-at-sjdccd.cc.ca.us}
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Hi, I am trying to find a good negative processor that will process the = following films: EM film (Kodak4489,SO163,etc.) Ektapan (4162) and Kodak 4127 (for SEM) i.e. 4 x 5 sheet film.
1. We want to go from dry film to dry film, thus do not want tank develop= ment. 2. We now have Mohr processors however in 3 yrs have not been able to = get rid of things like roller marks, etc. Believe me, we have tried = every solution that everyone has suggested. We are of course open to new = ones. 3. Does anyone know of other possibilities out there? possibly from our = photographic friends. 4. We need to specify something very quickly, but I haven't come up with = a solution.
Any help is appreciated. Thanks Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
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Thanks to everyone who took their time to respond to my inquiry about microscope user fees. It was a big help.
Best wishes,
Soumitra
Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
Dr. Nelson, I have FAXed you a short five page article I wrote describing VC, SAC, and EBIC. Remember, traditionally, EBIC refers to the measurement of currents generated in the specimen, usually a semiconductor with a PN junction at least, and the function of the electron beam is to excite hole-electron pairs in the sample. In this measurement, the primary electrons do not significantly contribute to the current measured. It sounds like your project for a connector company may not involve a semiconductor material as the sample, in which case the wiring connections of the sample, the current amplifier, the time constants, and the interpretation might be somewhat different. When I get a chance, I shall dig around for a few more references which may be of use. I hope the FAXed copy is legible. Jerry
jrnelson wrote: } } Jerome D. Schick, Ph.D. } Semiconductor Devices and Electron Microscopy } 26 Kuchler Drive } LaGrangeville, NY 12540 } Bus (914)223-7393 } FAX (914)227-2743 } jdschick-at-worldnet.att.net } } March 22, 1999 } } We are currently doing a very small R&D project for an electrical } contector company which involves SEM EBIC. I would love to read any } articles or notes that you may have on the subject. } } J. Roy Nelson, Ph.D } Material Testing Laboratory } Pennington, NJ 08534 } (609) 730-0575 } FAX 737-7119 } jrnelson-at-nj1.aae.com
I am working on a business project that requires an analysis of the microscopy market and am hoping someone can direct me to a good source of information. Defining Microscopy to include all techniques which employ a probe such as: photons (including x-rays), electrons, ions, mechanical and/or electromagnetic radiation to form a representation or characterization of the morphology, crystallography, elemental, chemical or electronic structure of any material in either physical and/or life sciences applications. The questions I am trying to answer are: How large is the microscopy community in the US and worldwide? What technology has the greatest number of users? (Optical microscopy, x-ray microscopy, scanning electron microscopy, transmission electron microscopy, atomic force microscopy, scanning tunneling microscopy, scanning ion microscopy, analytical electron microscopy, electron microprobe, x-ray energy dispersive spectroscopy, electron energy loss spectroscopy, electron diffraction, convergent beam diffraction, high resolution electron microscopy, high voltage electron microscopy, etc.) Thank you for your assistance. Dave Akerson Tektronix CPID 682-7471 800-825-6100, ext. 7471 david.l.akerson-at-tek.com {mailto:david.l.akerson-at-tek.com}
Last week, there was a posting regarding the preparation of buffy coats for the TEM. Unfortunately, I didn't save the message and I now have a student who would benefit from the protocol. If you saved the protocol (or are the author of the email) could you please forward it to me offline.
Thanks for your assistance!
Steve
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
Biology Dept., University of Saskatchewan http://www.usask.ca.biology
The individual will be responsible for the departemntal Electron Microscope facility, consisting of 2 Philips TEMs, an SEM and related vacuum and preparatory equipment. Duties will include alignment and maintenance of the instuments, assisting and training students, faculty and outside users in their use, and providing general electronics and instrumentation expertise to the Department.
Required qualifications: Grade XII, post-secondary technical qualifiactions in electronics, and five years of relevant experience, including experience in EM maintenance. A strong interest in computer technology is also desireable. Salary range $36,336 - $46,320 depending on qualifications, with good fringe benefits. Starting date May 1, 1999. Forward resume with names, addresses and phone numbers of references to:
Dr. L.C. Fowke, Head Department of Biology University of Saskatchewan Saskatoon, Sask., S7N-5E2 phone: (306) 966-4400
The posting is directed in the first instance to Canadian citizens and permanent residents. The University is committed to employment equity.
I would second Franks comments. As he points out not all "Forensic" labs do criminal case analysis for law enforcement agencies that you think of when the phrase comes up. I happen to work in one that does. To a great extent our work is not "gore" related. Though you do have to deal with it on a routine basis, the science is the interesting part not the "gore". After a few years you hardly notice it is there most of the time, the tissue on a bullet removed from a body, for example, (I guess that is what you'd classify as "gore") is just something you have to get past to get to the fibers that may tell you something when you turn them over the hair and fiber specialist.
I am a Firearm and Toolmark Examiner that also does some SEM work. As such I work mostly Homicide and other crimes aginst people type cases. Light microscopy makes up the majority of Firearms analysis (The bullets or cartridge case comparison to a given firearm) or Toolmark Analysis. Before they go to the comparison scope, stereo microscopy is the majority of the non comparative examination of bullets and cartridge cases, (though this information is still comparitive in nature). Bullets are looked at for trace evidence and GRC (General Rifling Charicteristics) on the stereo microscope then taken to the comparison scope. Most firearms comparison is carried out between 10 and 40 power on a forensic comparison microscope. The unknown bullet being mounted on one scope (stage) with a standard (known) bullet fired from the firearm in question on the other, the two fields of view being observed through the comparison bridge (though this is a highly intagrated unit in modern firearms scopes and thought of as a single scope). Stereo scopes are also used in examining clothing items for bullet holes, gunpowder, etc.. The SEM is a good tool for the examination of trace evidence, but the application of the SEM to the comparison of bullets is unnecessary and provides no additional information (the James E. Ray appeal claims not withstanding) beyond that seen on the stereo and comparison scopes if you are considering comparison to a firearm only. Only SEMs designed for direct comprison of two fields are sutable for bullet or toolmark comparison. Manipulation of captured images is dificult compared to the Comparison microscope and not considered appropriate without the real time control of the two images. As no properly trained firearms examiner would make an identification from a photograph you would also run into problems with acceptability in court if not using a comparison SEM (only a few exist that I am aware of). The SEM can be highly useful for examination of trace evidence on bullets.
LMs of various types plays a big part in trace evidence examinations, as can SEM/EDS. Blood and other physiologic fluid analysis (serology) relies on LM and other techniques but older methods are quickly being replaced by DNA analysis (this is out of may area). Guts? Our toxicologists do stomach contents for toxicological materials at times using some LM I think (also out of my area). Gun Shot Residue is one of the heavy uses of SEM in the Forensic Science Labs of police agencies (firearms examiners tend to stay clear of this work -do to our contamination from shooting and handling firearms all the time).
War Stories have their place but usually are only interesting if in answer to a question or from the speakers personal experience. Show them the interesting information they can learn with EM and maybe you'll get a different kind of "EEEEWWWW ."
I hope this fills in some of the gaps for you but don't think its really went where you wanted to go. You can probably get some "war storys" (we all have them) at Scanning '99 if your there, but the real information will be more interesting (to you and the students at the HS I suspect).
Jim Roberts Firearm and Toolmark Examiner
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Dear Paula:
I have read with some concern your request for forensic EM assistance.
I've just been asked to speak to a group of high school kids about EM, and how I got to where I am today (I'm not gonna tell them about all the little people I squashed along the way). Actually what I wanted to know is if there are any folks out there doing Forensic EM. You know analysis of blood, guts, bullets, things like that. You know that if I gross the kids out they'll think that EM is coolest thing since ice cubes. So if anybody does that sort of stuff or knows of anybody I can talk to about it, let me know. Cuz y'all know there's nothing better than gettin' a great big EEEEWWWW out of the young'uns.
I would be glad to talk to you about forensic applications of scanning electron microscopy as that is my job, however......you should understand "forensic EM" goes far beyond the common opinion of " analysis of blood, guts, bullets, things like that". An electron microscopist performing airborne particle analysis for possible occupational/environmental exposure to harmful particulates or the pathologist using electron microscopy to identify a pathogen/toxin induced lesion or foreign object are also performing forensic EM analyses. Forensic means "Of or used in legal proceedings or in public debate" (The American Heritage Dictionary, 2nd Edition). The investigative accountant analyzing data in the investigation of corporate illegal dealings for prosecution and the forensic scientist in the laboratory matching bloody fibers or a bullet found at a crime scene are both performing forensic analyses.
Is the point of your giving this talk to "gross them out" and generate " a great big EEEEWWWW out of the young'uns" or are you teaching to really stimulate some young minds and show them other areas of science that use microscopy for possible careers? High school students are not "young'uns" and you may have a profound impact on a few futures. You can either encourage and stimulate OR you can deter further interest altogether. Many of us were originally guided to our profession by a good speaker or teacher.
As one who both instructs and lectures extensively on forensic applications of electron microscopy, I have found the audiences (high schools, colleges, forensic science and microscopist societies) all enjoy the case history (not gore) from an investigation. The myriad of television shows on the subject speak to the popularity of this current "hot" topic. I'm sure the international emphasis on the "Crime of the Century" has contributed greatly to the recent popularity in forensic science and scientific evidence. You propose to SHOCK your audience with raw crime scene photos when the students are not used to seeing "the real thing". This is counter productive as they will be discussing the shock image(s) the whole time you are trying to explain the science/EM in the investigation. The "gore" is primarily what they will remember and take away with them.
If you really want to TEACH these high school students, show them how to identify objects from two places (exa. - particles such as soil samples from the "crime scene" with particles from a suspect's shoes - SEM/TEM/EDX) or how to find specific particles that confirm a scenario (exa. - specific lake diatoms in "lung fluids" indicating respiration of water from a lake in a possible drowning victim). You can make up your own "scenarios" and supply the "evidence" from the crime scene. Make up a class exercise "crime" and lead them through an investigation of the evidence by EM. Make your SEM/TEM/EDX lecture slides in advance and discuss possible conflicts/artifacts, etc.. You won't need "blood, guts, bullets, things like that" to impress and instruct the students. Leave that for "Hollywood" and the criminal investigator in the proper arena (the court room or instructing other forensic scientists).
You may want to attend the SCANNING 99 meeting in Chicago (April 11-14, 1999) where an all day short course (FORENSIC SCIENCE AND SCANNING MICROSCOPY) will be offered followed by a second day symposium (SCANNING MICROSCOPY APPLICATIONS IN FORENSIC SCIENCE). For information on that International Meeting, call Mary K. Sullivan (201) 818-1010 , web site { http://www.scanning-fams.org } .
Again, I offer my assistance with some other ideas if you are interested. Feel free to contact me off line or by DIRECT E-mail.
Sincerely,
S. Frank Platek Research Biologist/Electron Microscopist Forensic Chemistry Center US Food and Drug Administration 6751 Steger Drive, Cincinnati, OH 45237-3097 (513) 679-2700 , [FAX] (513) 679-2761 E-mail: fplatek-at-ora.fda.gov
DISCLAIMER: THE OPINIONS EXPRESSED ARE SOLELY MY OWN AND DO NOT REPRESENT THOSE OF THE U.S. FOOD AND DRUG ADMINISTRATION OR ANY OTHER AGENCY OF THE U.S. FEDERAL GOVERNMENT OR THE FOUNDATION FOR ADVANCES IN MEDICINE AND SCIENCE (FAMS, INC.).
Leica Microsystems, Diatome US, Electron Microscopy Sciences, and Bal-Tec announce another in a series of EM Specimen Preparation workshops. These seminars will focus on the following techniques:
Ultramicrotomy of Materials
Embedding of industrial samples Specimen trimming
Ultramicrotomy of hard materials Ultramicrotomy of polymers
Collection & handling of sections Staining of polymer sections
Low temperature ultramicrotomy
Ion Beam Milling of Materials
Ion Milling of plan view samples for TEM
Ion Milling of cross section samples for TEM
SEM preparation of interfaces using fast slope cutting technique
TEM / SEM preparation of temperature sensitive specimens
SEM / LM preparation of surface structures with ion milling
The format of our workshop is half day lecture and half day bench work. Samples will be supplied by the course instructors. Participants are encouraged to bring their own samples to work with as time allows.
Course Speakers & Instructors
Ultramicrotomy of Materials
Dr. Tom Malis CANMET Characterization Group Leader Materials Technology Laboratory Ottawa, Ontario
Mr. Bob Vastenhout DOW Chemical Polymer Microscopist Analytical Science Department Terneuzen, The Netherlands
Mr. Helmut Gnaegi Product Manager Microtomist Extaordinaire Diatome, Ltd., Switzerland
Ion Beam Milling
Dr. Wolfgang Gruenewald Bal-Tec Head of Applications Laboratory Liechtenstein
Mr. Arthur Buechel Bal-Tec Product Manager Liechtenstein
Hosted by: JoAn Hudson Clemson University Microscopy Facility Clemson, SC
When: Ion Milling June 7-8, 1999
Ultramicrotomy June 9-11, 1999
Tuition: Ion Milling $400.00
Ultramicrotomy $1,400.00
Ion Milling & Ultramicrotomy $1,700.00
Includes lodging at the lakefront Conference Center & Inn at Clemson University, continental breakfast and lunch daily, one group dinner, course supplies, and lab charges.
Contact: Mike Boykin, Leica Microsystems, Inc. 800-248-0665 X5092
I'm sorry to bother everyone, again, with a question about fixation of bacteria. I greatly appreciate the responses I received a few weeks ago. At one time Ryter-Kellenberger fixation was considered the standard. It produced an image with a relatively clear nucleoid containing fibrillar chromatin. Is this now considered to be an artifact of preparation?
John Wright West Desert Test Center (435) 831-3017
have you red the chapter about EBIC in the book of Ludwig Reimer?
Here is the reference:
Reimer, L: Scanning Electron Microscopy: Physics of Image Formation and Microanalysis. Springer Series in Optical Sciences Vol. 45 Chapter 7: Electron-Beam-INduced Current, Cathodoluminescence and Special Techniques p.272-312 (Springer-Verlag Berlin, Heidelberg, New York, Tokyo, 1985) ISBN 3-540-13530-8
Petra
} Hello, } } I am a neophyte to the microscopy world and am curious about EBIC and } Voltage Contrast. I am wondering if there are good resources on the } general mechanics of setting up the system. I also am having trouble } finding basic information/ uses for EBIC. The Goldstein et al book does } not delve too much into the topic of EBIC and am not sure of any other } good SEM resource books. } } Thanks } CP
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu Visit our WWW site! http://www.crpcu.lu/~wahlbrin
A phantastic opportunity to meet well-kmown scientists and an excellent chance to see and to compare the latest equipment for confocal and multiphoton fluorescence microscopy. Attend an excellent meeting and see the most recent technological developments.
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D FOCUS ON MICROSCOPY 1999
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
we would like to invite you to attend this years "Focus on Microscopy" conference.
Many of the well known scientists in this field will present their recent progress.
Here is a list of the plenary speakers: - G.J. Brakenhoff, Amsterdam - Christoph Cremer - Winfried Denk - Timothy Holmes - Anthony A. Hyman - Stefan Hell - Satoshi Kawata - Karsten K=F6nig - Andres Kriete - Ulrich Kubitscheck - Eric Manders - Colin Sheppard - Ernst H.K. Stelzer, Heidelberg - Kevin F Sullivan - Tony Wilson - Daniele Zink
The commercial exhibition offers an excellent overview. The modern instruments of all major microscope manufacturers as well as microscope accessories are on display. This is the 1999 event focussed on microscopy= in Europe.
Highlights of the commercial exhibition include:
Hamamatsu Photonics: ORCA Series - state of the art digital imaging =B7 Progressive scan interline transfer CCD with 1280x1024 pixel =B7 Lens-on-chip technology, TE cooling and optimized readout guarantee highest sensitivity =B7 Black & white cameras with 10-14 bit output =B7 Color cameras with RGB matrix filter or filterwheel =B7 Plug & play set incl. framegrabber and comfortable TWAIN driver avai= lable
L.O.T. Oriel GmbH & Co KG: Nanopositioning Unit and Fiber Laser
Molecular Probes: New fluorescent reagents for Cell Biology and Imaging highly - Fluorescent Alexa dyes - click here for more details
Nikon GmbH: Two compact confocal microscopes + fully automatic motorized research microscope
Olympus Optical Co. GmbH Confocal Laser-Scanning-Microscope with two-phot= on excitation
Omicron Vakuumphysik GmbH Scanning Near Field Optical Microscope (SNOM)
Visitron Systems GmbH: 2D/3D Fluorescence Imaging System based on Cooled Digital Camera System
Wallac Distribution GmbH: 1. EG&G Wallac LSR UltraView, confocal fluorescence microscope for "real time" images. It is the first presentation of this instrument in Germany. 2. EG&G Wallac Signifer, fluorescence microscope for recording images wi= th the time resolved fluorescence prinziple. 3. EG&G Berthold NightOWL, a universal imaging system for low level luminescence images.
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques = are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meet= ing point for developers and users working in these rapidly evolving fields a= nd play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic development= s in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
A phantastic opportunity to meet well-kmown scientists and an excellent chance to see and to compare the latest equipment for confocal and multiphoton fluorescence microscopy. Attend an excellent meeting and see the most recent technological developments.
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D FOCUS ON MICROSCOPY 1999
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
we would like to invite you to attend this years "Focus on Microscopy" conference.
Many of the well known scientists in this field will present their recent progress.
Here is a list of the plenary speakers: - G.J. Brakenhoff, Amsterdam - Christoph Cremer, Heidelberg - Winfried Denk, Murray Hill - Timothy Holmes, Watervliet - Anthony A. Hyman, Heidelberg - Stefan Hell, Goettingen - Satoshi Kawata, Osaka - Karsten K=F6nig, Jena - Andres Kriete, Giessen - Ulrich Kubitscheck, Muenster - Eric Manders, Amsterdam - Colin Sheppard, Sydney - Ernst H.K. Stelzer, Heidelberg - Kevin F Sullivan, La Jolla - Tony Wilson, Oxford - Daniele Zink, Munich
The commercial exhibition offers an excellent overview. The modern instruments of all major microscope manufacturers as well as microscope accessories are on display. This is the 1999 event focussed on microscopy= in Europe.
Highlights of the commercial exhibition include:
Hamamatsu Photonics: ORCA Series - state of the art digital imaging =B7 Progressive scan interline transfer CCD with 1280x1024 pixel =B7 Lens-on-chip technology, TE cooling and optimized readout guarantee highest sensitivity =B7 Black & white cameras with 10-14 bit output =B7 Color cameras with RGB matrix filter or filterwheel =B7 Plug & play set incl. framegrabber and comfortable TWAIN driver avai= lable
L.O.T. Oriel GmbH & Co KG: Nanopositioning Unit and Fiber Laser
Molecular Probes: New fluorescent reagents for Cell Biology and Imaging highly - Fluorescent Alexa dyes - click here for more details
Nikon GmbH: Two compact confocal microscopes + fully automatic motorized research microscope
Olympus Optical Co. GmbH Confocal Laser-Scanning-Microscope with two-phot= on excitation
Omicron Vakuumphysik GmbH Scanning Near Field Optical Microscope (SNOM)
Visitron Systems GmbH: 2D/3D Fluorescence Imaging System based on Cooled Digital Camera System
Wallac Distribution GmbH: 1. EG&G Wallac LSR UltraView, confocal fluorescence microscope for "real time" images. It is the first presentation of this instrument in Germany. 2. EG&G Wallac Signifer, fluorescence microscope for recording images wi= th the time resolved fluorescence prinziple. 3. EG&G Berthold NightOWL, a universal imaging system for low level luminescence images.
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques = are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meet= ing point for developers and users working in these rapidly evolving fields a= nd play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic development= s in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
David asks ... } } } I am working on a business project that requires an analysis of the } microscopy market and am hoping someone can direct me to a } good source of information. } ...
The best jump off point is the WWW-Virtual Library ...
http://www.ou.edu/research/electron/www-vl/ ...
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
It is possible to use a variety of other fixatives to preserve bacteria. However, we find that when all else fails, the R-K procedure nearly always gives a good fixation on organisms that otherwise look poorly preserved. The buffer is somewhat cumbersome to prepare since the veronal (sodium barbitol) is a controlled substance.
} I'm sorry to bother everyone, again, with a question about fixation of } bacteria. I greatly appreciate the responses I received a few weeks } ago. At one time Ryter-Kellenberger fixation was considered the } standard. It produced an image with a relatively clear nucleoid } containing fibrillar chromatin. Is this now considered to be an } artifact of preparation?
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I am looking for a procedure to do TEM of low density polyethylene films. I would like to stain the film so I can then microtome at room temperature. For high density films we use chlorosulfonic acid at 60C but this dissolves the low density material. I can cryo-microtome the low density films and then stain the sections with ruthenium tetroxide but this is giving only marginal results. I would really like to be able to room temperature microtome the material. Any help greatly appreciated.
------------------------------ Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
Need a simplified recipe for viewing circular DNA ~5000KB in a gamish of linear DNA. No special marker or complementary sequence available. Thanks.
*Disclaimer: Whatever... is not Tulane opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web:[ http://www.tmc.tulane.edu/ferminlab/] or [http://www.tmc.tulane.edu/imaging/] Internet: [cfermin-at-mailhost.tcs.tulane.edu] 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
I've received the following query and thought I'd post it so the various types of microscopists out there besides me might like to help this fellow out. Please reply directly to his eddress since he's not on the listserver.
Many thanks,
Dee Breger
hello:
my name is francisco sandoval, and I live in Guatemala in central america.
I am a photographer, and I am interested in microscope photography, but I do not know where to begin, could you help me with it.
I guess I do not understand the kind of microscope you are using. I do not know anything about microscopes..... If you do not mind helping me, tell me where to begin..... Thanks
Francisco Sandoval
kikophoto-at-geocities.com jfse30-at-hotmail.com
www.geocities.com/paris/jardin/4108/
____________________________________________________________________________ Automatic note: Sometimes I don't receive incoming emails (with no notice to the sender). If I don't respond to your message, please send it again! ____________________________________________________________________________ _ Dee Breger Manager, SEM/EDX Facility Lamont-Doherty Earth Observatory Route 9W Palisades NY 10964 USA
As a company that produces image processing systems (see disclaimer below), I would like to invite you to check out our website at http://www.soft-imaging.com or http://www.soft-imaging.de. We routinely deal with 12 to 16 bit gray scale images as most digital cameras supply them.
If you need further information, don't hesitate to contact us at one of the URLs or numbers below.
Thank you.
Michael Bode
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
} ---------- } From: Hendrik O. Colijn[SMTP:colijn.1-at-osu.edu] } Sent: Friday, March 19, 1999 6:16 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Image processing software } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Is anyone familiar with reasonably priced software programs that can } manipulate 16-bit gray scale images? While Adobe Photoshop can read } 16-bit } images, you have to change the images to 8-bit before you can run } filters } etc. Gatan's Digital Micrograph can work with 16-bit images, but its } price } is a little steep for equipping several computers. What are the } capabilities of other packages? I would like to use the software to } work } with diffraction patterns as well as images. } } TIA, } Henk } } Hendrik O. Colijn colijn.1-at-osu.edu } OSU Campus Electron Optics Facility (614) 292-0674 } "An optimist believes that we live in the best of all possible worlds. } A pessimist fears that this is true." }
I provide third party maintenance for EM in the midwest, so take the following as a somewhat biased view. I'd also like to apologize in advance for my typically long and meandering response. I don't pull punches, so perhaps I should start adding a disclaimer that the following are my opinions - if that's not enough, go ahead and sue, I don't have anything anyway.
An insurance company's primary usefulness is to large organizations that can off-load the responsibility for a large amount of equipment. Their customers will contract with them for all of their equipment, they handle the paperwork and arrange service. In these cases, a good portion of the cost savings can be in the customer's reduction in paperwork - one contract as opposed to hundreds. To do the service, they hire the manufacturer (don't know how many manufacturers go along with this, not all do) or an independant service provider (I have been approached a number of times, haven't done it yet) on an hourly basis to do the work, usually negotiating reduced rates. I am sure that, here in the early stages, they are underestimating their costs and will raise their rates as they gain a greater understanding.
A number of direct problems. Mainly, they will have a hard time guaranteeing response time. For any manufacturer or independant, contract customers come first. You are also at their mercy for who comes to work on your equipment, it may even be a different organization each time. From my point of view, you run a risk of never getting someone who has some long term interest or commitment in your needs, even if they use the original equipment manufacturer. The OEM will no longer bear any responsibility other than providing service when needed (and the more often the better for their bottom line when hourly).
Service used to be a loss-leader, a way for a company to increase their sales by creating good will and a positive corporate image. Question this, just count up the number of JEOLs sold in the last fifteen years. They made a conscious effort to provide exceptional service and sold like crazy on the basis of those efforts. A couple of decades ago, most companies found that there are profits to be made in service and have run their service accordingly and made the industry an increasingly tempting market for new manufacturers, third party service providers and the new insurance companies.
If you only have a few pieces of equipment, you'd do better to find a local independant. You'll probably find that an independant can be very flexible in contract terms. Consider asking them to remove the 'insurance' aspects of a service contract by removing parts coverage. You could still get PMs and emergency labor, but the service provider would not have to be escrowing a good chunk of money just in case an expensive part goes. That, of course, would require you to do so. There is some insurance value to the contract labor rates, but generally not much. A good technician can do a lot to prevent future service needs and it is a lot less traumatic to a small operation to spend extra time fixing an instrument than it is to replace that $10,000 whiz-bang that blew.
You can also, of course, handle your service on a billable basis with manufacturer or independent. Watch out for the manufacturers, though. Some like to charge a premium (I've seen over $300/hour) in order to encourage you to go under contract. Independents may play this game too, but not to that extent (I don't know of any who do). While I am obviously biased towards the independents, I truely think that supporting them is the only way to keep any real competition in this or any other industry.
Frankly, the insurance companies are hoping to become the big kid on the block in the future so that they can create their own terms with the manufacturers. Instrumental service rates are bad enough already, but this squeeze play could eventually hurt the industry the same way managed care and HMOs have hurt the medical industry. Manufacturers have income goals that will be met, one way or another. If the insurance companies squeeze out their service profits, manufacturer's prices on remaining contracts will go up which will then force more users to go to the insurance companies. Manufacturers will also have to increase the selling price for their equipment. We are merely adding another layer of for-profit companies to take a bite out of your pocket.
There is a potentially insidious effect of organizations like this. In claiming to reduce costs for a few, they wind up increasing costs for all. Rather than introducing new competition, they manipulate a market to the point where some existing competitors leave the market. They bring no new instruments to market, work counter to free enterprise by attempting to be the arbitor of pricing without actually providing the service and, as the market becomes saturated by them, will go into bidding wars between themselves reducing services in order to lower their costs. There will be some manufacturers that won't be able to stay competitive with the external pressure and others that won't want to bend to those pressures. Look to a future of greater cutbacks and mergers as these new players are accommodated in the market.
We are also already seeing another aspect of this. In attempting to cut costs, manufacturers may cut back their own sales efforts, relying on others instead. The EM markets are getting large enough and there are enough manufacturers that we may eventually see 'distributers' added on the front end, yet another layer of for-profit companies taking a piece of the pie. It is common for a company to enter a new national market through marketing arrangements. ISI has for a long time had direct sales presence in the US. While this is a company that has had its share of problems, their new marketing arrangement is a cutback in its sales force that is in large part due to its poor service history and poor profits in sales and service. While not precipitated by the insurance companies, it still may offer a view of their future effects.
It will not benefit any manufacturer to take part in these plans in the long run. Doing so would be giving up all control over a very important sector of their public relations, cut into their profits and resolve them to having other companies dictate their policies and pricing. Look for some manufacturers to resist because doing so allows them to distinguish themselves in the market by maintaining their own high level of service responsiveness and performance. However, as this snowballs we may well reach a critical mass where all manufacturers will have to participate in order to survive.
Good luck, whatever you choose.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com
-----Original Message----- } From: Tindall, Randy D. {TindallR-at-missouri.edu} To: 'microscopy-at-sparc5.microscopy.com' {microscopy-at-sparc5.microscopy.com}
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Stephen,
The following procedure has been adapted from the literature (I don't have the references with me):
The method that I normally use to image LLDPE is to first embed a strip of the film with Epofix epoxy using a silicone mold. After curing the epoxy, face-off the end of the specimen block to expose the film (assuming you want a cross-section) and then attach the block to a piece of double-sided tape on a glass microscope slide. The next step involves staining the specimen block with the vapor of a mixture of 0.2 g of ruthenium trichloride trihydrate and 10 ml of 5.25% sodium hypochlorite (regular household bleach). This produces a fairly aggressive RuO4 vapor. The staining mixture and the slide should be kept in a glass jar with a loosely fitting lid for 2 to 3 hours -at- room temp. Occasionally, the temp. needs to be elevated for higher density polyolefins. Do all of the staining in a fume hood!!
After staining is complete, remove the slide/specimen, rinse well and allow to dry. The specimen block can be trimmed and microtomed at room temperature. I normally orient the film parallel to the edge of the diamond knife. This helps prevents delamination from the epoxy.
I hope this procedure helps. I've been using this technique for several years with great success.
Gene Young Microscopy/Microanalysis Group Dow Chemical USA Freeport, TX
------------------------------ Original message:
I am looking for a procedure to do TEM of low density polyethylene films. I would like to stain the film so I can then microtome at room temperature. For high density films we use chlorosulfonic acid at 60C but this dissolves the low density material. I can cryo-microtome the low density films and then stain the sections with ruthenium tetroxide but this is giving only marginal results. I would really like to be able to room temperature microtome the material. Any help greatly appreciated.
------------------------------ Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
} Date: Tue, 23 Mar 1999 16:56:22 -0600 } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: photographer needs microscopy info } } } Dear collegues, } } I've received the following query and thought I'd post it so the various } types of microscopists out there besides me might like to help this fello } out. Please reply directly to his eddress since he's not on the listserver } } } Many thanks, } } Dee Breger } } hello: } } my name is francisco sandoval, and I live in Guatemala in central } america. } } I am a photographer, and I am interested in microscope photography, but } I do not know where to begin, could you help me with it. } } I guess I do not understand the kind of microscope you are using. I do } not know anything about microscopes..... If you do not mind helping me, } tell me where to begin..... Thanks } } Francisco Sandoval } } kikophoto-at-geocities.com } jfse30-at-hotmail.com } } _________________________________ } Francisco, I don't know if it is still available but the booklet by Eastman Kodak, Photography through the Microscope, is quite good. It isKodak Publication P-2 CAT 152 8371. Published in 1980
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
} I am looking for a procedure to do TEM of low density polyethylene films. I } would like to stain the film so I can then microtome at room temperature. } For high density films we use chlorosulfonic acid at 60C but this dissolves } the low density material. I can cryo-microtome the low density films and } then stain the sections with ruthenium tetroxide but this is giving only } marginal results. I would really like to be able to room temperature } microtome the material. Any help greatly appreciated.
I'm an etcher, not a stainer, but we do have some experience of staining materials in bulk. Possibly a room temperature chlorosulphonation might help. I've never used RuO4, but it might also work in bulk on a thin enough film. I'll have to ask a colleague, when he's in his office.
If you're interested in larger scale variations in morphology, then permanganic etching followed by reflection optical microscopy (Nomarski) is very useful.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Before you do anything, including cleaning the ball bearings, find out their history! The goop on the surface may be part of the story.
My advice: try light microscopy first. Reflected light DIC may tell you alot about the surface. Also try interferometry (I'll be back in the office Monday; call or contact me directly for further info). I had a similar project for a client some years ago with ball bearings for jet engines. If the height of the asperities was too tall, they would break off, leaving metallic residue in the raceway which caused the lubricant to polymerize and freeze up. Needless to say, not a desirable outcome for an airplane at 30,000+ feet!
If the bumps on the ball bearings are not the problem, then you might want to try SEM + EDS to see if there is some problem in the alloy.
Best of luck and let me know how things turn out.
Barbara Foster 125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
At 10:53 AM 3/4/99 -0500, Lesley S. Bechtold wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I've attached a recent announcement for a workshop as an example of an item "for sale." I believe that items for sale, whether services, supplies, or instruments all require the same kind of response you gave to my posting for "instruments available" recently. Please see that workshops for profit are stricken from the Listserver so that you are consistent with your own policy or, change your posting policy.
I believe that any of us who are in this field should be able to post any kind of useful informative item that would be of interest to those on the list. The availability of microscopes is a useful piece of info for someone who has a legitimate need for a used scope. There are a lot of people looking for microscope users and owners as a trusted resource rather than the manufacture's sales reps. Please consider this in the future.
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TEM & SEM Sample Preparation of Materials
A Two Part Workshop & Seminar
Leica Microsystems, Diatome US, Electron Microscopy Sciences, and Bal-Tec announce another in a series of EM Specimen Preparation workshops. These seminars will focus on the following techniques:
Ultramicrotomy of Materials
Embedding of industrial samples Specimen trimming
Ultramicrotomy of hard materials Ultramicrotomy of polymers
Collection & handling of sections Staining of polymer sections
Low temperature ultramicrotomy
Ion Beam Milling of Materials
Ion Milling of plan view samples for TEM
Ion Milling of cross section samples for TEM
SEM preparation of interfaces using fast slope cutting technique
TEM / SEM preparation of temperature sensitive specimens
SEM / LM preparation of surface structures with ion milling
The format of our workshop is half day lecture and half day bench work. Samples will be supplied by the course instructors. Participants are encouraged to bring their own samples to work with as time allows.
Course Speakers & Instructors
Ultramicrotomy of Materials
Dr. Tom Malis CANMET Characterization Group Leader Materials Technology Laboratory Ottawa, Ontario
Mr. Bob Vastenhout DOW Chemical Polymer Microscopist Analytical Science Department Terneuzen, The Netherlands
Mr. Helmut Gnaegi Product Manager Microtomist Extaordinaire Diatome, Ltd., Switzerland
Ion Beam Milling
Dr. Wolfgang Gruenewald Bal-Tec Head of Applications Laboratory Liechtenstein
Mr. Arthur Buechel Bal-Tec Product Manager Liechtenstein
Hosted by: JoAn Hudson Clemson University Microscopy Facility Clemson, SC
When: Ion Milling June 7-8, 1999
Ultramicrotomy June 9-11, 1999
Tuition: Ion Milling $400.00
Ultramicrotomy $1,400.00
Ion Milling & Ultramicrotomy $1,700.00
Includes lodging at the lakefront Conference Center & Inn at Clemson University, continental breakfast and lunch daily, one group dinner, course supplies, and lab charges.
Contact: Mike Boykin, Leica Microsystems, Inc. 800-248-0665 X5092
Hi. I was recently asked of a way to determine C film thickness. I = remember that using a polished brass specimen and observing the color is = one way. Does anyone have a source in the literature for this method? = Thanks.
______________________ Roberto Garcia Senior Analyst, Metallography North Carolina State University Analytical Instrumentation Facility Box 7531, Room 303 EGRC Raleigh, NC 27695-7531 rgarcia-at-unity.ncsu.com http://spm.aif.ncsu.edu/aif
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{META content=3D'"MSHTML 5.00.0910.1309"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT size=3D2} Hi. I was recently asked of a way to determine C = film=20 thickness. I remember that using a polished brass specimen and observing = the=20 color is one way. Does anyone have a source in the literature for this = method?=20 Thanks. {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D3} ______________________ {BR} Roberto Garcia {BR} Senior = Analyst,=20 Metallography {BR} North Carolina State University {BR} Analytical = Instrumentation=20 Facility {BR} Box 7531, Room 303 EGRC {BR} Raleigh, NC=20 27695-7531 {BR} rgarcia-at-unity.ncsu.com {BR} http://spm.aif.ncsu.edu/aif {/FONT= } {/DIV} {/BODY} {/HTML}
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I have a student who has done confocal on tetrahymena and now wants to move on to TEM. She is primarily interested in the microtubular arrays in the cilia and also within the body of the organism, especially at division. However, we also would like to get the best possible preservation of the entire organism.
Since these two needs may conflict, I would appreciate hearing from anyone with experience with this or similar organisms.
Possible approaches could include: a) using tannic acid to stabilize the microtubules (percent?, in all solutions or only primary fix?, best buffer system?, etc) b) using a combined glutaradyhde-osmium fix followed by straight osmium (recommended by Hayat.
Thanks in advance for the assistance.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
id HMDAAJC2; Wed, 24 Mar 1999 12:54:34 -0600 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" X-Sender: schattenh-at-pop.email.missouri.edu (Unverified) Message-Id: {v04020600b31ee56f600f-at-[128.206.215.129]}
Ph.D. graduate student or post-doctoral student (D.V.M. or M.D.) position available for research in the area of protozoal parasitology. Research will involve electron microscopy and cellular biology studies utilizing in vitro derived Sarcocystis spp. , Neospora spp. and Toxoplasma gondii parasites. Individual will work with a multi-disciplinary team at the University of Missouri-Columbia, in the Department of Veterinary Pathobiology and the Molecular Biology Program Electron Microscopy Core facility. Successful candidate should have background in tissue/cell embedding and processing for light and electron microscopy, and demonstrated proficiency in written and spoken English. Molecular biology and cell culture skills are helpful, but not a necessary requirement. U.S. citizenship is not required. For further information contact Dr. A.E. Marsh at ph: 573-884-2673, fax: 573-884-5414, or email: marshae-at-missouri.edu {mailto:marshae-at-missouri.edu} .
Try The American Mineralogist, Volume 58, pages 920-925, 1973. The role of Carbon Film Thickness in Electron Microprobe Analysis - Kerrick DM, Eminhizer LB amd Villaume JF.
Hi. I was recently asked of a way to determine C film thickness. I = remember that using a polished brass specimen and observing the color is = one way. Does anyone have a source in the literature for this method? = Thanks.
______________________ Roberto Garcia Senior Analyst, Metallography North Carolina State University Analytical Instrumentation Facility Box 7531, Room 303 EGRC Raleigh, NC 27695-7531 rgarcia-at-unity.ncsu.com http://spm.aif.ncsu.edu/aif
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD} {META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {STYLE} {/STYLE}
{META content=3D'"MSHTML 5.00.0910.1309"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT size=3D2} Hi. I was recently asked of a way to determine C = film=20 thickness. I remember that using a polished brass specimen and observing = the=20 color is one way. Does anyone have a source in the literature for this = method?=20 Thanks. {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D3} ______________________ {BR} Roberto Garcia {BR} Senior = Analyst,=20 Metallography {BR} North Carolina State University {BR} Analytical = Instrumentation=20 Facility {BR} Box 7531, Room 303 EGRC {BR} Raleigh, NC=20 27695-7531 {BR} rgarcia-at-unity.ncsu.com {BR} http://spm.aif.ncsu.edu/aif {/FONT= } {/DIV} {/BODY} {/HTML}
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Robert MacKay Department of Earth Sciences Dalhousie University Halifax, Nova Scotia, Canada B3H 3J5 Tel: 902 494-7087 Fax: 902 494-6889 e-mail rmackay-at-ac.dal.ca
} Subject: job opening } Priority: normal } X-mailer: Pegasus Mail for Windows (v2.42a) } Message-ID: {5586A5401D-at-dcsmserver.med.sc.edu}
} } Hi, } } We currently have a position open for a microscopist in the } University of South Carolina School of Medicine Instrumentation } Resource Facility. Primary responsibilities involve assisting } faculty, staff and students in the use of confocal and electron } microscopes and in digital preparation of images (primarily Adobe } Photoshop). Secondary responsibilities involve assistance a flow } cytometer and a BioRad phosphorimager. } } All current work in the facility is biological. Applicants should } have a masters degree or equivalent experience. The } position is listed as a Research Specialist II with a salary range } from $24,618-$35,081. Please reply directly to me at
} Price-at-med.sc.edu. } Robert L. Price } Director, Instrumentation } Resource Facility } USC School of Medicine } Garner's Ferry Road } Columbia, SC 29208 } Phone: 803-733-3393 } Fax:803-733-1533
I am one of the people involved in the "for sale" example that Mark feels to be equivalent to his "instruments available" posting, which, I must confess, I don't remember seeing, or Nestor's apparent striking of it from the Listserver. I have participated in 8 of these TEM specimen prep workshops for Leica and RMC over the years, plus I have taken part (with many others) in similar workshops or short courses that are university-based, with the famous Lehigh short courses being the most widely-known example. I think that there are 2 key points you are missing, Mark:
1) These workshops are not designed to make money. If they break even, the companies that organize them are ecstatic.
2) These workshops are highly educational, which is the justification used by the students attending to their superiors (and confirmed by attendee feedback). As a supervisor, I continually look for such workshops on the Listserver for professional upgrading and/or career changes for my scientists and technologists.
When I want a new (or used) research tool, I tend not to look to the Listserver, but talk to vendors or colleagues, walk the floor at the MSA commercial exhibition, etc, etc. Whether or not your posting meets Nestor's criteria is not up to me, but let's not confuse apples with oranges (or the difficulty in properly using a complex instrument with the initial purchase of the instrument).
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 613-992-8735 email: malis-at-nrcan.gc.ca
} ---------- } From: } "Mriglermas-at-aol.com"-at-Sparc5.Microscopy.Com[SMTP:"Mriglermas-at-aol.com"-at-Sparc } 5.Microscopy.Com] } Sent: March 24, 1999 10:49 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Cc: Mriglermas-at-aol.com } Subject: Fwd: TEM / SEM Sample Prep Workshop } } { {Message: TEM / SEM Sample Prep Workshop} } } Dear Friendly Sysop - Nestor: } } I've attached a recent announcement for a workshop as an example of an } item } "for sale." I believe that items for sale, whether services, supplies, or } instruments all require the same kind of response you gave to my posting } for } "instruments available" recently. Please see that workshops for profit } are } stricken from the Listserver so that you are consistent with your own } policy } or, change your posting policy. } } I believe that any of us who are in this field should be able to post any } kind } of useful informative item that would be of interest to those on the list. } The availability of microscopes is a useful piece of info for someone who } has } a legitimate need for a used scope. There are a lot of people looking for } microscope users and owners as a trusted resource rather than the } manufacture's sales reps. Please consider this in the future. } } } Thanks for your attention to this matter. } } Mark W. Rigler, Ph.D. } VP, MAS, Inc. }
} I'm sorry to bother everyone, again, with a question about fixation of } bacteria. I greatly appreciate the responses I received a few weeks } ago. At one time Ryter-Kellenberger fixation was considered the } standard. It produced an image with a relatively clear nucleoid } containing fibrillar chromatin. Is this now considered to be an } artifact of preparation? } } John Wright } West Desert Test Center } (435) 831-3017 }
Two of the more recent reviews by the same E. Kellenberger ("The bacterial nucleoid revisited" (1994) in Microbiol Rev 58:211-32; and "Functional consequences of improved structural information on bacterial nucleoids" (1991) in Res Microbiol 142:229-38) should answer your question in great detail. They also make a most enjoyable reading!
Briefly, the nucleoid, according to the results obtained with rapid freezing-freeze substitution, "is now more granular than fibrillar", thus, conceivably, reflecting the natural supercoiled state of the DNA. It has a "coralline" shape, and "the excrescencies reach far into the cytoplasm. Membrane contact is no longer excluded". Interestingly, but the liquid-crystalline form of the DNA also appears to be native in some cases...
The classic Ryter-Kellenberger method still makes a standard for a good chemical fixation. It is beneficial sometimes to add an aldehyde prefixation step or make other minor modifications, but treatment with AQUOEUS uranyl acetate before dehydration remains the most critical for good nucleoid preservation. Bacterial "chromosome" lacks the proteins that keep eukaryotic chromatin from collapse during dehydration, and only aqueous uranyl acetate, following conventional fixation, cross-links the nucleoid in that "liquid-crystalline" state.
I remember your original posting also generated discussion about delayed secondary fixation, or for how long the material can be left in glut. For fine ultrastructure work, I would not leave bacteria (as well as anything else) at any step longer than necessary. Even if the osmotic pressure is adjusted to minimize swelling/shrinking, you will most likely end up with a specific unpleasant granularity of the cytoplasm in your bacteria if you leave them in a glutaraldehyde fixative for too long. To split the protocol into two days, it is best to leave the material overnight in 70% ethanol in the refrigerator. But for just diagnostic, etc., purposes, it is, of course, O.K., and sometimes simply unavoidable, to store the samples for quite a while in a glutaraldehyde fixative in a refrigerator.
Please feel free to contact me directly if you can't find the journals or if other questions arise. Sincerely, Vlad.
Vladislav V. Speransky School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Phone: 207 581 2998 Fax: 207 581 2969 Email: vladis-at-maine.maine.edu
Theoretical stereology Mathematical morphology Advances in image=20 analysis, Modern techniques in microscopy /image acquisition=20 Quantitative fractography 3-D modelling and analysis dissemination of=20 stereology and image analysis applications=20
Conference language English, no translations anticipated
Conference program (proceedings will be distributed prior to the=20 conference)=20
No parallel sessions Invited lectures (25') Oral presentations (15')=20 Poster presentations combined with panel discussion=20
Deadlines=20
Preliminary registration, October 31, 1999=20 Second circular, December 31, 1999=20 Final papers, March 31, 2000=20 Final registration, May 31, 2000=20 Final circular with conference program, July 31, 2000=20
Correspondence:
STERMAT 2000
Institute of Materials Science Cracow University of Technology Al. Jana Paw=B3a II 37 31-864 Krak=F3w, Poland
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Roberto, We used a thin gold coating on the shiny side of household = aluminum foil. This works great for relative thicknesses. Russ
-----Original Message----- } From: Roberto Garcia [mailto:rgarcia-at-unity.ncsu.edu] Sent: Wednesday, March 24, 1999 11:06 AM To: MSA Microscopy
The accurate control of the thickness of evaporated carbon films.
Principle. When carbon is deposited onto gold, it forms interference colors that are well defined. They can be used to determine the thickness of the carbon.
The colors. If carbon is evaporated onto gold, as the thickness of the carbon increases, the color changes through the following sequence: gold, orange, red, blue, grey. The change of color from red to blue is particularly sharp and clear. The change of color from red to blue occurs when the thickness of the carbon is 24.0 nm +/- 0.5nm. This result was obtained by people at Balzers using a multibeam interference technique for calibration.
Details. 1 Take a glass slide (or any other suitable substrate) and evaporate onto it a layer of gold. The thickness is not critical as long as the gold is thick enough to give an opaque film that looks like gold. 2 Mount the slide in the same chamber with the specimen to be coated with carbon. the thickness of the carbon on the slide will be 24 nm so arrange the distance of the slide and the sample so that (by the inverse square law) the desired thickness on the sample will occur when the thickness on the slide is 24 nm. 3 Evaporate the carbon; stop the evaporation as the color changes form red to blue. If you are using a normal arc for the carbon evaporation, the light from the arc will allow you to see the colors. The bell jar will need to be reasonably clean.
Example. Suppose you need to deposit a carbon film of thickness T nm. Let d be the distance from the carbon arc to the gold slide; let D be the distance from the carbon arc to the specimen. Then [d/D]squared = T/24.
Reference: My thesis (1967).
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Reply to: RE: Thickness of carbon films. Re: Carbon Film Thickness When great accuracy of film thickness is not needed, we simply = place a small metal washer upon a glass slide near the "specimen area". after evaporation the step height between the shaded and coated areas is = measured optically on a bench interference microscope. Ours is a Zeiss = two beam unit that is good for 30 nanometer resolution, (+ or -), with no = physical contact with the film. It works for ANY reflective metallic film = and fairly well even on "dull" carbon coatings because it has three = different reference mirrors that can be quickly changed. Each mirror has a = different reflectivity that one merely tests to get reasonable = interference fringes which can also be photographed via polaroid or 35 mm. = film. Bernie Kestel Materials Science Division = Argonne National Laboratory 9700 South Cass Ave. Argonne, Il., 60439
E-mail {kestel-at-anl.gov} FAX: (630) 252-4289
Alwyn Eades wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
{HTML} {HEAD} {/HEAD} {BODY} {PRE = WIDTH=3D"132"} Reply to: RE: Thickness of carbon films.
{/PRE} {FONT FACE=3D"Geneva" SIZE=3D3 COLOR=3D"#000000"} Re: = Carbon Film Thickness {BR} When = great accuracy of film thickness is not = needed, we simply place a small metal washer = upon a glass slide near the "specimen = area". {BR} after evaporation the step = height between the shaded and coated areas = is measured optically on a bench interference = microscope. Ours is a Zeiss two beam unit = that is good for 30 nanometer resolution, = (+ or -), with no physical contact with = the film. It works for ANY reflective metallic = film and fairly well even on "dull" = carbon coatings because it has three different = reference mirrors that can be quickly changed. = Each mirror has a different reflectivity = that one merely tests to get reasonable = interference fringes which can also be photographed = via polaroid or 35 mm. film. {BR} Bernie = Kestel {BR} Materials Science Division {BR} = Argonne National Laboratory {BR} 9700 South = Cass Ave. {BR} Argonne, Il., 60439 {BR} {BR} = E-mail <kestel-at-anl.gov> FAX: = (630) 252-4289 {BR} {BR} Alwyn Eades wrote: {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >-----------------------------------------------------------------------= - {BR} >The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America {BR} They can be used to determine the thickness = of the carbon. {BR} > {BR} >The colors. {BR} >If = carbon is evaporated onto gold, as the thickness = of the carbon {BR} >increases, the color = changes through the following sequence: = gold, orange, {BR} >red, blue, grey. The = change of color from red to blue is particularly {BR} >sharp = and clear. The change of color from red = to blue occurs when the {BR} >thickness = of the carbon is 24.0 nm +/- 0.5nm. {BR} >This = result was obtained by people at Balzers = using a multibeam {BR} >interference technique = for calibration. {BR} > {BR} >Details. {BR} >1 Take = a glass slide (or any other suitable substrate) = and evaporate onto {BR} >it a layer of gold. = The thickness is not critical as long as = the gold is {BR} >thick enough to give an = opaque film that looks like gold. {BR} >2 Mount = the slide in the same chamber with the specimen = to be coated with {BR} >carbon. the thickness = of the carbon on the slide will be 24 nm = so arrange {BR} >the distance of the slide = and the sample so that (by the inverse square {BR} >law) = the desired thickness on the sample will = occur when the thickness on {BR} >the slide = is 24 nm. {BR} >3 Evaporate the carbon; = stop the evaporation as the color changes = form red {BR} >to blue. If you are using = a normal arc for the carbon evaporation, = the {BR} >light from the arc will allow = you to see the colors. The bell jar will {BR} >need = to be reasonably clean. {BR} > {BR} >Example. {BR} >Suppose = you need to deposit a carbon film of thickness = T nm. Let d be the {BR} >distance from = the carbon arc to the gold slide; let D be = the distance from {BR} >the carbon arc to = the specimen. Then [d/D]squared =3D T/24. {BR} > {BR} >Reference: = My thesis (1967). {BR} > {BR} >Alwyn Eades {BR} >Department = of Materials Science and Engineering {BR} >Lehigh = University {BR} >5 East Packer Avenue {BR} >Bethlehem {BR} >Pennsylvannia = 18015-3195 {BR} > Phone 610 758 4231 {BR} > Fax = 610 758 4244 {BR} > {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 = COLOR=3D"#0000FF"} {U} jae5-at-lehigh.edu {/U} {/FONT} {FONT FACE=3D"Geneva" = SIZE=3D1 = COLOR=3D"#000000"} {BR} > {BR} > {BR} > {BR} >RFC822 = header {BR} >----------------------------------- {BR} > {BR} > = Received: from dns2.anl.gov (dns2.anl.gov = [146.139.254.3]) by {BR} >horus.et.anl.gov = (8.6.11/8.6.11) with ESMTP id IAA26354 for = < {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} kestel-at-= horus.et.anl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} >; Thu, 25 {BR} >Mar = 1999 08:32:30 -0600 {BR} > Received: from = Sparc5.Microscopy.Com (sparc5.microscopy.com = {BR} >[206.69.208.10]) by dns2.anl.gov = (8.9.1a/8.6.11) with SMTP id IAA03123; Thu, = 25 Mar 1999 {BR} >08:32:29 -0600 (CST) {BR} > = Received: (from {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = daemon-at-localhost {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} ) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) {BR} >id HAA19493 for dist-Microscopy; = Thu, 25 Mar 1999 07:53:13 -0600 {BR} > Received: = from no_more_spam.com (Sparc5 [206.69.208.10]) = by {BR} >Sparc5.Microscopy.Com (8.6.11/8.6.11) =
for {Microscopy-at-Sparc5.Microscopy.Com} id QAA26644 (ESMTP). Thu, 25 Mar 1999 16:50:44 +0100 (MET) Received: by tnt1.chem.tue.nl with Internet Mail Service (5.5.2448.0) id {GXT3GKNM} ; Thu, 25 Mar 1999 16:50:44 +0100 Message-ID: {23285647B6CFD111B783002048083698174AC4-at-tnt1.chem.tue.nl} {Microscopy-at-Sparc5.Microscopy.Com}
Hallo Roberto,
In the first edition of "Electron Microprobe Analysis" of S.B.J. Reed (Cambridge University Press 1975) you can find the next table. for Carbon on polished brass
Thickness in nm Colour
15 Orange 20 Indigo red 25 Blue 30 Bluish green 35 Green blue 40 Pale green 45 Silver gold
We determined the thickness with our thin layer program and found out that the values were very good. The table is not found in newer editions of Reed's book.
Ir.Hans Heijligers Solid State and Materials Chemistry Lab. STO 2.45, Eindhoven University of Technology POBox 513 NL-5600 MB Eindhoven E-mail: H.J.M.Heijligers-at-TUE.NL Tel.: +31 (0)402473051 Fax.: +31 (0)402445619
Hi. I was recently asked of a way to determine C film thickness. I remember that using a polished brass specimen and observing the color is one way. Does anyone have a source in the literature for this method? Thanks.
______________________ Roberto Garcia Senior Analyst, Metallography North Carolina State University Analytical Instrumentation Facility Box 7531, Room 303 EGRC Raleigh, NC 27695-7531 rgarcia-at-unity.ncsu.com http://spm.aif.ncsu.edu/aif
I'm in the market for a good quality, but inexpensive, film scanner- e.g., for scanning 35mm slides into my PC. I've heard Nikon makes a good one (model LS2500), but it runs a hefty $2,500 or so (too expensive for me). Got any suggestions?
-- Steven J. Fliesler, Ph.D. Professor Dept. of Ophthalmology Saint Louis Univ. School of Med. 1755 S. Grand Blvd. St. Louis, MO 63104-1540 Phone: (314) 577-8259 Fax: (314) 771-0596 E-mail: Fliesler-at-slu.edu
I am thinking of purchasing a microwave tissue processor. I would appreciate your opinion as to the pro's and con's of this type of tissue preparation.
If you have access to an AFM, accurate measurement of C thickness takes about 4 minutes. If I'm looking at flat samples, I use a toothpick to remove a thin line o= f carbon. I then can bring the sample to the AFM and directly measure the height of the step betwwen the sample and the top of the carbon film. Alternatively, if I can't work directly on the sample I coat a plain glas= s slide (making sure it is the same distance from the arc as the sample) an= d measure the C thinckness on it. Hope this helps
Hello everyone, A friend of mine has informed me that there is a young boy in the south of England who has a terminal cancer and is close to the end of his suffering. Apparently, he wishes to enter the Guiness Book of Records for the largest collection of buisness cards. Could you all take the time to help boost this young man's collection. I was asked to tell everyone else to forward this information to as many other people as possible. His particulars are: Master Gary Richard, 30 Selby Road, Carshalton, Surrey, England, U.K.
Regards Martin Roe Macaulay Land Use Research Institute Aberdeen Scotland AB15 8QH U.K.
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---------------------- Original Message Follows ----------------------
Reply to: RE: Thickness of carbon films. Re: Carbon Film Thickness When great accuracy of film thickness is not needed, we simply place a small metal washer upon a glass slide near the "specimen area". After evaporation the step heigth between the shaded and coated areas is measured optically on a bench interference microscope. Ours is a Zeiss two beam unit that is good for 30 nanometer resolution, (+ or -), with no physical contact with the film. It works for ANY reflective metallic film and fairly well even on "dull" carbon coatings because it has three different reference mirrors that can be quickly changed. Each mirror has a different reflectivity that one simply tests to get reasonable interference fringes which can also be photographed via polaroid or 35 mm. film. Bernie Kestel Materials Science Division Argonne National Laboratory 9700 South Cass Ave. Argonne, Il., 60439
E-mail {kestel-at-anl.gov} FAX: (630) 252-4289
Alwyn Eades wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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We have a Cambridge S240 scanning electron microscope that is available, that needs a new lab home. The S240 was wrapped up recently when we unexpectedly received a field emission SEM. The S240 is a digital frame store SEM; has a LaB6 source; and a second backplate that adapts to a Microspec WDS spectrometer. The S240 also includes a mating Noran thin window EDS detector. The S240 was well maintained under service contract for 11 years, and still would be in use if the FESEM has not suddenly become available from another Raytheon facility.
We also can include a used Kevex 8000 EDS analyzer with the interfaces to the Noran detector. In addition, I know where that Microspec spectrometer can be obtained.
We would like to trade the Cambridge SEM/Noran detector, and if desired, the Kevex analyzer, for a new Noran Vantage upgrade to our Noran EDS analyzer. We would like to hear from anyone who is interested.
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Reply to: RE: Thickness of carbon films.
Re: Carbon Film Thickness When great accuracy of film thickness is not needed, we simply place a small metal washer upon a glass slide near the "specimen area". after evaporation the step height between the shaded and coated areas is measured optically on a bench interference microscope. Ours is a Zeiss two beam unit that is good for 30 nanometer resolution, (+ or -), with no physical contact with the film. It works for ANY reflective metallic film and fairly well even on "dull" carbon coatings because it has three different reference mirrors that can be quickly changed. Each mirror has a different reflectivity that one merely tests to get reasonable interference fringes which can also be photographed via polaroid or 35 mm. film. Bernie Kestel Materials Science Division Argonne National Laboratory 9700 South Cass Ave. Argonne, Il., 60439
E-mail {kestel-at-anl.gov} FAX: (630) 252-4289
Alwyn Eades wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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by darkwing.uoregon.edu (8.9.3/8.9.3) with SMTP id NAA13960; Thu, 25 Mar 1999 13:51:51 -0800 (PST)
Alwyn writes ...
} The accurate control of the thickness of evaporated carbon films. } } ... } } The colors: } If carbon is evaporated onto gold, as the thickness of the carbon } increases, the color changes through the following sequence: } gold, orange, red, blue, grey. The change of color from red to } blue is particularly sharp and clear. The change of color from } red to blue occurs when the thickness of the carbon is } 24.0 nm +/- 0.5nm.
We also find the red-} blue transistion the most easily recognized and the most consistent, even for a multiple-user facility ... and altho it may be considered a bit thick, the transistion is sharp enough to use for EPMA without need for coating standards and unknowns at the same time. However, for the sake of clarification ... the Mineralogy reference would imply "blue" is 24nm ... are you claiming the red-} blue transistion (i.e., "purple") is 24nm?? We have been writing our technique up as 22(+/-1)nm.
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Lawrence Kordon Nikon, Inc. Columbia, MD nikon-at-jagunet.com
"Steven J. Fliesler" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I'm in the market for a good quality, but inexpensive, film scanner- } e.g., for scanning 35mm slides into my PC. I've heard Nikon makes a } good one (model LS2500), but it runs a hefty $2,500 or so (too } expensive for me). Got any suggestions? } } -- } Steven J. Fliesler, Ph.D. } Professor } Dept. of Ophthalmology } Saint Louis Univ. School of Med. } 1755 S. Grand Blvd. } St. Louis, MO 63104-1540 } Phone: (314) 577-8259 } Fax: (314) 771-0596 } E-mail: Fliesler-at-slu.edu
Can anyone out there help us out with a current phone number for Dunaway - or with anyone else that might have replacement heaters for a diffusion pump?
Many thanks in advance
Gill Bond Department of Materials & Met. Eng. New Mexico Tech
Steve, It depends on the resolution you need. Microtek sells a scanner with a 35 mm film attachment (X6EL) for under $200. You can also get better ones for $600-1000 with a second scan bed designed for slides. If you only have a few slides, we have even taken slides apart and scanned them directly on a flat bed scanner with a transparency adapter.
Best regards,
Chris Baumann
"Steven J. Fliesler" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I'm in the market for a good quality, but inexpensive, film scanner- } e.g., for scanning 35mm slides into my PC. I've heard Nikon makes a } good one (model LS2500), but it runs a hefty $2,500 or so (too } expensive for me). Got any suggestions? } } -- } Steven J. Fliesler, Ph.D. } Professor } Dept. of Ophthalmology } Saint Louis Univ. School of Med. } 1755 S. Grand Blvd. } St. Louis, MO 63104-1540 } Phone: (314) 577-8259 } Fax: (314) 771-0596 } E-mail: Fliesler-at-slu.edu
Dear all, Does anyone have an old Hitachi H450 SEM lying around that they would like to donate or sell cheaply for parts? Eric Hines Microscopy Centre CSIRO Entomology Canberra.
Gil Bond wrote: =============================================== Can anyone out there help us out with a current phone number for Dunaway - or with anyone else that might have replacement heaters for a diffusion pump ? =============================================== The information for Duniway is the following: Duniway Stockroom Corporation 1305 Space Park Way Mountain View, California USA 94043 Toll Free: 800-446-8811 Phone: 650-969-8811 Fax: 650-965-0764 E-MAIL: info-at-duniway.com WEB: www.duniway.com
If they don't have it, one place that seems to always have the odd-ball item others don't have is the following:
TORR International, Inc. 12 Columbus Street New Windsor, NY USA 12553 Ph: 1-914-565-4027 Fax:1-914-561-7731 E-mail: torr.intl-at-juno.com WEB: www.torr.com Ask for Dr. Masud Naraghi
We have no interest in either of these firms, however we have done some amount of business as a satisfied customer with both.
Chuck
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Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Glenn Poirier and Bernard Kestel (both messages appended)=20 advocate very accurate means of determining C film=20 thickness. No doubt these means have some applications.=20 However, for most applications it is rather more convenient=20 to determine thickness at the time of coating with fairly=20 good accuracy. It is no use to an analyst to break the vacuum to determine=20 that another 3nm of carbon are required. For these reason=20 the polished brass slide method and the still more=20 convenient, auto-terminating thickness monitors are the=20 preferred means to determine coating thickness. Incidentally, for WDS/EDS I used indigo red - 20nm; its=20 enough C to prevent charging on flat specimens and absorbs=20 fewer light X-rays. Cheers Jim Darley ProSciTech Microscopy=20 PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Friday, March 26, 1999 4:28 AM, Glenn Poirier=20 [SMTP:glennp-at-eps.mcgill.ca] wrote: } =20 ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } =20 ---------------------------------------------------------- } -------------. } } } If you have access to an AFM, accurate measurement of C } thickness takes } about 4 minutes. } If I'm looking at flat samples, I use a toothpick to } remove a thin line of } carbon. I then can bring the sample to the AFM and } directly measure the } height of the step betwwen the sample and the top of the } carbon film. } Alternatively, if I can't work directly on the sample I } coat a plain glass } slide (making sure it is the same distance from the arc=20 as } the sample) and } measure the C thinckness on it. } Hope this helps } } Glenn } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} } Glenn Poirier Tel: (514) 398 6774 } MicroAnalysis Laboratrory Fax: (514) 398 4680 } Earth and Planetary Sciences email: glennp-at-eps.mcgill.ca } Rm. 238, 3450 University St.=09 } http://castaing.eps.mcgill.ca } Montr=E9al, Qc } H3A 2A7 } Millennium hand and shrimp } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}
When great accuracy of film thickness is not needed, we=20 simply place a small metal washer upon a glass slide near=20 the "specimen area". after evaporation the step height between the shaded and=20 coated areas is measured optically on a bench interference=20 microscope. Ours is a Zeiss two beam unit that is good for=20 30 nanometer resolution, (+ or -), with no physical contact=20 with the film. It works for ANY reflective metallic film=20 and fairly well even on "dull" carbon coatings because it=20 has three different reference mirrors that can be quickly=20 changed. Each mirror has a different reflectivity that one=20 merely tests to get reasonable interference fringes which=20 can also be photographed via polaroid or 35 mm. film. Bernie Kestel Materials Science Division Argonne National Laboratory
John, you are possibly right; a similar thought did occur to me before sending it to the list (what if it's bogus and is really someone trying to get marketing, company information, or even a non-cancer patient who is building up a collection for himself etc.) and I questioned my friend as to how reliable her information was. Of course she could not guarantee that this wasn't a hoak but in the absence of any evidence to the contary I was prepared to take this more or less at face value and I thought well what if it is true? It's not much of an effort to help someone out and nothing has been lost in doing so even if it turns out not to be genuine.
Regards
Martin Roe Macaulay Land Use Research Institute Aberdeen Scotland AB15 8QH
John Mansfield wrote: } } I hate to pour cold water on this, but it looks like another of } } those net myths.
Steven writes ... } } } I'm in the market for a good quality, but inexpensive, film scanner- } e.g., for scanning 35mm slides into my PC. I've heard Nikon makes a } good one (model LS2500), but it runs a hefty $2,500 or so (too } expensive for me). Got any suggestions? } } ...
I believe you mean the LS-2000 ... and it can be found for less $$. For example, http://www.pricewatch.com will show you many places for which it can be purchased for near $1600. There are lesser expensive scanners and I've seen a comparison of the Nikon with the new HP Photosmart scanner with will only leave you with why would you pay so much more for the Nikon. Still, the Nikon comes with "dust removal" software which works very well, and you also have an option for 4X and 16X multiple scans for removing LED noise in the dense areas of the slide or negative. Both the Nikon and new HP will deliver 48bit files to image editors which can handle that color depth (e.g., Photoshop).
I agree with Jim Darley when he says=20 =93However, for most applications it is rather more convenient=20 to determine thickness at the time of coating with fairly=20 good accuracy.=94 What I find hard to understand is the assumption he and other contributor= s to this thread have made that the interference color method allows you to evaporate carbon of only one thickness. The brass (or in the method I gave, gold) test sample has always the same thickness of carbon but the thickness of carbon on the sample can be what you like. The idea is that the brass/gold test piece is placed at a distance from the source which i= s different from the distance between the source and the sample to be coate= d. Then the thickness on the sample can be calculated from the inverse squa= re law. When I first used this technique, I used it to apply a coating only= 1 nm thick.
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Dear Steven, I have the top model Hewlett-Packard flat bed scanner and that comes with a 35 mm slide scanning attachment that works very well. Cost ~ $500. You wrote:
} } I'm in the market for a good quality, but inexpensive, film scanner- } e.g., for scanning 35mm slides into my PC. I've heard Nikon makes a } good one (model LS2500), but it runs a hefty $2,500 or so (too } expensive for me). Got any suggestions? } } -- } Steven J. Fliesler, Ph.D. } Professor } Dept. of Ophthalmology } Saint Louis Univ. School of Med.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Ph.D. graduate student or post-doctoral student (D.V.M. or M.D.) position available for research in the area of protozoal parasitology. Research will involve electron microscopy and cellular biology studies utilizing in vitro derived Sarcocystis spp. , Neospora spp. and Toxoplasma gondii parasites. Individual will work with a multi-disciplinary team at the University of Missouri-Columbia, in the Department of Veterinary Pathobiology and the Molecular Biology Program Electron Microscopy Core facility. Successful candidate should have background in tissue/cell embedding and processing for light and electron microscopy, and demonstrated proficiency in written and spoken English. Molecular biology and cell culture skills are helpful, but not a necessary requirement. U.S. citizenship is not required. For further information contact Dr. A.E. Marsh at ph: 573-884-2673, fax: 573-884-5414, or email: marshae-at-missouri.edu {mailto:marshae-at-missouri.edu} .
A colleage at our university needs servicing on an ISI Alpha 9 SEM. The problem is related to the manually valved vacuum system. The hand-turned knob for valving does not appear to be actually moving the vacuum cylinder (plunger) up and down. Either it has come off of the main shaft or the vacuum cylinder (plunger) is jammed.
Does anyone service these instruments commercially? Any directions available for opening the vacuum system (for example, to change the diffusion pump oil)? Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
My name is Randy Anderson and I am a senior at Northern Il University. I am currently finishing the second semester of a TEM/SEM independent study. The reason that I am writing is to find out how I can apply for an E.M. certification and to see how the best way to look for a job position in the E.M. field when I graduate in the summer of 99. Thank you for taking the time to answer my questions as the information will be a valuable assest.
I've put copies of my Reichert Zetopan manuals on http://users.skynet.be/sky95421/
Availlable manuals ? Zetopan.zip Instruction manual for the Zetopan.Large research Microscope. In Englisch= . 1 298 kb. Contains one page from an older Osram-catalogue regarding much used bulbs for older European microscopes.
? Zetopol.zip Instruction manual for the Zetopan-Pol .Large Polarization Microscope. In French. 487 kb.
? reflight.zip Instruction manual for the Universal polarization illuminator for opaque objects. In French. 179 kb.
? micflash.zip Instruction manual for the Universal microflasch equipment for Zetopan. I= n German. 523 kb.
? MS140.zip Instruction manual for the MS 1.40 multisystem condenser. In German. 319 = kb.
? Binolux.zip Instruction manual for the Binolux III twin lamp unit. In Englisch. 1 063 kb.
uri-at-watson.ibm.com has devised a procedure to treat these files when you = are using Unix (thanks Uri=85)
Nigel Browning schrieb: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } {bold} "Analyzing Materials Interfaces at Atomic Resolution" {/bold} } } } } There will be a Materials Science symposium at Scanning 99 entitled } "Analyzing Materials Interfaces at Atomic Resolution". Scanning 99 is } being held at the Hyatt Regency O'Hare in Rosemont, IL from April 11-14 } 1999. The "Analyzing Materials at Atomic Resolution" symposium is } scheduled for Tuesday April 13th. The details of the conference can be } found at http://www.scanning.org or can be requested from Mary } Sullivan (e-mail: scanning-at-fams.org, tel:201-818-1010). Registration } for members of the Midwest Microscopy and Microanalysis Society is at } the reduced rates of $150 (regular $ 235) for the whole conference or } $50 (regular $ 95) for a single day (all attendees of the MMMS } symposium held at UIC last May are members of MMMS). } } } } } {bold} Speakers {/bold} } } } } 9.00 {bold} M. Haider-CEOS GmbH, Germany {/bold} } } "Towards sub-Angstrom resolution by correction of spherical } aberration" } } } 9.30 {bold} O. Krivanek-University of Washington {/bold} } } "Towards sub-Angstrom electron probes by Cs-corrected STEM." } } } } 10.00 Break } } } } 10.30 {bold} E. M. James-University of Illinois at Chicago {/bold} } } "Atomic resolution scanning transmission electron microscopy on the } 200kV FEGTEM" } } } 11.00 {bold} S. J. Pennycook-Oak Ridge National Lab {/bold} } } "Probing the Origin of Interfacial Properties by STEM" } } } 11.30 {bold} D. A. Muller-Lucent Technologies {/bold} } } "The End of the Roadmap for Silicon Dioxide: The Electronic Structure } of Hyper-Thin Gate } } Oxides at the Atomic Scale" } } } 12.00 {bold} D. B. Williams-Lehigh University {/bold} } } "Atomic-Resolution X-ray Microanalysis in the AEM" } } } } 12.30 Lunch } } } } 2.00 {bold} L. D. Marks-Northwestern University {/bold} } } "Picometer structure determination using Electron Diffraction" } } } 2.30 {bold} J. M. Gibson -University of Illinois at } Urbana-Champaign {/bold} } } "Statistical Measurement of Electron Scattering Fluctuations in } Amorphous } } Materials - A new Structural Tool" } } } } 3.00 Break } } } } 3.30 {bold} M. Gajdardziska-Josifovska-University of Wisconsin at } Milwaukee {/bold} } } "Quantitative surface microscopy and diffraction over the length } scales: Morphology and } } crystallography of polar oxide surfaces. " } } } 4.00 {bold} M. Tanaka-NRIM, Tsukuba, Japan {/bold} } } "Nano-behavior of Small Metal Particles in the Electron Beam" } } } } } } } } } } } } ___________________________________________________________________________ } } } Nigel D. Browning, PhD } } Assistant Professor } } University of Illinois at Chicago } } Department of Physics } } 845 West Taylor Street, } } Chicago } } IL 60607-7059. USA } } } Tel: 312-413-8164 } } Fax: 312-996-9016 } } } } http://interface.phy.uic.edu } } } ___________________________________________________________________________ } } }
I work on virtually everything in EM but have not worked on that model. Is that one of ISI's integrated vacuum valves, i.e. a hand operated valve with a number of individual positions for backing, roughing, etc. (usually just marked 0, 1, 2)? Or is it an individual valve with a single function?
I suspect that it is one of their integrated models, similar to manual and electrically operated models common to many of their instruments. They do come completely apart and have a very simple mechanism. There is probably a spring or taper pin that connects the hand turned portion to the central shaft. In some models, the shaft is visible from the top. If the shaft is not turning with the knurled hand piece then that pin has sheared off. While the pin can be easily driven out and replaced, this would also be a good time to rebuild the valve as there might be a problem inside that caused the pin to shear (these pins are often used a 'fuses' to prevent excessive force being applied to a mechanism).
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com
-----Original Message----- } From: John J. Bozzola {bozzola-at-siu.edu} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
Linda asks ... } } Dear listservers: } I would like to know which brands/models of dye sub printers do people } find give the best quality images. Thanks. Linda Chicoine }
A timely question ... 6-10 months ago photographic quality and trouble-free dye-subs were $5k-$10k printers ... plus $3/page. While you can buy a dye-sub today for { $700, it is still very much a case of how well the printer's software works with your application and OS and the quality of its color profile. I believe the consensus at one time was Kodak and Fujitsu for photographic and color quality. Today Alps should be considered in any decision because it has broken the price barrier to allow it to be considered along with color ink jets. I've been keeping an eye on lists and newsgroups without a good feeling for how well Alps supports its printers (... tech support, and ICC profiles ..) ... but the demo print they sent me was definitely excellent quality. If I were you I'd indicate which computer platform you use, and with what applications ... someone may reply with specific problems associated with less expensive dye-subs.
We have a M=E4rzh=E4user MRC-3 which is used to control stage movement and focusing by a joystick. The equipment is capable of being controlled from a computer but due to the need to write Basic instructions most users are put off this option. A similar problem exists using NIH Image macros. All very sad for the stereologists who would like to be able to have random sampling, montages, etc. The main problem is we use a Power Macintosh for the image processing side of things so would like to find a stage control program compatable with a Mac. At this point things grind to a halt, has anyone any knowledge of a Mac compatable stage control program with a user friendly interface?
Thanks
Andrew McNaughton
____________________________________________________________________________= __ Andrew McNaughton South Campus Electron Microscope Unit C/-Department of Anatomy and Structural Biology School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
We have great luck with the Codonics NP1600. However, Ii have recently talked with a Tektronix rep who tells me that they are geting out of the dye sub market and focusing on color laser and solid ink technologies. According to him, with the current color laser printers on the market, you can approach dye sub quality for between 2 and 3 k but the cost per sheet is } $.30 where a Codonics print costs $1.90.
Regards,
Chris Baumann
Linda Chicoine wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear listservers: } I would like to know which brands/models of dye sub printers do people } find give the best quality images. Thanks. Linda Chicoine
You might consider the Fuji Pictrography. We have one here and it delivers very high quality prints. I believe the price is competitive with dye-subs.
Mohan Kalyanarman
Materials Characterization Catalyst Technology Group Mobil Technology Company PO Box 480 Paulsboro, NJ 08066 609-224-3989 (ph) 609-224-3608 (fax) mohan_kalyanaraman-at-email.mobil.com
My company's R&D group is interested in visualizing several of our = polymers. The M&M '99 session, "Developments in Scanned Probe = Microscopy of Polymers", will be helpful. However, we would like some = basic/general information prior to the meeting. Any suggestions of = review articles, textbooks, etc. would be greatly appreciated.
In advance, thanks!
Janet H. Woodward, Ph.D. Corporate Technical Specialist - Microscopy & Microbiology Buckman Laboratories 1256 N. McLean Street Memphis, TN 38108 (901) 272-6408 jhwoodward-at-bbuckman.com
My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly) PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration: Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr
6. Gap Juctions in Drosophila PBS = phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} DW = distilled water wash buffer = 0.5 M NaCl and 0.05% Tween-20 in PBS
Fixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C) antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. “Differential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.” Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. “Gap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.” Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. “Gap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.” Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. “Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.” Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. “The 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).” Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. “Electron Microscope Immunolocation of Gap Junctions in Drosophila.” Tissue & Cell 1989;21(6): p 835-39.
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I am thinking of purchasing a microwave tissue processor. I would appreciate your opinion as to the pro's and con's of this type of tissue preparation.
Laura Robles
Hi Laura. I have been using my Pelco microwave for over a year now and I am very happy with it. I can process tissue in 2 hours instead of 2 days. It is especially useful for dehydration and infiltration. I would highly recommend getting one. It will pay for itself in the amount of time saved very quickly. The only con is that you have to experiment to find the best parameters for your tissue since there are few published protocols. Try it!
Does anyone have tips for dimpling soft metal alloys for TEM sample prep? I'm having difficulty dimpling an aluminium composite alloy. It appears that the abrasive (2-4 um cubic BN slurry) is getting embedded into the surface of the alloy. This then attacks the dimpling wheel (phosphor bronze).
Thanks, Wharton
++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
Central Michigan University Electron Microscope Facility Supervisor Biology Department
Central Michigan University is looking for an electron microscopy technician to supervise teaching/research EM facility. Assists in teaching introductory TEM and SEM classes. Solicits externally funded contracts. Required qualifications include education equivalent to Bachelor's degree; two years qualifying work experience; working knowledge of the principles and techniques of transmission and scanning electron microscopy, ultramicrotomy, vacuum evaporation, critical point drying and biological specimen preparation; general knowledge of solid-state electronics; proficiency with TERM, SEM, EDS, digital imaging and light microscopy. Experience with routine maintenance of EM and optical microscopes desired.
Review of applications will begin on May 1, 1999, and will continue until the position is filled. Please send resume materials to Central Michigan University, Human Resources/Staff, Rowe 109, Mt. Pleasant, MI 48859. CMU provides flexible benefits, an excellent retirement program with tax deferred investment options, tuition waiver for employee and family, and competitive salaries in an environment committed to excellence and customer service.
-- Amy B. Gambrell Phone: 517.774.3339 Human Resources/Staff Fax: 517.774.3256 109 Rowe Hall E-mail: Amy.Beth.Gambrell-at-cmich.edu Central Michigan University http://www.sps.cmich.edu Mt. Pleasant, MI 48859
To find out about exciting job opportunities at CMU, click here: http://www.sps.cmich.edu/jobs.html
My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly) PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration: Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr
6. Gap Juctions in Drosophila PBS = phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} DW = distilled water wash buffer = 0.5 M NaCl and 0.05% Tween-20 in PBS
Fixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C) antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. “Differential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.” Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. “Gap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.” Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. “Gap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.” Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. “Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.” Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. “The 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).” Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. “Electron Microscope Immunolocation of Gap Junctions in Drosophila.” Tissue & Cell 1989;21(6): p 835-39.
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My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly) PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration: Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr
6. Gap Juctions in Drosophila PBS = phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} DW = distilled water wash buffer = 0.5 M NaCl and 0.05% Tween-20 in PBS
Fixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C) antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. “Differential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.” Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. “Gap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.” Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. “Gap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.” Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. “Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.” Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. “The 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).” Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. “Electron Microscope Immunolocation of Gap Junctions in Drosophila.” Tissue & Cell 1989;21(6): p 835-39.
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Hey people, Anyone have any experience with the new RMC crx cryounit that can attach to a Riechert E? It is supposedly much cheaper than Leica's but is it as good? Does anyone have any general experience with RMC. Thanks.
I am interested in finding out if there is any facility that has an ESEM Tensile Stage with Peltier capabilities for structure/function studies using ESEM.
SPM of Polymers is one of the hottest new trends in microscopy. I am in the middle of writing a review of "What's New in Microscopy at PITTCON" (May, 1999, American Lab) and this topic was well represented by new offerings from several companies. I'd suggest you visit the websites for
1. Digital Instruments - they have lots of great app notes on this topic
2. ThermoMicroscopes (previously known as Park Scientific and Topometrix) and look for information on the new Explorer PolymerSystem SPM (integrated microthermal analysis and pulsed force mode imaging)
3. JEOL - their new SPM has a full hot/cold stage which runs from 130K to 800K, for watching thermal transitions.
Other relevant technology I found at Pittcon included the new thermal stage in Philips' ESEM, the new Continuum microscope for full microscopy imaging (Including DIC) integrated with FTIR from SpectraTech, and a number of interesting systems for Raman/confocal from Renishaw and Instruments SA.
Hope this "preview" was helpful.
CAVEAT: Neither Barbara Foster nor MME has any commercial interest in any of these instruments.
Best regards,
Barbara Foster
Consortium President
Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118
MME is America's first national consortium providing
customized on-site workshops in all areas of
microscopy, sample preparation, and image analysis.
At 10:29 AM 3/29/99 -0600, Janet H. Woodward wrote:
} } } }
{excerpt} My company's R&D group is interested in visualizing several of our polymers. The M&M '99 session, "Developments in Scanned Probe Microscopy of Polymers", will be helpful. However, we would like some basic/general information prior to the meeting. Any suggestions of review articles, textbooks, etc. would be greatly appreciated.
My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Fixation: 1% paraformaldehyde-0.1M PB Sectioning: 30 mm Rinsing: PBS (24 hr) Blocking: BSA-PBS (30 min) 1st Incubation:(4 C) antibody+[BSA-PBS] {1:1000} (12 hr) Washing: (4 C) PBS (12 hr) 2nd Incubation: (4 C) Au conj. anti-mouse IgG+[BSA-PBS] {1:20} (12 hr) Post Fixation:=09 1% osmium tetroxide-0.1 M PB (1 hr) Dehydration:=09 Embedding: Epon 812 Ultra-thin Sectioning:=09 80 nm Mounting: mesh grids Total Time: 61.5 hr =09 2. Human Smooth Muscle =09 PB =3D phosphate buffer Au conj. =3D gold conjugated (post-embedding) {1993} =09 Fixation: 0.5% glutaraldehyde- 0.175 M cacodylate buffer Dehydraton:=09 Embedding: (4 C) LR White Polymerization:=09 cold+UV light (?? hr) Ultra-thin Sectioning:=09 parallell to plane of c-slip Mounting: formvar, 200 Ni mesh grids Blocking: 0.01 M glycine+1% NFDM+0.5% gelatin+0.1 M PB (?? hr) 1st Incubation: (R.T.) rabbit anti-Cx43 serum+[NFDM+gelatin+PB] {1:750} (1 hr) Washing: (entensive) PB 2nd Incubation: (R.T.) Au conj. goat anti-rabbit IgG+[NFDM+gelatin+PB] {1:10} (1hr) Washing: =09 Fixation: (brief) 2% glutaraldehyde+PB Rinsing: water Drying: air Staining: uranyl acetate and Reynold's lead citrate Total Time: ?? hr =09 3. Stratum Granulosum Cells of Mouse Follicles=09 PB =3D phosphate buffer PBS =3D phosphate buffered saline Au conj. =3D gold conjugated (pre-embedding) {1993} =09 Fixation: 1% paraformaldehyde- 0.1 M PB (2-4 hr) Sectioning: 80 mm Rinsing: PB Blocking: PBS containing 1% BSA & 0.05% sapoinin (30 min) 1st Incubation: (4 C)=09 anti-Cx43 rabbit serum+[PBS containing 1% BSA & 0.05% sapoinin] {1:200} (28-40 hr) Washing: PBS (5 hr) 2nd Incubation: 5 nm Au conj. Antibody+[PBS containing 1% BSA & 0.05% sapoinin] {1:3} (22-24 hr) Washing: PBS (4-6 hr) Fixation: sodium cacodylate buffer (pH 7.4, 0.1 M) + 2.5% glutaraldehyde (1.5 hr) Post Fixation:=09 1% osmium tetroxide (1.5 hr) Dehydration: ethanol & propylene monoxide Embedding: Epon 812 Ultra-thing Sectioning:=09 Mounting: formvar-film coated single-slot grids Staining: uranyl acetate and lead citrate solutions Total Time: 64.5 - 82.5 hr =09 4. Rat Heart Gap Junction Membranes=09 PBS =3D phosphate buffered saline (pre-embedding) {1990}
1st Incubation: (R.T.)=09 poly-L-lysine coated 96-well plates (unbouding of membranes??) (14 hr) Blocking: PBS containing 5% BSA (1 hr) 2nd Incubation: 25-to-100 fold dilution of preimmune or immune serum (1 hr) ---or--- 2nd Incubation:=09 80-115 mg ml-1 affinity purified antibodies Washing: PBS 3rd Incubation:=09 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10} (1 hr) Washing: PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration:=09 Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 15+ hr =09 5. Heart Gap Junctions=09 PBS =3D phosphate buffered saline (pre-embedding) {1989}
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: =095 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly)=09PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration:=09 Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr =09 6. Gap Juctions in Drosophila=09 PBS =3D phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} =09 DW =3D distilled water wash buffer =3D 0.5 M NaCl and 0.05% Tween-20 in PBS
Fixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C)=09 antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. =93Differential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.=94 Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. =93Gap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.=94 Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. =93Gap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.=94 Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. =93Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.=94 Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. =93The 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).=94 Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. =93Electron Microscope Immunolocation of Gap Junctions in Drosophila.=94 Tissue & Cell 1989;21(6): p 835-39.
My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Fixation: 1% paraformaldehyde-0.1M PB Sectioning: 30 mm Rinsing: PBS (24 hr) Blocking: BSA-PBS (30 min) 1st Incubation:(4 C) antibody+[BSA-PBS] {1:1000} (12 hr) Washing: (4 C) PBS (12 hr) 2nd Incubation: (4 C) Au conj. anti-mouse IgG+[BSA-PBS] {1:20} (12 hr) Post Fixation:=09 1% osmium tetroxide-0.1 M PB (1 hr) Dehydration:=09 Embedding: Epon 812 Ultra-thin Sectioning:=09 80 nm Mounting: mesh grids Total Time: 61.5 hr =09 2. Human Smooth Muscle =09 PB =3D phosphate buffer Au conj. =3D gold conjugated (post-embedding) {1993} =09 Fixation: 0.5% glutaraldehyde- 0.175 M cacodylate buffer Dehydraton:=09 Embedding: (4 C) LR White Polymerization:=09 cold+UV light (?? hr) Ultra-thin Sectioning:=09 parallell to plane of c-slip Mounting: formvar, 200 Ni mesh grids Blocking: 0.01 M glycine+1% NFDM+0.5% gelatin+0.1 M PB (?? hr) 1st Incubation: (R.T.) rabbit anti-Cx43 serum+[NFDM+gelatin+PB] {1:750} (1 hr) Washing: (entensive) PB 2nd Incubation: (R.T.) Au conj. goat anti-rabbit IgG+[NFDM+gelatin+PB] {1:10} (1hr) Washing: =09 Fixation: (brief) 2% glutaraldehyde+PB Rinsing: water Drying: air Staining: uranyl acetate and Reynold's lead citrate Total Time: ?? hr =09 3. Stratum Granulosum Cells of Mouse Follicles=09 PB =3D phosphate buffer PBS =3D phosphate buffered saline Au conj. =3D gold conjugated (pre-embedding) {1993} =09 Fixation: 1% paraformaldehyde- 0.1 M PB (2-4 hr) Sectioning: 80 mm Rinsing: PB Blocking: PBS containing 1% BSA & 0.05% sapoinin (30 min) 1st Incubation: (4 C)=09 anti-Cx43 rabbit serum+[PBS containing 1% BSA & 0.05% sapoinin] {1:200} (28-40 hr) Washing: PBS (5 hr) 2nd Incubation: 5 nm Au conj. Antibody+[PBS containing 1% BSA & 0.05% sapoinin] {1:3} (22-24 hr) Washing: PBS (4-6 hr) Fixation: sodium cacodylate buffer (pH 7.4, 0.1 M) + 2.5% glutaraldehyde (1.5 hr) Post Fixation:=09 1% osmium tetroxide (1.5 hr) Dehydration: ethanol & propylene monoxide Embedding: Epon 812 Ultra-thing Sectioning:=09 Mounting: formvar-film coated single-slot grids Staining: uranyl acetate and lead citrate solutions Total Time: 64.5 - 82.5 hr =09 4. Rat Heart Gap Junction Membranes=09 PBS =3D phosphate buffered saline (pre-embedding) {1990}
1st Incubation: (R.T.)=09 poly-L-lysine coated 96-well plates (unbouding of membranes??) (14 hr) Blocking: PBS containing 5% BSA (1 hr) 2nd Incubation: 25-to-100 fold dilution of preimmune or immune serum (1 hr) ---or--- 2nd Incubation:=09 80-115 mg ml-1 affinity purified antibodies Washing: PBS 3rd Incubation:=09 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10} (1 hr) Washing: PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration:=09 Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 15+ hr =09 5. Heart Gap Junctions=09 PBS =3D phosphate buffered saline (pre-embedding) {1989}
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: =095 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly)=09PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration:=09 Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr =09 6. Gap Juctions in Drosophila=09 PBS =3D phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} =09 DW =3D distilled water wash buffer =3D 0.5 M NaCl and 0.05% Tween-20 in PBS
Fixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C)=09 antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. =93Differential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.=94 Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. =93Gap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.=94 Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. =93Gap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.=94 Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. =93Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.=94 Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. =93The 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).=94 Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. =93Electron Microscope Immunolocation of Gap Junctions in Drosophila.=94 Tissue & Cell 1989;21(6): p 835-39.
My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly) PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration: Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr
6. Gap Juctions in Drosophila PBS = phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} DW = distilled water wash buffer = 0.5 M NaCl and 0.05% Tween-20 in PBS
Fixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C) antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. “Differential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.” Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. “Gap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.” Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. “Gap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.” Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. “Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.” Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. “The 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).” Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. “Electron Microscope Immunolocation of Gap Junctions in Drosophila.” Tissue & Cell 1989;21(6): p 835-39.
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We are a non-profit botanical garden conducting plant anatomy research. We are looking for donations or second hand low-cost microscopy equiptment. We can pay for the shipping of the following items:
Compound research/student style microscope
We are looking for a Riechert Sliding Microtome & knives from anyone/institution wishing to dispose of one.
We are also looking for non-disposible blades for a Spencer 820 rotary microtome
& just the INSTRUCTIONS for a Sensaur microtome knife sharpener: Aloe - Cat # V60041; Model # SH 67-R; Serial # 1107; 115 Vac 50/60 C 5A
Please reply to my email: crow-at-aloha.net Thanks very much in advance
"The light which puts out our eyes is darkness to us. Only that day dawns to which we are awake. There is more day to dawn. The sun is but a morning star."
Try one, two, three or all of the following: 1. Use a stainless steel wheel instead of a bronze wheel; 2. Decrease the balance load; 3. Change slurry (size, abrasive particles etc. etc....) 4. If I were you, I would also completely skip the dimpling procedure and try a Grind Holder (such as Gatan's) or Tri-pod (such as south Bay Tech's) and put the specimen on ion-mill right after obtaining {20um thick foil.
Hope the above helps.
-cy, Rodelee-to-be
On Mon, 29 Mar 1999, Wharton Sinkler wrote:
} Does anyone have tips for dimpling soft metal alloys for TEM sample prep? } I'm having difficulty dimpling an aluminium composite alloy. It appears } that the abrasive (2-4 um cubic BN slurry) is getting embedded into the } surface of the alloy. This then attacks the dimpling wheel (phosphor } bronze). } } Thanks, } Wharton } } ++++++++++++++++++++++++++++++++++++++++++++++++++ } Wharton Sinkler } Argonne National Laboratory West } P. O. Box 2528 } Idaho Falls, ID 83403 } Tel: (208) 533-7724 } FAX: (208) 533-7863 } } mailto:wharton.sinkler-at-anlw.anl.gov } }
Politeness would suggest that you enter something a little more informative in your subject field!
To just say "I AM NOT SPAM" makes you look suspiciously like spam, but also somehow manages to suggest that you think that, compared to YOUR important message, others may be spam.
rtch
} Hello, } } My name is Aaron Rea. I am working on an undergraduate research project at } Southwest Missouri State University (Springfield, MO USA). I am starting work } with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying } to develop a specimen preparation protocol for the immunolocalization of Cx43 } within this cell line. I have outlined the prep protocols from 6 papers that } utilized immunoEM below. I am looking for any suggestions or recommendations } on which protocol might yield the greatest success. Any other thoughts or } ideas would be greatly appreciated. Thanks to all who responded back in } October when I began this project!! Please reply offlist at: } aaronrea-at-hotmail.com }
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Hello, could you please post my message below to the list with the title of the subject being, "HELP! ImmunoEM Prep for Cx43 Localization". Thank you for your time.
Sincerely, Aaron Rea 439 E. Madison #7 Springfield, MO 65806 aaronrea-at-hotmail.com ---------------- Hello,
My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly) PBS =46ixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration: Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr
6. Gap Juctions in Drosophila PBS =3D phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} DW =3D distilled water wash buffer =3D 0.5 M NaCl and 0.05% Tween-20 in PBS
=46ixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C) antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. =ECDifferential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.=EE Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. =ECGap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.=EE Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. =ECGap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.=EE Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. =ECBiochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.=EE Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. =ECThe 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).=EE Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. =ECElectron Microscope Immunolocation of Gap Junctions in Drosophila.=EE Tissue & Cell 1989;21(6): p 835-39.
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Barbara Foster says: } Uri, Let me see what I can dig out. I'm headed out on an extended } business trip... will be back around 3/15. Could you send me a reminder?
Barbara, since my attempts to contact you via e-mail failed so far, I decided to post the reminder here, hoping that it will reach you.
Do you (or anybody else on this list) have any Zetopan manuals, especially for Phase Contrast, Anoptral Contrast, transmitted light Interference Contrast, Fluorescence, Zetopan-POL? [Needless to say, I need those.]
Thank you. -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
} My company's R&D group is interested in visualizing several of our } polymers. The M&M '99 session, "Developments in Scanned Probe } Microscopy of Polymers", will be helpful. However, we would like some } basic/general information prior to the meeting. Any suggestions of } review articles, textbooks, etc. would be greatly appreciated.
Janet,
It strikes me that it is better not to limits oneself to SPM. This technique is powerful, but limited in its application. Etching and staining can often do better - it really depends which polymers you're lloking at. If you can get hold of "Atlas of Polymer Morphology" by Arthur E. Woodward (distributed in USA by Oxford University Press, New York, ISBN 0-19-520758-0) this would give you a lot of ideas.
If staining interests you, the following article is superb. However, the author is out of reprints, so it might be better to use some form of library loan, if you don't take the journal.
TI: Reflections on the use of microtomy for materials science specimen preparation AU: Plummer_HK NA: FORD MOTOR CO,RES LAB,MAIL DROP 3028,SRL,DEARBORN,MI,48121 JN: MICROSCOPY AND MICROANALYSIS, 1997, Vol.3, No.3, pp.239-260 IS: 1431-9276 DT: Review
If you are into polyethylene or polypropylene, and also some blends, etching might be useful. You can see a few pictures on my home page by following the link "Picture Gallery" and also on:
http://www.reading.ac.uk/~spshosir/gallery.htm
from a colleague. If you want to follow this further, please let me know.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
} Janet H. Woodward, Ph.D. Corporate Technical Specialist - Microscopy & } Microbiology Buckman Laboratories 1256 N. McLean Street Memphis, TN } 38108 (901) 272-6408 jhwoodward-at-bbuckman.com
I was having problems with my e-mail account yesterday and wasn't sure if I was getting my message through. I now have the problem figured out and this will not happen again. I am truly sorry for any inconvenience or annoyance this caused. A special thanks to those who responded with Cx43 protocol ideas!
Sincerely,
Aaron Rea Get Your Private, Free Email at http://www.hotmail.com
***************************************************** Electron Microscopy/Image Analysis Technician
An NIH-funded position is available immediately for an electron microscopist to study the molecular structure and function of muscle. Projects involve determination of the 3D molecular structure of actin and myosin filaments and the structural changes underlying contraction. Our main approaches involve electron microscopy (negative staining, cryo-EM) and 3D image analysis (helical and tomographic reconstruction) combined with biochemical, immunological and molecular biological methodology. Candidates should be motivated individuals with a good background in molecular level electron microscopy and/or image processing. Our laboratory is equipped with Philips CM120 cryo, CM10 and EM400 TEMs, Gatan cryoholder and TV system, ancillary EM equipment (freeze plungers, freeze fracture, evaporators, etc), optical diffractometer, scanning densitometer, Silicon Graphics image processing workstations, etc. Applicants should provide a CV and names and addresses of 3 references to Dr Roger Craig, Department of Cell Biology, University of Massachusetts Medical School, Worcester MA 01655. Tel: (508) 856 2474; Fax: (508) 856 6361; Email: Roger.Craig-at-ummed.edu. *****************************************************
Dear Wharton, I have successfully dimpled Al2O3-Al MMC (metal matrix composite) samples, using the VCR Dimpler. The dimpling abrasive is 3 um diamond paste diluted in water and the wheel is steel. In the notes, this problem of embedding the abrasive in a soft work piece is addressed and recommends using a softer abrasive or a diamond tool. You might contact VCR at: 650-875-1000 for advice. You wrote:
} } Does anyone have tips for dimpling soft metal alloys for TEM sample prep? } I'm having difficulty dimpling an aluminium composite alloy. It appears } that the abrasive (2-4 um cubic BN slurry) is getting embedded into the } surface of the alloy. This then attacks the dimpling wheel (phosphor } bronze). } } Thanks, } Wharton } } ++++++++++++++++++++++++++++++++++++++++++++++++++ } Wharton Sinkler } Argonne National Laboratory West
No financial interest, just a happy customer. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Barbara Foster says: } I have a general manual which outlines all the transmitted light functions. } Unfortunately, there is only a small amount on Anoptral Phase --- they } refer to a "separate manual" for that. There are also other accompanying } materials which discuss Fluorescence.
I am interested in the Fluorescence materials. The general manual - I got it, thanks to Yvan and Wayne.
} Also, Yvan Lindekens has posted his copy of the Zetopan manual at } {http://users.skynet.be/sky95421} . I am having trouble with my unzip files } so have not been able to open his manual to compare it to mine, but suggest } you see what is there.
I was probably the first who got Yvan's copy of the manual (:-). It's a pretty good one, in case anybody cares to know.
After you unzip it, you get one large TIFF file. So now you need software that (a) can deal with multi-page TIFF's, or (b) can convert TIFF to something the printers like better [such as PostScript]. See my comments on Yvan's Web site.
Yvan, please get in touch with me off-line - I need to figure out how to scan a few manuals myself, and put them on your Web site. -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
The Listserver system went bizzare today in synchronism with my attempts to check for viruses. As a result, a number of of mail messages were rejected.
I believe all is well again. Those of you that were rebuffed please accept my apology.
My name is Birna Gu=F0bj=F6rnsd=F3ttir working at Icelandic Fisheries Laboratories as a food scientist. This field is quite new to me and I am looking for information how I can prepare wood samples, both wet nad dry, for SEM. Any information would be greatly appreciated.
My name is Birna Gu=F0bj=F6rnsd=F3ttir working at Icelandic Fisheries Laboratories as a food scientist. This field is quite new to me and I am looking for information how I can prepare wood samples, both wet nad dry, for SEM. Any information would be greatly appreciated.
Birna wrote: } My name is Birna Gudbjoernsdottir working at Icelandic Fisheries } Laboratories as a food scientist. This field is quite new to me and I am } looking for information how I can prepare wood samples, both wet nad dry, } for SEM. Any information would be greatly appreciated.
Dear Birna,
If you wish to keep the wood wet, you will have to use an environmental SEM (ESEM), in which you can examine the sample with a pressure above 5 torr (the equilibrium pressure of water vapour just above 0 degree C) in the specimen chamber. If you can accept that the wood slowly dries, you may use a low-vacuum SEM (LVSEM) where you can work with a pressure of up to 1-2 torr. Both ESEM and LVSEM allows the examination of electrical insulators.
In the conventional high vacuum SEM you cannot have wet wood. Dry wood may be examined, but then the surface must be made conductive e.g. by sputtering gold onto the sample.
Best regards, Joergen.
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
I have been using Syquest 230 EZ Flier dirves on my TDL12 8000/Delta system for some time. Other than the reliability problem of the Syquest hardware and the posibility of little future support from Syquest, they work well and are rather faster than the bernoulliis. A "sort-of" special cable is needed to interconnect the drives. The interface instructions simply address the setup of DMON (nowdays that would be called a driver, I guess). The disks are formatted rt11 and instead of 230MB, hold 44Mb, but it was an overall improvement. I would guess that similar, more current external SCSI-1 drives could also be made to work.
First, thanks to everyone who responded to my question on fixation of cells grown on chambered Permanox slides. As soon as I try the suggested procedures (the cells crashed!), I will post a summary of responses to the list.
My question today has to do with the presence of "pepper" on TEM sections. We process rather large tissue blocks (4X5 mm) of nervous tissue that we have dissected from perfused animals (5% glut. in PB is routine perfusate). Post-fixation is in osmium tetroxide in PB and all buffer rinses are done in PB. We dehydrate using an ethanol series, transition through propylene oxide (although we are experimenting with ethanol with no succes so far) and infiltrate/embed in PolyBed 812. The pepper shows up most obviously on the cytoskeletal elements of nerves, mitochondria, and collagen. I have tried pretreating the sections with 0.5-1.0 N HCl or 1% EDTA (Mollenhauer, 1987)prior to staining with uranyl acetate/lead citrate.....no luck. I can visualize the pepper on unstained sections. I seem to recall that this may have something to do with phosphate buffer, but the details are hazy. I would appreciate any suggestions.
Birna Guđbjörnsdóttir wrote: =================================================== My name is Birna Guđbjörnsdóttir working at Icelandic Fisheries Laboratories as a food scientist. This field is quite new to me and I am looking for information how I can prepare wood samples, both wet nad dry, for SEM. Any information would be greatly appreciated. =================================================== Assuming you would have just a conventional SEM at your disposal, we have generally had good results in our own laboratory, when the need has arisen in the past, to examine wood, to dry it with critical point drying. A second (but we believe inferior) approach is to use the HMDS drying protocol , which tends to be favored by those without access to a CPD unit. Whatever the drying method, since the sample itself will be nonconductive, it should be made conductive via the application of a thin layer of gold, or for higher resolution work, chromium or (more recently) osmium (metal).
Information about CPD and HMDS and also metal coaters can be found on our website given below or the websites of the other major manufacturers and suppliers of sample preparation equipment and consumable supply items for SEM laboratories.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Does anyone involved in Electron Microscopy know where I can obtain = spare glass scoring wheels for an LKB 2178 Knife Maker II. Any helpful = advise would be welcome as our machine requires a replacement scorer = ASAP.
Dr Terry Robertson (PhD) Senior Research Fellow Department of Pathology University of Western Australia Nedlands 6907
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
I am interested in finding out if there is any facility that has an ESEM Tensile Stage with Peltier capabilities for structure/function studies using ESEM.
by majestic.tcs.tulane.edu (8.9.3/8.9.3) with SMTP id PAA21132; Wed, 31 Mar 1999 15:13:53 -0600 (CST) Message-Id: {199903312113.PAA21132-at-majestic.tcs.tulane.edu}
Dear Colleagues: Sometime ago I posted a request for information on microscopy fees. I received several wonderful information packages and thank those who contributed. Before submitting a proposal internally at Tulane, I would like to build a table with a comparison among several institutions. Thus, I need information on fees charged as shown in the sample below. Additional information on structure and function of morphology cores and microscopy coresis welcome as well. You do not have to be specific, and I will not disclose your name or associate it with the information outside. I will only use your name inside Tulane if you authorize it. Thus far not a single institution can recoup costs from fees. Thus, what I need to demonstrate is that such fact is not just an idea, and for that I need your input.
} Our facility charges $25.00 per hour for a Hitachi H7000 TEM and will be } charging $35.00 for a new TEM with digital capabilities. Technical help is } 35.00 per hour, an inverted multi-photon confocal is $25 per hour and a 3 } laser upright confocal with nomarski is $15 per hour. We offer two hours of } training at no charge. Use of the darkroom which covers chemicals and the } service on our processor is $10 per hour...Unfortunately we can't charge } } what it really cost to operate the facility or we'd have no business--so } } the university subsidizes us.
When all is done, I will summarize information received (leaving out names and institutions unless specifically asked to leave in the posting) and post for MSA users. Thanks in advance for your help.
*Disclaimer: Whatever... is not Tulane opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web:[ http://www.tmc.tulane.edu/ferminlab/] or [http://www.tmc.tulane.edu/imaging/] Internet: [cfermin-at-mailhost.tcs.tulane.edu] 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
We are trying to study the microstructure of cold drawn 25 =B5m gold = wires. We meet the difficulty in etching the gold wires to reveal the grain structure in SEM and optical observations. Could somebody give us some suggestions on how to etch the wires.
Thanks in advance!
Best regards,
Kun Li
Kun Li, Ph. D Institute of Materials Research and Engineering BLK S7, Level 3 Office: BLK S13, #02-13d National University of Singapore Lower Kent Ridge Road Singapore 119260
Birna wrote: My name is Birna Gu=F0bj=F6rnsd=F3ttir working at Icelandic Fisheries Laboratories as a food scientist. This field is quite new to me and I am looking for information how I can prepare wood samples, both wet nad dry, for SEM. Any information would be greatly appreciated.
Dear Birna,
for conventional SEM your samples have to be dry. If your sample is already dry, just sputter with gold. For wet wood it is depending on the age of the the wood and the quality you want. Maybe air drying is possible? If not you have to perform critical point drying or freeze drying.
Best wishes from Anne Heller
Dr. Anne Heller Arbeitsgruppe Elektronenmikroskopie Institut fuer Botanik (210) Universitaet Hohenheim Garbenstr.30 D-70593 Stuttgart Tel.0049-711-459-2180 Fax 0049-711-459-3355
hi everybody ! What is specific to surface profile imaging technique in HRTEM ? Is it classical HRTEM used on surface imaging and profile only means that HRTEM images are used to deduce the structure (=profile) of the surface (=edge) of the microcrystal in a classical way (structural models + simulations) or is there a specific technique ? Thanks B. DOMENGES CRISMAT - ISMRA Boulevard du Marechal Juin 14050 CAEN CEDEX France E-mail : domenges-at-crismat.ismra.fr Tel. : (33) 02 31 45 26 32 Fax. : (33) 02 31 95 16 00
Sandra: Pardon a copied reply, but this is a common problem with phosphate buffer fixation. I posted the following reply to a similar request in September '98.
Sally, about 15 years ago somebody published the reason for this Os "pepper". Simply, for this to occur, some chemically unbound Os, GA and phosphate must remain in the tissue. If any one of these is missing you will not get this frustrating artefact.
I suppose this is one major reason for the popularity of cacodylate buffer. It makes the very thorough rinsing process which is required between GA and OS redundant; simply any unbound GA no longer matters.
Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Thursday, April 01, 1999 5:10 AM, Sandra Perkins [SMTP:skperkin-at-vt.edu] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } Hi- } } First, thanks to everyone who responded to my question on } fixation of cells } grown on chambered Permanox slides. As soon as I try the } suggested } procedures (the cells crashed!), I will post a summary of } responses to the } list. } } My question today has to do with the presence of "pepper" } on TEM sections. } We process rather large tissue blocks (4X5 mm) of nervous } tissue that we } have dissected from perfused animals (5% glut. in PB is } routine perfusate). } Post-fixation is in osmium tetroxide in PB and all buffer } rinses are done } in PB. We dehydrate using an ethanol series, transition } through propylene } oxide (although we are experimenting with ethanol with no } succes so far) } and infiltrate/embed in PolyBed 812. The pepper shows up } most obviously } on the cytoskeletal elements of nerves, mitochondria, and } collagen. I have } tried pretreating the sections with 0.5-1.0 N HCl or 1% } EDTA (Mollenhauer, } 1987)prior to staining with uranyl acetate/lead } citrate.....no luck. I } can visualize the pepper on unstained sections. I seem to } recall that this } may have something to do with phosphate buffer, but the } details are hazy. } I would appreciate any suggestions. } } Thank you very much! } } Sandy Perkins } }
Hi Terry: I found scoring wheels available at a local glass supplier. You may have to purchase a glass cutter and then remove the wheel. The wheels I bought had a smaller diameter, but they worked well for many years. The wheel diameter does not affect any calibration since the clamping position and pressure are determined by they weight of the head.
If needed, the axle could be replaced by part way grinding and then breaking a piece from the shaft of a small drill. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Thursday, April 01, 1999 9:51 AM, Terry Robertson [SMTP:terryr-at-cyllene.uwa.edu.au] wrote:
} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } } Does anyone involved in Electron Microscopy know where I } can obtain spare glass scoring wheels for an LKB 2178 } Knife Maker II. Any helpful advise would be welcome as } our machine requires a replacement scorer ASAP. } } Dr Terry Robertson (PhD) } Senior Research Fellow } Department of Pathology } University of Western Australia } Nedlands 6907 } } Phone 618 9346 2935 } Fax 618 9346 2891 } Mobile phone 618 0415 986531 } email terryr-at-cyllene.uwa.edu.au } }
Sandra, Regarding your problem with pepper on sections, I have seen that phenomenon in sections that had not been buffer washed enough after glut. Your blocks are very large and so may require longer and more requent washing.If the glutaraldehyde is not completely washed out it will form a ppt with osmium. You might also check the ph of your buffer. It should be around 7.4 for mammalian tissue.
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
Subject: Polymers SPM -----Original Message-----From: george sibbald {geos-at-goldrush.com} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com} , Barbara Foster {mme-at-map.com} Date: Tuesday, March 30, 1999 6:45 PMSubject: Polymers SPM Barbara Although Molecular Imaging was not at Pittcon, MI also has products for polymer several with significant technical advantage. eg: Temperature, environmental control, MAC Mode, "Pulse Force" and Force spectroscopy. George Sibbald -----Original Message-----From: Barbara Foster {mme-at-map.com} To: Janet H. Woodward {jhwoodward-at-buckman.com} ; microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com} Date: Monday, March 29, 1999 5:37 PMSubject: Re: SPM of Polymers------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.Dear Janet,SPM of Polymers is one of the hottest new trends in microscopy. I am in the middle of writing a review of "What's New in Microscopy at PITTCON" (May, 1999, American Lab) and this topic was well represented by new offerings from several companies. I'd suggest you visit the websites for 1. Digital Instruments - they have lots of great app notes on this topic2. ThermoMicroscopes (previously known as Park Scientific and Topometrix) and look for information on the new Explorer PolymerSystem SPM (integrated microthermal analysis and pulsed force mode imaging)3. JEOL - their new SPM has a full hot/cold stage which runs from 130K to 800K, for watching thermal transitions.Other relevant technology I found at Pittcon included the new thermal stage in Philips' ESEM, the new Continuum microscope for full microscopy imaging (Including DIC) integrated with FTIR from SpectraTech, and a number of interesting systems for Raman/confocal from Renishaw and Instruments SA.Hope this "preview" was helpful.CAVEAT: Neither Barbara Foster nor MME has any commercial interest in any of these instruments.Best regards,Barbara FosterConsortium PresidentMicroscopy/Microscopy Education ...Educating microscopists for greater productivity.125 Paridon Street Suite 102 Springfield, MA 01118PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.comVisit our web site {http://www.MME-Microscopy.com/education} *************************************** ***************MME is America's first national consortium providingcustomized on-site workshops in all areas ofmicroscopy, sample preparation, and image analysis.At 10:29 AM 3/29/99 -0600, Janet H. Woodward wrote: } } } } My company's R&D group is interested in visualizing several of our polymers. The M&M '99 session, "Developments in Scanned Probe Microscopy of Polymers", will be helpful. However, we would like some basic/general information prior to the meeting. Any suggestions of review articles, textbooks, etc. would be greatly appreciated.In advance, thanks!Janet H. Woodward, Ph.D.Corporate Technical Specialist - Microscopy & MicrobiologyBuckman Laboratories1256 N. McLean StreetMemphis, TN 38108(901) 272-6408 {mailto:jhwoodward-at-bbuckman.com} jhwoodward-at-bbuckman.com { { { {
Several years ago I bought some at the local hardware store that fit our LKB 7801A, they were the standard replacements for commercial window glass cutters. If I remember rightly they were only about a dollar each.
Terry Robertson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anyone involved in Electron Microscopy know where I can obtain spare glass scoring wheels for an LKB 2178 Knife Maker II. Any helpful advise would be welcome as our machine requires a replacement scorer ASAP. } } Dr Terry Robertson (PhD) } Senior Research Fellow } Department of Pathology } University of Western Australia } Nedlands 6907 } } Phone 618 9346 2935 } Fax 618 9346 2891 } Mobile phone 618 0415 986531 } email terryr-at-cyllene.uwa.edu.au
--
Richard J. Mount Auditory Science Laboratory, Department of Otolaryngology & Brain and Behaviour Division/Research Institute The Hospital for Sick Children Toronto, Ontario, Canada (416) 813-6551; Fax (416) 813-8456 http://www.sickkids.on.ca/HSCWeb/Otolaryngology/Otoalias/Earhome1.htm
I am looking for a software package or algorithm to produce 3D images from stereo-pair images (LM or SEM). From the 3D images, we want to extract information such as distance, height, angles, volumes, etc. Any suggestions would be warmly welcome!!
My Safety Officer wants all our mechanical pumps vented outdoors. The pumps are Sargent-Welch, Alcatel, and Edwards and are used to back TEMs, film desiccators, evaporators, etc. Initially I thought this was a good idea, having done it simply in past lives by drilling a hole in a wall, ganging the outlet hoses, and presto - out the bad smoke went.
However the B&G engineering consultant got hold of the project (yes now it's a 'project') and now EVERY pump must get its own line up to the roof. This means each pump's discharge is directed to a run of 1-inch copper pipe with three to five 90-degree bends over a total vertical length of about 20'. (Luckily I am on the top floor.) There is also talk about inserting a clean-out or filter at each feed-through for dealing with accumulated oil.
I'm concerned that the pumps are not designed for backpressure on the outlet side coming from the 20' column of air plus the resistence from the 90-degree bends and filter.
Could this setup affect the (1) efficiency or (2) overall life of the pumps? Am I being overly cautious?
I'd appreciate feedback from the List (including manufacturers) re the feasiblity of this approach, and the pump specs - I haven't been able to find anything about discharge 'load' tolerances.
Thanks, you can reply offline and if there is sufficient interest I'll summarize responses for the List.
Ann Hein Lehman EM Facility Mgr Trinity College Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-exchange.cc.trincoll.edu
Postdoctoral Position to Study the Microstructure and Chemistry of Catalyst Materials
The Materials Analysis User Center (MAUC) at the Oak Ridge National Laboratory in Oak Ridge, Tennessee, is seeking a postdoctral candidate to perform research using primarily electron microscopy and surface chemistry to characterize the structure and chemistry of catalysts and other materials. This position will support Department of Energy research programs aimed at reducing both gaseous and particulate emissions from gasoline and diesel engines. The researcher will work with others who are producing and performance testing advanced catalyst formulations. The researcher will be expected to use high resolution electron microscopy and other tools to relate structure at the atomic level to performance. Although a small portion of the research will be proprietary, most will not, and the researcher will be expected to present and publish results. The successful candidate must have a PhD (in materials science, surface chemistry, or a related discipline), experience characterizing catalyst materials using TEM and/or STEM; and should have a strong computer background (the MAUC is entirely digital). The researcher should also have experience with other analytical tools such as FEG-SEM, FEG-SAM, XPS, or electron microprobe, and experience characterizing a wide range of other non-catalyst materials. The researcher will work with many DOE contractor, industrial and university researchers, including some as a part of DOE-ORNL user programs; thus he or she should enjoy collaborating with others. This position is for one year, extendable to two.
ORNL, a multipurpose research laboratory managed by Lockheed Martin Energy Research Corporation for the U.S. Department of Energy, is an equal opportunity employer committed to building and maintaining a diverse work force.
Please send curriculum vitae and bibliography to:
Ted Nolan Manager, Materials Analysis User Center High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 Oak Ridge, TN 37831-6064
Dear Mr. Li, Although I don't have experience etching gold myself, the chemical recommended in the Vander Voort book for gold is aqua regia, a mixture of HNO3 and HCl. Either a 60% HCl, 40% HNO3 or 60 ml. HCl, 20 ml. HNO3, used boiling, is recommended. Always use aqua regia fresh, in a fume jood and never cap it, as it evolves gas. We had a nasty explosion here from sommeone who screwed the cap on a two liter bottle of aqua regia. The MSDS os the separate acids do not prepare one for this hazard. You wrote:
} Hi, } } We are trying to study the microstructure of cold drawn 25 =B5m gold= wires. } We meet the difficulty in etching the gold wires to reveal the grain } structure in SEM and optical observations. Could somebody give us some } suggestions on how to etch the wires. } } Thanks in advance! } } Best regards, } } Kun Li } } Kun Li, Ph. D } Institute of Materials Research and Engineering Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
} I am looking for a software package or algorithm to produce 3D images from } stereo-pair images (LM or SEM). From the 3D images, we want to extract } information such as distance, height, angles, volumes, etc. Any suggestions } would be warmly welcome!! } } James Ma } 3M Building 201-1E-15 } St. Paul, MN 55144 } Some years ago we were interested in doing similar things. You would expect the algorithms to have been all worked out for problems like aerial photography, remote mapping of Mars and Venus, etc. etc., and indeed, for those applications they have been. I worked, with colleagues, with Meemong Lee, then at JPL, to try to apply her programs to our images, but we could not make them work. More recently I discussed this with a friend, Tom Parr, who is a specialist in satellite imagery at The Analytical Services Corporation (TASC) in Reading, Mass. He suggested that the problem arises in extensive assumptions made by the software about the characteristics of the topology under investigation. Planetary surfaces tend to be relatively smooth, and the software has been optimise to take advantage of this characteristic. His thought was that to apply the algorithms to SEM images (in our case, of fracture surfaces in polymers) would require a re-working of the code (by someone who knew what they were doing, of course!!). Needless to say, we didn't try this!
If there is anything avaliable that would help in this problem, we, too, would still be most interested in hearing.
It has been pointed out to me that I phrased my comments regarding the header a bit harshly.
I'm sorry.
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Does anyone have some advice for a novice DIC user. I have an older Leitz Diaplan that is supposed to have DIC, but it doesn't look as good as I think it should.
Can anyone suggest a good test subject for adjusting the DIC and does anyone have some good practical advice/experience in adjusting the DIC system.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I have been trying all week to get in touch with the VCR company. I have three different voice mail messages with three different people there, including Mr. Carlino. Does anybody have any ideal where or what they are doing? I have one of their dimplers and I am trying rather desperately to order the 3i dimpling tool. Any help out there? Anybody by any chance have an extra one of them? I could either purchase it directly from you or when I finally am able to order one from VCR, I can replace it. Any help will be appreciated.
Thank you.
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
Good Afternoon, Just recently, I was asked to manage an imaging core facility which actually needs to be designed and pulled together to include all imaging equipment now in the department with visions of future acquisitions. At this point in time, the major challenge is getting service for a Zeiss TEM 10C, vintage 1986 which has an X-Ray analyzer attached and a JEOL 35C SEM. Neither of these instruments is under contract. There is also a Phillips 201C, now housed in a different building, which is under contract and has been since its purchase in 1974. It is in mint condition, but because it is no longer very productive, the powers that be want to drop the contract. My question to you is, do any of you have independent service engineers servicing your scopes whom you would be willing to recommend? Or do you know of any such engineers whom I might contact? I will be most grateful for any information you might be able to provide.
Sincerely, Sandra Zane Sandra F. Zane, EM Tech. sfzane-at-email.uncc.edu Dept. of Biology, UNC-Charlotte Ph.(704)547-4051 9201 University City Blvd. Fax (704)547-3128 Charlotte, NC 28223
Our facility is going to be replacing an aging Ilford print processor soon. I have been looking at different company's products and am interested in hearing from anyone on the list who would have experience with Colex B&W print processors. In particular, I am looking at the Colette Pro model. I am not familar with this company. I am also considering the Ilford 2650.
If anyone has experience, good or bad, I would like to hear from you.
Thank you,
Jean Ross Central Microscopy Research Facility University of Iowa 85 EMRB Iowa City, IA 42242 319-335-8142 Fax 319-335-6710
We produce an image processing system called "analySIS", and one of the available modules (called "Stereo") provides the measurements you mention in your email. It also does anaglyphic images, surface rendering (heightmapped, textures, illumination), has a very fast computation algorithm, and can perform the calculations with sub-pixel accuracy.
If you need more information, please visit our website at:
http://www.soft-imaging.com
or contact me through email.
Thanks.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
} ---------- } From: } "jma2-at-mmm.com"-at-sparc5.microscopy.com[SMTP:"jma2-at-mmm.com"-at-sparc5.micros } copy.com] } Sent: Thursday, April 01, 1999 8:15 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Stereoscopy software } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } } } I am looking for a software package or algorithm to produce 3D images } from } stereo-pair images (LM or SEM). From the 3D images, we want to extract } information such as distance, height, angles, volumes, etc. Any } suggestions } would be warmly welcome!! } } James Ma } 3M Building 201-1E-15 } St. Paul, MN 55144 } } }
Hello, My first response would be to query whether the safety office really knows what he desires to vent with this very expensive venting project. Usually the most noxious part of a roughing pump exhaust is the particulate or oil mist. If you are not pumping something dangerous like a poison gas or something equally dreadful, I see no reason to go to the expense. The exhaust filters which have been designed for use on your pumps usually do a fine job at trapping any problem vapors. There should be no problem with back pressure. If someone finds me in error, I'm sure we will hear about it.
Good luck.
Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. Number 499, Post Office Box 19400 Austin, Texas 78760 Phone: 512/282-5507 FAX 512/280-0702
An alternate source for your electron microscope maintenance needs.-----Original Message----- } From: Lehman, Ann {Ann.Lehman-at-exchange.cc.trincoll.edu} To: Microscopy-at-Sparc5.Microscopy.Com {Microscopy-at-Sparc5.Microscopy.Com}
I would say you have nothing to fear. Typically, a roughing pump should be fitted with an oil mist filter. The back pressure generated by the filter would "swamp" any from the piping you described. My answer is predicated on pumping down a reasonably sized, sealed vacuum chamber with a rotorary vane pump or similar. After the initial few gulps of gas, the volume of gas moved (at STP) is very tiny. If your vacuum chamber is the size of a small room and the initial pumpdown is by something like a roots blower, that is different... {g}
Dear Ann, I can sympathize with your problems. I run a plastic sump pump hose from each of my rotary pumps out to the window. If you have ever watched an untrapped rotary pump, you will notice that there is almost no air movement out of the pump after the initial ten seconds of pumping. There should be no problem with long runs, bends, etc. if there is nothing but simple atmospheric pressure to oppose the air movement. A simple drain at the first bend up should drain any condensed oil, or a wad of steel wool in the line. Use as large a diameter pipe as you can get them to run. One solution I've seen is to put a commercial car oil filter onto the pump exhaust. If this didn't bother it, neither should your copper line. You wrote:
} Dear Listers, } } My Safety Officer wants all our mechanical pumps vented outdoors. The pumps } are Sargent-Welch, Alcatel, and Edwards and are used to back TEMs, film } desiccators, evaporators, etc. Initially I thought this was a good idea, } having done it simply in past lives by drilling a hole in a wall, ganging } the outlet hoses, and presto - out the bad smoke went. } } However the B&G engineering consultant got hold of the project (yes now it's } a 'project') and now EVERY pump must get its own line up to the roof. This } means each pump's discharge is directed to a run of 1-inch copper pipe with } three to five 90-degree bends over a total vertical length of about 20'. } (Luckily I am on the top floor.) There is also talk about inserting a } clean-out or filter at each feed-through for dealing with accumulated oil. } } I'm concerned that the pumps are not designed for backpressure on the outlet } side coming from the 20' column of air plus the resistence from the } 90-degree bends and filter. } } Could this setup affect the (1) efficiency or (2) overall life of the pumps? } Am I being overly cautious? } } I'd appreciate feedback from the List (including manufacturers) re the } feasiblity of this approach, and the pump specs - I haven't been able to } find anything about discharge 'load' tolerances. } } Thanks, you can reply offline and if there is sufficient interest I'll } summarize responses for the List. } } Ann Hein Lehman } EM Facility Mgr } Trinity College } Hartford, CT 06106
Best of luck! Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
To: Janet Woodward Quesant also did not exhibit its range of AFMs at Pittcon, but would like you to be aware of the fact that polymer imaging can also be done with ambient AFMs, such as ours, costing much less than the more specialized systems. Much depends on the specifics of your application. Good luck, Thomas Pelnar Quesant Instrument Corporation
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Our Coolwell "SE-Style" Chiller hooked to our TEM is on the fritz. An answering machine at the Coolwell phone number says they went out of business and refers one to Litron (or Lintron?). A call to them reveals that all they bought from Coolwell was their name and "marketing strategy". Apparently the marketing strategy is to not produce replacement parts for Coolwell chillers. So we are on our own. Does anyone have a wiring diagram, schematics, specs (such as type and amount of coolant), and/or advice for a SE Style Chiller they could share with me? Our campus refrigeration guy thinks he can fix it but he would like some help on the unit design.
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
I am a technical recruiter, and I am looking for help with a job opening
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I applaud the decision to vent the exhaust. I know of no EPA or other regulation to do so, but the exhaust of mechanical pumps on initial pumpdown is something I have always recommended my customers be rid of. I hope that you are also taking measures to move the pumps into separate rooms or acoustic enclosures to reduce their noise output.
Working with EMs often means spending many hours at a time living with these problems. While occasional exposure can be acceptable, the constant drone of pumps and exposure to pump oil vapors can only be detrimental.
The setup you detail presents no problem to operation of the pumps. Air is very compressable and the volume of air present in the exhaust lines means that there will be only a negligible pressure increase due to the various bends.
A clean-out filter is unnecessary. Proper design of the exhaust stack should provide for a low point where accumulated oil can be either be drained or returned to the pump. This requires less than 90 degree bends in the stack so that any condensed oil can flow back to the pump or a drain. If the exhaust provides for a return to the pump, you have to be aware that there might also be other condensed materials, primarily water, included. Best bet would be to have a slightly greater than 90 degree bend close to the pump followed by a vertical section forming a trap. At that second bend, a drain should be included that would allow for the drainage of all fluids trapped there. Any such scheme would of course require the regular emptying of the trap.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com
-----Original Message----- } From: Lehman, Ann {Ann.Lehman-at-exchange.cc.trincoll.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
VayTek (http://www.vaytek.com ) has been working on something like this. I know of someone who has done it on an SGI computer but I don't know if they have commercialized it yet - I'll try to contact them anad see.
Bill Miller
At 12:58 PM 4/1/99 -0500, Tony Garratt-Reed wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A Jeol JEM-4000EX high-resolution TEM in my former colleague's lab needs a TV rate imaging or digital imaging camera system. Your information about suitable types of the camera system and related manufacturers or vendors is highly appreciated.
The names of the manufactures or vendors that provide diamond saw or wiresaw for cross-sectional sample preparation are also needed. Thank you in advance!
Cheers,
Jian-Guo Zheng
Dr. Jian-Guo Zheng Materials Science and Engineering Department of Engineering The University of Liverpool Ashton Street Liverpool, L69 3GH United Kingdom Telephone: +44-151-794-4671(office)/5382(lab) Fax: +44-151-794-4675 Email: J.G.Zheng-at-Liverpool.ac.uk
The steps for setting up "good" DIC are simple: 1. Start with setting up Koehler on a well-behaved sample (a cheek cell smear is great for this; make sure that you have the correct thickness coverslip - a number 1 1/2 or 0.17mm thick -- not diameter) 2. If you have separate polarizer and analyzer, cross them so that you get the blackest background possible 3. Insert the DIC prism and tune so that the background is soft gray. This should give you the best 3D impression and contrast.
I wrote a very extensive article on DIC which appeared in the April 1988 (!) issue of American Lab and will send you a Xeroxed copy. There is also a detailed discussion in "Optimizing Light Microscopy for Biological and Clinical Laboratories". Details are available on our website (see signature bar).
Hope this is helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 10:58 AM 4/1/99 -0800, Jon Krupp wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Terry, } We purchased a new scoring wheel about a year ago from Reichert-Jung (at } the time; heaven knows who owns anybody anymore) for our Knifemaker II } which, I believe is the Reichert update(?) of your LKB Knifemaker. Check } your Reichert-Jung reps for details but I think the parts are all the } same, though the part numbers have changed (probably to protect the } innocent). The details are -- Part 16 706 699 - Scoring Wheel Assembly } Complete. It was simple to install and now works as it did "in the } beginning." Good luck. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Winston W. Wiggins, Supervisor 01 Apr1999 4:50 PM } CRC-Electron Microscopy Lab Ofc: 704-355-1267 } Carolinas Medical Center Lab: 704-355-7220 } P.O. Box 32861 Fax: 704-355-7648 } Charlotte, NC 28232-2861 USA Eml: WWiggins-at-Carolinas.org } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } -----Original Message----- } From: Terry Robertson [SMTP:terryr-at-cyllene.uwa.edu.au] } Sent: Wednesday, March 31, 1999 6:51 PM } To: 'Microscopy-request' } Subject: Glass Scoring Wheel for LKB 2178 Knife Maker II } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone involved in Electron Microscopy know where I can obtain spare } glass scoring wheels for an LKB 2178 Knife Maker II. Any helpful advise } would be welcome as our machine requires a replacement scorer ASAP. } } Dr Terry Robertson (PhD) } Senior Research Fellow } Department of Pathology } University of Western Australia } Nedlands 6907 } } Phone 618 9346 2935 } Fax 618 9346 2891 } Mobile phone 618 0415 986531 } email terryr-at-cyllene.uwa.edu.au } }
South Bay Technology produces both wire saws and diamond wheel saws for T= EM cross section preparation (among many other uses). You can see additiona= l information on all of our products on our web site at:
www.southbaytech.com.
You may also have an interest in our new Ion Milling System which feature= s patented, new Low Energy Gun Technology (LEG). Please contact me off-lin= e for more information.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy. Message text written by "Dr J. G. Zheng" } ----------------------------------------------------------------------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Dear All,
A Jeol JEM-4000EX high-resolution TEM in my former colleague's lab needs = a TV rate imaging or digital imaging camera system. Your information about suitable types of the camera system and related manufacturers or vendors = is highly appreciated.
The names of the manufactures or vendors that provide diamond saw or wiresaw for cross-sectional sample preparation are also needed. Thank you=
There is an immediate opening for an experienced Scanning Electron Microscopy (SEM) Research Technician at Exxon Research and Engineering Company's Corporate Research Laboratory in Clinton, New Jersey. The position initially is on a temporary basis driven by an immediate need. Initial duration of the position is 4 to 12 months.
As a member of the Advanced Characterization Group at Corporate Research, this position will involve the operation of the state-of-the-art JEOL FESEM, and JEOL Analytical SEM instruments with associated Energy Dispersive and Wavelength Dispersive Spectrometers. The position will involve the creative application of high resolution SEM imaging and elemental characterization of samples related to a wide range of projects at Exxon's Corporate Research Laboratory. The position will also involve general laboratory operations, sample preparation, SEM maintenance and computer analysis of the SEM data (both image analysis and spectral analysis).
Previous SEM experience and experience with high vacuum systems and computers (DOS, Windows, and Unix) is required. Successful candidates should have experience in chemistry, physics or material science with a Baccalaureate degree or equivalent experience. Since a number of projects are simultaneously in progress, it is essential for the researcher to be very organized and to pay attention to detail and accuracy in reporting results.
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Being the Diaplan Microscope utilized the individual objective prisims it is not a easy task tuning to achieve the preferred DIC image. To do the tunning, you must loosen the three little set screws on the 10mm collar located above the objective. Then with polarizers crossed and the matching condenser prisim in place you should rotate the objective to achieve the three demensional appearance. The best position will allow to have an even flatfield image throughout the entire field. If this does not appear to be the case then you do not have the correct orientations. You can also remove the eyepiect and look down the tube to see an interference patter. Typically when you are aligned the pattern which appears as a black line should be orientated on a forty five degree angle from two oclock to eight oclock. Once you have these prisims orientated be sure to lock the little set screw on the objective. FYI this setup with objective and prisim together will not allow you to use the same optics for typical brightfield applications should that be desired. Usually back then because of the nature of the beast (Diaplan) you had to have a interchangeable nosepiece when going from one illumination method to another. With todays infinity corrected systems and the technological developments in prisims it is easy to use a microsscope with all the latest gizmos and not have to take anything off the unit.
If you would like, I have a old instruction manual called "Interference Contrast T for the Leitz Diaplan". I would be more then happy to copy it sometime this week and send it along.
Let me know... Good Luck!
Also, a good sample to use is a section of unstained tissue about five microns or less. If you have access to someone working in a clinical pathology lab you may want to ask them.. to get this for you..I may have one or two laying around. If I do I will send it along with the manual.
C. Passione -----Original Message----- } From: Jon Krupp {jmkrupp-at-cats.ucsc.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
You should not have to go through all this trouble! Leitz/Leica have a great reputation for quality and those prisms should have been aligned "at the factory".
The tuning I was referring to was the compensator which adjusts the "pol color" in the background.
Also, I am surprised to hear that you could not pull out the prism/compensator from the back focal plane of the objective and move the condenser to the brightfield setting to re-establish good imaging for brightfield.
Jan Hinsch, if you are listening .... any comments?
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 07:21 AM 4/3/99 -0500, Cono Passione wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I beleive the previous Leica/Leitz microscopes of the fixed focal length vintage almost alway utilized individual prisims that were permanent unless you unscrewed the objective then physically remove the 10mm prisim. You then would be able to put the objective back in place on the nosepiec to utilize its BF capabilities. To return to the DIC mode you would have to put the DIC prisim back in place and realign over again. The prisim in place trying to do BF techniques will be distorted and very difficult to resolve anything. This is from past experience. If someone else has developed a system to remove the prisims on a slider type set the Diaplan or laborlux series I would be intereset in knowing more about it. I also beleive the older Zeiss mciroscopes utilized an individual prisim that was pused in and out on a forty five degree angle right above the objective being used at any particular time. Some of these prisims were capable of matching up with Leitz objectives with various combinations.
Thank You
C. Passione -----Original Message----- } From: Barbara Foster {mme-at-map.com} To: Cono Passione {iami-at-nauticom.net} ; Microscopy-at-Sparc5.Microscopy.Com {Microscopy-at-Sparc5.Microscopy.Com} ; Jon Krupp {jmkrupp-at-cats.ucsc.edu}
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Kun Li wrote the following: =================================================== We are trying to study the microstructure of cold drawn 25 µm gold wires. We meet the difficulty in etching the gold wires to reveal the grain structure in SEM and optical observations. Could somebody give us some suggestions on how to etch the wires. =================================================== While many laboratories have the capability, very few encounter the kinds of samples that would actually benefit from the application of "sputter etching". This is the process of reversing the polarity of a normal gold (sputter) coater, or having it operate in "etch" mode, whereby instead of sputtering gold onto the sample, you are sputtering (in this case, gold) off of the sample. Assuming you are working with polished cross-sections (but the outside surface of the wires should be able to be studied this way as well), you should be able to end up revealing details of the microstructure.
We have not done this in-house ourselves but we have been told by others they use this approach, for example, on gold alloys used in dentistry.
The second approach is to do this by reactive plasma etching. It is a different physics so the effect should not be expected to be the same. Again we have not done this ourselves in-house, but we have customers who claim to have done it. I am not real clear what would be the optimum etching gas. We ourselves found that we could eventually remove gold from a gold coated SEM sample using only oxygen (for 20 minutes). Others have used Ar and CF4. I have myself never quite been able to understand why oxygen would remove the gold layer from a gold coated SEM sample.
Is one approach "better" than the other approach? I don't know. What we would predict is that if the two approaches were compared, sputter etching vs. reactive plasma etching, there should be differences. But then again, the physics is different when comparing the two approaches so this should not be surprising.
Finally you might wonder about why chemical etching would not be suggested for this kind of application. I am not sure whether it would or would not work, but when I have heard people discussing doing it, they seem to be referencing some pretty terrible and nasty reagents, the main one being aqua regia. Therefore the interest in alternatives to wet chemical etching.
Disclaimer: SPI Supplies manufactures both sputter coaters with etch and reactive plasma etchers and we therefore have a vested interest in seeing these techniques used more widely. There is nothing "special" about the SPI etch mode, the same physics would be at work in anyone's sputter coater so long as it has the ability to have its polarity reversed and operate in "etch" mode.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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I am looking for a used Zeiss 10C. Please let me know if you have one for sale or if you know of anybody who does. Thank you very much. Peter Jordan, EMSI
I'd like to get from somewhere or somebody a manual on SEM Hitachi S-4100 in English. Copying and shipment charge is definitely to be compensated.
Dmitri. P.S. Please, send me at least some ideas, where to get it from!
__________________________________________ Dmitri V. Sokolov, Doctor Course student Research Center for Interface Quantum Electronics, Hokkaido University, North 13, West 8, Kitaku, Sapporo 060, Hokkaido, Japan Phone 81-11-706-7174 Fax 81-11-716-6004 http://www.geocities.com/SiliconValley/Campus/1314 AOL Instant Messenger FalconDot ICQ 9418072 __________________________________________
As sometimes happens, some people find they can't get money to attend courses at the last minute while others don't hear about them until it is too late to apply.
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Three late cancellations at the UBC Live-Cell course mean three opportunities for those of you who have ten days free this June.
We expect to have an international faculty of 15, and on-site equipment that includes 2-photon systems from Olympus (and maybe Leica). There will also be single-photon confocals from Bio-Rad, Olympus, Noran, Nikon, Zeiss and Yokogawa/EG&G, deconvolution set-ups from API, AutoQuant/Universal-Imaging, Improvision, Intelligent-Imaging and Vaytek all set up for "live-cell" work. In addition, there will be was laser tweezers, microinjection and sundry other delights.
These systems are not just to look at but to use for over 45 hours of organized 2D and 3D live-cell laboratory sessions, plus 20 hours of evening sessions for group live-cell projects.
Although the eight "Groups-of-3" have already been set up and have chosen their "individual projects," we are able to accept 3-2 more students as long as their backgrounds will fit into one of these groups.
which has the rest of the story, including a preliminary program.
Then fill out the Registration Form from the site to tell us about you, Fax it to me ASAP and we will see if we can fit you in. (I will be away the week of April 9-18)
Hope that you can join us in Vancouver this June 16-27.
Jim Pawley, Organizer **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
I am looking for a used carbon coater for our SEM teaching lab. If you have one for sale or if you know of anybody who does, please let me know. Thanks!
Mei-Zhen Dang Research Associate Department of Physics University of Ottawa
We use the Hitachi S4100 on many occasions during our "in house" courses.= =
We do not have the Hitachi manual but we have developed a "quick guide" t= o the instrument for our clients, that I can e-mail as an attachment if yo= u wish? The trainers also use a mechanical alignment guide that we produce= , that is also available to you if you wish?
Sorry I cannot act today but with ester and all that lawns to cut etc.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
This young man has contacted me twice in regards to interviewing a person involved in nanotechnology. If you can help please respond to him directly with the answers to the questions at the bottom of the page.
Mr. Nano {nanoteknologist-at-hotmail.com}
Thanks
Susanne Pignolet Brandom MicroWorld 978-456-3100
} Hi, my name is Michael Lee, and I am a high school student attending } Bellevue High. I am doing a project about a career in nanotechnology. } As part of the project, I need to interview someone with general } questions about a career in this field. I was wondering if you would be } able to help me by letting me interview you, or could refer me to } someone who I would be able to interview. If I would be } able to interview you, please let me know what type of contact would be } most convenient for you (email, phone, etc). I would really appreciate } it, thank you. } } -Michael Lee
-----Original Message----- } From: Mr. Nano {nanoteknologist-at-hotmail.com} To: spb-at-mwrn.com {spb-at-mwrn.com}
} ---------- } From: Earnhart, James P. } Sent: Monday, April 05, 1999 7:25 AM } To: Ingber, Bruce F. } Subject: RE: Mech Pump Discharge Backpressure } } Only to say that if the pumps were designed to be able to } operate in the described manner then the manufacturers would have specs on } how and what type of ventilation system to install under different } applications, i.e. extraction type if there is more than 10' of pipe to } outside, or more than 2 bends totaling more than 90 degrees. Also, if } discharging the "bad air" outside then EPA standards must be met...such as } installing "scrubber" systems to insure no oil or certain hydrocarbons are } discharged into the air and eventually being introduced into the ground } water. Best thing is to install a factory offered oil recovery exhaust } filter system such as the one we spoke about on the coating system. And } to check with OSHA as to the "hazards Imposed" by the Vacuum Pump vapors. } If they say they are too hazardous for normal laboratory environments then } we may explore the possibility of routing the exhaust into existing "Fume } Hoods" in the laboratories. That is assuming they have the necessary } environmental "cleaning" systems built into them. } } Bottom line is I doubt that without a lot of engineering the } pumps will be able to be operated properly with the exhaust "routed" } through any type of piping more than a few feet long. Meaning that if you } hook any lines, more than a few feet long without bends or any that have } more than 90 degrees of bends, to remove the fumes to any of the pumps, } they won't work properly without a lot of engineering to eliminate any } "back pressure" that may be caused. } } ---------- } From: Ingber, Bruce F. } Sent: Thursday, April 01, 1999 3:43 PM } To: Earnhart, James P., Electronic Technician } Subject: FW: Mech Pump Discharge Backpressure } } Any ideas? } Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124-4305
} ---------- } From: Lehman, } Ann[SMTP:Ann.Lehman-at-exchange.cc.trincoll.edu] } Sent: Thursday, April 01, 1999 9:56 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Mech Pump Discharge Backpressure } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } Dear Listers, } } My Safety Officer wants all our mechanical pumps } vented outdoors. The pumps } are Sargent-Welch, Alcatel, and Edwards and are used } to back TEMs, film } desiccators, evaporators, etc. Initially I thought } this was a good idea, } having done it simply in past lives by drilling a } hole in a wall, ganging } the outlet hoses, and presto - out the bad smoke } went. } } However the B&G engineering consultant got hold of } the project (yes now it's } a 'project') and now EVERY pump must get its own } line up to the roof. This } means each pump's discharge is directed to a run of } 1-inch copper pipe with } three to five 90-degree bends over a total vertical } length of about 20'. } (Luckily I am on the top floor.) There is also talk } about inserting a } clean-out or filter at each feed-through for dealing } with accumulated oil. } } I'm concerned that the pumps are not designed for } backpressure on the outlet } side coming from the 20' column of air plus the } resistence from the } 90-degree bends and filter. } } Could this setup affect the (1) efficiency or (2) } overall life of the pumps? } Am I being overly cautious? } } I'd appreciate feedback from the List (including } manufacturers) re the } feasiblity of this approach, and the pump specs - I } haven't been able to } find anything about discharge 'load' tolerances. } } Thanks, you can reply offline and if there is } sufficient interest I'll } summarize responses for the List. } } Ann Hein Lehman } EM Facility Mgr } Trinity College } Hartford, CT 06106 } v. 860-297-4289 } f. 860-297-2538 } e. ann.lehman-at-exchange.cc.trincoll.edu } } }
} I applaud the decision to vent the exhaust... } While occasional exposure can be acceptable, the constant drone } of pumps and exposure to pump oil vapors can only be detrimental. } Not to mention that droplets of condensed oil vapor could contami- nate the specimens. Yours, Bill Tivol
One of our students has been given the use of a Model ZF Coulter counter but we cannot find the conversion chart which we have been told is necessary to have, to convert the machine readings to a cell count. Does anyone on the list have this same machine and could you perhaps FAX the chart to us at 617-638-5591? We are located in Boston, MA. Are instructions for this device hard to come by?
* Subject: Wanted Leitz Parts...... * Date: Mon, 05 Apr 1999 09:34:37 -0400 * From: Joseph Passero {jp-at-spacelab.net} * To: Microscopy-at-Sparc5.Microscopy.Com * * I am looking for the following Leitz parts for use with the Zernike * 402a phase contrast condenser. * * An auxiliary focusing (magnifier) with the knurled ring for focusing on * the phase ring, Leitz part number 513 123 * * A pair of centering key for the annular stops of the condenser. * * PHACO objectives * * If you have these or any other Leitz items you would like to * sell, please tell me what you have with a price. * * Thank You * Joseph Passero * jp-at-spacelab.net
-----Original Message----- } From: Ingber, Bruce F. {bingber-at-commserver.srrc.usda.gov}
} } Only to say that if the pumps were designed to be able to } } operate in the described manner then the manufacturers would have specs on } } how and what type of ventilation system to install under different } } applications, i.e. extraction type if there is more than 10' of pipe to } } outside, or more than 2 bends totaling more than 90 degrees. Also, if } } discharging the "bad air" outside then EPA standards must be met...such as } } installing "scrubber" systems to insure no oil or certain hydrocarbons are } } discharged into the air and eventually being introduced into the ground } } water. Best thing is to install a factory offered oil recovery exhaust } } filter system such as the one we spoke about on the coating system. And } } to check with OSHA as to the "hazards Imposed" by the Vacuum Pump vapors. } } If they say they are too hazardous for normal laboratory environments then } } we may explore the possibility of routing the exhaust into existing "Fume } } Hoods" in the laboratories. That is assuming they have the necessary } } environmental "cleaning" systems built into them.
If you are worried about back pressure you can put a fan in the lines to put negitive pressure at the outlet of the vacuum pumps.
Gordon
Gordon Couger gcouger-at-couger.com www.couger.com/gcouger Stillwater, OK 405 624-2855 GMT -6:00
A few months ago I posted a request for a used SEM to purchase for a customer of mine and I was grateful for the range of replies I received. We managed to find an instrument that I felt was a perfect fit for their needs and that was within their budget, while the university that was selling it got the price they wanted and a quick sale. Thanks to all who helped along the way.
Well, I've got another request from our neighbors south of the border. A pathology lab is being setup from scratch, and one of the requirements is for an SEM, no mention of an EDS or WDS system. While they would like a later model instrument, their needs are for a basic SEM. Any and all replies are welcome, please email the specifics to me, I'll compile them and present the responsible parties with the data. While I'm not likely to travel again to Mexico to install and service an SEM (did it once before - long trip, language problems), I may come and deinstall the instrument.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com
Thanks to those who responded with information on upgrading the computer on our BioRad Confocal 600 with COMOS
Our new computer is 8gig HD with CD, zip, internet card and all the bells and whistles. Apparently there is a limitation with the old program having trouble with hard drives that are too big, so we partitioned ours. Otherwise, it appears to have no trouble accessing any of the drives. We have a Panasonic WORM, that we are setting up on the old computer as a separate station - along with a 5 1/4, 3 1/2, and Zip so that whatever walks in the door can be transferred to a more recent fad of data storage. We try to keep a lot of older equipment/computer technology around and working so others can retrieve their data after they've suddenly been aroused from dormancy. e.g. we had a researcher come in with data stored on a WORM drive (she was working with the government) and we were the only ones she found(she states)with one still in service. Similar story with the 5.25 drive.
Now, refurbishing the detector head and laser.... Oh boy, I can hardly wait.
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu ********************************************
DearListers, I have a GATAN (model#613) single tilt , LN cooled side entry cold stage for a Phillips 300 or 400 TEM available. I'm inquiring to see if there is any interest in purchasing it? Rosemary
Hi everyone, I'm training to be a tech here in SEM and Confocal microscopy. I am in need of a method to clean a leaf from an ocean plant to then be viewed with SEM. Any help for this rookie would be greatly appreicated. Thanks Lee Ann Baldridge SEM &Confocal Facility IUSD Indpl., IN lhadley-at-iusd.iupui.edu 317-274-5142
Our lab is looking to buy a belt grinder for heavy grinding of mounted cross-sections (metallographic cross-sections). The one quote I got was for over $5,000 for a single belt with coolant system (1/3 hp motor). The dual belt system was almost $9,000. This seems quite pricey for what you get. Is there any other options available to us?
Mark
---------------------------------------- Mark C. Biesinger, Research Scientist Surface Science Western The University of Western Ontario London, Ontario, Canada Tel: (519) 661-2173, Fax: (519) 661-3709 http://www.uwo.ca/ssw/
Members of the list: I seem to remember that a company existed that made disposable microtome knives. The knives were about the size of a regular knife, only thinner. The company seemed to go by something like "American Scientific Products." Is this company still in business, or are there satisfactory substitutes for students doing basic slide making?
George W. Smith
********************************* George W. Smith, Ph.D. Associate Professor of Biology Department of Biological Sciences Union College Schenectady, NY 12308 (518)388-6245 smithg-at-union.edu *********************************
Dear Users, Someone sent me a message asking for help using Molecular Probes Alexa dye reagents. I no longer have this message as our email file server crashed before I could respond and several days messages were lost. Recovery of damaged disk is underway. Would the person who asked about this please re-send their message. However, I would like to know more of the specifics of your problem. We have had no problem substituting these reagents for like reagents in a variety of cell type systems and tissues?
Images & Info at http://www.molbio.princeton.edu/confocal {http://www.molbio.princeton.edu/confocal}
Joe Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University jgoodhouse-at-molbio.princeton.edu {mailto:Jgoodhouse-at-molbio.princeton.edu}
I believe the item you are looking for was made by Scienctific Products; they were item # M7200 and came 12 to the box. A quick scan of the catalog didn't show them anymore, but maybe they are still around. I have several old (from the '70's) boxes of of them you are welcome to have if you wish. For my Histology class, I use instead special holders for single edge razor blades; cheap, easy, no resharpening and most importantly, very short exposed edge to prevent bloody knuckles. I'm not sure where they come from, but if you are interested I will try to find the source.
For a project on cartilage regeneration in dogs, I would like to be able to determine the origin of the new cartilage cells. I know that one way would be a classic pulse-chase experiment with tritiated thymidine to label dividing cells, but I'm wondering whether there might non-radioactive methods that could tell me the same thing. I'd prefer not to have to inject radioactive solutions (even tritium) into these animals. Any ideas?
Gary Radice 804-289-8107 Department of Biology 804-289-8233 (FAX) University of Richmond gradice-at-richmond.edu Richmond VA 23173
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After reading the excellent ideas already presented about this subject, it occurred to me that there may be one other method that would work. First, it is no longer necessary to use things like aqua regia to polish gold. My paper in Ultamicroscopy 25 (1988) 351-354 describes an electropolish for gold using certain chemicals dissolved in methanol and butyl cellosolve at -50 degrees C. It is often possible to develop a nice etch by simply using an electropolishing solution at a lower voltage. In this case I would suggest dropping the voltage from 150 to perhaps 70 volts. A temperature of 0 C. may also be cold enough. The good thing about this is that by controlling the time the current is "on", the amount of etching can be controlled. The electrolyte does not attack gold when the current is off. This solution is easy to rinse off the specimen.
Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439
for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 7 Apr 1999 07:49:02 -0700 (PDT) X-SMTP: helo gaugler.calweb.com from gaugler-at-calweb.com server -at-gaugler.calweb.com ip 207.173.132.32 user=Pgauglr Message-Id: {4.1.19990407073131.009d1100-at-pop.calweb.com} X-Sender: gaugler-at-pop.calweb.com X-Mailer: QUALCOMM Windows Eudora Pro Version 4.1
I would appreciate hearing from and about independent companies or individuals who take down, move and set up SEMs. Specifically, I would like to receive quotes for moving an Amray 1830 from TX to Sacramento CA. This would involve disconnecting the system, locking down the turbo pump, crating, moving, and re-install at my location.
I have a quote from Amray but they do not handle moving. I am wondering if this is standard practice or if there are reputable folks who can handle the whole job from start to finish via one contact. I would anticipate that someone who knows about this specific SEM model would be preferred.
Anticipated timeframe for start of project is in about 1-2 months from now.
Dear George, Several suppliers of disposable microtome knives come to mind:
Electron Microscopy Sciences 321 Morris Road Ft. Washington, PA 19034 1-(800) 523-5874 (web site address: http://www.emsdiasum.com.)
C.L. Sturkey, Inc. 824 Cumberland Street Lebanon, PA 17042 1-(800) 274-9446 FAX (717) 274-9442
Sturkey offers free samples of their knives. Our experience in recent years has been with Sturkey knives, but other brands may be prefered depending on your application. Best regards, Henry **************************************************************************** Disclaimer: I have no vested interest in the firms mentioned above. ****************************************************************************
Henry Eichelberger, EM Facility Manager Department of Biological Sciences Binghamton, University
Try C.L. Sturkey... They should be on the net......
C YA -----Original Message----- } From: George Smith {smithg-at-union.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
I would just like to support and reinforce some of the many comments recently made about getting copies of older instruction manuals. It would be a real service to our community if a master compilation of these manuals, with a suitable index and regular updates, could be put together. I wish that I had the time and the resources to volunteer for this task but I don't at the moment. Hope somebody else does.
I am trying to lift the carbon coating off a metallic film. The C is less than 10nm thick. My best results so far involved aqua regia vapors, then dipping the sample in DI water, and finally picking up the C on grids as it floats on the water. However, one of the components of the metal film under the carbon is Pt, and a large number of small Pt particles stay on the carbon. Could someone recommend a Pt etch?
} Job Description } There is a TEM microscopist position open for a materials scientist at } Intel Malaysia. The primary job responsibility is to provide technical } consultation and leadership in applying materials/surface analysis & } microscopy techniques to solve day-to-day engineering and manufacturing } problems, including operation of a JEOL 2010F TEM. This individual is also } expected to develop and proliferate materials & surface analysis } capabilites within Intel Malaysia; to recommend and set up new analytical } techniques; and to coach & mentor junior-level failure analysis engineers. } } Requirements } Candidates should have a Ph.D. in Materials Science, Surface Science, } Physics, Chemistry, Thin Film Engineering or equivalent. Successful } candidates will have a strong background in microelectronic materials & } process engineering (silicon and/or packaging materials). You will have } studied and applied advanced bulk, thin film and surface characterization } techniques intensively to problems relevant to microelectronics } processing, packaging and/or failure analysis during your graduate } education and/or research/industrial experience. You should have extensive } hands-on TEM experience. Knowledge & hands-on experience in applying XPS } & TOF-SIMS in analyzing organic contaminants would be an added advantage. } } You should be highly motivated, self-directed, effective working } independently or in a team. Good problem solving skills, interpersonal, } verbal & written communication skills are necessary. Must be able to } impart your knowledge/skill to junior-level engineers. } } Interested individuals please contact: } Kian Sin Sim } Intel Technology Sdn. Bhd. } 11900 Penang, Malaysia. } Tel: ++ 604-859-6477 } Fax: ++ 604-859-6749 } kian.sin.sim-at-intel.com
Hi, We need to replace the stereo scope on a Reichert-Jung Ultracut E (purchased in 1986, Type 70-17-04, Fabr NR 396313). Something is messed up in the lens system making it impossible to focus. We sent it to Leica to be repaired and they couldn't fix it. I'd like to hear from you if you have a stereo scope for sale that would work on this microtome. The stereo scope label says: Stereo Star ZOOM Reichert 0.7X TO 4.2X 570
thanks, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
We are a training company that has been operating for the past 18 years teaching SEM, TEM and EDX in many of the English speaking countries of th= e world. As part of our "in house" course procedures we have produced our own 1 to 4 page instruction sheets to help our clients. These "quick guides" now number well in excess of 100, even with some of them covering= a range of microscopes with almost identical operating panels. This means = we have data going back to instruments produced in the late 80's which, provided the demand is not too excessive, we are pleased to offer via attachments to e-mail. We also have basic maintenance procedures for man= y models.
Hope this may help those who are in trouble with the older instrumentatio= n?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
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Hi,
can somebody help me with a protocol for embedding plant material in LR white. The material is fixed in PFA and kept in 70% EtOH at -20 C.
Bo
-- Dr. Bo Johansen Associate Professor Botanical Institute University of Copenhagen Gothersgade 140 DK-1123 Copenhagen K Denmark
Is there a quantitative method for determining the metal grain size when metal shadowing a biological TEM specimen?
Shadowing is new to me and I'd like to find the best conditions for TaW shadowing in our shadowing unit. To that end, I was wondering if there was something more quantitative than eyeballing TEM images of the shadowed samples?
Hello everyone. I would like to know if there is a quick write-up on the = web that explains how to prepare thin sections of rocks and sand for = microscopy. I need to know specifically what type of epoxy is used to = affix the specimen to the microscope slide. Thanks.
______________________ Roberto Garcia Senior Analyst, Metallography North Carolina State University Analytical Instrumentation Facility Box 7531, Room 303 EGRC Raleigh, NC 27695-7531 rgarcia-at-unity.ncsu.com http://spm.aif.ncsu.edu/aif
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD} {META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {STYLE} {/STYLE}
{META content=3D'"MSHTML 5.00.0910.1309"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT size=3D2} Hello everyone. I would like to know if there is a = quick=20 write-up on the web that explains how to prepare thin sections of rocks = and sand=20 for microscopy. I need to know specifically what type of epoxy is used = to affix=20 the specimen to the microscope slide. Thanks. {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D3} ______________________ {BR} Roberto Garcia {BR} Senior = Analyst,=20 Metallography {BR} North Carolina State University {BR} Analytical = Instrumentation=20 Facility {BR} Box 7531, Room 303 EGRC {BR} Raleigh, NC 27695-7531 {BR} {A=20 href=3D"mailto:rgarcia-at-unity.ncsu.com"} rgarcia-at-unity.ncsu.com {/A} {BR} {A=20 href=3D"http://spm.aif.ncsu.edu/aif"} http://spm.aif.ncsu.edu/aif {/A} {/FON= T} {/DIV} {/BODY} {/HTML}
I've called their number (973) 882 6611 to confirm, and it appears true that Coolwell has folded up. However, one can still leave a voice mail message at that number for Frank Haze, and presumably he can be helpful.
We have an early analog SE-111A, which we have substantially modified as various parts have malfunctioned or worn out. Despite its variable reliability, in my opinion, it's still the best chiller design out there, and our unit easily keeps the STEM coolant temperature within 0.10 of 700F.
I have the operating manual as well as the "pilot circuit" and the "power circuit" diagrams; however, Coolwell has refused to release the refrigeration system diagram, treating it as a trade secret. I will be glad to scan these and e-mail them to any interested party. Please, direct your requests to:
rwafu-at-bsco.com Valdemar Furdanowicz Bethlehem Steel Research Labs G-165 Bethlehem, PA 18016
-----Original Message----- } From: Bob Wise [mailto:wise-at-vaxa.cis.uwosh.edu] Sent: Thursday, April 01, 1999 6:35 PM To: Microscopy-at-sparc5.microscopy.com
To all,
Our Coolwell "SE-Style" Chiller hooked to our TEM is on the fritz. An answering machine at the Coolwell phone number says they went out of business and refers one to Litron (or Lintron?). A call to them reveals that all they bought from Coolwell was their name and "marketing strategy". Apparently the marketing strategy is to not produce replacement parts for Coolwell chillers. So we are on our own. Does anyone have a wiring diagram, schematics, specs (such as type and amount of coolant), and/or advice for a SE Style Chiller they could share with me? Our campus refrigeration guy thinks he can fix it but he would like some help on the unit design.
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
} From Microscopy-request-at-sparc5.microscopy.com Thu Apr 8 13:57:42 1999 } } From: "Roberto Garcia" {rgarcia-at-unity.ncsu.edu} } To: "MSA Microscopy" {Microscopy-at-Sparc5.Microscopy.Com} } Subject: Geologic Thin Specimen } Date: Thu, 8 Apr 1999 11:10:01 -0400 } } } Hello everyone. I would like to know if there is a quick write-up on the } web that explains how to prepare thin sections of rocks and sand for = } microscopy. I need to know specifically what type of epoxy is used to = } affix the specimen to the microscope slide. Thanks. } } ______________________ } Roberto Garcia } Senior Analyst, Metallography } North Carolina State University } Analytical Instrumentation Facility } Box 7531, Room 303 EGRC } Raleigh, NC 27695-7531 } rgarcia-at-unity.ncsu.com } http://spm.aif.ncsu.edu/aif } } Robert, We have had good success with Epotek 301 (Epoxy Technology, Inc. 978-667-3805) for both thin sections and thick sections (slabs) for cathodoluminescence studies. With cathodoluminescence it is important that the epoxy can stand up reasonably well under the electron beam bombardment if it is hit directly (e.g., in a void) and also that it not outgas excessively and harm the vacuum.
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology "A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
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We would like to label artificial Xenopus oocyte nuclei after sticking them on glass. So far, we tried to centrifuge the nuclei to a glass cover slip, fix it with formaldehyde, briefly extract with triton, label them and flat embed in Epon. The problem is they do not stick very well. So far, we tested naked glass, polylysine coated glass and carbon coated/glow-discharged glass. After the centrifugation, there is usually plenty of nuclei, but with the processing, we are loosing many if not all.
Any suggestions would be greatly appreciated.
Regards,
-- Michal Jarnik, Ph.D. Fox Chase Cancer Center Electron Microscopy Facility 7701 Burholme Ave. Philadelphia PA 19111 Tel. 215-728-5675 Fax 215-728-2412
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} We would like to label artificial Xenopus oocyte nuclei after sticking } them on glass. So far, we tried to centrifuge the nuclei to a glass } cover slip, fix it with formaldehyde, briefly extract with triton, label } them and flat embed in Epon. The problem is they do not stick very well. } So far, we tested naked glass, polylysine coated glass and carbon } coated/glow-discharged glass. After the centrifugation, there is usually } plenty of nuclei, but with the processing, we are loosing many if not } all. } } Any suggestions would be greatly appreciated.
You might try "silane", 3-aminopropyltriethoxysilane to be precise. Get it from Sigma (cat. no. A-3648). 1. Wash slides thoroughly with hot, soapy water. Rinse very well, final 2 rinses with distilled. water. 2. Dry in an oven. 3. Dip slides in 2% silane in actone for 10 sec., then 2 rinses of acetone, 1 min. each. 4. Air dry in a vertical position.
You might also take a look at Stain Technology 62: 27-33 and 93-99, 1987. Two very interesting papers by S. Fink.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
Just testing yet another filter I've configured. This one is supposed reject all Email which has attachments. This is the most common way that virus's are transmitted by Email as attached documents.
Of course, NONE of you attach documents to postings on the Microscopy Listserver right? After all that is part of our rules.
Remember VCF cards that some WWW browsers attach to your Email are also attachments so you had better make sure your browser is configured so as not to attach those annoyances.
} Mime-Version: 1.0 } Date: Thu, 8 Apr 1999 17:59:16 -0600 } To: microscopy-at-Sparc5.Microscopy.Com } From: "Nestor J. Zaluzec" {zaluzec-at-Sparc5.Microscopy.Com} } Subject: Administrivia: Testing Virus Filter } } Colleagues.... } } Just testing yet another filter I've configured. This one is supposed } reject all Email which has attachments. This is the most } common way that virus's are transmitted by Email as attached } documents. } } Of course, NONE of you attach documents to postings on the } Microscopy Listserver right? After all that is part of our } rules. } } Remember VCF cards that some WWW browsers attach to } your Email are also attachments so you had better make } sure your browser is configured so as not to attach those } annoyances. } } } Nestor } } }
Just testing yet another filter I've configured. This one is supposed reject all Email which has attachments. This is the most common way that virus's are transmitted by Email as attached documents.
Of course, NONE of you attach documents to postings on the Microscopy Listserver right? After all that is part of our rules.
Remember VCF cards that some WWW browsers attach to your Email are also attachments so you had better make sure your browser is configured so as not to attach those annoyances.
It's taken me a while, but I think that I now have a running version of the Email attachment filter I was working on last week, a few of you will recall a batch of rejected mail messages on 3/30. which was part of my early testing.
Local testing is now completed and I have put this filter on-line as of ~ 7 pm CST on the Full Microscopy Listserver. Please bear with me as I'm sure there will be a few glitches along the way. Especially the first few days of full operation.
This filter scans each Email message for an attachment. If an attachment is found (and I can't guarantee the filter will catch all of them) then the Email posting will be REJECTED and an explainatory message is sent to the poster. Attachments are one of the most common way that computer virus's are transmitted by Email especially they are embedded in various documents. Removing the attachment will generally allow your message through, unless of course your Email triggers yet another different "warning flag".
Of course, NONE of you attach documents to postings on the Microscopy Listserver right? After all that is part of our rules which you all have received on subscription and have read completely. The only attachments we will see are from JUNK mailers...right?
Just a final warning, you should all appreciate the fact that VCF cards that some WWW browsers "append" to Email messages are also "attachments". The filter does not differentiate attachments. So I run the risk of getting a handful of you annoyed at me. Obviously the simple solution is for those of you who may be affected to reconfigure your browser/mailer not to send VCF cards. (Netscape is particuliarly bad in this regard) so WWW users beware!
In the long run, I think it is better to protect the larger group of you and catch the grief I will get from those few who don't realize that VCF cards are attachments and consider my rejection of their mail an afront on their "virtual" personality. .......
Cheers.... Nestor Your Friendly Neighborhood SysOp.
----------------------------------------
PS. Just for those of you that are curious the filter has logged 255 messages as potential SPAM/JUNK mail since it was started August of 98.
In other words about 8 message per week trigger the filter. About 3/4 of those caught are true JUNK mail. It also inadvertently catches a few valid postings which after the author contacts me as per the instructions the messages are allowed through, albeit a day or so later.
Right...... time for a cold beer and some dinner. G'night all
Ritchie Sims wrote: } } } Date: Wed, 07 Apr 1999 15:54:35 -0700 } Hi, Earl } } Just off the top of your head, how much would you guess that a } competent person might be able to disassemble a JEOL 840 in a US } city, and crate it suitably for international despatch? } } Just to help my budgeting } } cheers } } Ritchie } } } Hi all, } } } } There are a number of qualified technicians, some of whom have worked } } for Amray that can handle this type of job. The Amray 1830 is a } } relatively simple SEM requiring about 8 hours maximum to dis-assemble } } and about 16 hours to re-assemble. I don't recommend crating the system. } } Instead, we usually ship via "padded air-ride van". Shipping costs are } } charged according to weight and distance. Maximum costs for shipping and } } SEM coast to coast has been about $3,000.00. Shipping costs from Texas } } to Calfornia should be considerably less. } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
Hi Ritchie,
A Jeol 840 has quite a bit more cables. It would take a good tech about 12 hours (max) to properly dis-assemble and then another four hours to supervise the crating. The costs for crating is about $800.00 USD. The labor for 16 hours is about $2000.00 USD plus travel time & travel expenses.
Actually, a rather complicated subject. In regards to your overall question, anyone is capable of arranging the shipping. That is a simple matter. Many independents, however, may be reluctant to accept the responsibility. If something goes wrong, you might end up with multiple lawsuits - you sue the contractor, who sues the shipper, who counter-sues the contractor, etc. It generally is easier for an single independent to contract for the de-installation and re-installation without regard to the shipping. In that case, you have a, hopefully, expert and unbiased third opinion as to the condition of the instrument both before and after the actual shipping. In court, that third opinion may well carry the day as definitive.
If the independent has arranged for the shipping, they will be seen as having a biased interest in the overall operation. The lines between the preparation for freight, the delivery of freight and the receiving of freight will be blurred and any court may appropriately spread the blame for a problem between the independent and the freight company. In other words, there will be no way to definitivily identify the source of a problem. In the worst case, a court may not be able to assign blame to any identifiable source.
An elecron microscope is a sensitive instrument, subject to many mechnical and electronic variables. A major move of an instrument like this should involve an objective assestment of the instrumental operation before the move and an objective assesment of the instrumental operation after the move. Any problems can then be identified to the source.
In all honesty, I have yet to see any problem in moving any SEM. However, considering the potential losses involved in moving a recent model SEM, these problems should be kept in mind.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com
-----Original Message----- } From: Dr. Gary Gaugler {gaugler-at-calweb.com} To: MSA listserver {Microscopy-at-sparc5.microscopy.com}
a database containing user manuals for a range of older popular microscopes would be a very good idea, especially since the purchase of a second hand microscope is usualy the only way for an amateur microscopist to get hold of a decent microscope at a (more or less) reasonable price. That's why I have put some Reichert Zetopan manuals on my website.
In the best case scenario this service should be easy accesible and free if at all possible. If that isn't possible a small fee could be asked to cover the costs.
But it isn't simple:
I don't think that the large manufacturers would like to support this idea, as it isn't in their short-term intrests to support users of second-hand equipment, they rather like to sell new gear (one example: I know from a very reliable German source, that Zeiss has, after the "wende", destroyed large stocks of spare parts for older "Eastern-German Zeiss manufactureres"). I think they see it the wrong way: I can't imagine much amateurs who spend the exorbitant prices the large manufactureres ask for their products, at least I wouldn't...
And, unfortunally, as far as I know, the manufacturers are the owners of the copyrights of their manuals, so we're stuck here, that would be the first problem to be solved...
After that: finding someone to coordinate the project, finding the manuals and scan those (can't be much of a problem I suppose), finding a server to host the documents and finding some people to do investigations regarding brands, models, serial numbers... to match manuals and models/versions...
I would like to volunteer for such a project...
Hello Royal microscopical society (England), Microscopy Society of America, German microscopy clubs, Micscape Magazine, manufacturerers...?
Yvan Lindekens.
-----Oorspronkelijk bericht----- Van: donald j marshall {dmrelion-at-world.std.com} Aan: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} Datum: donderdag 8 april 1999 1:26 Onderwerp: manuals
} I would just like to support and reinforce some of the many comments } recently made about getting copies of older instruction manuals. It would be } a real service to our community if a master compilation of these manuals, } with a suitable index and regular updates, could be put together. I wish } that I had the time and the resources to volunteer for this task but I } don't at the moment. Hope somebody else does. } } Don Marshall } } }
We're a film production company and need to shoot some footage through a stereo microscope -- stereoscopically. Would someone be so kind as to point us to a source for a scope to which *two* videocameras could be attached, one for the right field and one for the left? Please reply to JPWELT-at-aol.com.
Hi Bo, We have for sometime been using LRW for Immuno-staining for plant tissue. We have based our protocol on that of Kathryn A. Vandenbosch and a good reference is in Chapter 5 Immunogold Labelling in Electron Microscopy of Plant Cells eds.J.L.Hall & C.Hawes,Academic Press 1991. Our protocol uses medium grade LRW, it is often benificial after fixation to add some Ruthenium Red to the buffer wash to stain the tissue as later its refractive index will be the same as the resin and specimens are easily lost. AS you specimens are in 70% Alc already dissolve the stain in water and use that to dilute 100% Alc to 70%Alc to stain the tissue. If you have any problems obtaining the reference i will send you our protocol.
I would suggest using a pulse of BrdU. We use it routinely as a marker for cell proliferation. It is a non-radioactive thymidine analog. We use Dako's anti Brdu and DAB as the chromagen. Feel free to call me for specifics.
-- Begin original message --
} For a project on cartilage regeneration in dogs, I would like to be able to } determine the origin of the new cartilage cells. I know that one way would } be a classic pulse-chase experiment with tritiated thymidine to label } dividing cells, but I'm wondering whether there might non-radioactive } methods that could tell me the same thing. I'd prefer not to have to inject } radioactive solutions (even tritium) into these animals. Any ideas? } } Gary Radice 804-289-8107 } Department of Biology 804-289-8233 (FAX) } University of Richmond gradice-at-richmond.edu } Richmond VA 23173 -- End original message --
best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**Suppose you were an idiot... And suppose you were a member of Congress ... But I repeat myself.-Mark Twain**
Just take the Stereo microscope. and have two exact cameras, and two exact camera to microscope eyepiece adapters.
Ed Sharpe
}
Hello -
We're a film production company and need to shoot some footage through a stereo microscope -- stereoscopically. Would someone be so kind as to point us to a source for a scope to which *two* videocameras could be attached, one for the right field and one for the left? Please reply to JPWELT-at-aol.com.
I do not know of any write ups on the web for preparation of thin sections but Allied High Tech (I work for Allied High Tech) has a quick curing epoxy (5-10 min.) called Epoxy Bond 110. We also have extensive experience preparing thin sections, samples for Metallography, SEM and TEM samples. I am located in Raleigh and would like to work with you and help you develop your applications.
We also have a precision polisher, the MultiPrep system. It has a vertical spindle that prevents unwanted faceting. The spindle is calibrated perpendicular to the abrasive and the specimen is calibrated parallel to the abrasive. If a specific angle is required it can be set too. The MultiPrep system also allows you to monitor the amount of material that has been removed from the specimen in real time. The system is ideal for parallel polishing, preparation of thin section, SEM and TEM specimens.
If you would like additional information about the MultiPrep system or any of Allied's products please contact me off-line. I may be reached via email or at the phone number listed below. All of Allied's products are also on our web site. The address is also below.
Regards,
Ed Hirsch
At 11:10 AM 4/8/99 -0400, Roberto Garcia wrote:
} } } }
{excerpt} Hello everyone. I would like to know if there is a quick write-up on the web that explains how to prepare thin sections of rocks and sand for microscopy. I need to know specifically what type of epoxy is used to affix the specimen to the microscope slide. Thanks.
Hello I am looking for a provider of single Crystal cristal viewing screens ( ma= de of YAG), that are used as a pick up screens in image capture system to TEM. any suggestion are welcome thanks =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D Fernando D. Balducci Laboratorio de Microscopia Electr=F3nica Facultad de Ingenier=EDa - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
Ingber, Bruce F. wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } ---------- } } From: Earnhart, James P. } } Sent: Monday, April 05, 1999 7:25 AM } } To: Ingber, Bruce F. } } Subject: RE: Mech Pump Discharge Backpressure } } } } Only to say that if the pumps were designed to be able to } } operate in the described manner then the manufacturers would have specs on } } how and what type of ventilation system to install under different } } applications, i.e. extraction type if there is more than 10' of pipe to } } outside, or more than 2 bends totaling more than 90 degrees. Also, if } } discharging the "bad air" outside then EPA standards must be met...such as } } installing "scrubber" systems to insure no oil or certain hydrocarbons are } } discharged into the air and eventually being introduced into the ground } } water. Best thing is to install a factory offered oil recovery exhaust } } filter system such as the one we spoke about on the coating system. And } } to check with OSHA as to the "hazards Imposed" by the Vacuum Pump vapors. } } If they say they are too hazardous for normal laboratory environments then } } we may explore the possibility of routing the exhaust into existing "Fume } } Hoods" in the laboratories. That is assuming they have the necessary } } environmental "cleaning" systems built into them. } } } } Bottom line is I doubt that without a lot of engineering the } } pumps will be able to be operated properly with the exhaust "routed" } } through any type of piping more than a few feet long. Meaning that if you } } hook any lines, more than a few feet long without bends or any that have } } more than 90 degrees of bends, to remove the fumes to any of the pumps, } } they won't work properly without a lot of engineering to eliminate any } } "back pressure" that may be caused. } } } } ---------- } } From: Ingber, Bruce F. } } Sent: Thursday, April 01, 1999 3:43 PM } } To: Earnhart, James P., Electronic Technician } } Subject: FW: Mech Pump Discharge Backpressure } } } } Any ideas? } } } Bruce F. Ingber } Biologist- Electron Microscopy } USDA-ARS, SRRC } 1100 Robert E. Lee Blvd. } New Orleans, LA 70124-4305 } } (504) 286-4270; fax (504) 286-4419 } bingber-at-nola.srrc.usda.gov } } } ---------- } } From: Lehman, } } Ann[SMTP:Ann.Lehman-at-exchange.cc.trincoll.edu] } } Sent: Thursday, April 01, 1999 9:56 AM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Mech Pump Discharge Backpressure } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } Dear Listers, } } } } My Safety Officer wants all our mechanical pumps } } vented outdoors. The pumps } } are Sargent-Welch, Alcatel, and Edwards and are used } } to back TEMs, film } } desiccators, evaporators, etc. Initially I thought } } this was a good idea, } } having done it simply in past lives by drilling a } } hole in a wall, ganging } } the outlet hoses, and presto - out the bad smoke } } went. } } } } However the B&G engineering consultant got hold of } } the project (yes now it's } } a 'project') and now EVERY pump must get its own } } line up to the roof. This } } means each pump's discharge is directed to a run of } } 1-inch copper pipe with } } three to five 90-degree bends over a total vertical } } length of about 20'. } } (Luckily I am on the top floor.) There is also talk } } about inserting a } } clean-out or filter at each feed-through for dealing } } with accumulated oil. } } } } I'm concerned that the pumps are not designed for } } backpressure on the outlet } } side coming from the 20' column of air plus the } } resistence from the } } 90-degree bends and filter. } } } } Could this setup affect the (1) efficiency or (2) } } overall life of the pumps? } } Am I being overly cautious? } } } } I'd appreciate feedback from the List (including } } manufacturers) re the } } feasiblity of this approach, and the pump specs - I } } haven't been able to } } find anything about discharge 'load' tolerances. } } } } Thanks, you can reply offline and if there is } } sufficient interest I'll } } summarize responses for the List. } } } } Ann Hein Lehman } } EM Facility Mgr } } Trinity College } } Hartford, CT 06106 } } v. 860-297-4289 } } f. 860-297-2538 } } e. ann.lehman-at-exchange.cc.trincoll.edu } } } } } }
Bruce, Unless your lab is at a partial vacuum and you are venting your pump to atmosphere, backpressure is a moot point. The gas flow is miniscule except for initial roughing.
Runs, elbows, diameters, restrictions are all critical on the INTAKE side of the pump, but have no significant effect on the outlet side of the pump.
We used LR White in order to embedd immobilised root protoplasts in alginate beads.
After dehydration try 96% EtOH 2x10 min, 100% EtOH 2x10 min and infiltrate with LR White, 3x1 hr at room temperature, transfer to gelatine capsules. Polymerization takes place at 60 C, 48 hr.
Gary.
On Thu, 8 Apr 1999, Bo Johansen wrote:
} Hi, } } can somebody help me with a protocol for embedding plant material in LR } white. The material is fixed in PFA and kept in 70% EtOH at -20 C. } } Bo } } -- } Dr. Bo Johansen } Associate Professor } Botanical Institute } University of Copenhagen } Gothersgade 140 } DK-1123 Copenhagen K } Denmark } } Tlf: + 45 35 32 21 57 } email: boj-at-bot.ku.dk } http://www.bot.ku.dk/staff/boj.htm
Dear Sir/Madame, Could you please tell me the going rate of using a TEM (dollars/hour) with and without a technician? Thank you in advance, Toni Milroy
I have two unrelated projects with a common element - I need to be able to get particles dispersed nicely across an SEM stub (and one of them on a TEM grid, as well). One is biological, the other material.
Tiny particles tend to clump as the solution they are in dries. In one case a client is trying to look at fungal spores, and in another someone is trying to look at tiny metal needles. I would love to hear all your expert suggestions, since this comes up often!
Snow in San Jose? It's in the low 80s F, sunny, a bit windy here on Oahu.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Here is a possible cheap digital solution for you. I think that you will have to go through the eyepieces to get stereo. The following message was from a rep about a digital camera that could be inserted in the eyepiece. Two might give you what you want. Call and find out. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
--------message follows-----
Dear Mr. Walck:
Thank you for your interest in the Microrder Electric Eyepiece.
To use the Microrder our customers just take out their normal optical eyepiece and drop in our Electric Eyepiece which fits into any DIN standard 23 mm tube. The Microrder connects to any PC to allow for live previews, capturing, storing, editing, and emailing of still images or movies. An adapter is available that allows the Microrder to fit in a 30.5 eyepiece such as that normally found with an inspection microscope.
Features of the Microrder Electric Eyepiece:
-704 x 556, 24 bit color digital camera for $279US (mail-in rebate of $50) -included software that lets you preview, capture, email, and crop and edit you image or video in real-time on your computer -1 micron per pixel resolution with a 10X objective lens -the ability to image the central half of the field of view that you would see in your normal optical eyepiece -the ability to save audio with your images and even send AudioCards which are a still image with an audio track attached -inplace activation compatibility so that you can operate our software and Electric Eyepiece inside of most desktop software applications like Word, Excel and Powerpoint -connects to standard parallel port -powered by keyboard or mouse connector -30 day money back guarantee -1 year warranty -free camera driver upgrades, monthly newsletter and special discounts for Microrder owners during sale events
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Phone: (650) 299-9794 Fax: (650) 366-7760 Email: info-at-imicrovision.com URL: http://www.imicrovision.com Digital Microscopy Solutions featuring the Microrder Electric Eyepiece
---------- } From: "ICEMANCINE-at-aol.com"-at-Sparc5.Microscopy.Com To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hello -
We're a film production company and need to shoot some footage through a stereo microscope -- stereoscopically. Would someone be so kind as to point us to a source for a scope to which *two* videocameras could be attached, one for the right field and one for the left? Please reply to JPWELT-at-aol.com.
Dear Tina, } } Tiny particles tend to clump as the solution they are in dries.
If the solution wets the particles, that will certainly be true; however, if not, and the particles are attracted to the grid/formvar/ carbon, etc., then the solution should not drag them together. The solution to the solution is to find one which has the properties needed; for the metal particles, try to make the grid hydrophyllic (assuming the needles are wet by water) and use a nonpolar solvent such as petro- leum ether, and if the fungal spores have a static charge, the same may work for them. If the fungal spores are hydrophobic, try a bare formvar coat and ethanol (C-coat afterwards). You may have to fiddle with the grid-solvent combinations a lot, so good luck.
} In one } case a client is trying to look at fungal spores, and in another someone } is trying to look at tiny metal needles. I would love to hear all your } expert suggestions, since this comes up often! } } Snow in San Jose? It's in the low 80s F, sunny, a bit windy here on Oahu. } We are actually having springlike weather in Albany (~60 F, no snow) in spite of the dire predictions based on La Nina. Yours, Bill
Fernando D. Balducci wrote: ============================================== I am looking for a provider of single Crystal cristal viewing screens ( made of YAG), that are used as a pick up screens in image capture system to TEM. any suggestion are welcome =================================================== SPI Supplies has offered YAG screens for some number of years, in different dimensions and different thicknesses. Quite a bit of technical and other information about these screens is available on our website URL http://www.2spi.com/catalog/scintill/spi-tem-yag.html
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Dear Robert, } } Two related items cause me to respond to the list. } 1. How do we know what we see through the microscope is real? } 2. Alan Sokal and the "Science Wars." } } There is a group of social scientists who are known as social } constructionists. Of this group there are those known as } deconstructionists. And of this group there are those who are engaged in } the "Science Wars". These individuals more or less advocate that what } scientists do is a construction of their mind and not necessarily an } expression of nature. Some continue to argue that the actions or } constructions of scientists is done in part to get money to play around } in the lab. Some of these people ask that federal funding be curtailed } for these constructions of people called scientists. In some way the } arguments remind one of vegans attacking medical research. } They are partially correct--the half-truth is that everything we do is filtered through our individual perceptions--but they leave out the part of science which insists that all discoveries must be reproducible by an independent observer. This limits science in a way that art, for example, is not, but it gives the scientific method more power and generality than the other "theories" which the deconstructionists compare it to. It is clear to any experimental scientist that scientific observations are, at best (which is how *we* do them), approximations to reality, and that theories are also approximations, since only limited evidence is available to test them. It is true that without funding no scientist can function for very long, so it follows that part of the motivation for our activities is to obtain funding. However, each of us can say with confidence that science is a unified body of knowledge, even though much of that knowledge eludes us, and that the entirety of scien- tific knowledge has benefitted mankind far more than the cost to produce it (can the deconstructionists make the same claim?).
} Alan Sokal wrote an article call Transgressing the Boundaries:... and it } was accepted and published in the journal Social Text. This journal with } a circulation of about 800 is a leading journal for social } constructionists and a place where debates about the science wars have } been taking place. Sokal, an avowed leftist physicist, wrote } Transgressing as a hoax and immediately proclaimed so when the Social } Text article was published. His purpose was to show that the Science } Wars advocates were on thin intellectual grounds. You may wish to read } several recent articles of Academe (an AAUP publication) that tries to } put this all into focus. } } So back to the microscope. Those of us who do microscopy know that much } of what we look at is a construction. The tissue is dead, chemically } altered, stained, dehydrated, infiltrated, and sectioned into to little } pieces... AT BEST. Clearly we are constructing what we hope is a correct } interpretation of nature. Akin to walking through a graveyard and trying } to guess what really happened in the living lives of the people under the } tombstones.
Archeologists, in fact do essentially this. There is nothing in- herently wrong about making inferrences about reality based only on a severely modified part of that reality. As long as we realize the con- sequences of the modifications and do not over-interpret our results, we will be on reasonable safe ground. However phenomena such as the micro- trabecular network, which was shown to be an artefact, should serve to remind us of the pitfalls waiting for the unwary.
} We also know in the best sense of Popper that we are self } doubting and trying to better (disprove) much of what was published } before. These social constructionists do not seem to understand any of } this. Just like I don't understand why people watch soap operas on } daytime television. } Can one get funding by watching soap operas?
} So the question put forth in the original post is a very important one. } How do we tell a public what we see through the microscope is real?
We don't. We tell them that we see a more-or-less reasonable representation of reality which we continually strive to understand and improve, and we tell them that there are limitations to how close any particular method can come to reality (this is why some of us do cryo- EM on unstained material, which also has its limitations).
} It } is a kind of "Daddy, why is the sky blue?" question. As microscopists we } have a responsibility to address the question and help the public } understand what we see is "real" and represents nature.
The "" around real are well deserved. Only by being aware of the representational and limited aspects of science will we really help the public understand what we do.
} And yes it is a } construction of sorts but that is what science is all about: Humans' } feeble attempt to reconstruct the beauty of nature... but not in the } sense of the deconstructionists. } Agreed.
} I recognize this is an unusual thread for this microscopy list, but this } is an important issue because it can dramatically affect the funding that } many of us share. } Not to mention the little detail that without serious considera- tion of the nature of the scientific process, we will not get far in our search for the nature of reality. Yours, Bill Tivol
I was searching the Internet to find some information about STM (Scanning Tunneling Microscopy). This is about to write down a technical paper covering the physic apsects involved in STM.
Anyone could send me good links? Any link / good information in spanish?
{ {Tiny particles tend to clump as the solution they are in dries.} }
I find that a drop of poly-L-lysine in the suspension works wonders for most particle types and allows for a quite even distribution on filmed TEM grids. I have used it mostly on non-biological samples.
I use the standard 0.1% solution offered by EM supply companies. I add one drop of that (from a pasteur pipette) to 1 ml of suspension. It spreads well on all TEM grid films. Would be worth trying on SEM mount surfaces.
Am interested in hearing how it works.
Ted Dunn Maui, Hawaii
The surf is up and it is so beautiful here just now - Spring skies and brisk trade winds.
We are trying to prepare SrTiO3 (001) single crystal TEM samples by chemical means. Does anyone know a good solution to use?
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Generally the customer wants one source for responsibility. The questions about objectivity by using a third party only complicates the matter. When I was a customer using and maintaining SEMs and other equipment, we subcontracted different portions of the job to the lowest quailified bidder. Thereafter, no one person assummed responsibility for the entire job and when things went wrong all the subcontractors pointed "their collective fingers" at each other and ultimately at us (the customer) for not co-ordinating things properly.
When we move an SEM we use subcontractors that we have experience with for crating and moving. We don't have a vested interest in using these subcontractors (commissions or kickbacks) other than the fact that we have used them before and trust them.
I also take pictures of the SEM and have the movers watch me take these pictures. I FEDEX the pictures to the customer so their receiving department has a record of the condition of the SEM when it arrives.
In the eighteen years we have service SEMs and moved them I have had only one problem: moving a JEOL IC 845. It was stored in a warehouse for one week before shipping. Unfortunately, in transporting the SEM around the warehouse, the movers dropped the column. Questions arose about the SEM being packed properly. The insurance company (Yes I always insure the shipment) tried to "relieve themselves of responsibility". The insurance company said that the shipment was not secured properly as Scanservice did not have the experience to pack the equipment. Of course the insurance company had no idea what was required to pack an SEM, much less what an SEM is. I made one phone call to the moving company (United Van Lines) and assured them that if they wanted our continued business they would re-imburse us for the SEM. Within one day, upper management at United called and assured me that they would pay any and all damages no matter what the insurance company stated. Within one week, we were re-imbursed for our services, the customer was fully re-imbursed and all are happy.
I continue to do business with United even though they are at times slightly higher.
I shutter to think what would happen if we had several other parties involved in dis-assembly crating, moving, uncrating, and re-assembly.
The SEM is a comlicated piece of equipment but after 25 years experience it complexity seems much more trivial. Any system is the sum of it's components. When I first repaired SEMs, they were considered very "Hi tech". The "Hi tech" machines of today will soon be mundane. Look at computers. We are not reluctant to assume total responsibility for the SEM move.
This is not to say that you are wrong. May independents may or may not be willing to accept full responsibility for the move. We will as long as the customer understands that he needs to use sub-contractors that we recommend.
Earl Weltmer Scanservice Corporation
Allen R. Sampson wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Actually, a rather complicated subject. In regards to your overall } question, anyone is capable of arranging the shipping. That is a simple } matter. Many independents, however, may be reluctant to accept the } responsibility. If something goes wrong, you might end up with multiple } lawsuits - you sue the contractor, who sues the shipper, who counter-sues } the contractor, etc. It generally is easier for an single independent to } contract for the de-installation and re-installation without regard to the } shipping. In that case, you have a, hopefully, expert and unbiased third } opinion as to the condition of the instrument both before and after the } actual shipping. In court, that third opinion may well carry the day as } definitive. } } If the independent has arranged for the shipping, they will be seen as } having a biased interest in the overall operation. The lines between the } preparation for freight, the delivery of freight and the receiving of } freight will be blurred and any court may appropriately spread the blame for } a problem between the independent and the freight company. In other words, } there will be no way to definitivily identify the source of a problem. In } the worst case, a court may not be able to assign blame to any identifiable } source. } } An elecron microscope is a sensitive instrument, subject to many mechnical } and electronic variables. A major move of an instrument like this should } involve an objective assestment of the instrumental operation before the } move and an objective assesment of the instrumental operation after the } move. Any problems can then be identified to the source. } } In all honesty, I have yet to see any problem in moving any SEM. However, } considering the potential losses involved in moving a recent model SEM, } these problems should be kept in mind. } } Allen R. Sampson } Advanced Research Systems } 317 North 4th. Street } St. Charles, IL 60174 } PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com } } -----Original Message----- } } From: Dr. Gary Gaugler {gaugler-at-calweb.com} } To: MSA listserver {Microscopy-at-sparc5.microscopy.com} } Date: Wednesday, April 07, 1999 9:31 AM } Subject: SEM moving } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I would appreciate hearing from and about independent companies } } or individuals who take down, move and set up SEMs. Specifically, } } I would like to receive quotes for moving an Amray 1830 from TX } } to Sacramento CA. This would involve disconnecting the system, } } locking down the turbo pump, crating, moving, and re-install at my } } location. } } } } I have a quote from Amray but they do not handle moving. I am } } wondering if this is standard practice or if there are reputable folks } } who can handle the whole job from start to finish via one contact. } } I would anticipate that someone who knows about this specific SEM } } model would be preferred. } } } } Anticipated timeframe for start of project is in about 1-2 months } } from now. } } } } gary g. } } } } my fax is 916.791.8186 } } telco is 916.791.8191 } } } } } }
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Dear Robert, } } Two related items cause me to respond to the list. } 1. How do we know what we see through the microscope is real? } 2. Alan Sokal and the "Science Wars." } } There is a group of social scientists who are known as social } constructionists. Of this group there are those known as } deconstructionists. And of this group there are those who are engaged in } the "Science Wars". These individuals more or less advocate that what } scientists do is a construction of their mind and not necessarily an } expression of nature. Some continue to argue that the actions or } constructions of scientists is done in part to get money to play around } in the lab. Some of these people ask that federal funding be curtailed } for these constructions of people called scientists. In some way the } arguments remind one of vegans attacking medical research. } They are partially correct--the half-truth is that everything we do is filtered through our individual perceptions--but they leave out the part of science which insists that all discoveries must be reproducible by an independent observer. This limits science in a way that art, for example, is not, but it gives the scientific method more power and generality than the other "theories" which the deconstructionists compare it to. It is clear to any experimental scientist that scientific observations are, at best (which is how *we* do them), approximations to reality, and that theories are also approximations, since only limited evidence is available to test them. It is true that without funding no scientist can function for very long, so it follows that part of the motivation for our activities is to obtain funding. However, each of us can say with confidence that science is a unified body of knowledge, even though much of that knowledge eludes us, and that the entirety of scien- tific knowledge has benefitted mankind far more than the cost to produce it (can the deconstructionists make the same claim?).
} Alan Sokal wrote an article call Transgressing the Boundaries:... and it } was accepted and published in the journal Social Text. This journal with } a circulation of about 800 is a leading journal for social } constructionists and a place where debates about the science wars have } been taking place. Sokal, an avowed leftist physicist, wrote } Transgressing as a hoax and immediately proclaimed so when the Social } Text article was published. His purpose was to show that the Science } Wars advocates were on thin intellectual grounds. You may wish to read } several recent articles of Academe (an AAUP publication) that tries to } put this all into focus. } } So back to the microscope. Those of us who do microscopy know that much } of what we look at is a construction. The tissue is dead, chemically } altered, stained, dehydrated, infiltrated, and sectioned into to little } pieces... AT BEST. Clearly we are constructing what we hope is a correct } interpretation of nature. Akin to walking through a graveyard and trying } to guess what really happened in the living lives of the people under the } tombstones.
Archeologists, in fact do essentially this. There is nothing in- herently wrong about making inferrences about reality based only on a severely modified part of that reality. As long as we realize the con- sequences of the modifications and do not over-interpret our results, we will be on reasonable safe ground. However phenomena such as the micro- trabecular network, which was shown to be an artefact, should serve to remind us of the pitfalls waiting for the unwary.
} We also know in the best sense of Popper that we are self } doubting and trying to better (disprove) much of what was published } before. These social constructionists do not seem to understand any of } this. Just like I don't understand why people watch soap operas on } daytime television. } Can one get funding by watching soap operas?
} So the question put forth in the original post is a very important one. } How do we tell a public what we see through the microscope is real?
We don't. We tell them that we see a more-or-less reasonable representation of reality which we continually strive to understand and improve, and we tell them that there are limitations to how close any particular method can come to reality (this is why some of us do cryo- EM on unstained material, which also has its limitations).
} It } is a kind of "Daddy, why is the sky blue?" question. As microscopists we } have a responsibility to address the question and help the public } understand what we see is "real" and represents nature.
The "" around real are well deserved. Only by being aware of the representational and limited aspects of science will we really help the public understand what we do.
} And yes it is a } construction of sorts but that is what science is all about: Humans' } feeble attempt to reconstruct the beauty of nature... but not in the } sense of the deconstructionists. } Agreed.
} I recognize this is an unusual thread for this microscopy list, but this } is an important issue because it can dramatically affect the funding that } many of us share. } Not to mention the little detail that without serious considera- tion of the nature of the scientific process, we will not get far in our search for the nature of reality. Yours, Bill Tivol
Funny how we were just talking about vacuum pump oil vapours and how to deal with them... Last Tuesday I had a guy in the lab to change the locks (long, unrelated story), including the lock on the door right behind our ESEM. Our vacuum lines pass through a cut-out on this door to the pumps located in a big warehouse-like area. We have the usual little filters on the pumps, but as we all know, you can usually smell a little something in the air anywhere near them. While changing the lock, the guy was standing very close to the pumps for perhaps 10 minutes or so, and said he could smell something "oily". Well, I just found out that later that day, he became quite ill, with apparently a toxic reaction to a substance or substances unknown, and was hospitalized for a few hours, though he later, apparently, recovered completely. We're not absolutely sure that breathing our oil vapours for a few minutes is actually what caused the reaction, but it appears to be the only possible toxin the man was in contact with that day ( or at least that's the story). It's possible that the gentleman just had an unusual sensitivity to the vapours; I know at times I've had lots more exposure than he did, and have personally never gotten so much as a headache. One thing bothers me a bit, though; we use Alcatel 102, an oil specially made for ESEM's, with their increased water throughput, and I wonder if this stuff might be a bit less user-friendly than the more common types. Has anyone else out there ever seen, or heard of, a toxic reaction to this or any other pump oil? (I have a funny feeling that I haven't heard the end of this incident yet...)
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B2Y 4A2
What brands and types of razor blades are used as a disposable microtome knive (paraffin, manual rotary microtome)? What are their limitations regarding section thickness for non-problematic animal and plant tissues?
Alvaro: The links from our site include a section with 11 STM=20 sites. No doubt those sites in turn would have links to=20 most significant STM sites available. Cheers Jim Darley
ProSciTech Microscopy=20 PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, April 10, 1999 10:19 AM, =C1lvaro Hern=E1ndez=20 Tortosa [SMTP:aht-at-mx3.redestb.es] wrote: } } Hi everybody! } } I was searching the Internet to find some information } about STM } (Scanning Tunneling Microscopy). This is about to write } down a technical } paper covering the physic apsects involved in STM. } } Anyone could send me good links? Any link / good } information in spanish? } } Thank you, } } Alvaro. }
In a message dated 4/10/99 6:43:54 PM EST, gbza40-at-udcf.gla.ac.uk writes:
{ { Not to mention the little detail that without serious consideration of the nature of the scientific process, we will not get far in our search for the nature of reality. Yours, Bill Tivol } }
I thoroughly enjoyed your posting. Thanks.
The word REALITYcan be interchanged with TRUTH. I don't believe that in a wider sense the scientific process necessarily has anything to do with one's ability to know Truth.
This is my first posting to this listserver. I am in position to purchase a Jeol 840A sem in good condition equipped with a Kevex 8000 EDS detector and need to know its approximate value on the used market. Responses can be made directly to me or via the listserver.
The ion pump on our JEOL 2010 (LaB6) has just about died and we were wondering what sort of lifetimes people were getting for pumps on similar (JEOL 2000 series) machines. Our samples, though biological are not changed often and even so, with the configuration of the machine ie differential pumping etc. we would not expect the ion pumps to have to work terribly hard. We have had the microscope for around 5 years and would like to know if that is about normal for these pumps.
We were also curious as to what people did when the pumps finally fell over. As far as we know there are 3 options - replace with new pumps, replace with reconditioned pumps or replace the filaments in the existing pumps.
What have people found to be the most "cost effective" solution?
Thanks for any help with this.
Colin Veitch
Instrumentation Scientist Fibre Structure & Function Group CSIRO Wool Technology PO Box 21 BELMONT Vic 3216 Australia
Tel: +61 (0)3 5246 4000 Fax: +61 (0)3 5246 4811
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Wool Technology on +61 3 5246 4000.
I was shown a very simple technique for dispersing dry particles on a SEM stub by Dave Gittens in PGT(UK).
Take a sheet of paper & staple it in a funnel shape. Place a small amount of the powder in the funnel & hold the stub ( with adhesive ) at the end. Use compressed air/N2 to propel the powder out of the funnel onto the stub. It may take a bit of trial & error to get the level of dispersion right, but it should give a very even dispersion without many particles touching. Obviously this should be carried out in a fume hood for safety.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Tina Carvalho {tina-at-pbrc.hawaii.edu} To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com}
The lifetime of the ion pump of our Jeol 2000FX was 5 years. Officially, JEOL says that the lifetime of a recondioned pump is three years and a new pump five years. Since the microscope is 10 years old, it was the second ion pump to be changed. We choose to replace the pump by a reconditioned pump. This solution save about 50% of the price of a new pump. In fact, this solution was prefered because the reconditioning consists in changing the storing elements and the result is a new pump. The only risk resides in the possibility of getting a very old pump and in this case some soldering can be defective, but it is very rare. Anyway, the pump is always tested before installation and the risk is quasi null.
Eric LEROY Dr. Laboratoire de Chimie Metallurgique des Terres Rares UPR 209 - CNRS Groupe des Laboratoires de Thiais 2-8, rue Henri Dunant 94320 THIAIS cedex
Hi Colin, Tina, Using a compressed air stream, or any moving air for that matter, is a good way to classify particles. I'm not saying it cannot be done, just that I would be very uneasy about a proper sampling. Dispersing particles is not an easy process, although it can seem that way. It would be nice if there were some simple solution, but unfortunately there isn't. We have found the best results by dispersing in an organic solvent with a dissolved polymer carrier and casting films on either water or cleaved mica. Even this technique has it's drawbacks and limitations. Particle material, density, shape, size, size distribution, surface energy and tribo properties and all have affects on dispersability. In other words the best technique for your particles must be determined by you by trials and more trials. Good luck, Russ Gillmeister, Xerox
-----Original Message----- } From: Colin Reid [mailto:creid-at-tcd.ie] Sent: Monday, April 12, 1999 1:27 AM To: MSA.Microscopy.Com
Hi Tina,
I was shown a very simple technique for dispersing dry particles on a SEM stub by Dave Gittens in PGT(UK).
Take a sheet of paper & staple it in a funnel shape. Place a small amount of the powder in the funnel & hold the stub ( with adhesive ) at the end. Use compressed air/N2 to propel the powder out of the funnel onto the stub. It may take a bit of trial & error to get the level of dispersion right, but it should give a very even dispersion without many particles touching. Obviously this should be carried out in a fume hood for safety.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Tina Carvalho {tina-at-pbrc.hawaii.edu} To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com}
In my former position as the TEM Technical Specialist at NYS College of Ceramics for 6 years, I was the lucky one who had to make the same decision about the sputter ion pump (SIP) on our JEOL 2000FX. The microscope was installed in early 1987 and the SIP died in May 1994. I elected to have it reconditioned which resulted in no problems other than the amount of time it took since we did not have a service contract on the TEM. The reconditioned SIP was working up to the time I left that position (Aug. 1998) and is still working as far as I am aware. Since ion pumps are classified as solid entrainment type pumps, their lifetime depends on their pumping history, i.e. operating pressure. The SIP on that particular JEOL 2000FX was typically operating in the mid to high 10-7 Torr range in standby mode, which resulted in a lifetime of 7.5 years. The same operating condition has been maintained for the reconditioned SIP and it is currently coming up on 5 years this summer. Also, the cost for reconditioning was ~$1020 + shipping (US-1994 dollars). Good luck!
David
* * * * * * * * * * * * * * * David T. Hoelzer, Ph.D. Metals and Ceramics Division Oak Ridge National Laboratory Bldg. 5500, Mail Stop 6376 P. O. Box 2008 Oak Ridge, Tennessee 37830 (423) 574-5096 {Work} (423) 574-0641 {Fax}
Colin asks ... } } } Hi All, } } The ion pump on our JEOL 2010 (LaB6) has just about died and we were } wondering what sort of lifetimes people were getting for } pumps on similar (JEOL 2000 series) machines. ...
5 years should be considered a better than average lifetime. We had a problem replacing the ion pump on our 6300 SEM, because it was a JEOL pump, which has since stopped supporting (... at least they didn't have one on their shelf when we asked, and instead wanted to re-furbish the existing pump elements ...). Since the JEOL's IP HV connector was a non-standard type, we then decided to go with the same capacity and an established 3rd party and a standard connector. It could be that your IP has a standard connector, and that all you have to do is replace the elements. It would be also just as cost effective to have JEOL or a 3rd party service refurbish your elements. Most cost effective would be to extend the pump's lifetime by baking it out, but not all elements can be baked effeciently or at all. And, yet another option, is to try and find a replacement element which can accommodate an "in situ" bake-out heater, altho more probably, this may mean replacing the entire pump.
I have more info around here somewhere, but can't easily find it ... but I did find the 3rd party refurbishing and replacment services via searching the wwweb with "metacrawler.com".
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Hi Colin, I'm replying to the List because my system thinks your address is invalid (due to two "-at-" symbols?).
I inherited a TEM with an IGP that was less than 5yrs old. It was used in a rapid-turnaround pathology lab. Both the samples and the film were cycled very quickly through the system. (In fact, this scope would frequently crash because the film was used up so quickly, the auto-sequence for pumping the column was interrupted.) No one used the cold trap. The IGP became exhausted and had to be replaced. Luckily we had covered the IGP as a rider on a service contract.
The reason for its early demise was attributed partly to the biological specimens that essentially put a lot of water into the vacuum system but mostly to the fact that the scope and film desiccator were backfilled with impure (wet) nitrogen. (Room air would have been even worse.) The nitrogen we used was run-of-the-mill 'standard' nitrogen, and the service contract provider wanted to nullify the IGP rider coverage because of it. Lucky for us, since the rider did not specify what TYPE of nitrogen to use, we were covered. Once the pump was replaced and we switched to UHP (ultra-high purity) nitrogen for backfilling, the system performed fine.
I was not there long enough to complete another 5yr study; however, I now have another IGP system and I do specify UHP nitrogen. I also use the cold trap. Someone else may add to this, but I was told this delivers any specimen-derived water vapor to the vacuum system in a slow, controlled fashion as the trap heats up (rather than all at once).
Hope this helps.
Ann Hein Lehman Trinity College Hartford CT
-----Original Message----- } From: "colin.veitch-at-dwt.csiro.au"-at-Sparc5.Microscopy.Com [mailto:"colin.veitch-at-dwt.csiro.au"-at-Sparc5.Microscopy.Com] Sent: Monday, April 12, 1999 1:02 AM To: microscopy-at-Sparc5.Microscopy.Com
Hi All,
The ion pump on our JEOL 2010 (LaB6) has just about died and we were wondering what sort of lifetimes people were getting for pumps on similar (JEOL 2000 series) machines. Our samples, though biological are not changed often and even so, with the configuration of the machine ie differential pumping etc. we would not expect the ion pumps to have to work terribly hard. We have had the microscope for around 5 years and would like to know if that is about normal for these pumps.
We were also curious as to what people did when the pumps finally fell over. As far as we know there are 3 options - replace with new pumps, replace with reconditioned pumps or replace the filaments in the existing pumps.
What have people found to be the most "cost effective" solution?
Thanks for any help with this.
Colin Veitch
Instrumentation Scientist Fibre Structure & Function Group CSIRO Wool Technology PO Box 21 BELMONT Vic 3216 Australia
Tel: +61 (0)3 5246 4000 Fax: +61 (0)3 5246 4811
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Wool Technology on +61 3 5246 4000.
I need to pump this group for information (sorry, it slipped out).
The recent discussion of ion pump lifetimes has generated some talk here about the lifetime of turbo pumps. Does anyone have figures for turbos? For example, say one has a TP with magnetic bearings and runs the pump continuously, at what point would one expect some sort of problem due to normal wear and tear? I assume that metal fatigue would be a factor to consider here. Thanks.
While the following list is by no means complete, the following come to mind:
Leica Olympus Triangle Biomedical Accuedge
Ther are several more out there. All of them have their advocates in the histology community.
All these blades will work fine with paraffin processed tissue.
-- Begin original message --
} } Hi all, } } What brands and types of razor blades are used as a disposable microtome } knive (paraffin, manual rotary microtome)? What are their limitations } regarding section thickness for non-problematic animal and plant tissues? } } Thanks in advance for any input! } } Yvan Lindekens.
-- End original message --
best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**Suppose you were an idiot... And suppose you were a member of Congress ... But I repeat myself.-Mark Twain**
there is a short communication in "Powder Technology" from 1971 that describes a eutectic camphor-napthalene mixture that can be used to deagglomerate small particles for observation as individuals. i have used this technique with some success in the past. what it amounts to is dispersing the particles by "kneading" them into the eutectic mixture and then subliming the mixture away to leave only the particles. this avoids the pulling effect of a drying liquid dispersant and helps keep the particles apart.
hope this helps.
ref: Powder Technology, 5 (1971/72) p.377-9.
b-
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
716-275-3058 716-244-4936(fax)
"You may get to the top of the ladder of success only to find its been leaning against the wrong wall" A. Raime
I am experimenting with phosphorous doped silicon. The goal is to determine dopant concentrations in the doped regions between the source/gate and gate/drain. I believe that with the correct recipe, the doped silicon will etch at a faster rate than the undoped regions, revealing thickness fringes when view in the TEM in the WBDF configuration. Thus far I have not obtained the desired results, ie, the etchants seem to attack all areas equally. Do you know of a wet chemical etch recipe which would achieve this result?
i've got a perfectly good noran light element detector that i'd like to keep and plug into a new low-cost rear end electronics and computer system. i've spoken with some vendors (that shall remain nameless) that cannot interface easily to the detector. are there any vendors that can do this and provide qual/quant, digital imaging, and stage control for a reasonable price?
(this may not be of general interest so 'reply' may be most appropriate)
thx b-
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
716-275-3058 716-244-4936(fax)
"You may get to the top of the ladder of success only to find its been leaning against the wrong wall" A. Raime
If the wick is regularly replaced and the operating conditions are clean, the TP should last many years. I have a Balzers 240 that is 11 years old and still works perfectly. It uses a TP120 controller.
JEOL typically sells a JEOL 840 for about $40,000.00 installed with a one year warranty. This is without the EDS. On the used market the 840's have been selling for about $35,000.00 to $45,000.00 with an EDS but not the EDS is not guaranteed working. In other words the EDS and related hardware is given gratis. About half the time I have found them to be in working order as the detectors have been left at room temperature for an unspecified time. I would be heisitant to purchase a Kevex 8000 series as reliability has been an issue with this series because of the wirewrap mother boards. Check with Kevex (now Noran) for the details.
Hope this helps.
Earl Weltmer
Cochran wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi All, } } This is my first posting to this listserver. I am in position to } purchase a Jeol 840A sem in good condition equipped with a Kevex 8000 } EDS detector and need to know its approximate value on the used market. } Responses can be made directly to me or via the listserver. } } Thanks a lot for your time and assistance. } } Ray Cochran
In a message dated 4/9/99 6:14:32 PM US Mountain Standard Time, walck-at-ppg.com writes:
} microscopy-at-sparc5.microscopy.com
We have a most ancient AO microtome with cryostat and a dull ugly blade in a wood box. It would be neat if we could find one of the holders that held the razor blades ... any floating out there??! Ed Sharpe
Turbo pump life time is like asking how long is a piece of string. We = have many pumps on various systems that go for a very long time ( 8 to = 10 years) however we have also had those that go in one year for no = apparent reason.( plus one month just so that they are out of warranty )
However most pumps, from whichever manufacturer, seem to last pretty = long and are very reliable if treated with respect. One of the main = killers of turbos seems to be oxidation and fatigue of the metal = components. Some of the EM's, when switched off, vent the backing line = and thus the turbo. This means that the turbo parts, which are designed = to be under vacuum all the time, oxidise quicker than originally = planned.=20 So my advise is to ensure that the turbo is always on or at least under = vacuum. If you need to leave the system "off" over holiday periods, it = is better to simply unplug the turbo controller and leave the rotary = pump running. This will keep the system under vacuum without the danger = of having a power failure or such like, kill the turbo whilst no one is = around. Servicing the turbo oil wick is also recommended on a regular basis and = our experience shows that once a year is sufficient.=20 But believe me, I would much rather have a turbo system to any diff. = pump or Ion pump system.=20
Cheers Luc Harmsen Anaspec, South Africa Technical support on microscopy. Tel + 27 (0) 11 476 3455 Fax + 27 (0) 11 476 7290 anaspec-at-icon.co.za
-----Original Message----- } From: "JBozzola-at-aol.com"-at-sparc5.microscopy.com = [SMTP:"JBozzola-at-aol.com"-at-sparc5.microscopy.com] Sent: Monday, April 12, 1999 7:03 PM To: Microscopy-at-sparc5.microscopy.com
I need to pump this group for information (sorry, it slipped out).=20
The recent discussion of ion pump lifetimes has generated some talk here =
about the lifetime of turbo pumps. Does anyone have figures for turbos? = For=20 example, say one has a TP with magnetic bearings and runs the pump=20 continuously, at what point would one expect some sort of problem due to =
normal wear and tear? I assume that metal fatigue would be a factor to=20 consider here. Thanks.
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This is really cool!
PREMIUM CHANNELS........Descrambled!
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On Mon, 12 Apr 1999 JBozzola-at-aol.com-at-Sparc5.Microscopy.Com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I need to pump this group for information (sorry, it slipped out). } } The recent discussion of ion pump lifetimes has generated some talk here } about the lifetime of turbo pumps. Does anyone have figures for turbos? For } example, say one has a TP with magnetic bearings and runs the pump } continuously, at what point would one expect some sort of problem due to } normal wear and tear? I assume that metal fatigue would be a factor to } consider here.
John,
I have run a Seiko-Seiki Maglev on our JEOL4000 without any problems for 8 or 9 years. It does not give any noticable vibration or magnetic field problem (we resolve 0.25nm) although I do use a Balzers vibration isolator as I was pretty paranoid when I first fitted it in place of the SIP. We have had a few (2 or 3) severe vacuum crashes when it suddenly pumped air and hit the safety bearings but it has survived OK. The reason we replaced the SIP is that the microscope has been modified to include an apertured gas reaction cell so the pump often sees a pretty poor vacuum (10-5 mbar) of H, He, N, O, CO, Ar and various hydrocarbons for several hours at a time. However, the STP can still pull 10-7 mbar on the column given time to clear gasses out of the column properly.
Unfortunately, I have no financial interest in any of the companies mentioned.
Ron
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
Robert, it is a far from easy task to get 'real' numbers out of this. The etch you should use is (roughly) 0.5% HF acid (usual conc., i.e. 48%) in Nitric acid (again, 'usual' conc. - 50%???). At room temperature this will give you the desired effect in a few seconds on a prepared TEM specimen. Au or diamond grids will stop Cu being dissolved from the grid and being re-deposited on your sample. UV light is supposed to be needed for one polarity of dopant, but I can't remember which. I did see one paper a couple of years ago that said that the thickness of the TEM specimen had an effect, so they etched just one side - before doing the second polish to make a thin specimen, and ion milling from the unetched side (I can probably dig this reference out if you like). Ron Anderson from IBM told me they do it at about -100 degrees C in a dark tank for a few minutes; the solution should be a kind of 'slush' like the iced drinks you can get. He said this slowed down the etch rate and ! gave better uniformity and more re In my opinion the experimental variability in this method is likely to be much worse than the FE-SEM method, where you get doping contrast due to work function changes. I'm also not sue whether the TEM method is primarily sensitive to the active or inactive dopant concentration. In either case you will need a planar sample for SIMS to give you a calibration curve.
Cheers,
Richard Beanland
} --------------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } --------------------------------------------------------------------. } } } I am experimenting with phosphorous doped silicon. The goal is to determine } dopant } concentrations in the doped regions between the source/gate and gate/drain. } I } believe that with the correct recipe, the doped silicon will etch at a faster } rate than the undoped regions, revealing thickness fringes when view in the } TEM in the WBDF } configuration. Thus far I have not obtained the desired results, i.e., the } etchants seem to attack all areas equally. Do you know of a wet chemical } etch recipe which would achieve this result? } } Thank you, } } Robert Pomrenke } bpom-at-aol.com
Try Conneaut Lake Scientific in Conneaut Lake, Pa. for what you are looking for.
Just to let you and others know that Mike Gemble the owner/operator has a warehouse full of used microscopes and microscope related items such as preparation equipment and accessories.... He can be reached at myroscope-at-aol.com or his phone is 814-382-1604... If anyone is interested and passing through Norhtwest Pennsylvania, it would be a good place to stop and browse if you are into this type of equipment... Also his web site is www.conneautlakesci.com Check it out!
Good Luck!
C YA C. Passione -----Original Message----- } From: "COURYHOUSE-at-aol.com"-at-sparc5.microscopy.com {"COURYHOUSE-at-aol.com"-at-sparc5.microscopy.com} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
I am looking for sources to obtain a quality "student" grade microscope for my son, who is 10 years old. Can you or the MSA offer an recommendations or guidance?
I operate and service our Philips CM12. The +15 volt power supply in the remote racks has failed, mainly in the area of the 2 MJH-16010 transistors.
The designation for this supply is PE 1130/00 and I require this manual to troubleshoot the supply rather than paying out 1000's of dollars to have the it replaced by Philips.
If anyone has this manual (or circuit diagrams) I would be very appreciative of a copy.
You can contact me offline if you wish, or at the numbers below.
Thanks in advance
Fred Pearson
******************************************************* Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
Has anyone received information about recertification? I have not received anything and I'm a bit concerned that my certification will expire even while working in the field.
JBozzola-at-aol.com-at-Sparc5.Microscopy.Com wrote: } } } I need to pump this group for information (sorry, it slipped out). } } } } The recent discussion of ion pump lifetimes has generated some talk here } } about the lifetime of turbo pumps. Does anyone have figures for turbos? For } } example, say one has a TP with magnetic bearings and runs the pump } } continuously, at what point would one expect some sort of problem due to } } normal wear and tear? I assume that metal fatigue would be a factor to } } consider here. } Dear John, The HVEM has 2 Balzers TPU330's, one on the accelerator and the other on the column. The accelerator is always kept at ~3*10^-7 torr, and the pump is run at ~2/3 speed in standby mode. The column is at ~7*10^-6 torr and is occasionally aired and pumped out. We have had the reccommended bearing changes at 2-year intervals (the wicks are also changed at that time) and we change the oil twice a year. We have had no problems at all with the pumps--in contrast to the old TMPs we replaced them with many years ago. Yours, Bill Tivol
SUMMARY:(To anyone interested in the details, all replies follow.)
Thanks to all for your feedback (including phone calls). I was surprised by the volume, but that helped justify to others here my concern about venting in general.
The overall view was that I had nothing to worry about in terms of pump performance and lifetime, although there were a couple of horror stories. Many responders vent to their fume hood air-handling systems, or other positive-pressure exhaust systems. What happens to the 'bad air' once discharged outside was a real concern - I have left that one to the engineers. Other concerns included handling condensation especially water coming down from the roof, and maintenance of filters.
What I did was have them use 45-degree angles where possible and include a T-cleanout at the wall for each pump.
Thanks again-- Ann Lehman
--------------------------------------------------- THE QUESTION: Dear Listers,
My Safety Officer wants all our mechanical pumps vented outdoors. The pumps are Sargent-Welch, Alcatel, and Edwards and are used to back TEMs, film desiccators, evaporators, etc. Initially I thought this was a good idea, having done it simply in past lives by drilling a hole in a wall, ganging the outlet hoses, and presto - out the bad smoke went.
However the B&G engineering consultant got hold of the project (yes now it's a 'project') and now EVERY pump must get its own line up to the roof. This means each pump's discharge is directed to a run of 1-inch copper pipe with three to five 90-degree bends over a total vertical length of about 20'. (Luckily I am on the top floor.) There is also talk about inserting a clean-out or filter at each feed-through for dealing with accumulated oil.
I'm concerned that the pumps are not designed for backpressure on the outlet side coming from the 20' column of air plus the resistence from the 90-degree bends and filter.
Could this setup affect the (1) efficiency or (2) overall life of the pumps? Am I being overly cautious?
I'd appreciate feedback from the List (including manufacturers) re the feasiblity of this approach, and the pump specs - I haven't been able to find anything about discharge 'load' tolerances.
Thanks, you can reply offline and if there is sufficient interest I'll summarize responses for the List.
Ann Hein Lehman EM Facility Mgr Trinity College Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-exchange.cc.trincoll.edu
--------------------------------------------------- THE ANSWERS:
My guess is that this will be hard to judge. We have a much saner system with the pump exhausts fed into the air handling system (which takes exhaust air from our home in the second sub-basement to the 46th floor where it is released). When we put a filter on the exhaust, we notice no adverse effects, and as the filter becomes full of oil, the back-pressure must rise. I'd think that the air column--bends and all-- would be a small contribution compared to the filter. I would definitely reccommend having some sort of oil trap on the line, since otherwise oil would accumulate and potentially cause problems. Good luck. Yours, Bill Tivol --------------------------------------------------- Yes, please summarize. I assume you are using an exhaust filter now and this presents some impedance to the exhaust. Presumably with the copper pipe
exhause, the filter would be removed so this would save a little on the overall impedance.
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
--------------------------------------------------- If you are worried about back pressure you can put a fan in the lines to put negitive pressure at the outlet of the vacuum pumps.
Gordon Couger gcouger-at-couger.com www.couger.com/gcouger Stillwater, OK 405 624-2855 --------------------------------------------------- Although we do not manufacture these pumps we do supply the range as backing pumps for our Range of Preparation Equipment, Sputter Coaters, Carbon Coaters etc, etc. We would always recommend an Oil Mist Filter and always provide one, these can be of a type which remove the oil and just vent into the air or you can have an inline type where they remove the oil and have an outlet position which you can vent to air.
In general terms the single and dual oil filled rotary vacuum pumps will withstand back pressures of at least 1/4 of a bar i.e. approx. 4 p.s.i. which equates to 8 feet head of water back pressure. So you will have no problem with your 20 feet of air.
Trust this is of help. On behalf of Emitech Ltd. --------------------------------------------------- Unless your lab is at a partial vacuum and you are venting your pump to atmosphere, backpressure is a moot point. The gas flow is miniscule except for initial roughing.
Runs, elbows, diameters, restrictions are all critical on the INTAKE side of the pump, but have no significant effect on the outlet side of the pump.
Ken Converse owner, Quality Images Delta, PA 717-456-5491 717-456-7996 fax --------------------------------------------------- I think you are being overly cautious. The static pressure inside your 20' of pipe is, of course, the same as the pressure outside. The only worry would be dynamic pressure as you pumped down the column, dessicator, or whatever. A length of 1" pipe can handle an enormous amount of air, and you only pump at maximum rate for a few moments. After all, a small-to-medium rotary pump has a capacity of, say, 5 cu. ft. per minute, which corresponds to 8,640 cu. in per minute, or 144 cu. in pr second. Very roughly, the cross-section of your pipe is 1 sq. in., so the velocity is about 150 in/sec, or 13 ft/sec, 10 mph. The pressure drop to get this velocity will be quite small. When the chamber or whatever is evacuated, the gas flow is negligible.
The whole design plan seems like overkill, though. Does the design engineer have an interest in the plumbing company that would install the pipework? Our system at MIT vents into ducts left over from a hood formerly installed in the area, so the fan, ductwork to the roof, etc., were all in place.
tonygr-at-MIT.EDU --------------------------------------------------- I don't think back pressure is an issue but something to consider: under the right conditions the ID of the cool Cu tubing (in the building) is condensation water from outside. We know the penalty for filling our pumps with water... The work around is to exhaust into a duct of a forced air exhaust system (like a chemical fume hood). Also I recommend that at your end of the exhaust line that a "T" fitting be used & you connect to the RA. Extend the lower side of the "T" ~6 inches & terminate with a ball valve. That way if anything comes down the pipe it goes into the trap rather than your pump.
& the big one: I'd be sure the F&E (B&E) group understands the liability they expose themselves to. Unknown to us till the pump croak, our F&E guys modified our exhaust line & poured water into our Fomblin full pump.
good luck, Bruce Brinson Rice U. --------------------------------------------------- I would say you have nothing to fear. Typically, a roughing pump should be fitted with an oil mist filter. The back pressure generated by the filter would "swamp" any from the piping you described. My answer is predicated on pumping down a reasonably sized, sealed vacuum chamber with a rotorary vane pump or similar. After the initial few gulps of gas, the volume of gas moved (at STP) is very tiny. If your vacuum chamber is the size of a small room and the initial pumpdown is by something like a roots blower, that is different... {g}
Woody White McDermott Technology --------------------------------------------------- My first response would be to query whether the safety office really knows what he desires to vent with this very expensive venting project. Usually the most noxious part of a roughing pump exhaust is the particulate or oil mist. If you are not pumping something dangerous like a poison gas or something equally dreadful, I see no reason to go to the expense. The exhaust filters which have been designed for use on your pumps usually do a fine job at trapping any problem vapors. There should be no problem with back pressure. If someone finds me in error, I'm sure we will hear about it.
Good luck. Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. Number 499, Post Office Box 19400 Austin, Texas 78760 Phone: 512/282-5507 FAX 512/280-0702 --------------------------------------------------- I can sympathize with your problems. I run a plastic sump pump hose from each of my rotary pumps out to the window. If you have ever watched an untrapped rotary pump, you will notice that there is almost no air movement out of the pump after the initial ten seconds of pumping. There should be no problem with long runs, bends, etc. if there is nothing but simple atmospheric pressure to oppose the air movement. A simple drain at the first bend up should drain any condensed oil, or a wad of steel wool in the line. Use as large a diameter pipe as you can get them to run. One solution I've seen is to put a commercial car oil filter onto the pump exhaust. If this didn't bother it, neither should your copper line.
mager-at-interchange.ubc.ca --------------------------------------------------- we have 21 mechanical pumps in a total of twelve rooms in our lab. This includes pumps for specimen prep equipment, electron microscopes and desiccators much like with your situation. We remodeled the labs in 1992 and installed a new exhaust system to remove oil mists. We had previously installed some exhaust systems in the late 1970s because we had a post doc MD/Ph.D. here who insisted that breathing oil mist vapors was not healthy. I have accumulated the oil since 1992 (it condenses out into one liter containers above our pumping stations). Today I measured the total volume accumulated in all rooms. It was just a little more than one liter. This is a long time to accumulate oil, but it may be useful information for you. There are pumps on the roofs of the respective buildings which house the equipment referred to above. All roof pumps are at least 30 feet away from the instrument pumps.
We used to use the oil mist traps but we found they required a lot of maintenance with that many pumps. Cleaning them requires a lot of time and if you replace them every few months, the cost adds up quickly.
I assume that most of our accumulated oil is SUPERGRADE A OIL, the type we use for our mechanical pumps. This is produced by Inland Vacuum Industries in Churchville, NY, according to the MSDS sheets I have. These sheets also indicate the following: "VENTILATION US Gov't 8 hr TWA limit for exposure to oil mists is 5 mg per cubic meter."
I assume that some of the accumulated oil I collected is diffusion pump oil (Santovac 5). This is a polyphenyl ether and is made my Monsanto. The Permissible Exposure Limit (PEL) is also 5Mg/cubic meter for this oil.
I like our system because it is maintenance free for us. Our university air conditioning mechanics maintain the pumps on the roof.
John.Wheatley-at-ASU.Edu --------------------------------------------------- The biggest problem with that sort of a set-up is that the water vapor will condense in the exhaust line and after the pump is turned off flow back into the pump ruining it in a short time. You will need a trap for the condensation and you will have to empty it on a regular basis. The long exhaust line will not add to the efficiency of your pumps.
Hope that helps, Peter Jordan, EMSI --------------------------------------------------- I applaud the decision to vent the exhaust. I know of no EPA or other regulation to do so, but the exhaust of mechanical pumps on initial pumpdown is something I have always recommended my customers be rid of. I hope that you are also taking measures to move the pumps into separate rooms or acoustic enclosures to reduce their noise output.
Working with EMs often means spending many hours at a time living with these problems. While occasional exposure can be acceptable, the constant drone of pumps and exposure to pump oil vapors can only be detrimental.
The setup you detail presents no problem to operation of the pumps. Air is very compressable and the volume of air present in the exhaust lines means that there will be only a negligible pressure increase due to the various bends.
A clean-out filter is unnecessary. Proper design of the exhaust stack should provide for a low point where accumulated oil can be either be drained or returned to the pump. This requires less than 90 degree bends in the stack so that any condensed oil can flow back to the pump or a drain. If the exhaust provides for a return to the pump, you have to be aware that there might also be other condensed materials, primarily water, included. Best bet would be to have a slightly greater than 90 degree bend close to the pump followed by a vertical section forming a trap. At that second bend, a drain should be included that would allow for the drainage of all fluids trapped there. Any such scheme would of course require the regular emptying of the trap.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com --------------------------------------------------- All four of our mechanical pumps, those on vacuum evaporator, glow discharge, film dessicators, and TEM, have been fitted with oil mist eliminator filters on the exhaust line. I suppose there may still be some noxious fumes emitted especially at initial evacuation but our noses don't detect them. One less expensive alternative to the piping exhaust that your Safety Officer proposes might be to mount an appropriate exhaust filter on one of the pumps and have the Safety Officer monitor the output with an appropriate sensing device and compare to unfiltered output.
However, if the Safety Office budget is going to pay for the piping project...
Just a couple of thoughts. Don Gantz Boston Univ Med School --------------------------------------------------- Nothing personal, but your safety and facilities people are NUTS! But, to give you some ideas:
1) at BNL in the Light Source, the rotary-pumps exhausts are attached to a flexible corrugated-hose. This hose has a slight negative pressure. 2) Where possible, I attach rotary-pump exhaust to the "house vacuum". But, you have to monitor the "house vacuum", incase the compressor is turned off. 3) Some of our Edward's pumps have large oil-filters on the exhaust. We get these filters at the local car-parts store. They are cheaper and more massive than the ones the vacuum companies sell.
hasta, Jim jquinn-at-dol1.eng.sunysb.edu --------------------------------------------------- I have two suggestions. If these are low volume pumps 3/4" copper is probably enough for each pump. If two or more pump lines are joined then the size should be increased to maintain a cross sectional area equal to the the 3/4" time the number of pumps. Yes, traps should be incorporated into the lines to catch condensed oil and water. If the line is going out the roof you must prevent water from entering the line and it may need to be insulated to prevent water condensation on the inside only if you pump significant water.
Good Smokin, Russ RGillmeister-at-sdms.usa.xerox.com --------------------------------------------------- I think venting the MPs to the outside is a good idea. We did this in our lab by connecting the exhausts to the nearest hood using a copper pipe and rubber connectors. For our Hitachi, which has three MPs, our maintenance department built a large wooden box with a removable plexiglas lid. The lid has a large, flexible aluminum air duct connected to it. The whole thing exhausts via a hood. We can really smell the difference! So far we have had no problems.
Good luck Jordi.Marti-at-alliedsignal.com --------------------------------------------------- Only to say that if the pumps were designed to be able to operate in the described manner then the manufacturers would have specs on how and what type of ventilation system to install under different applications, i.e. extraction type if there is more than 10' of pipe to outside, or more than 2 bends totaling more than 90 degrees. Also, if discharging the "bad air" outside then EPA standards must be met...such as installing "scrubber" systems to insure no oil or certain hydrocarbons are discharged into the air and eventually being introduced into the ground water. Best thing is to install a factory offered oil recovery exhaust filter system such as the one we spoke about on the coating system. And to check with OSHA as to the "hazards Imposed" by the Vacuum Pump vapors. If they say they are too hazardous for normal laboratory environments then we may explore the possibility of routing the exhaust into existing "Fume Hoods" in the laboratories. That is assuming they have the necessary environmental "cleaning" systems built into them.
Bottom line is I doubt that without a lot of engineering the pumps will be able to be operated properly with the exhaust "routed" through any type of piping more than a few feet long. Meaning that if you hook any lines, more than a few feet long without bends or any that have more than 90 degrees of bends, to remove the fumes to any of the pumps, they won't work properly without a lot of engineering to eliminate any "back pressure" that may be caused.
--------------------------------------------------- (This is not a direct response, but a related one - In reply, I would add that the lab I used to work in had its air intake sited exactly where the delivery trucks would park and idle their engines for hours while unloading. The fumes made several people sick - nausea & headaches. So it is possible your guy suffered a hypersensitive reaction. --Ann Lehman)
Funny how we were just talking about vacuum pump oil vapours and how to deal with them... Last Tuesday I had a guy in the lab to change the locks (long, unrelated story), including the lock on the door right behind our ESEM. Our vacuum lines pass through a cut-out on this door to the pumps located in a big warehouse-like area. We have the usual little filters on the pumps, but as we all know, you can usually smell a little something in the air anywhere near them. While changing the lock, the guy was standing very close to the pumps for perhaps 10 minutes or so, and said he could smell something "oily". Well, I just found out that later that day, he became quite ill, with apparently a toxic reaction to a substance or substances unknown, and was hospitalized for a few hours, though he later, apparently, recovered completely. We're not absolutely sure that breathing our oil vapours for a few minutes is actually what caused the reaction, but it appears to be the only possible toxin the man was in contact with that day ( or at least that's the story). It's possible that the gentleman just had an unusual sensitivity to the vapours; I know at times I've had lots more exposure than he did, and have personally never gotten so much as a headache. One thing bothers me a bit, though; we use Alcatel 102, an oil specially made for ESEM's, with their increased water throughput, and I wonder if this stuff might be a bit less user-friendly than the more common types. Has anyone else out there ever seen, or heard of, a toxic reaction to this or any other pump oil? (I have a funny feeling that I haven't heard the end of this incident yet...)
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B2Y 4A2
Does anyone know of any short courses given in New Jersey related to=20 Failure Analysis, Metallurgy, Metallography, Etc=2E? If so, I would=20 greatly appreciate the information=2E =20 TIA, =20 Leslie Link e-mail: Leslie=2ELink-at-us=2Egtc=2Eboc=2Ecom
I have an OLD razor blade holder with plastic anti-roll plate. There is IEC engraved on it. Could this be what you are looking for? Lilith ------------------------------------------------------ Lilith Ohannessian-Barry National Research Council Institute of Biological Sciences CANADA Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca ---------- } From: "COURYHOUSE-at-aol.com"-at-sparc5.microscopy.com To: microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------.
In a message dated 4/9/99 6:14:32 PM US Mountain Standard Time, walck-at-ppg.com writes:
} microscopy-at-sparc5.microscopy.com
We have a most ancient AO microtome with cryostat and a dull ugly blade in a wood box. It would be neat if we could find one of the holders that held the razor blades ... any floating out there??! Ed Sharpe
Check with your local Mattel toy store for the recently released "Play X3 Digital Video Microscope" with a list price of $99. This is a joint venture with Intel and is CMOS based. It is designed to let children of all ages display images from the microscope on a PC at both still and video rates, print images and email them. Mattel/Intel will also use similar technology for their Me2Cam. Naturally the results will not compare with the $100,000. instrumetns we use in research or will they? Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
I know you all have been through this subject before, so I thought I'd ask what month's archives would be best to search for your prior discussions on the good,bad and ugly of film scanners.
I'm responsible for near 60 years of 4x5 negatives from our R&D work here at TIMET, some of the oldest are 4x5 TEM and view camera negs but the majority are Polaroid negatives.
Perhaps, someone archived just the appropriate responses?
Any help or discussion would be greatly appreciated.
I am experimenting with phosphorous doped silicon. The goal is to determine dopant concentrations in the doped regions between the source/gate and gate/drain. I believe that with the correct recipe, the doped silicon will etch at a faster rate than the undoped regions, revealing thickness fringes when view in the TEM in the WBDF configuration. Thus far I have not obtained the desired results, ie, the etchants seem to attack all areas equally. Do you know of a wet chemical etch recipe which would achieve this result?
I was wandering if anyone could help me out as i am new to this technique. i have been retorgradely labeling cells in the spinal cord with CTB-HRP in rats and then staining with TMB and DAB reactions. however this has not been highly succesfull..any suggestions? i think the problem might be in the injections rather than the staining from what i gather under EM i should be looking for a black pepper like precipitate and some form of tungstate crystals? any suggestions on how to get dendritic staining? Thanks Jennifer Wilson
Jennifer Wilson Neurosciences Loeb Research Institute Ottawa Civic Hospital 751 Parkdale Avenue Ottawa, Ontario, K1Y 4E9 Canada Tel;( 613) 798-5555 x6387
Robert, I do not know of any selective etchant for this process, although they may exist. Stains can be used to identify n vs. p regions using optical microscopy, although this gives no quantitative data. I would probably do some C-V measurements (Capacitance-Voltage) to get at the doping. Or if these are big devised, a 4-point probe would do the trick. -good luck -andrew ___________________________________ I am experimenting with phosphorous doped silicon. The goal is to determine dopant concentrations in the doped regions between the source/gate and gate/drain. I believe that with the correct recipe, the doped silicon will etch at a faster rate than the undoped regions, revealing thickness fringes when view in the TEM in the WBDF configuration. Thus far I have not obtained the desired results, ie, the etchants seem to attack all areas equally. Do you know of a wet chemical etch recipe which would achieve this result?
Thank you,
Robert Pomrenke bpom-at-aol.com
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_ 1:15 =F1 2:00 : " Scanning Probe Microscopy - From Birth to Adolescence=EE, H. Kumar Wickramasinghe, IBM T.J. Watson Research Center, Yorktown Heights, NY.
_ 2:00 =F1 3:15 : "Strategy and Tactics of (Microprobe) Analysis=EE - Pau= l =46. Hlava, Sandia National Labs, Albuquerque, NM. --------------------------------------------------------------------------- ------- =46or more information please contact either:
Phil Flaitz or Evan Slow (914) 892-3094 (201) 760-2524 flaitz-at-us.ibm.com ess-at-feico.com
by styx.services.ou.edu (8.9.1/8.9.1) with ESMTP id RAA19617 for {microscopy-at-sparc5.microscopy.com} ; Tue, 13 Apr 1999 17:19:22 -0500 (CDT) Message-ID: {3713C2B2.5CF8BD6B-at-ou.edu}
We have inherited an OLD RCA TEM minus the HVPS. I have what I think is the serial number (the ID plate is probably on the HVPS). We have no intention of trying to make it operational (we will clean it up and put it in static display), but I would like to find out the model number, about how old it is, and whatever else I can dig up on it. I used an old RCA 4 (I think) back in about '72 and that was a modern instrument compared to this one. I know RCA quit making TEM's a LONG time ago, but can anybody point me in the right direction to begin getting some info on this scope.
Thanks, Bill -- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
} I am looking for sources to obtain a quality "student" grade microscope for } my son, who is 10 years old. Can you or the MSA offer an recommendations or } guidance? } Bill -
You'll find detailed advice on what to buy, plus a list of suppliers, on the Project MICRO website (URL below).
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Howdy Folks, } } I know you all have been through this subject before, so I thought I'd ask } what month's archives would be best to search for your prior discussions on } the good,bad and ugly of film scanners. } } I'm responsible for near 60 years of 4x5 negatives from our R&D work here at } TIMET, some of the oldest are 4x5 TEM and view camera negs but the majority } are Polaroid negatives. } } Perhaps, someone archived just the appropriate responses? } } Any help or discussion would be greatly appreciated. } } Bill } TIMET
Since the 2000FX was referred to in recent discussions regarding sputter ion pump lifetimes, I thought it pertinant to add a few items that can have an effect on the life of the pump with this instrument and others as well.
Of course the basis for pump life has alot to do with how tight the chamber it pumps on. If ultimate pressure is near 1 X 10 -5 Pa you can safely assume the gun/column area is reasonably tight. An order of magnitude or so higher and you need to find the vacuum leak (or replace the pump). A simple leak rate check will verify gun/column integrity.
Here at ASU we got 9 & 1/2 years use from the orginal ion pump. The reason why has alot to do with the above. However there are other reasons to consider. One being we have religiously used the cold trap (or ACD). If this is cold prior to sample insertion, the vacuum recovery time after insertion is remarkably quicker as the admitted gases are trapped immediately and thus saved from the ion pump. When baking the ACD at the end of the day, the pumping configuation is changed (in ACD HEAT mode)to where the ion pump is shut off as the trap is heated. A normally closed valve opens and the trapped contamination is released and diffusion- pumped from the system. Incorporating this system of trapping, heating, and diffusion pumping contamination from the microscope will extend the life of the pump as well as keep the contamination rate minimal in the beam/specimen interaction area.
Bob Roberts Arizona State University Center for Solid State Science PSA-213 Tempe, Arizona 85287-1704
Bill Chissoe III wrote: ================================================== We have inherited an OLD RCA TEM minus the HVPS. I have what I think is the serial number (the ID plate is probably on the HVPS). We have no intention of trying to make it operational (we will clean it up and put it in static display), but I would like to find out the model number, about how old it is , and whatever else I can dig up on it. I used an old RCA 4 (I think) back in about '72 and that was a modern instrument compared to this one. I know RCA quit making TEM's a LONG time ago, but can anybody point me in the right direction to begin getting some info on this scope. ================================================ If it is not an EMU-4 (e.g. A, B, or C), then it would have been produced by RCA in early 1969 or before, that being the time they introduced the EMU 4-A . That means it probably would be an EMU3 A, B, C,....or H. The first production of the EMU3 series would have started in the early 1960's (if I remember correctly).
You might want to contact Prof. Arthur Smith of West Chester University. He has some number of these older RCA EMU-3's in operation which he uses in conjunction with some very effective courses on EM maintenance. His e-mail address is asmith2-at-wcupa.edu
He and his students service all of these ancient RCA TEMs and Prof. Smith probably is the leading remaining authority on anything related to these old instruments.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Fred Pearson wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello Everyone: } } I operate and service our Philips CM12. The +15 volt power } supply in the remote racks has failed, mainly in the area of the 2 } MJH-16010 transistors. } } The designation for this supply is PE 1130/00 and I require this manual to } troubleshoot the supply rather than paying out 1000's of dollars to have } the it replaced by Philips. } } If anyone has this manual (or circuit diagrams) I would be very } appreciative of a copy. } } You can contact me offline if you wish, or at the numbers below. } } Thanks in advance } } Fred Pearson } } ******************************************************* } Fred Pearson } Brockhouse Institute for Materials Research } McMaster University } 1280 Main St. West } Hamilton, Ontario } Canada L8S 4M1 } } email: eoptics-at-mcmaster.ca } phone: (905) 525-9140 ext. 24609 } fax: (905) 521-2773 } ******************************************************** Fred,
You may also use a number of alternative sources for spare parts. In this particular case I recommend switching power supply made by ETA POWER, LTD. Power supply part # 618-FHP15SX, rated 15V 32A (original Philips one rated 15V 25A). You can order it from MOUSER ELECTRONICS, (800)346-6873. Order number 33R42, price- $495, warranty- 3 years (not bad). I used ETA supplies more than once with great success. These are fully protected top of the line units. It is possible, of course, to use much less expensive replacement if you do not care about warranty. Contact me off line shall you have any questions.
Vitaly Feingold SIA, Inc. (770)232-7785 office (770)605-6105 mobile
Polaroid SprintScan 45 is probably the best for 4x5 negs or chromes. It is much more compatable with a Mac than with a PC however. It requires a simple slow speed narrow SCSI bus.
gary g.
At 12:33 PM 4/13/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
RCA actually was, I believe, the leading manufacturer of TEMS in America in the early days.
Ed Sharpe
} Subj: old tem } Date: 4/13/99 5:48:16 PM US Mountain Standard Time } From: wchiss-at-ou.edu (Bill Chissoe) } Reply-to: {A HREF="mailto:wchiss-at-ou.edu"} wchiss-at-ou.edu {/A} } To: microscopy-at-sparc5.microscopy.com (Microcoscpy List Server) } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have inherited an OLD RCA TEM minus the HVPS. I have what I think is } the serial number (the ID plate is probably on the HVPS). We have no } intention of trying to make it operational (we will clean it up and put } it in static display), but I would like to find out the model number, } about how old it is, and whatever else I can dig up on it. I used an old } RCA 4 (I think) back in about '72 and that was a modern instrument } compared to this one. I know RCA quit making TEM's a LONG time ago, but } can anybody point me in the right direction to begin getting some info } on this scope. } } Thanks, } Bill } -- } ============================================================= } Bill Chissoe III } Electron Microscopist } University of Oklahoma } 770 Van Vleet Oval } Norman, Ok. 73019 } E-mail: wchiss-at-ou.edu Ph. (405)325-4391 } ============================================================= } } } } } } ----------------------- Headers -------------------------------- } Return-Path: {Microscopy-request-at-sparc5.microscopy.com} } Received: from rly-yb03.mx.aol.com (rly-yb03.mail.aol.com [172.18.146.3]) } by air-yb01.mx.aol.com (v59.4) with SMTP; Tue, 13 Apr 1999 20:48:16 -0400 } Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) } by rly-yb03.mx.aol.com (8.8.8/8.8.5/AOL-4.0.0) } with SMTP id UAA10462; } Tue, 13 Apr 1999 20:48:04 -0400 (EDT) } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) } id RAA12287 for dist-Microscopy; Tue, 13 Apr 1999 17:23:07 -0500 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by Sparc5. } Microscopy.Com (8.6.11/8.6.11) with SMTP id RAA12281 for " } MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 13 Apr 1999 17:22:35 -0500 } Received: from hermes.services.ou.edu (hermes.services.ou.edu [129.15.2.121]) } by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA12272 for { } microscopy-at-sparc5.microscopy.com} ; Tue, 13 Apr 1999 17:22:22 -0500 } Received: from styx.services.ou.edu by hermes.services.ou.edu with ESMTP; } Tue, 13 Apr 1999 17:19:23 -0500 } Received: from ou.edu (botchiss.bio.ou.edu [129.15.38.205]) } by styx.services.ou.edu (8.9.1/8.9.1) with ESMTP id RAA19617 } for {microscopy-at-sparc5.microscopy.com} ; Tue, 13 Apr 1999 17:19:22 -0500 ( } CDT) } Message-ID: {3713C2B2.5CF8BD6B-at-ou.edu} } Date: Tue, 13 Apr 1999 16:18:26 -0600 } From: Bill Chissoe {wchiss-at-ou.edu} } Reply-To: wchiss-at-ou.edu } Organization: University of Oklahoma } X-Mailer: Mozilla 4.06 [en] (Win95; I) } MIME-Version: 1.0 } To: Microcoscpy List Server {microscopy-at-sparc5.microscopy.com} } Subject: old tem } Content-Type: text/plain; charset=us-ascii } Content-Transfer-Encoding: 7bit } Errors-to: Microscopy-request-at-sparc5.microscopy.com } }
I have found that SEMs decrease in value following a curve like a piano off a cliff.
A JEOL 840 or other similar SEM is probably worth (going price) about $10K-$15K. Just about any other SEM that is 7-9 years old is also worth about the same amount. SEMs older than that are of course worth less. "Worth" is an interesting word in this context.
Most companies depreciate their capital equipment over 8-9 years. Thus, you would see SEMs offered that are 8-9 years old. According to the company, the equipment has been depreciated to zero value. Therefore, the salvage value, if greater than zero, might actually cost them money in recouping depreciation amounts. consequently, one sees offers to "take this thing away" or some such freebee.
The flip side is that if a person has a SEM that is working and is on-line, that makes a big difference between one that is not or is not covered by a maintenance contract. It is not unusual for an organization to stop maintenance on a SEM several years before the depreciated life is up. Then, it is caveat emptor.
Please send me a picture of this I will check the archives.
One curious TEM RCA made actually was a small unit that sat atop the bench I saw one in a microbiology for nurses manual I have. Does anyone out there have one of those? I would be interested in it for the museum. thanks, Ed Sharpe
We have inherited an OLD RCA TEM minus the HVPS. I have what I think is the serial number (the ID plate is probably on the HVPS). We have no intention of trying to make it operational (we will clean it up and put it in static display), but I would like to find out the model number, about how old it is, and whatever else I can dig up on it. I used an old RCA 4 (I think) back in about '72 and that was a modern instrument compared to this one. I know RCA quit making TEM's a LONG time ago, but can anybody point me in the right direction to begin getting some info on this scope.
Thanks, Bill -- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
----------------------- Headers -------------------------------- Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: from rly-yb03.mx.aol.com (rly-yb03.mail.aol.com [172.18.146.3]) by air-yb01.mx.aol.com (v59.4) with SMTP; Tue, 13 Apr 1999 20:48:16 -0400 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by rly-yb03.mx.aol.com (8.8.8/8.8.5/AOL-4.0.0) with SMTP id UAA10462; Tue, 13 Apr 1999 20:48:04 -0400 (EDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA12287 for dist-Microscopy; Tue, 13 Apr 1999 17:23:07 -0500 Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id RAA12281 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 13 Apr 1999 17:22:35 -0500 Received: from hermes.services.ou.edu (hermes.services.ou.edu [129.15.2.121]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id R
That is amazing!!! Keep the software and the camera part, throw away the microscope and use on any larger microscope. I was considering getting one of the Mattel Barbie cameras for $69 to gut for the active electronics. Actually I was going to get 2 of them one for permeant mounting to a microscope and the other I was going to kluge into one of my old Nikon F bodies...... Ed Sharpe
} Subj: Student Microscopes } Date: 4/13/99 3:03:09 PM US Mountain Standard Time } From: mishot-at-itsa.ucsf.edu (Larry) } To: microscopy-at-sparc5.microscopy.com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Check with your local Mattel toy store for the recently released "Play X3 } Digital Video Microscope" with a list price of $99. This is a joint venture } with Intel and is CMOS based. It is designed to let children of all ages } display images from the microscope on a PC at both still and video rates, } print images and email them. Mattel/Intel will also use similar technology } for their Me2Cam. Naturally the results will not compare with the $100,000. } instrumetns we use in research or will they? } Larry D. Ackerman } Lily & Yuh Nung Jan Laboratories } Howard Hughes Medical Institute } UCSF, Box 0725, Rm U226 } 533 Parnassus Ave. } San Francisco, CA 94143 } } (415) 476-8751 FAX (415) 476-5774 } mishot-at-itsa.ucsf.edu }
Just a reminder please post messages to the server to the address
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do not post messages to
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That addrress (listserver-at-msa.microscopy.com) is the administrative site for problems and/or subscribe/unsubscribe requests. All mail to that address comes directly to me, not to the general mailing list.
I always forward mail that arrives there incorrectly and most of this is do to USERS simply sending to the wrong address. I don't mind occassional errors, but lately ALOT of you have been posting to the wrong address. It just creates more work for me and delay's your messages. So please check your address books etc.. and make sure you have the addresses correctly recorded.
I would like to get a new Critical Point Drier. Would you be kind enough to share your opinions on and experiences with different models ? So far, I am in favour for a EMS 850 CPD, but nobody here knows anything about this model.
You are welcome to write me off-line if you would like me to treat your reply confidential.
Yours sincerely
PETER FUNCH Assistant Professor, Ph.D.
Dept. of Zoology - Institute of Biological Sciences University of Aarhus Universitetsparken - Building 135, DK-8000 Aarhus C - Denmark Phone: + 45 8942 2764 - fax: + 45 8612 5175 E-mail: peter.funch-at-biology.aau.dk *************************************************************
Bill, Why not hook it up and get it running, at least to some demonstrable extent. One of the worst things about renovating an older instrument is finding a place to put it. If you have a location, I recommend pursuing the necessary facilities for operation. Put your graduate students to work. A great opportunity for them. Jerry ______________________ Jerome D. Schick, Ph.D. Semiconductor Devices and Electron Microscopy 26 Kuchler Drive LaGrangeville, NY 12540 Bus (914)223-7393 FAX (914)227-2743 jdschick-at-worldnet.att.net
COURYHOUSE-at-aol.com-at-Sparc5.Microscopy.Com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have inherited an OLD RCA TEM minus the HVPS. I have what I think is } } the serial number (the ID plate is probably on the HVPS). We have no } } intention of trying to make it operational (we will clean it up and put } } it in static display), but I would like to find out the model number, } } about how old it is, and whatever else I can dig up on it. I used an old } } RCA 4 (I think) back in about '72 and that was a modern instrument } } compared to this one. I know RCA quit making TEM's a LONG time ago, but } } can anybody point me in the right direction to begin getting some info } } on this scope. } } } } Thanks, } } Bill } } -- } } ============================================================= } } Bill Chissoe III } } Electron Microscopist } } University of Oklahoma } } 770 Van Vleet Oval } } Norman, Ok. 73019 } } E-mail: wchiss-at-ou.edu Ph. (405)325-4391 } } ============================================================= } } } } } }
ALL ARE WELCOMED TO ATTEND CMS members-please let everyone know because our listserver is down and this will be the only announcement before you receive the newsletter! } "SPRING DINNER MEETING: APRIL 27" } } This year we have decided that instead of the winter conference we would } have an additional dinner meeting. It is with great pleasure that we } have Dr. Jennifer Lippincott-Schwartz talking to use this spring at } second dinner meeting about membrane trafficking and protein transport } in the cell using GFP tagged proteins. } } Place: Tia Queta } 4839 Del Ray Ave. } Bethesda, MD } 301-654-4443 } } Date: April 27, 1999 (Tuesday) } } Time: 6:00 - 6:30 appetizers } 6:30 - 7:30 dinner: } red snapper,tenderloin and chicken } 7:30 - 8:30 speaker } } Cost: $25/person, $15/student } } RSVP by April 24 to: Jenny Hinshaw } 301-594-0842 } jennyh-at-helix.nih.gov } } Directions: From 495 take 355 or Wisconsin Ave exit off } 495 and head south toward Bethesda. After you pass NIH follow the } directions below. } } From NIH take Wisconsin Ave south to Woodmont Ave. Take a right } onto Rugby (soon after you veer right onto Woodmont) and then an } immediate left onto Del Ray. The restaurant is down a block on the } right. If the street parking is filled, they have valet parking for $3 } or public parking further down (a block) on Del Ray on the right. Also } there is public parking on Woodmont right before you turn onto Rugby. } } } } }
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i do very low volume CPD work, and have had good luck with a samdri pvt-3b manually operated system. it was cheap and seems to be pretty rugged, and most importantly, easy to operate.
good luck
b-
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
716-275-3058 716-244-4936(fax)
"You may get to the top of the ladder of success only to find its been leaning against the wrong wall" A. Raime
Lately there has been a lot of responses about Scanners for TEM negatives and they seem to have all been centering around the flat-bed type. Does anybody have any comments about drum scanners, for example the JEI eX4, or the Imacon Inc. FlexTight Precision II? I know that typically drum scanners are more expensive as a rule, but do any of you use these? We are considering something along those lines. We currently have the LeafScan45, but it is feeling it's age and we want to be prepared for it's eventual end.
Thanks in advance for your comments.
Peggy Bisher.
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
} ...the Imacon Inc. FlexTight Precision II?...do any of you use these?...
A collaborator and I had been using an ancient Perkin-Elmer scanning microdensitometer (circa late 70's) which provided 2500 dpi and gave good images, but we grew weary of its slowness (~30 min/ for a 1" x 1" scan) and lack of support. We compared the output of the Imacon with that of the Perkin-Elmer and a Polaroid Sprintscan and were quite pleased with the Imacon, so my collaborator bought one. He has since moved to Richmond, and as I have not heard otherwise, I presume that the Imacon is performing well.
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Larry -
I don't have a "local toy store" here on the coast, but I need to check this out immediately for Project MICRO. Can you supply me with any kind of contact info? Have you seen this thing?
Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
We have just had a cold stage installed on our SEM capable of obtaining near -200C. Although our intentions are primarily for cathodo-luminescence and would enjoy discussing other issues and techniques, this e-mail primarily addresses sample preparation and mounting standard thinsections on the cold stage.
The current specimen hold offers too little area for even a 1" round TS, so we have in mind machining a platform of polished brass. The question here is with respect to the glass slide and holder. That is, is there a viscous medium which will hold the TS in place and provide good thermal conductivity? ... and at the same time be easily cleaned for care of the carbon coat.
If a thinsection were to be made from scratch with the cold stage in mind, is there an especially thermally conductive epoxy?
TIA and cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
You will see more on CMOS cameras. There was an ad, in Advanced Imaging I believe, which I can't find now, that offered CMOS cameras for about $50 in bulk.
In the meantime, www.supercircuits.com offers CCD "spy" cameras for about $150.
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
We have a Reichert-Jung 2050 microtome and it needs service. Can anyone provide me a phone number or an email address of a service company in or near Oklahoma state so that I can contact them. Thanks
X.S. Ding Plant Biology Division The Noble Foundation Ardmore, OK 73401 Tel. 580 223-5810
I am upgrading from this 1600T to an 1830 and have priced this workhorse to sell. It is a turbo pump unit with ion pump to handle W or LaB6 emitters (both heads are included). A vibration isolation platform is also included. The scope is on-line and operational. You can see it listed on LabX at:
http://www.labx.com/aref.cfm?ad=19348
with an advertised price higher than my current value.
There are numerous pictures at http://www.gaugler.com/1600T
Dear Colleagues: } } The Core EM Facility at Dana Farber Cancer Institute ( Boston, MA, } USA) has the following equipment available immediately: } } 1. JEOL JEM-100 CXII transmission electron microscope, Side-entry } w/power supply, pumps, and water chiller. } } 2. JEOL JEM-100 CXII transmission electron microscope, Top entry } w/power supply, pumps, and water chiller. } } 3. JEOL JSM-35 CF scanning electron microscope } } 4. Reichert-JUng Ulrtacut E ultramicrotome w/FC4D cryochamber } attachment, Nitrogen tank, vibration-free table. } } 5. MT 5000 Sorvall ultramictotome with cryoattachment. } } 6.Ladd critical point dryer } } 7.Polaron SEM coating system } } 8.Durst Laborator s -45 special enlarger system } } 9.Agfa 3700 printing machine } } All items are in excellent working conditions. Please contact } Dr.Yuhui Xu by email. No telephone call please. } } Regards, } } Yuhui Xu, MD,PhD } EM Core, DFCI, } Boston, MA 02115 }
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} ... in a container eventually on its way to Taiwan as scrap mental
Hey, great! Does Taiwan really take this stuff? I have file drawers and file drawers full of scrap mental I ought to get rid of. Don't think any of it's actually toxic, but some of it was really dumb.
Now if I could only rid my brain of all those beer and soap jingles from the 60's and 70's life would be grand indeed.
MME offers customized, on-site workshops in these areas. Give me a call or email me directly and I can provide details.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 01:52 PM 4/13/99 -0500, leslie.link-at-US.GTC.BOC.COM"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Bottom line: there is no comparison. While the two sound similar in functionality, the optical and electronic resolution and contrast are not even in the same universe. Then we could talk about image size, transfer speed, etc.
We have a unique and challenging situation rising to critical level here: As microscopists, we need to know how our instruments work (do you know that most pathologists, for example, don't even know what Koehler illumination is, yet alone how to do it?!) as well as the intricacies of our own scientific disciplines. Now we also need to become fluent in the electronics behind cameras and all the hardware/software issues involved in computer technologies. Sorry guys, but as much as the manufacturers are trying to make things simple and "transparent to the user", we need at least a basic understanding of the principles behind each of the components in our systems. I am in the process of writing an article on "Microscopy for the Next Millenium" and the research for that article has pointed up this need, in spades!f
For those of you who could use a refresher on basic principles in microscopy and digital imaging, a number of these issues are addressed in "Optimizing Light Microscopy". Details are on our website.
As always, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 12:11 PM 4/13/99 -0700, Larry wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
MME offers customized, on-site workshops in these areas. Give me a call or email me directly and I can provide details.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 01:52 PM 4/13/99 -0500, leslie.link-at-US.GTC.BOC.COM"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ---------- } From: McLean, Dorrance } Sent: Thursday 715, 24199997 8:37 AM } To: 'Microscopy Listserver' } Subject: TEM Samples of Titania } } Dear All, } } I'm in need of some advice regarding a sample I've been asked to prepare. } The sample is Ti O2 and I have been trying to get a good thin sample for } viewing in the TEM. } } So far I've had a lot of cracking during disc cutting and dimpling. I've } learned that it's better to thin the sample to about 300 microns prior to } using the disc cutter and I'm getting good results with that. I then thin } the sample to around 150 microns using diamond films. (only from the } backside as there is a film on the sample.) I have also found from trial } and error that the Bronze wheel on the dimpler causes too much damage to } the sample so I've tried thinning with the Felt polishing wheel and } diamond paste but it's hard to measure the dimple progress. I have } stopped thinning at around 30-40 microns as the thinner samples have all } cracked on the dimpler. } } I have managed to get two samples into and out of the PIPS. Of course the } "perfect" sample took a suicide leap from between the jaws of my tweezers } just as I was loading it into the specimen holder of the microscope...so } I'll never know just how perfect it was. The second sample was amorphous } at the thin areas and I suspect that (1.) I may not have removed all the } glue from the sample, or (2.) maybe the sample was too thick and sputter } was redeposited onto the sample. } } Either way I'm stumped and I would sure welcome some advise. } } Thanks for your time. } Dorrance } }
} From Microscopy-request-at-sparc5.microscopy.com Thu Apr 15 13:57:33 1999
} Date: Thu, 15 Apr 1999 13:26:59 -0400 } To: Larry {mishot-at-itsa.ucsf.edu} , {microscopy-at-Sparc5.Microscopy.Com} } From: Barbara Foster {mme-at-map.com} } Subject: Re: Student Microscopes } } Hi, } } Bottom line: there is no comparison. While the two sound similar in } functionality, the optical and electronic resolution and contrast are not } even in the same universe. Then we could talk about image size, transfer } speed, etc. } } We have a unique and challenging situation rising to critical level here: } As microscopists, we need to know how our instruments work (do you know } that most pathologists, for example, don't even know what Koehler } illumination is, yet alone how to do it?!) as well as the intricacies of } our own scientific disciplines. Now we also need to become fluent in the } electronics behind cameras and all the hardware/software issues involved i } computer technologies. Sorry guys, but as much as the manufacturers are } trying to make things simple and "transparent to the user", we need at } least a basic understanding of the principles behind each of the component } in our systems. I am in the process of writing an article on "Microscopy } for the Next Millenium" and the research for that article has pointed up } this need, in spades! } } For those of you who could use a refresher on basic principles in } microscopy and digital imaging, a number of these issues are addressed in } "Optimizing Light Microscopy". Details are on our website. } } As always, } Barbara Foster } }
Barbara, I couldn't agree with you more. I work in the cathodoluminescence area and am amazed to see how many potential users understand all about color videos and associated computer wizardry but when you start to talk about the beam power hitting the sample and possible implications of heating and sample deterioration, they don't know what I'm talking about. And when you bring up beam power per unit area (beam power density) it can really draw a blank. It's too easy to accept that everything is fine just because a pretty picture emerges. The same thing occurs in mass spectroscopy. A set of notes on Fundamentals of Mass Spectroscopy that I wrote back in the mid 1980s make almost no mention of computers but they do discuss the details of electron impact and why one often uses a 70 electron volt energy for the electron beam, etc.; Many users are astonished that they might need to worry about occasionally checking or, heaven forbid, setting such variables. But almost universally when people do see this basic physics information available in a readable format in these notes, they are very receptive and seize upon it.
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology "A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
I have not been able to track down the Mattel microscpe yet. It is not on their website, the local Toys R Us store does not have it and the Mattel Merchandising Manager does not return my calls. I assume this is another example of publicity preceding product. I saw press releases in the San Francisco Examiner newspaper some time ago (?March) and last week in the new issue of Advanced Imaging. When I locate the actual product I'll post more information.
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
Absolutely no disc cutter. Nor or at most just a little bit dimpler. A disc-grind holder will work. Specifically, you grind any irregular piece down to as close to a thickness of {=20um as possible with the help of the holder. Break it using, for example, a hammer. Pick up a decent piece or several pieces with maximum diameter(s) {3mm, and glue it(them) to a(several) single-slot grid(s) using, say, regular SEM carbon paint or silver paint. And then put the specimen on your PIPS miller. Usually, you should mill for {10 hours. } 10 hour milling time may quite possibly indicate a preparation failure. There is another method called small-angle-cleavage method you may want to try. But that's a territory of Dr. Walck Scott ("Walck. Scott D." {walck-at-ppg.com} ). You can contact him directly. Good luck.
-cy
On Thu, 15 Apr 1999, McLean, Dorrance wrote:
} Date: Thu, 15 Apr 1999 12:07:33 -0600 } From: "McLean, Dorrance" {dmclea-at-sandia.gov} } To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} } Subject: FW: TEM Samples of Titania } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } ---------- } } From: McLean, Dorrance } } Sent: Thursday 715, 24199997 8:37 AM } } To: 'Microscopy Listserver' } } Subject: TEM Samples of Titania } } } } Dear All, } } } } I'm in need of some advice regarding a sample I've been asked to prepare. } } The sample is Ti O2 and I have been trying to get a good thin sample for } } viewing in the TEM. } } } } So far I've had a lot of cracking during disc cutting and dimpling. I've } } learned that it's better to thin the sample to about 300 microns prior to } } using the disc cutter and I'm getting good results with that. I then thin } } the sample to around 150 microns using diamond films. (only from the } } backside as there is a film on the sample.) I have also found from trial } } and error that the Bronze wheel on the dimpler causes too much damage to } } the sample so I've tried thinning with the Felt polishing wheel and } } diamond paste but it's hard to measure the dimple progress. I have } } stopped thinning at around 30-40 microns as the thinner samples have all } } cracked on the dimpler. } } } } I have managed to get two samples into and out of the PIPS. Of course the } } "perfect" sample took a suicide leap from between the jaws of my tweezers } } just as I was loading it into the specimen holder of the microscope...so } } I'll never know just how perfect it was. The second sample was amorphous } } at the thin areas and I suspect that (1.) I may not have removed all the } } glue from the sample, or (2.) maybe the sample was too thick and sputter } } was redeposited onto the sample. } } } } Either way I'm stumped and I would sure welcome some advise. } } } } Thanks for your time. } } Dorrance } } } } } }
Whoever finds one of these new wunderkund toys-- please post an image on your website so we all can observe what kind of image the next generation of microscopists is seeing.....
} I have not been able to track down the Mattel microscpe yet. It is not on } their website, the local Toys R Us store does not have it and the Mattel } Merchandising Manager does not return my calls. I assume this is another } example of publicity preceding product. I saw press releases in the San } Francisco Examiner newspaper some time ago (?March) and last week in the } new issue of Advanced Imaging. When I locate the actual product I'll post } more information. } } } Larry D. Ackerman } Lily & Yuh Nung Jan Laboratories } Howard Hughes Medical Institute } UCSF, Box 0725, Rm U226 } 533 Parnassus Ave. } San Francisco, CA 94143 } } (415) 476-8751 FAX (415) 476-5774 } mishot-at-itsa.ucsf.edu
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Chairman, Educational Outreach subcommittee promoting access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
I have a Brinkmann AS2 microscope I would like to know more about. It appears to be Leitz or a copy of one. Every thing on it has matching serial numbers and the oculars are marked 10x pl. It looks like it was made in the late 50's or early 60's. Talking to Brinkmann they say it was made for them by Zeiss. Talking to Ziess they say they never made scopes for anybody but themselves. It does say made in Germany.
The field is very flat and I don't see any distortion on the edge so I expect the pl mean plan optics. The oil immersion lens is a 160 100/0/1.3.
Pictures or it are at www.couger.com/auction/scope.
I bought the scope because I have a triocular head that fits it.
If anyone could help me with more information about the scope or copy of a manual I would pay unreasonable copying and shipping cost.
Thanks Gordon
Gordon Couger gcouger-at-couger.com www.couger.com/gcouger Stillwater, OK 405 624-2855 GMT -6:00
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Actually, the brand and model of SEM has some degree of influence over the depreciated value of the scope. For example: Hitachi: 500, 520, 570 are boat anchors. Hitachi: 800, 900 are FE scopes and are more modern. Hitachi: 2960 and 3000+ are really more modern and can include the Nature option and are very good scopes.
A JEOL 840 series is good.
Philips? maybe a XL40FE. But as with any field emission scope, one must be willing to put up with the extra grief of FE over LaB6. Yet, there is an image intensity benefit. It is a tradeoff.
gg
At 10:03 PM 4/15/99 , you wrote: } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } The point you make here, of course, is quite correct. Our fifteen year old } Model 840, set up and working, in our laboratory, has far more value to us } than what is showing on our books. } } With regard to the depreciation of AMRAY instrumentation, it sounds like } those instruments might be depreciating faster than say, JEOL or Philips } equipment. } } Chuck } -------- REPLY, Original message follows -------- } } } Date: Thursday, 15-Apr-99 06:17 PM } } } } From: Dr. Gary Gaugler \ Internet: (gaugler-at-calweb.com) } } To: Garber, Charles A. \ Internet: (cgarber-at-2spi.com) } } } } Subject: Re: Value of SEM } } } } At 12:12 AM 4/15/99 , you wrote: } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } Hi Gary: } } } } } } I think that you have to make some distinction between the a) depreciated } } } value vs. b) market value. There two can be very much different. } } } } } } We depreciate most of our equipment over five years, our computers we } } } depreciate even faster. After five years it has zero value on our books. } } } } But if you wanted to purchase it from us, we would seek to charge you } full } } } market value. And if it meant there was some recovery of earnings, then } we } } } would pay taxes on those recovered earnings. } } } } } } But that itself have meaning only if the company has earnings on which } taxes } } } have to be paid. If a firm is operating at a loss, then when they } recover } } } those funds, they do not pay any taxes. And even a highly profitable } } } company like DuPont or IBM often times try very aggressively to sell } their } } } depreciated equipment for as much as they can get for it, and to whatever } } } degree they might recover assets, they pay their taxes, but are still } better } } } off than they would be had they just given it away. } } } } } } I do know however of certain companies that will give away equipment } rather } } } than selling it, under the premise that if someone is injured at some } later } } } date, they would then have less exposure to those claims. } } } } } } Chuck } } } } } } Each company probably has a different cost/benefit scenario and thus a } } different view of the salvage value of a SEM at the end of the } depreciation } } period. Nevertheless, my point was that SEMs drop in value really fast. } And } } by about 6-8 years, they are near worthless in monetary terms. The cost } of } } takedown, setup and moving overshadow the actual cost of the machine. I } just } } bought an Amray 3600LEAP that is 1.5 years old for $150K when the new } price is } } $422K. I also bought an 8 year old Amray 1830 for $8K. In each case, the } cost } } of take down and setup by Amray is about $7K. The cost of moving is about } } $3500 each. So for the older unit, the handling and shipping is more than } cost } } of the instrument. } } } } } } Cheers, } } Gary Gaugler, Ph.D. } } } } -------- REPLY, End of original message -------- }
I am about to do some estimates of our SEMs resolution under different conditions. I am intending on using gold particles of varying size for this task. I intend to analyse the change in intensity as the beam traverses a particle. I am worried about the subjective nature of such a test and would like to find a more objective procedure. Can anyone advise on other methods?
Question: How can serial sections be picked up onto a formvar/carbon coated large slot grid? So far I have had problems with wrinkling of sections, sections drying down onto metal instead of remaining in the slot window.
Message somehow bounced back. I am re-sending it. Sorry if you receive a duplicate. A few more words:
If you just want to mill one side of the sample on your PIPS, the other side does need to be protected from re-deposition with a 3mm thin disc of mica (you can use your disc cutter his time). Mica has numerous layers. Peel a thin piece from the top with a pair of good twizers. Overall, Preparing a plan view specimen of this kind of material should not be as difficult as preparing Chinese foods.
Regarding the handling of the prepared specimens, great care is of course the 1st thing I can mention. There are some other tricks. But please excuse me for not being capable of describing them in English. You will get some sense after a while. Also, if possible try a specimen holder with a specimen screw cap instead of a specimen clamp. Good luck. Dont get too much frustrated.
-cy
---------- Forwarded message ----------
POSTDOCTORAL POSITION IN INTERFACE PHYSICS
DEPARTMENT OF PHYSICS, UNIVERSITY OF ILLINOIS AT CHICAGO
{smaller} A postdoctoral research associate position is currently available in the Interface Physics Group at the University of Illinois at Chicago (UIC) to perform atomic scale analysis of interfaces and defects in semiconductor heterostructures. The successful candidate is expected to work closely with the MBE and microfabrication groups at UIC investigating a wide variety of technologically relevant systems (including II-VI, III-nitride, III-V, SiO/SiGeO, SiOxNy/SiGeOxNy and magnetic multilayers). =20
Research in the Interface Physics Group focuses on the use atomic resolution imaging and analytical techniques in electron microscopy, coupled with theoretical simulations, to determine the structure-property relationships at internal interfaces on the fundamental atomic scale. Current research programs involve ceramics, high-Tc superconductors and optoelectronic/high-power semiconducting materials and devices. The experimental facilities to perform this research are comprehensive: a JEOL 2010 Field-Emission STEM/TEM featuring a 1.4=C5 probe size, "drift free" stage, high-angle annular dark-field detector (Z-contrast), Gatan Imaging Filter, and Noran EDS; a VG HB501A Field-Emission dedicated STEM with EDS, EELS and Auger spectrometers; a JEOL 3010 conventional TEM with digital imaging capabilities and EDS; a JEOL 6320 Field-Emission SEM with EDS; and a JEOL JXA733 microprobe. In addition to the electron microscopes, specimen preparation facilities include a Gatan Duo-mill, Fischione precision ion-mill, SouthBay plasma cleaner and Leica Ultramicrotome.=20 The Interface Physics Group has a Silicon Graphics R10000 Power Indigo workstation with the Molecular Simulations' Cerius 2 package incorporating the CASTEP pseudopotential code. The physics department has additional workstations and access to the UIC Convex Exemplar Supercomputer and the National Center for Supercomputing Applications at UIUC. =20
Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound background in the relevant materials issues and an ambition to be part of a developing program pushing at the frontiers of interface physics. Please send a resume and publication list to Professor Nigel D. Browning at the address below. Prior experience in STEM or TEM is essential. =20 However, consideration will be based on the candidates overall potential for success in the field and applicants with prior experience in related fields are encouraged to apply. Positions are for one year initially, normally renewed for a second year with possibilities existing for further years. Salary is commensurate with experience.=20 UIC is an equal opportunity employer.
I have spoken with an executive from Mattel who said that the X3 microscopes will not be out on the market before the fall of 1999.
Carlton Bowers TECH TREK Mobile Research Laboratory (937) 222-2934
Subj: Re: Student Microscopes
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I have not been able to track down the Mattel microscpe yet. It is not on their website, the local Toys R Us store does not have it and the Mattel Merchandising Manager does not return my calls. I assume this is another example of publicity preceding product. I saw press releases in the San Francisco Examiner newspaper some time ago (?March) and last week in the new issue of Advanced Imaging. When I locate the actual product I'll post more information.
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
by hms1.med.harvard.edu (Post.Office MTA v3.5.2 release 221 ID# 0-58538U3500L600S0V35) with ESMTP id edu; Fri, 16 Apr 1999 10:42:50 -0400
Dear Colleagues:
I have received quite a few inquires regarding to our posting about the used equipment available at Dana Farber Cancer Institute. The following is the information I could gather about those items:
I. The side entry TEM (JEOL 100 CXII) was purchased in 1988 ( purchase price $145,500). This is equipped with all standard features with additional specimen rotation and tilt correction device. The top entry scope was purchased a little earlier. The JSM-35 CF scanning electron microscope was purchased in 1982 ( purchase price of $75,900). All three scopes have been maintained under service contracts directly with JEOL. II. The Polaron sputter coater was purchased in 1983 (purchase price $6,180) with film thickness monitor, gold/palladium annular target, digital temperature indicator, and specimen holders.
I do not have information as to the exact time and price when the other items were purchased other than their model numbers. The Reichert microtome was purchased probably about 7-8 years ago. The MT5000 is probably 4-5 years older. As I said, they are all in excellent working condition.
As I understood from our administration, the institute would like to sell these equipment although it is also possible some of these items will be donated to non-profit organizations if no bids are received. Final decision will be made by our administration. Since I do not have pricing information, please feel free to make your reasonable offers and I will forward them to the institute. I will speak to our administration about possible arrangements for the demonstration of the equipment if necessary.
Thank you for your inquiries. Please feel free to email me if you have further questions. Best regards,
There is a press release with a brief description and a poor picture of the unit at: http://www.hottoynews.com/mattel1.htm Scott
} } I have not been able to track down the Mattel microscpe yet. It is not on } their website, the local Toys R Us store does not have it and the Mattel } Merchandising Manager does not return my calls. I assume this is another } example of publicity preceding product. I saw press releases in the San } Francisco Examiner newspaper some time ago (?March) and last week in the } new issue of Advanced Imaging. When I locate the actual product I'll post } more information. } }
-------------------note: new mailing address---------------------- Scott Wight e-mail: scott.wight-at-nist.gov NIST 222/A113 W voice: 301-975-3949 100 Bureau Dr STOP 8371 | fax: 301-417-1321 Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
Dear Gary, I certainly beg to differ. I would buy a Hitachi S-570 over a JEOL 840 of the same age any day. I did when I evaluated them new in 1986. The S-570 is still running perfectly and still meets spec easily. I have two S-570s, one recently purchased used. All the old Hitachi's run forever. You wrote:
} } Actually, the brand and model of SEM has some degree of influence over } the depreciated value of the scope. For example: } Hitachi: 500, 520, 570 are boat anchors. } Hitachi: 800, 900 are FE scopes and are more modern. } Hitachi: 2960 and 3000+ are really more modern and can include the Nature option } and are very good scopes. } } } A JEOL 840 series is good. } } Philips? maybe a XL40FE. But as with any field emission scope, one } must be willing to put up with the extra grief of FE over LaB6. Yet, there } is an image intensity benefit. It is a tradeoff. } } gg } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
I have made it a speciality of mine to study the performance of SEM all over the world. Running courses the first thing I need to know is how go= od is the instrument that I am using? For this you need a reliable standard=
that really tests the microscope. Any gold on carbon that I have used ha= s never been up to the task. It is quite possible to take photographs of this type of specimen and find that there is no difference in instrument performance from say 10 to 15kV, there must be a difference!
For this reason I developed a test specimen of my own which I have used f= or the past 18 years. The specimen is made up of polystyrene latex spheres that are allowed to dry from a liquid to form a solid white block. Durin= g the drying process, provided the latex preparation is free from contaminants, the spheres will deposit in an array that is of a square packing in one direction and hexagonal in the other. If a piece of the solid material is fractured, by pricking with a fine point, this opens up=
the internal structure of the compacted material, displaying the two type= s of array.
Having a very well defined structure the hexagonal arrays make a very goo= d subject for judging the performance of a scanning electron microscope. A= ny hexagonal area on the specimen is comparable with another set in the same=
orientation. Another advantage of this specimen is that the latex are of= a specific size which may act as an inbuilt calibration. In most cases of performance monitoring the operator simply needs to take a test picture a= t a specific magnification and use a comparative process to judge performance. The latex are a nominal 0.24um but when compacted in an arr= ay they are visibly reduced to about 0.2um.
Should the sample become damaged it is easily recovered by re coating or once again pricking it with a pin to open up new areas and then re coatin= g.
Making A Test Specimen
To convert the latex specimen into a high resolution test specimen a meta= l coating is required. A sputter coating will make the specimen conducting=
but further coats will build a sub structure on the surface of the sphere= s. The sub structure may be used for high resolution performance monitoring= . =
The level of sub structure desired will depend upon the capabilities of t= he instruments to be investigated. For instruments with a conventional tungsten source multi coating the latex with gold is satisfactory. For more advanced instruments the finer coating of gold-palladium may be more=
desirable.
The coating procedure depends upon the efficiency of the coater being use= d. Sputter coaters that use relatively high voltages (1 to 3kV) will requir= e the following procedure.
i. Set the coater at a 5cm target to specimen distance. ii. Sputter at 20mA, 1kV for one minute, wait one minute and repeat t= he process. iii. Coat for 4 one-minute periods and then check the specimen in the microscope. iv. If you need more coats, because you cannot see the metal, repeat the "coat and wait" procedure until the structure is satisfactory.
The more metal you put down the coarser the structure will be on the spheres. Low levels of coat will require better operating techniques in order to resolve the coat. Do not expect a conventional (W hairpin) instrument to be able to resolve less than 4 coats.
If you have a modern coating unit, which will be much more efficient at putting down the coat, use 10mA for 30 seconds per coat. Experimenting with coating procedures will enable you to tune the coating parameters an= d coating time to obtain the exact specimen that you require. For field emission instruments a gold-palladium coat, if carefully applied, will gi= ve you grains in the region of 8nm and a spacing of less than 1nm.
Operating Procedures
If you intend to push yourself and your microscope to near its limits the= re are some basic operating procedures that will be required. Firstly the specimen must be placed in the instrument and the high voltage must be switched on for at least 45 minutes prior to trying to work at high magnifications (} 30,000X). This period is required for the high voltage tank, and hence the high voltage, to reach stability. After this period the heat gained by the components is equal to the heat lost through the walls of the tank and the high voltage will be at its most stable.
Whilst stability is being achieved move the specimen to a short working distance ( {5mm) and set the instruments alignment to the best of your ability. Find areas of the specimen that are in the hexagonal array and flat to your view. A slight tilt of the array is not as good for comparison as is a perfectly flat surface; you may need to tune your specimen tilt slightly.
Hope this helps make your own and try it?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Chris asks ... } } } I am about to do some estimates of our SEMs resolution under } different conditions. I am intending on using gold particles } of varying size for this task. I intend to analyse the change } in intensity as the beam traverses a particle. I am worried } about the subjective nature of such a test and would like to } find a more objective procedure. Can anyone advise on other } methods? } } Chris Walker
The best method, or least subjective, would be to do a line scan perpendicular to a knife edge. This allows a measurement of signal intensity a function of distance, and you'd be able to quantifiably compare your different parameters by comparing the distance between two points ... e.g., at 20% maximum and 80% maximum.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I have two older SEM/EDS systems for sale. Where do I advertise? How do I get the word out most effectively? Any suggestions welcome. Is there a Web site where one can post SEM's for sale?
Dear Listers, For those who may be interested in upgrading an existing EDX system to a Windows NT, PC-based system, check out the Quartz Imaging web site for their lastest announcement: www.quartzimaging.com. I've tried a demo system and it's excellent.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
I would once again like to tap the collective expertise (and my apologies for not thanking each of you each time you help me out)!
A researcher is trying to fix isolated mitochondria (rat brain, I think) that have been subjected to different treatments just before isolation. He expects to see different degrees of swelling. Being extremely conservative about data, I need to make sure the swelling we see is not due to osmotic stress, among other artifacts. Does anyone have a favorite recipe for isolated mitochondria? So far 2.5% glut in 0.1M cacodylate, postfixation with 1% OsO4 in cacodylate, and en bloc uranyl acetate in the 50% ethanol dehydration step seems adequate, but not great.
Thank you all in advance!
Aloha, Tina
No surf today. **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
If I understand you correctly, you want to measure the resolution of the image, i.e., the finest detail that can be resolved with the microscope, not its reproducibility or linearity.
Perhaps you can take a clue from the way this is done on TEMs:
On a TEM you acquire an image of an amorphous material. The Fourier Transform of the image is then either calculated or produced by using an optical diffractometer. This FT then shows the frequency distribution of the image. At the center you have the low frequency information, i.e., the slowly varying brighness changes, and the further out you go, the higher the frequencies. In a TEM, this information is modulated with the contrast transfer function, leading to rings of intensity. By including a little bit of crystalline material with a known lattice constant, this FT image can be calibrated and the radius determined, at which the information (or intensity in the FT image) disappears. This radius is the inversly proportional to the smallest distance that can be resolved.
For an SEM you will need a sample that shows structure or features around the expected resolution limit. Then take an image and calculate the FT. This is best done on a computer. Make sure, that the pixel size is at least a factor of two smaller than the expected resolution or you will see artifacts from pixelation.
The images from the SEM will not show the rings from the CTF, but by doing a radial integration of the FFT you should be able to determine the point, where the intensity in the FFT has gone down below a threshold. You can then use this graph as a measure of your resolution.
Good luck.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
} ---------- } From: Chris Walker[SMTP:chris.walker-at-physics.org] } Sent: Friday, April 16, 1999 1:30 PM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: SEM: Resolution tests } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } I am about to do some estimates of our SEMs resolution under different } } conditions. I am intending on using gold particles of varying size for } this } task. I intend to analyse the change in intensity as the beam } traverses a } particle. I am worried about the subjective nature of such a test and } would } like to find a more objective procedure. Can anyone advise on other } methods? } } Chris Walker }
Dear Chris, The best way to test resolution is to take your best picture at each operating condition and see how small a feature you can resolve. That is the definition of resolution. I actually examine the Polaroid negative with a 10X magnifyer and measure the smallest gap between gold particles on a carbon substrate that I can see (at about 50,000 or 100,000X mag.). Divide by the mag to get the resolution in nm. You need a sample with a variety of gold particle sizes on it. You wrote:
} } I am about to do some estimates of our SEMs resolution under different } conditions. I am intending on using gold particles of varying size for this } task. I intend to analyse the change in intensity as the beam traverses a } particle. I am worried about the subjective nature of such a test and would } like to find a more objective procedure. Can anyone advise on other } methods? } } Chris Walker } Best of luck, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Hi Sheila R. I happen to publish Microscopy Today, a publication which is sent 10 times a year to over 8,400 microscopists in the U.S. - each of which who has requested a no cost subscription. I do have an used equipment section - charging a modest $25 plus a dollar a word, with a $50 minimum. I also publish employment opportunities - also with modest rates. Kindly advise by return email should you be interested in this service. Others might comment to you on any value they may feel with my pub. Best, Don Grimes, Microscopy Today
Dear All: I have a question on the problem of Kevex EDS system which I have here since November, 1997. The system is with a thin window detector and attached to the Topcon SEM. The problem is that sometimes the peak position in a spectrum is found shifted around 300 eV downside from the right position. It happens anytime, even after calibration, or after acquiring data from standard specimen for the qualitative or quantitative analysis. The higher the energy is, the larger the amount of shifting is. The peak width stays thin but sometimes become broad. When I change the bias voltage of the detector from 500 to 450 or 400 V, the shifting is getting worse or better, without specific direction. Service man from the Kevex has been trying to solve the problem for seven or eight months, but still they could not find the source of problem. It could be FET in the detector, preamp, analyzer, cable, plug, or whatever. The most difficult thing is that the shifting problem does not happen all the time. It disappears all of a sudden and does not occur for a few days, and shows up again, driving me and service men crazy. Does any of you have experienced this kind of shifting problem? Could you figure out the cause of the problem? It is really a nuisance and I I want to get rid of it ASAP. Any information will be appreciated. Sincerely, Jondo Yun, Ph.D.Department of Inorganic Materials Engineering Electron Microscopy LaborotaryCenter for Instrumental AnalysisKyungnam University449 Weolyeong-dong, Masan, 631-701, Korea82-551-249-2697 (tel)82-551-248-5033 (fax)jdyun-at-hanma.kyungnam.ac.kr
It looks like something in the feedback loop is flaky. The fact that changing the detector bias sometimes affects the shift makes it look like something in the front end. The most obvious cause would be something wrong with the feedback capacitor in the JFET package, maybe a dust fiber flipping back and forth under the influence of charging. The feedback capacitor is on the order of 0.05 picofarads, so it wouldn't take much of a piece of lint to change the gain by 300 eV.
best regards mark
Mark W. Lund, PhD VP Engineering } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"This is a YOUNG business...How can I tell you what YOUR job is when I don't know what MINE is?" --Pogo
Jundo wrote:
Dear All: I have a question on the problem of Kevex EDS system which I have here since November, 1997. The system is with a thin window detector and attached to the Topcon SEM. The problem is that sometimes the peak position in a spectrum is found shifted around 300 eV downside from the right position. It happens anytime, even after calibration, or after acquiring data from standard specimen for the qualitative or quantitative analysis. The higher the energy is, the larger the amount of shifting is. The peak width stays thin but sometimes become broad. When I change the bias voltage of the detector from 500 to 450 or 400 V, the shifting is getting worse or better, without specific direction. Service man from the Kevex has been trying to solve the problem for seven or eight months, but still they could not find the source of problem. It could be FET in the detector, preamp, analyzer, cable, plug, or whatever. The most difficult thing is that the shifting problem does not happen all the time. It disappears all of a sudden and does not occur for a few days, and shows up again, driving me and service men crazy. Does any of you have experienced this kind of shifting problem? Could you figure out the cause of the problem? It is really a nuisance and I I want to get rid of it ASAP. Any information will be appreciated. Sincerely, Jondo Yun, Ph.D.Department of Inorganic Materials Engineering Electron Microscopy LaborotaryCenter for Instrumental AnalysisKyungnam University449 Weolyeong-dong, Masan, 631-701, Korea82-551-249-2697 (tel)82-551-248-5033 (fax)jdyun-at-hanma.kyungnam.ac.kr
Mattel had made an error in my email address which delayed the following press release:
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**Microsoft and Windows are either registered trademarks or trademarks of Microsoft Corporation in the U.S and/or other countries ***Intel is a registered trademark and Intel Play is a trademark of Intel Corporation
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Hi, I usually don't prepare the slot grids with the formvar/carbon film in the hole before picking up sections. Instead, I precoat the slot grids with formvar - making them somewhat hydrophobic - and then pick up the sections. The sections float in the hole. They are then laid down on formvar in holes of a "bridge" to dry down. The bridge is a bent aluminum structure that has holes drilled in it that a sheet of formvar is laid over. One collects the formvar floating on water with the bridge rather than laying grids on it. Once dry, the grids with sections can then be laid on the formvar suspended in the bridge holes, and carefully removed later (without tearing the formvar on the grid - takes some practice).
The bridges can be made, or ordered from one of the usual EM vendors.
On Fri, 16 Apr 1999 08:19:22 -0600 ckuzmiak-at-chuma.cas.usf.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Colleagues ..... } } This one is out of my area of expertise... Please reply direct to } the sender... } } Email: ckuzmiak-at-chuma.cas.usf.edu } Name: Carolyn Kuzmiak } School: USF } } Question: How can serial sections be picked up onto a formvar/carbon } coated large slot grid? So far I have had problems with wrinkling of } sections, sections drying down onto metal instead of remaining in the } slot window. } } --------------------------------------------------------------------------- } }
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu ********************************************
Dear sir i am Dr s.k. jatty associated with the r&d division of the cement group of The A.C.C. ltd, mumbai, india working on the field of cement and concrete microscopy.
I would like to have some information about the laboratories and institutios all obver world working inthis particular field. Hope u will reply positively.
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Tina Carvalho wrote: } } I would once again like to tap the collective expertise (and my apologies } for not thanking each of you each time you help me out)! } } A researcher is trying to fix isolated mitochondria (rat brain, I think) } that have been subjected to different treatments just before isolation. } He expects to see different degrees of swelling. Being extremely } conservative about data, I need to make sure the swelling we see is not } due to osmotic stress, among other artifacts. Does anyone have a favorite } recipe for isolated mitochondria? So far 2.5% glut in 0.1M cacodylate, } postfixation with 1% OsO4 in cacodylate, and en bloc uranyl acetate in the } 50% ethanol dehydration step seems adequate, but not great. } Dear Tina, We have had good luck with high-pressure freezing. We have examined unstained, frozen-hydrated mitos on the 400 kV instrument here. Although this procedure requires instruments not available at all facilities, I expect to get minimal artefact. Since we are a NIH resource, investigators can use our facilities for free, so if your colleague wants to try this, but doesn't have access to the requisite equipment, (s)he can apply to use our resource by accessing our web site, www.wadsworth.org/bmirr Yours, Bill Tivol
In a message dated 4/19/99 6:10:52 AM Hawaiian Standard Time, { { } Email: ckuzmiak-at-chuma.cas.usf.edu } Name: Carolyn Kuzmiak } School: USF, writes: } } Question: How can serial sections be picked up onto a formvar/carbon } coated large slot grid? So far I have had problems with wrinkling of } sections, sections drying down onto metal instead of remaining in the } slot window. } }
A technique I have used is to pick the secions out of the water trough using a thin loop (I made mine of .008 gold/paladium wire for no better reason than that is what I had in the lab!). The loop should have a diameter of less than 3mm. 2mm works well.
If the section ribbon is teased away from the knife edge the loop can be lowered from the top and will pick up a "film" of water plus sections.
A slot grid, with a formvar/carbon support film is used. The grid needs to be held firmly during the transfer process of the secions from the loop. I actually use an old Hitachi specimen holder where the grid is secured on top of a 3mm dia peg with a retaining cap. This kind of setup can easily be made in the workshop. The grid can also be held by the adhesive applied to "gridsticks" (available from EM supply companies).
The loop is lowered onto the filmed surface of the grid and the water drop with sections transfers readily to the support film.
Treating the filmed grids with poly-L-lysine beforehand also helps. ( Apply a drop of .01% poly-L-lysine to the filmes surface of the grid. After 5 mins blot off with filter paper applied to edge of grid. Allow grid to dry at room temperature or more quickly in an oven at 60 degrees centigrade.)
Does anybody have ever used BALTEC RES 100 for TEM (or SEM) specimen preparation? any comment ? I am carrying out a comparison with PIPS (Gatan) and LAMP 1010 (Fischione) in order to select the equipement to be purchased by my institute. We need it to prepare inorganic samples for TEM observations, mainly Materials Science research activities.
Any kind of information will be very useful. Thank You all in advance, Edoardo Bemporad
Does anybody have ever used BALTEC RES 100 for TEM (or SEM) specimen preparation? any comment ? I am carrying out a comparison with PIPS (Gatan) and LAMP 1010 (Fischione) in order to select the equipement to be purchased by my institute. We need it to prepare inorganic samples for TEM observations, mainly Materials Science research activities.
Any kind of information will be very useful. Thank You all in advance, Edoardo Bemporad
-----Original Message----- } From: THE A.C.C. - RCD - THANE [mailto:rd3-at-bom3.vsnl.net.in] Sent: Monday, April 19, 1999 4:34 PM To: Microscopy-at-Sparc5.Microscopy.Com
Dear sir i am Dr s.k. jatty associated with the r&d division of the cement group of The A.C.C. ltd, mumbai, india working on the field of cement and concrete microscopy.
I would like to have some information about the laboratories and institutios all obver world working inthis particular field. Hope u will reply positively.
kindly mail me " jattysk-at-hotmail.com"
thanking you with regards
Dr s.k.jatty
Dr. Jatty,
I suggest contacting the International Cement Microscopy Association, 1206 Coventry Lane, Duncanville, TX 75137, USA. Membership is free, and meetings are held in the spring once a year at a North American city. This organization covers all aspects of cement and concrete microscopy.
George Polkowski Lab R&D Center Saudi Aramco Dhahran, Saudi Arabia
1. Measuring resolution by measuring the wave-form across an edge is=
not very constructive in the real world. Designed to monitor spot size,=
with the assumption that the resolution attainable is similar to the smallest spot, this technique is messy and in no way does it develop in a= n operator for the correct procedures to attain maximum performance.
Visual indications of performance or lack of it are very important when operating at the limits of the instrument at a particular kV. Using "normal" images for performance testing helps the operator develop a feel=
for the instrument, thereby sensing that they could use a smaller spot size, or a higher emission current, a better astigmatism correction or better focus. Not only do you test the instrument but you also test and develop the operator. Reasons why the South African Microscopical Societ= y are looking at a basic "quality" procedure for EM nation wide.
2. The TEM method of using an optical diffractometer is very interesting. I have used it many times for TEM and it would work for SEM=
negatives, one problem is that the biggest users of SEM are in industry a= nd few will have access to a diffractometer.
Just a few thoughts on an interesting but to many rarely discussed question, or is it?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Does anybody have ever used BALTEC RES 100 for TEM (or SEM) specimen } preparation? any comment ? I am carrying out a comparison with PIPS (Gatan) } and LAMP 1010 (Fischione) in order to select the equipement to be purchased } by my institute. We need it to prepare inorganic samples for TEM } observations, mainly Materials Science research activities. } } Any kind of information will be very useful. Thank You all in advance, } Edoardo Bemporad
Dear Edoardo Bemporad,
we use the BAL-TEC RES 100 about one year and mainly for TEM sample preparation of semiconductor heterostructures (SiC/Si, AlN/Si, ...). But I have no experiences with the Gatan and the Fischione equipment. What kind of information are useful for you about the RES 100?
Dr. Jatty, The International Cement Microscopy Association has a web site at http://www.cemmicro.org/ Greg
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
Our lab is removing the basement membrane from sheets of tissue containing Bruch's membrane. We want to verify that the basement membrane has been removed. We would like to do this by labeling pieces for the presence of basement membrane. Positive label would indicate unsuccessful removal. Is there a specific label for basement membrane that would not label underlying material such as the collagen of Bruch's membrane?
I have following recipe to mix the four components for EPON 812. For 25grs we use 11.556 g Epikot 7.125 g Dodecanylsuccinateanhydride, 6.275 g Methylnodicanhydride and 0.375 g DMP-30 (2,4,6 Triphenol)
Well, anyone who uses other formulas to mix Epon or do you agree with my
formula? I would be happy to receive other propositions to mix Epon or any further information on this stuff. It is the first time I try Epon while using the 3-component ARALDITE normally.
I am looking to buy a used EDX specimen holder for a JEOL 1200EX or EXII. If you have one that you haven't used and are willing to part with, please e-mail me at: jfb-at-uidaho.edu
On Mon, 19 Apr 1999 DUNNTEM-at-aol.com-at-Sparc5.Microscopy.Com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } In a message dated 4/19/99 6:10:52 AM Hawaiian Standard Time, { { } } Email: ckuzmiak-at-chuma.cas.usf.edu } } Name: Carolyn Kuzmiak } } School: USF, writes: } } } } Question: How can serial sections be picked up onto a formvar/carbon } } coated large slot grid? So far I have had problems with wrinkling of } } sections, sections drying down onto metal instead of remaining in the } } slot window. } } } } A technique I have used is to pick the secions out of the water trough using } a thin loop (I made mine of .008 gold/paladium wire for no better reason than } that is what I had in the lab!). The loop should have a diameter of less than } 3mm. 2mm works well. } } If the section ribbon is teased away from the knife edge the loop can be } lowered from the top and will pick up a "film" of water plus sections. } } A slot grid, with a formvar/carbon support film is used. The grid needs to be } held firmly during the transfer process of the secions from the loop. I } actually use an old Hitachi specimen holder where the grid is secured on top } of a 3mm dia peg with a retaining cap. This kind of setup can easily be made } in the workshop. The grid can also be held by the adhesive applied to } "gridsticks" (available from EM supply companies). } } The loop is lowered onto the filmed surface of the grid and the water drop } with sections transfers readily to the support film. } } Treating the filmed grids with poly-L-lysine beforehand also helps. ( Apply a } drop of .01% poly-L-lysine to the filmes surface of the grid. After 5 mins } blot off with filter paper applied to edge of grid. Allow grid to dry at room } temperature or more quickly in an oven at 60 degrees centigrade.) } } Hope this helps. } } Ted Dunn } Maui, Hawaii } Hi,
Very good ideas! I do it mostly the same way, but I don't use a bridge. It also works if you do not coat the "pick-up grid" with formvar. In order to hold the final, filmed- with- formvar grid steady when transferring the section from the water drop, the grid can be clamped into a reverse forceps (I have 5 of them). The grids in the five forceps can be laid down or put into a holder after the water is drawn off with a filter paper (or not - the grids can just be allowed to dry). One has to try it out, because depending on the embedment and the specimen one method may be more liable to cause wrinkling than another. Another very helpful idea is to use boat water which has been treated by sitting in one of the Pella cups meant for this purpose. Using this water prevents the sections to pedal around in the boat and mixed up. If the sections are irreplaceable, etc and are due to make you rich and famous, then another method may be in order. Trim a HUGE block face. Section one section at a time, and pick up each section seperately on a seperate grid. Tedious, but won't we do to become rich and famous? Bye, Hildy Crowley {hcrowley-at-du.edu}
Our laboratory is planning to buy a print processor. Can anyone give us a recommendation? We would like something simple like the ancient Ilford type which is fast and easy. We can fix prints at a later date seperately. I would so much appreciate anyone's comments with their hands-on experience.
I agree that it would be very unusual for an SEM site to have an optical bench for optical diffractometry. But as I mentioned in my posting, this should also work by acquiring the image on a computer (everybody has digital image acquisition on the SEM, right? If not, contact us. So much for a plug. See disclaimer below) and calculating the Fourier Transform of the image. Then you have to do the radial integration and you should be able to judge the resolution. Using optical diffraction is better as it includes more information from a negative, so for doing it on a computer, you want an image as big as possible (min. 2K x 2K). Whether this also works with images scanned from Polaroids I don't know.
I hope, this helps.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
} ---------- } From: Steve Chapman[SMTP:PROTRAIN-at-CompuServe.COM] } Sent: Tuesday, April 20, 1999 12:55 AM } To: American } Subject: SEM Resolution } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } The few comments posted pose interesting points. } } 1. Measuring resolution by measuring the wave-form across an edge } is } not very constructive in the real world. Designed to monitor spot } size, } with the assumption that the resolution attainable is similar to the } smallest spot, this technique is messy and in no way does it develop } in an } operator for the correct procedures to attain maximum performance. } } Visual indications of performance or lack of it are very important } when } operating at the limits of the instrument at a particular kV. Using } "normal" images for performance testing helps the operator develop a } feel } for the instrument, thereby sensing that they could use a smaller } spot } size, or a higher emission current, a better astigmatism correction or } better focus. Not only do you test the instrument but you also test } and } develop the operator. Reasons why the South African Microscopical } Society } are looking at a basic "quality" procedure for EM nation wide. } } 2. The TEM method of using an optical diffractometer is very } interesting. I have used it many times for TEM and it would work for } SEM } negatives, one problem is that the biggest users of SEM are in } industry and } few will have access to a diffractometer. } } Just a few thoughts on an interesting but to many rarely discussed } question, or is it? } } Steve Chapman } } Senior Consultant E.M. } Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. } Tel & Fax 44 (0)1844 353161 } Web Site - http://ourworld.compuserve.com/homepages/protrain } For Consultancy and Courses in Electron Microscopy World Wide } }
Senior Research Technician*- Oklahoma Medical Research Foundation = Imaging Facility =20 The newly formed Imaging Core Facility at the Oklahoma Medical Research = Foundation is seeking a research technician with a BS/BA degree in life = sciences (minimum) and a background in preparatory methods for light = and/or electron microscopy. Candidates should be able to carry out = standard specimen fixation, embedding, sectioning and staining = techniques. Prior experience in mammalian tissue processing and some = knowledge of histological methods would be helpful but are not required. = Salary will be commensurate with experience. The successful applicant = will receive further training on the TEM, laser scanning confocal = microscope and other instruments. Please submit a Curriculum Vitae and = names and addresses of three references to:
Stephen Fields, Ph.D. Oklahoma Medical Research Foundation Program in Molecular and Cell Biology 825 NE 13th St. Oklahoma City, OK 73104 405-271-7245 (office) 405-271-3153 (fax) steve-fields-at-omrf.ouhsc.edu
*Applicants must be capable of a full range of body positions and = movements including, but not limited to, standing, walking, sitting, = climbing, bending/stooping, squatting, twisting, and reaching. Moreover, = fine motor skills and sense of touch are necessary to carry out the many = procedures associated with the job, some of which could be dangerous = without full or nearly full motor and sensory attributes, including = hearing and vision.=20
I am looking into options for visible microspectrophotometry, and would appreciate your suggestions. A system could be added to one of our microscopes (Olympus BX60 or BH2), or adapted to our Nicolet FT-IR spectrometer/Spectra-Tech microscope.
Recommendations from satisfied users of either option, or another, will be appreciated.
The following are comments that I gave to Edoardo Bemporad with several items added. It thought that it might be useful to this crowd since there is not a lot of experience out there with the RES100 ion mill.
I assume that you would like my recommendations with respect to the Bal-Tec ion mill. I like it very much. I am getting very good samples. There are things that the RES 100 can do that the other ion mills can not. In particular, the RES100 can coat non-conducting samples with a sputter coating quite easily and can also etch samples. These last two things become very important if you have a reasonably good SEM. I have also done a little ion polishing of SEM samples that were examined in a high resolution SEM with good results.
The capability of imaging the sample for alignment is very significant for me because most of my samples are cross sections of glass samples and they fluoresce under the mill. With every sample, I can routinely check the ion mill alignment if I want to. The RES100 has the ability to capture the digital image of the fluorescence and superimpose it on the image. This allows you to see when shadowing of the center of the sample occurs due to the rim of the dimple. This then defines the minimum milling angle that you can use.
I have found that the alignment of the guns are extremely good over time. I can say that I have not had the same experience with the limited use that I have had with a PIPS system. I use the RES100 to sputter coat carbon onto my glass samples so that I can use them in a FEG-TEM. (If I coat them with my normal evap carbon coater, they contaminate because it is a dirty diffusion pumped system.) It could be set up to sputter coat other materials quite easily.
There are areas for improvement in the mill and Bal-Tec is very receptive to good suggestions. They have made a number of corrections or improvements suggested by me.
The low-angle stage works well, but I have dropped some samples while milling. I have found that this is less of a problem if I am careful and make sure that my samples are less than 100 um in total thickness. (Around 90 um works very well.) Bal-Tec is aware of this and as I understand it are looking at alternative designs for the low angle stage. We also had some problems with the rotation motor early on, but they did a redesign of the motor mount that fixed the problem and have had no problems since.
I bought the RES100 because at the time, it was only one of two instruments that had the ability to terminate with a Faraday cup. This feature works well. However, I find that the image processing termination feature of the RES100 is much more sensitive and fine tunable than the PIPS. Again, they made a few modifications to this portion of the software after some comments by me and others that significantly improved it. It would be able to set up silicon, for example, when it becomes optically transparent in the red. It looks like it is very possible to use image processing to terminate optically transparent samples. I showed them some image processing steps that could do this, but I don't know if they will implement it or not. I thought that they were interested in this.
Programming the ion milling is fairly straightforward. I would like to the ion milling history that a sample goes through recorded better. It is also not easy to perform milling from the substrate side on both sides of the sample like the PIPS instrument does. It is doable, but only with one gun at a time. The best way to do it is by cycling the same gun on top and bottom of the sample and rocking the sample back and forth (without rotation) with an angle of 30-35. Since you can't yet loop the program, you have to brute force it by copying two lines of the program and pasting them many times.
There is a learning curve to the use of the instrument that is a little longer than the other ion mills that I have used. I think that this is so because the design and philosophy of the instrument is different than others.
I have used the PIPS instrument and am familiar with the Fischione instrument.
Given the choice between the three instruments, I would buy the RES100 again if I had to make the decision tomorrow because of its versatility. However, I understand that the ion mill South Bay Technology sells (IV-3?) now has the Faraday termination option and I would like to check that and compare it to the RES100.
Overall, I am quite pleased with my system and the service that I received from Bal-Tec.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
BookWhere? is a powerful software package that runs on the Microsoft Windows Operating System and allows the user to search remote databases for published material of interest.
Link to this site is available on www.coleoptera.org in the directory {Software house} .
Hundreds of library catalogues and other types of information databases can be searched via the Internet using BookWhere? The software comes with a pre-configured list of available hosts and databases making access to this universe of information easy and convenient.
The Essential Research Tool
SINGLE INTERFACE: This single interface is your window on global libraries. BROADCAST SEARCHING: Fast, direct, simultaneous access to remote databases. COMPREHENSIVE SOURCES: Select library catalogues from built in lists organized by name, location or type. HYPERLINK SEARCHES: Hyperlink to a second level search directly from retrieved records. ANALYZE RESULTS: Customize the analysis window for grouping records by various keys. Jump to a subset instantly. EASY TO USE: Easy-to-use Windows-based interface. EXPORT RECORDS: Supports ALL popular bibliographic citation management packages. Transfer records directly to ProCite and Reference Manager. MARC RECORDS: Supports the display and export of MARC records for use in library automation systems
link to this site is available on www.coleoptera.org directory {Software house} .
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptiste (surname) of Bejsak (Bayshark)-Collorado-Mansfeld
http://www.coleoptera.org Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., P.O.Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Michael Reiner wrote I have following recipe to mix the four components for EPON 812. For 25grs we use 11.556 g Epikot 7.125 g Dodecanylsuccinateanhydride, 6.275 g Methylnodicanhydride and 0.375 g DMP-30 (2,4,6 Triphenol)
Well, anyone who uses other formulas to mix Epon or do you agree with my
formula? I would be happy to receive other propositions to mix Epon or any further information on this stuff. It is the first time I try Epon while using the 3-component ARALDITE normally.
Dear Michael Reiner,
for Epon 812 with WPE 157 we use
52,3 g Epon 23,0 g MNA 23,0 DDSA 1,5g DMP30 or 2,5 g BDMA
best wishes Anne Heller
Dr. Anne Heller Arbeitsgruppe Elektronenmikroskopie Institut fuer Botanik (210) Universitaet Hohenheim Garbenstr.30 D-70593 Stuttgart Tel.0049-711-459-2180 Fax 0049-711-459-3355
We calculate for each bottle of Epon 812(or EMBed 812). Most commercially available epoxy resins sold today give the "Weight for Epoxy Equivalent" of the Epon component. The "WPE" is listed on the bottle for each batch you buy. This value for Epon 812 usually varies between 140-180. If the same resin consistency is to be maintained, formulation of the resin must change to compensate for the varying WPE's of the resin. This can be done by varying the amount of hardener(anhydride) For example, the weight of the anhydride used can be determined from the following formula:
weight of anhydride= (weight of resin/WPE) x anhydride equivalent x anhydride/epoxy ratio
WPE is determined from label on bottle. anhydride equivalent = molecular weight of DDSA (Dodecenyl Succinic Anhydride) = 226.0 NMA (Nadic Methyl Anhydride)=178
Ratio of anydride/epoxy = ratio of molar concentrations of anhydride to epoxy Luft's original 1961 mixture that gave optimal cutting qualities for Epoxy 812 (also known as Epon or EMBed 812) was 0.7 A/E.
Example: If you wish to use 100 grams of Epon resin with a WPE of 160, how much DDSA and NMA would you need to achieve an A/E of 0.7?
DDSA weight = (100/160) x 266 x 0.7 = 116.4 grams NMA weight = (100/160) x 178 x 0.7 = 77.87 grams
You would combine 100 grams of Epon with 116.4 grams of DDSA and then 77.87 grams NMA with 100 grams of Epon. (note that you end up with 200 grams of Epon at the end) Mix these well and add DMP-30 at a 1-3% ratio (we usually use 1%) And of course, we cut the amount to fit in a 30 ml syringe for storage and ease of use.
Good luck. john
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu ********************************************
On Tue, 20 Apr 1999 16:32:52 +0200 Michael Reiner {a2811111-at-smail.Uni-Koeln.DE} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear microscopists: } } I have following recipe to mix the four components for EPON 812. } For 25grs we use } 11.556 g Epikot } 7.125 g Dodecanylsuccinateanhydride, } 6.275 g Methylnodicanhydride } and 0.375 g DMP-30 (2,4,6 Triphenol) } } Well, anyone who uses other formulas to mix Epon or do you agree with my } } formula? } I would be happy to receive other propositions to mix Epon or any } further information on this stuff. } It is the first time I try Epon while using the 3-component ARALDITE } normally. } } Thank you for hints and tips, } keep trying ... } } Michael Reiner }
Sorry, I forgot to add the Epon/araldite info. If you are using an Epon/araldite formulation, go by the amounts listed in the kit. If you lost this, you can contact any of the suppliers and they are usually kind enough to mail that info. I tend to stick with my "usual and customary" amount for this (as I am a naturally lazy person):
Embed 812: 25 grams or 25 ml Araldite 502: 16 grams or 15 ml DDSA: 48 grams or 55 ml DMP 30: 1.6 gram or 1.5-2 ml
I'm sure that there are others that have a more precise or different rendering of this formula.
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu ********************************************
Hi all! I could do with a bit of serious technical advice from someone with experience in preparing diffraction crystals. Basically, it goes like this. I've pillaged the diffracting crystals from an old Phillips spectrometer that we have sitting in the basement. Inside were LiF, LiF220, Ge, PET and TAP on their nice Al backing plates attached with dollops of silicon (?) rubber sealant. Because of the exorbitant price of replacing diffraction crystals for our JEOL 'probe, particularly TAP at ca. R40,000 each, I thought that it might be a cunning ploy to remove these from their backings and cut them to size. I could get several and if they work that would be great. Not only would it sort out our TAP problem but also extend our choice of crystals to use. I have located a diamond wire saw with which I could do the cutting. However, there are certain other things that spring to mind for which help from those who actually prepare these things would be greatly appreciated. I appreciate that those who produce these for the JEOL 'probes would rather I bought new ones from them. Sorry guys, but our lab can't afford it with the current state of the Rand. 1) Is the diamond saw good enough for this kind of work? 2) TAP appears particularly fragile, is there a way to minimise flaking? 3) I guess lubricant should be avoided, but if it is possible to use one, what should be used for which crystal type?
Any help would be greatly appreciated. Cheers, Malc. Dr MP Roberts Department of Geology Rhodes University Grahamstown 6140 South Africa Tel: +27 46 6038316 Fax: +27 46 6229715 ******************************* "If God had meant birds to fly, he would have given them engines" Anon.
For those of you who have not yet used microspectrophotometry, it is a great way to add optical and chemical information to our microscopy imaging. Spectroscopy and Microscopy are converging technologies; that convergence was strongly visible at this year's PITTCON meeting. (For details, write me or see the May issue of American Lab, "Focus on Microscopy" column)
But back to Jamie's questions: Leica and Zeiss both make fully integrated UV-Vis microspectrophotometers, with the necessary software for spectral manipulation. In terms of add-ons, there are now 2-3 companies, but I would have to do research to dig them out. The only one I have current data on is a company run by Felix Brogna called Optical Technologies. They are located in Hawthorne, NY (just north of White Plains) and can be reached at 914-592-1900. While I know that there are at least 2 other companies, I cannot find their data in my current files.
Optical Technologies has two devices: a manual spectrometer (you dial in the wavelengths) and a computer controlled one (via RS232 cable). Both sell for around $10K or less (prices may have risen since I last spoke to them) but they provide good value for money. As I remember, they also had a focusable pinhole to eliminate stray light.... a very desirable feature.
If you have not had much experience in microspectrophotometry, might I suggest Horst Piller's book Microscope Photometry, ISBN 3-540-08094-5; Spring Verlag, publisher, 1977. If you can't get it on the open market, call Irv Toplin at Zeiss and see if he can track down a copy either here in Thornwood or in Germany. I used to be one of two microspectrometry field specialists with Zeiss and found this book valuable. It stresses the importance of a stablized light source (if not available from your manufacturer, see Mel Dekker at Optiquip), aperturing, optical and statistical errors, etc.
Also interesting: A book edited by Michael Morris and put out by Marcel Dekker. My copy is not here today, so I can't give you the complete information but I think the title is Spectroscopic and Microscopic Imaging of the Chemical State. While the first chapter on microscopy is a bit weak (Dr. Morris and I have discussed that), the rest of the book is great.
Caveat: I have no commercial interest in any of these products, only an interest in promoting better use of microscopy in general.
Hope that all this helpful.
Best regards
Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 03:56 PM 4/20/99 -0400, James Martin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Malc asks ... } } } Hi all! } I could do with a bit of serious technical advice } from someone with experience in preparing diffraction crystals. } ... } I've pillaged the diffracting crystals from an old Phillips } spectrometer that we have sitting in the basement. ...
I have to wonder if these crystals could be used with the JEOL Rowland circle?? ... a focussing issue, whereby the xtal surface curvature is focussed on the opposite side of the circle.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Hi Richie, Quartz & Silice is now a part of the French conglomerate Saint-Gobain.
best regards, mark
Mark W. Lund, PhD VP Engineering } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"This is a YOUNG business...How can I tell you what YOUR job is when I don't know what MINE is?" --Pogo
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear microscopists: } } I have following recipe to mix the four components for EPON 812. } For 25grs we use } 11.556 g Epikot } 7.125 g Dodecanylsuccinateanhydride, } 6.275 g Methylnodicanhydride } and 0.375 g DMP-30 (2,4,6 Triphenol) } } Well, anyone who uses other formulas to mix Epon or do you agree with my } } formula? } I would be happy to receive other propositions to mix Epon or any } further information on this stuff. } It is the first time I try Epon while using the 3-component ARALDITE } normally. } } Thank you for hints and tips, } keep trying ... } } Michael Reiner } } } Hi, When trying something new with embedding media, it might be best to go to a formulation which has been existence for 30 years - the formulations by Luft. These formulations have been used for millions of embedments with success. Look in any TEM textbook and you will find them. The "medium hard" formulation is a favorite of pathologists, because it is versatile and will embed many samples satisfactorily. By the way - It is not necessary to weigh out epoxides. Use disposable syringes for measuring accurately. You will not be able to detect a difference in block properties between monomers carefully weighed and carefully measured. I have data to support this. Hildy Crowley {hcrowley-at-du.edu}
I have a Hitachi 2500C, S/N 32001622-5303, with a Large Stage C and I am looking for a computer controllable stage controller. Do you have one that is available or do you know where I can obtain one. I would prefer an RS 232 interface but that is not absolutely necessary. I am also looking for a video converter to put the picture on the WEB like an Web Cam. I also have a Sigma system # 20400, customer #8294-01. I would also like to put the X-ray maps on the web in real time. I have also sent this message to Hitachi and Kevex and will be posting it on some other Web pages.
Dennis Coad Boeing Huntsville Central Labs Lead MS: JW-56 (256) 461-2976
} -----Original Message----- } From: ckuzmiak-at-chuma.cas.usf.edu [SMTP:ckuzmiak-at-chuma.cas.usf.edu] } Sent: Friday, April 16, 1999 10:19 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: How can serial sections be picked up } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Colleagues ..... } } This one is out of my area of expertise... Please reply direct to } the sender... } } Email: ckuzmiak-at-chuma.cas.usf.edu } Name: Carolyn Kuzmiak } School: USF } } Question: How can serial sections be picked up onto a formvar/carbon } coated } large slot grid? So far I have had problems with wrinkling of sections, } sections drying down onto metal instead of remaining in the slot window. } } -------------------------------------------------------------------------- } - [MORETZ,DR,ROGER TX BIPUS]
Jumping in a little late, the technique I use is not totally unlike the that described by Hildegarde Crowley, but may be sufficiently so to justify the response.
I use the reverse action, anti-capillary forceps--the #5 style to hold the slotted grid, pre-coated with formvar and carbon. Using the anti-capillary forceps upside down allows you to hold the grid at an angle. This additional angle provides greater flexibility in bringing the grid under the sections, attaching the first section to the support film and drawing the grid and ribbon onto the grid along the length of the slot. Repulsion of the grid and the ribbon can be eliminated or at least controlled by zapping each grid (just prior to immersion in the boat) with a Zerostat. I have also used a demagnetizing coil (purchased from Ladd many, many years ago) to reduce the repulsive interactions. The other thing I have used to control placement of the ribbon and to ensure attachment to the grid over the slot is the use of a lash to manipulate the ribbon. Those are difficult to come by unless you have long lashes.
This technique sounds (and I guess it is) tedious, but I have used it to successfully section over 500um through an amyloid plaque--over 700 grids in all, and the only sections lost were when the forceps tips perforated a slot during staining!
Thanks to all of you who replied to my query about differentiating apotosis from necrosis in TEM. I really appeciate it! The replies are attached below. In addition to the references give, I also found useful Voume 46 of Methods in Cell Biology; Cell Death, edited by Lawrence .M. Schwartz and Barbara A. Osborne.
As it turns out, the cells I was looking at are neither apototic nor necrotic, but now appear to have a strange disorder. But that's another story...
The replies:
Differentiating apoptosis and necrosis morphologically is based primarily upon nuclear changes, although there are characteristic cytoplasmic changes as well. In general (note the wiggle words), necrotic cells swell and lyse, whereas apoptotic cells shrink and fragment. Chromatin in apoptotic cells forms electron-dense crescents at the nuclear envelope, then breaks up. Apoptotic cells fragment into "apoptotic bodies" that may contain bits of chromatin. A good place to see characteristic ultrastructural changes of apoptosis is in lymphoid tissues, where lymphocytes die via apoptosis and then are phagocytosed by resident macrophages (the ones that are sometimes called "tingible body macrophages" because of the staining properties of the apoptotic cell remnants in them.)
Differenting apoptosis from necrosis is a tricky deal. Apoptotic cells may undergo secondary necrosis, during which they swell and lyse. So, just because you see necrotic cells doesn't mean that they didn't die apoptotically. Like everything else, it's complicated; there is a continuum of change with apoptosis and necrosis at opposite poles and a lot of stuff in between!
There are lots of good reviews on this topic. The Aug 28, 1998 issue of Science had a special section on apoptosis, and on page 1302, there is a series of three electron micrographs of neurons undergoing apoptosis. One of the first reviews of the subject contains the best collection of micrographs I've found - "Cell death: the significance of apoptosis" in the International Review of Cytology 68:251-306, 1980. Another good review was in Amer J of Pathol 146:3-15, 1995. The title is "Apoptosis, oncosis, and necrosis: an overview of cell death."
Jane Fagerland
There's lots of stuff on t.e.m. of apoptosis in vertebrate cells (it's very popular in HIV, cancer, inflammation response etc) and much of it indicates visible nuclear changes but relative stability of cytoplasm compared with necrosis.
There was a review article (as a good starting point): Microscopical Study of Cell Death via Apoptosis by S. Verhaegen in MIcroscopy and Analysis, January 1998 pp5-7
But you could do a reference or citation 'trawl' on the authors: Kerr, J.F.; Wyllie, A.H. or Currie
Malcolm Haswell
In response to your question, I did some necrosis versus apoptosis questions concerning a mycobacterium ulcerans toxin question we had here, and two papers that were particularly helpful distinguishing the two were from Scanning Microscopy Vol. 10, No. 1, 1996 pages227-237,by E. Falcieri, et al, Different Approaches to the study of Apoptosis, and in the same journal, by Dini et al, pages 239-252, an article entitled Recognition and Phagocytosis of Apoptotic Cells.
Beth Fischer
Apoptosis: the molecular basis of cell death - Current Communications in Cell and Molecular Biology 3. L. D. Tomei and F. O. Cope editors. Cold Spring Harbor Laboratory Press 1991
Cell Death in Biology and Pathology. I.D. Bowen and R.A. Lockshin ed. Chapman and Hall publs.
John (Keoni) Hardy
The most "conventional" way to detect apoptosis is to look for DNA fragmentation using in situ hybridization probes. I would not recommend that you go down that path unless you really need this technique to work in your lab. There are many in situ probes available to detect apoptotic cells if you do decide this is what you want. The DNA fragmentation that occurs during apoptosis will produce "patterns" in the nuclear chromatin which researchers have used to identify apoptotic cells. However, this has the same pitfalls as other morphologic characterizations (mostly in proving this is what you are looking at).
You might try looking for ways to detect cytoplasmic cytochrome c or activated caspases (not easy yet).
Here are some reviews to read: Baker et al, 1996 Oncogene 12:1-9. Lincz, 1998 Immuno.and Cell Bio. 76:1-19. Van Engeland et al, 1998 Cytometry 31:1-9 Solary et al 1998 Cell Bio. and Toxicol 14:121-132. Green & Reed 1998 Science 281:1309-1312 Dickson 1998 Trends Biotechnol 16:339-342 Cai et al 1998 Biochem. Biophys Acta 1366:151-165. O'Brian 1998 J. Gen virol 79:1833-1845. Granville et al 1998 Lab Invest 78:839-913 Kuan and Passaro 1998 Arch Surg 133:773-775.
Paul Webster
Aloha, Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
We have a Debben stage on our Hitachi 2460N. It works well for us and has a RS-232 interface. Ours is interfaced to a Link ISIS EDS system. I would not care to program such a thing by myself. (I am getting too old for it.)
I would be interested in what you hear about putting the video on the web. I have played around a little with it myself, but keep running out of time. Other responsibilities keep calling.
The x-ray maps could be put on line in pseudo real time if the Kevex saves files to a standard imaging format and supports an FTP server (i.e., it is standard Windows 95 OS). We use Microsoft's Personal Web Server the WarFTP FTP server to offer up files in particular directories. We save the files in a format that others can view and/or retrieve, and they are available as soon as we hit the Save button. You can check us out at WWW.MARL.IASTATE.EDU and see for yourself. BTW, JPG and GIF formats are best for this applications. Your clients would need to set up a helper application to view TIFF files, plus the files get big, fast.
At 11:30 AM 4/21/1999 -0500, you wrote: } } I have a Hitachi 2500C, S/N 32001622-5303, with a Large Stage C and I am } looking for a computer controllable stage controller. Do you have one that } is available or do you know where I can obtain one. I would prefer an RS 232 } interface but that is not absolutely necessary. I am also looking for a } video converter to put the picture on the WEB like an Web Cam. I also have a } Sigma system # 20400, customer #8294-01. I would also like to put the X-ray } maps on the web in real time. I have also sent this message to Hitachi and } Kevex and will be posting it on some other Web pages. } } Dennis Coad } Boeing Huntsville } Central Labs Lead } MS: JW-56 } (256) 461-2976
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
The Northern California Society for Microscopy will hold a Biomaterials Symposium on May 6th at Roche Biosciences in Palo Alto. A complete program is attached for those interested in attending. The symposium will begin at 1:00 p.m.
A social hour is scheduled for 5:45 followed by a Mexican Fiesta dinner featuring chicken and beef fajitas and cheese enchiladas, black beans and Spanish rice. The cost of the dinner is $15.00 for members and $7.50 for students. Reservations should be made by May 3rd. Contact Greg Lum to make your reservation. Telephone (415) 338-1091 or email glum-at-sfsu.edu
Photo contest: Remember to bring a micrograph for consideration for the cover of our directory 'Scope'. The NCSM council will act as judges and the winner will receive $100.00
Vendors are invited to bring brochures and other information describing their products. Vendors, please remember to bring a one-sheet advertisement to be included in the new 'Scope'.
All attendees are encouraged to bring a buddy to the meeting.
The Northern California Society for Microscopy will hold a Biomaterials Symposium on May 6th at Roche Biosciences in Palo Alto. A complete program is attached for those interested in attending. The symposium will begin at 1:00 p.m.
A social hour is scheduled for 5:45 followed by a Mexican Fiesta dinner featuring chicken and beef fajitas and cheese enchiladas, black beans and Spanish rice. The cost of the dinner is $15.00 for members and $7.50 for students. Reservations should be made by May 3rd. Contact Greg Lum to make your reservation. Telephone (415) 338-1091 or email glum-at-sfsu.edu
Photo contest: Remember to bring a micrograph for consideration for the cover of our directory 'Scope'. The NCSM council will act as judges and the winner will receive $100.00
Vendors are invited to bring brochures and other information describing their products. Vendors, please remember to bring a one-sheet advertisement to be included in the new 'Scope'.
All attendees are encouraged to bring a buddy to the meeting.
The Northern California Society for Microscopy will hold a Biomaterials Symposium on May 6th at Roche Biosciences in Palo Alto. A complete program is attached for those interested in attending. The symposium will begin at 1:00 p.m.
A social hour is scheduled for 5:45 followed by a Mexican Fiesta dinner featuring chicken and beef fajitas and cheese enchiladas, black beans and Spanish rice. The cost of the dinner is $15.00 for members and $7.50 for students. Reservations should be made by May 3rd. Contact Greg Lum to make your reservation. Telephone (415) 338-1091 or email glum-at-sfsu.edu
Photo contest: Remember to bring a micrograph for consideration for the cover of our directory 'Scope'. The NCSM council will act as judges and the winner will receive $100.00
Vendors are invited to bring brochures and other information describing their products. Vendors, please remember to bring a one-sheet advertisement to be included in the new 'Scope'.
All attendees are encouraged to bring a buddy to the meeting.
The Northern California Society for Microscopy will hold a Biomaterials Symposium on May 6th at Roche Biosciences in Palo Alto. A complete program is attached for those interested in attending. The symposium will begin at 1:00 p.m.
A social hour is scheduled for 5:45 followed by a Mexican Fiesta dinner featuring chicken and beef fajitas and cheese enchiladas, black beans and Spanish rice. The cost of the dinner is $15.00 for members and $7.50 for students. Reservations should be made by May 3rd. Contact Greg Lum to make your reservation. Telephone (415) 338-1091 or email glum-at-sfsu.edu
Photo contest: Remember to bring a micrograph for consideration for the cover of our directory 'Scope'. The NCSM council will act as judges and the winner will receive $100.00
Vendors are invited to bring brochures and other information describing their products. Vendors, please remember to bring a one-sheet advertisement to be included in the new 'Scope'.
All attendees are encouraged to bring a buddy to the meeting.
The Northern California Society for Microscopy will hold a Biomaterials Symposium on May 6th at Roche Biosciences in Palo Alto. A complete program is attached for those interested in attending. The symposium will begin at 1:00 p.m.
A social hour is scheduled for 5:45 followed by a Mexican Fiesta dinner featuring chicken and beef fajitas and cheese enchiladas, black beans and Spanish rice. The cost of the dinner is $15.00 for members and $7.50 for students. Reservations should be made by May 3rd. Contact Greg Lum to make your reservation. Telephone (415) 338-1091 or email glum-at-sfsu.edu
Photo contest: Remember to bring a micrograph for consideration for the cover of our directory 'Scope'. The NCSM council will act as judges and the winner will receive $100.00
Vendors are invited to bring brochures and other information describing their products. Vendors, please remember to bring a one-sheet advertisement to be included in the new 'Scope'.
All attendees are encouraged to bring a buddy to the meeting.
The Northern California Society for Microscopy will hold a Biomaterials Symposium on May 6th at Roche Biosciences in Palo Alto. A complete program is attached for those interested in attending. The symposium will begin at 1:00 p.m.
A social hour is scheduled for 5:45 followed by a Mexican Fiesta dinner featuring chicken and beef fajitas and cheese enchiladas, black beans and Spanish rice. The cost of the dinner is $15.00 for members and $7.50 for students. Reservations should be made by May 3rd. Contact Greg Lum to make your reservation. Telephone (415) 338-1091 or email glum-at-sfsu.edu
Photo contest: Remember to bring a micrograph for consideration for the cover of our directory 'Scope'. The NCSM council will act as judges and the winner will receive $100.00
Vendors are invited to bring brochures and other information describing their products. Vendors, please remember to bring a one-sheet advertisement to be included in the new 'Scope'.
All attendees are encouraged to bring a buddy to the meeting.
The Northern California Society for Microscopy will hold a Biomaterials Symposium on May 6th at Roche Biosciences in Palo Alto. A complete program is attached for those interested in attending. The symposium will begin at 1:00 p.m.
A social hour is scheduled for 5:45 followed by a Mexican Fiesta dinner featuring chicken and beef fajitas and cheese enchiladas, black beans and Spanish rice. The cost of the dinner is $15.00 for members and $7.50 for students. Reservations should be made by May 3rd. Contact Greg Lum to make your reservation. Telephone (415) 338-1091 or email glum-at-sfsu.edu
Photo contest: Remember to bring a micrograph for consideration for the cover of our directory 'Scope'. The NCSM council will act as judges and the winner will receive $100.00
Vendors are invited to bring brochures and other information describing their products. Vendors, please remember to bring a one-sheet advertisement to be included in the new 'Scope'.
All attendees are encouraged to bring a buddy to the meeting.
The Northern California Society for Microscopy will hold a Biomaterials Symposium on May 6th at Roche Biosciences in Palo Alto. A complete program is attached for those interested in attending. The symposium will begin at 1:00 p.m.
A social hour is scheduled for 5:45 followed by a Mexican Fiesta dinner featuring chicken and beef fajitas and cheese enchiladas, black beans and Spanish rice. The cost of the dinner is $15.00 for members and $7.50 for students. Reservations should be made by May 3rd. Contact Greg Lum to make your reservation. Telephone (415) 338-1091 or email glum-at-sfsu.edu
Photo contest: Remember to bring a micrograph for consideration for the cover of our directory 'Scope'. The NCSM council will act as judges and the winner will receive $100.00
Vendors are invited to bring brochures and other information describing their products. Vendors, please remember to bring a one-sheet advertisement to be included in the new 'Scope'.
All attendees are encouraged to bring a buddy to the meeting.
The Northern California Society for Microscopy will hold a Biomaterials Symposium on May 6th at Roche Biosciences in Palo Alto. A complete program is attached for those interested in attending. The symposium will begin at 1:00 p.m.
A social hour is scheduled for 5:45 followed by a Mexican Fiesta dinner featuring chicken and beef fajitas and cheese enchiladas, black beans and Spanish rice. The cost of the dinner is $15.00 for members and $7.50 for students. Reservations should be made by May 3rd. Contact Greg Lum to make your reservation. Telephone (415) 338-1091 or email glum-at-sfsu.edu
Photo contest: Remember to bring a micrograph for consideration for the cover of our directory 'Scope'. The NCSM council will act as judges and the winner will receive $100.00
Vendors are invited to bring brochures and other information describing their products. Vendors, please remember to bring a one-sheet advertisement to be included in the new 'Scope'.
All attendees are encouraged to bring a buddy to the meeting.
Michael Reiner wrote ================================================ I have following recipe to mix the four components for EPON 812. For 25grs we use 11.556 g Epikot 7.125 g Dodecanylsuccinateanhydride, 6.275 g Methylnodicanhydride and 0.375 g DMP-30 (2,4,6 Triphenol)
Well, anyone who uses other formulas to mix Epon or do you agree with my formula? I would be happy to receive other propositions to mix Epon or any further information on this stuff. It is the first time I try Epon while using the 3-component ARALDITE normally. ================================================ The trade name Epon® is owned by Shell Chemical. Until approximately 1970, Shell produced a product called Epon 812 and was widely used through EM-land . But at about that date, Shell announced the discontinuation of Epon 812. Shell continues to make other Epon resin grades but unfortunately, none seem to have value for applications in EM.
There is now a range of Epon "substitutes". Most of the major EM suppliers of consumables and supplies have their own "brand" of this substitute. However, they do not come from the same source. And they might come close to approximating the original Epon (in some respects some of them might be superior in terms of infiltration) but they are not identical, either with respect to the original Epon or with regard to the variety of different "substitutes".
So I make this comment, which I hope is not considered out of place, because if indeed someone did have some of the original Epon 812 left in bottles (some claim to still be working off a stock pile they purchased nearly thirty years ago), it may or may not be performing as it would have in 1970. And since workers today are probably using "substitutes", if precise formulaes are going to be stated to the fourth and fifth significant figures , then probably it would be appropriate to state precisely which "substitute" is being used for the resin component.
In our own laboratory at least, we are constantly changing the formula where we are seeking to end up with a hardness more appropriate for specific samples. So in our case, the "formula" can vary by some considerable amount
WASHINGTON (AP) - A single-celled microbe large enough to be seen with the naked eye has been found by researchers sampling ocean dredgings in the South Atlantic.
The bacterium - as big as the period at the end of this sentence - is the largest ever identified.
The microbe, discovered near Namibia, lives by absorbing sulfur and nitrates, and it swells as the chemicals are stored inside its cell walls, researchers report in a study published today in the journal Science.
The biggest of the bacteria is 0.75 millimeter, according to a report by Science.
Researchers at the Max Planck Institute for Marine Microbiology, who made the discovery, said the microbes form chain-like colonies that tend to glow from the absorbed nitrates.
``They look like a thin string of pearls,'' said the scientists, who named the new microbe Thiomargarita namibiensis, which means ``Sulfur Pearl of Namibia.''
``If the largest Thiomargarita was a blue whale,'' Science said in a statement, ``then an ordinary bacterium would be a bit smaller than a newborn mouse.''
In this analogy, the largest previously known bacterium ``would be about as big as a lion,'' about 100 times smaller than Thiomargarita, Science reports. The previous record holder was Epulopiscum fishelsoni, a microbe found in the gut of surgeonfish.
Max Planck researchers said the bacteria live in an environment with high levels of hydrogen sulfide, conditions that are toxic to most other forms of life.
The scientists said Thiomargarita is found in great concentrations in Namibian coastal sediments that contain high levels of toxic sulfide.
If we could use these in dumps and what have you it could do a lot.
Gordon
Gordon Couger gcouger-at-couger.com www.couger.com/gcouger Stillwater, OK 405 624-2855 GMT -6:00
CMP Cient=EDfica and Philips are holding a joint colloquium on Microscopy an= d Materials Analysis in Madrid on 26th May 1999. The colloquium is aimed at all Microscopists and analysts working on both materials and biological applications. The object is to discuss new techniques and instrumentation that will aid us in characterisation, and technologies that can speed up sample preparation.
Working languages will be Spanish & English.
=46or more details see www.cmp-cientifica.com or e-mail me direct.
MAMAS will be meeting at NIST on Tuesday, May 18, 1999 from 10:30am until 3:00pm. There will be coffee and doughnuts and two speakers. All are welcome. See below for details.
Hope to see you there, -- John Henry
John Henry J. Scott NIST Microanalysis Research Group http://www-sims.nist.gov/Division/Contacts.html
ANNOUNCEMENT ------------
Mid-Atlantic Microbeam Analysis Society (MAMAS) Meeting
at the National Institute of Standards and Technology (NIST) Gaithersburg, MD
on
Tuesday, May 18, 1999 10:30am -- 3:00pm
Lecture Room D, Administration Building
10:30am: Coffee and Doughnuts
10:45am: Dave Wollman, NIST (Boulder) "Microcalorimeter EDS with 3 eV Energy Resolution"
Noon: Lunch
1:15pm: Jan S. Iwanczyk and Bradley E. Patt, Photon Imaging, Inc. "High Count Rate and High Energy Resolution Silicon Drift Detectors for X-Ray Microanalysis"
directions to the NIST campus can be obtained from the NIST website at: http://www.nist.gov/public_affairs/maps/nistmaps.html
for more information, contact John Henry Scott (NIST): (301) 975-4981 (301) 417-1321 FAX email: johnhenry.scott-at-nist.gov
Hello to everyone, A colleague just called me with a very interesting question. Does anyone know of a way to assess the possible leakiness of blood vessels in skin samples from archived tissue. This tissue was not formalin fixed, so antigenic sites should be fine.
Ramin Rahbari PARKE-DAVIS Pharmaceutical Research Pathology and Experimental Toxicology 2800 Plymouth Road Ann Arbor, MI 48105 Office (734) 622-3383 Fax (734) 622-5001 Ramin.Rahbari-at-WL.COM
The discussion regarding WPE values for the Epon substitues was basically correct. WPE values used to be very important for creating blocks of a standard set of mechanical and chemical properties during the time when Shell Epon WPE values ranged between 142 and 165. The same formulation used without taking these differences into account made enormous difficulties in the 60's or as long as labs used the Shell product. If anyone today is buying a substitute which varies significantly from batch to batch in WPE values, I urge them to change suppliers immediately. Not all substitutes are created identically. Some contain dilutents, plasticicers, Araldite, etc. All this is proprietary information. It cannot be said that one substitute is better than any other. It is important to stick to one substitute, learn its characteristics, and make adjustments. Should anyone have a problem with the mechanical or chemical characteristics of an 812 substitue, please contact me at {hcrowley-at-du.edu} . I have an entire notebook full of formulations based on WPE values dating from a time when I was faced with an extremely difficult embedding problem. I may have some valuable information. Bye, Hildy Crowley University of Denver Denver, CO
I have a broken Varian 30 L/s ion pump that needs repair or replacement/exchange. Anyone have any experience with reputable firms that perform this service? Would appreciate hearing about sources.
I have question concerning cleaning of white ceramic insulator inside electron gun of TEM JEOL 2000FX. As a consequence of electric charges thereare brown traces on the surface of insulator. Does anybody has experience how to remove these traces and what cleaning solution (e.g. aceton, methanol or something else) to use. Thank you very much.
Yours sincerely
Peter Makroczy
Dept. of Mat.Science
Technical University of Kosice
Slovak Republic
makroczy-at-tuke.sk
-- Peter Makroczy Technical University of Kosice Department of Materials Science Park Komenskeho 11 042 00 Kosice Slovak Republic E-mail: makroczy-at-tuke.sk Tel.: +421 95 602 25 40 Fax.: +421 95 633 27 23
On Fri, 23 Apr 1999 08:47:25 +0200 Peter Makroczy {makroczy-at-tuke.sk} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear microscopists, } } I have question concerning cleaning of white ceramic insulator inside } electron gun of TEM JEOL 2000FX. As a consequence of electric charges } thereare brown traces on the surface of insulator. Does anybody has } experience how to remove these traces and what cleaning solution (e.g. } aceton, methanol or something else) to use. Thank you very much. } } Dear Peter,
We have successfully cleaned these by rubbing with alumina powder (Al2O3) in alcohol as a paste on a tissue or cloth. However, it is important that you remove all traces of the alumina powder when you have finished.
Unless it is a very small mark which is easily accessible we would completely dismantle the gun and work on the insulator alone. After cleaning off the insulator marks wipe and blow off any excess powder with a clean gas supply (N2 from a cylinder not compressed air which usually has oil in it). Then wash it in alcohol in an ultrasonic bath changing the alcohol at least 3 times.
If there are a lot of discharge marks or they are very bad then grit blast it to clean it instead of rubbing with alumina paste. Wash it thoroughly afterwards as described.
Sometimes it is not possible to clean out very deep discharge tracks and the insulator needs to be replaced.
If you are dubious about carrying this out I know that JEOL(UK) clean customers guns by this method when reconditioning them. I am also fairly certain that they would recondition yours (but I don't know the cost).
Good luck, Ron
---------------------- Ron Doole Dept. of Materials, University of Oxford, Parks Road, Oxford. OX1 3PH. ron.doole-at-materials.ox.ac.uk
Never found any solvent for my similar stains. On my Etec, they have to be rather bad to cause a problem. Anyway, I did clean the glazed porcelain insulator (to a degree) using 1/4 micron diamond paste and a small, high speed felt wheel ("Dremel Tool"). Followed with UT in detergent/warm water, water rinse, isopropanol rinse, blow dry w/N2. This was the first serious cleaning in 15 years. Never could determine any performance difference - only cosmetic.
} From Microscopy-request-at-sparc5.microscopy.com Thu Apr 22 20:02:53 1999 } } Date: Thu, 22 Apr 1999 16:33:28 -0700 } To: MSA listserver {Microscopy-at-Sparc5.Microscopy.Com} } From: "Dr. Gary Gaugler" {gaugler-at-calweb.com} } Subject: Ion pump repair sources? } } ---------. } } } I have a broken Varian 30 L/s ion pump that needs repair } or replacement/exchange. Anyone have any experience } with reputable firms that perform this service? Would } appreciate hearing about sources. } } Cheers, } Gary Gaugler, Ph.D. } } Gary, From your email address, I assume you are located in the US. I think that Duniway stockroom has an excellent reputation in this area. They can be contacted at 1-800-446-8811 or at www.duniway.com. They are located in Mountain View, CA but with the excellent UPS and Fedex and other services we enjoy nowadays it makes little difference where they are, once we have gone to the trouble of boxing up the item. Suggest you post a summary of your responses.
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology "A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
Hello everyone. I'm looking for a diamond scribing attachment for old Zeiss compound scopes that was used to score areas on slides. It attached to the objective ring like a lens, and when you found the object of interest, you swung this scribe around and turned it to etch a circle at that spot. The one I have used was in Mel Fuller's lab at UGA, but it disappeared when he retired (sound familiar?). It could be adjusted for any diameter circle. It may not necessarily be specific for a Zeiss: that was the scope I used it on. I can't remember the labels or markings on it now. It was extremely useful for flat-embedded material that was to be cut out and glued to blank BEEMs for sectioning.
If you know where I can find one, let me know how to purchase it.
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu ********************************************
The matter of cleaning parts from the vacuum systems of electron microscopes, including ceramic insulators and the interiors of electron gun chambers, is discussed in some detail on pp. 69 - 75 of my book 'Vacuum Methods in Electron Microscopy' (see: http://swww.2spi.com/catalog/books/book48.html and http://www.bookshop.co.uk/portland/).
A procedure we have found to be satisfactory for cleaning the guns of TEMs operating at accelerating voltages up to 200 kV is as follows: Rub the interior of the gun chamber and the ceramic insulator with lint-free, grease-free cloth pads that have been dipped in a thick slurry of reagent grade isopropyl alcohol and either finely ground calcium carbonate powder or the common 0.05 micrometer grade of aluminum oxide metallographic polishing powder. After all areas are cleaned in this way they are wiped repeatedly with clean pads moistened only with isopropyl alcohol to remove the polishing powder. Then all surfaces, corners,and joints are blasted with a clean, dry gas blaster to remove any residual particles. The insulator and interior of the gun should be heated with a clean hair dryer to remove as much residual surface moisture as possible just before the gun is reassembled.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
As I recall, the first commercial model TEM produced by RCA was called the model EMB. (Model EMA was an experimental model that was never sold) I believe this model was introduced sometime around 1940. I took my first electron micrographs on the second one of these instruments sold. It was located in the laboratory of Prof. Robley Williams, in the Physics Department here at the U. of Mich. There is a brief discussion of this model, and a picture of it on p. 205 of the First Edition of Cecil Hall's book, 'Introduction to Electron Microscopy'. McGraw-Hill, 1953.
According to Hall, RCA introduced it's model EMU microscope in 1944. A picture of this instrument is shown on p.206 of the above book. The RCA model EMU2 looked the same as the EMU, shown in this picture. I know, because i did a lot of work on one of these instruemnts in the laboratory of Dr. Thomas Francis, in the School of Public Health here at the Univ. of Mich.
After the EMU2 model, and starting with model EMU3, RCA changed over to an more user-friendly design which was carried through in the EMU4 instruemnts with little modification. A picture of an RCA EMU3 microscope is shown on p.179 of the Second Edition of Hall's book (1966). I don't know exactly when the EMU3 microscopes were introduced, but it was sometime in the early 1950s. RCA went out of the business of producing electron microcopes in 1969. Again, I know, because I was President of EMSA that year, and had the sad task of dealing with this at our annual meeting that year. I believe the current model at that time was the EMU4, so it was probably introduced sometime shortly after 1966.
This is the best info I can come up with off-hand. Somewhere I have a page from an RCA bulletin showing all their instruments and giving dates of production, but I don't seem to be able to find it among the clutter in my office. However, If you want more detailed info, I'll go through my files at home and see if I can find it there. However, It is likely that you have an EMU or EMU2 model (they look alike). I'd be very surprised if any of the EMBs are still in existance.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Has anyone made a biprism for a TEM using a drawn SiO_2 coated with Au?
I am looking for ideal fibre thickness, heat source that was used (which gases & flame temperature) and for any tips on how to make them. The current suggestions that I have heard are an H_2 and O_2 torch to get the temperature up to aroung 2000 K, but these gases can be quite hazardous to work with.
I would really appreciate any advice.
Thanks, Jon
-- ***************************************** Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
I experienced last year the same problem with the electron gun of my JEOL 2000FX. The brown traces are provoqued by the discharges inside the freon gas. To clean these traces, don't use any solution because the ceramic is porous and after opening the gun, you will have to purge several times the atmosphere of the gun because of the oxygen and the water vapor contained in the air. For this reason it is not recommended to put solution on the ceramic. To clean the ceramic, we used a stick of ceramic like an eraser. After removing the main part of the brown traces, we cleaned with a tissue with alcohol and put the gun into a furnace at 100=B0C for some hours. Then we remount the gun, after we had cleaned the metallic chamber from the traces of discharges (white spots on the steel) with POL paste. You should fill the atmosphere of the gun with nitrogen and purge two or three times and finally fill the gun chamber with freon or SF6. To evacuate the gun there is a valve behind the microscope near the pressure gauge of the gun. You can find the position to use if you look into the manual. This is to clean the ceramic part that is in the freon atmosphere (were the polarisation resistances are located) if you want to clean the ceramic part inside the column, it is more complicated because you don't have access to all electrodes.
That's all! But for us this cleaning was not sufficiant and we had to exchange the gun that was fortunatelly under contract.
Eric LEROY Dr. Laboratoire de Chimie Metallurgique des Terres Rares UPR 209 - CNRS Groupe des Laboratoires de Thiais 2-8, rue Henri Dunant 94320 THIAIS cedex
Dear Peter, I would recommend you ask the JEOL service engineers first. The only time I tried to clean the insulator on a TEM gun, I tried Wenol metal polish first. This removes some of the diffusion pump oil deposit, but did not get off the brown marks, so I next tried a diamond paste (6 micron). That did work. Remove this thoroughtly with several washes of clean ethanol. You wrote:
} } Dear microscopists, } } I have question concerning cleaning of white ceramic insulator inside } electron gun of TEM JEOL 2000FX. As a consequence of electric charges } thereare brown traces on the surface of insulator. Does anybody has } experience how to remove these traces and what cleaning solution (e.g. } aceton, methanol or something else) to use. Thank you very much. } } } Yours sincerely } } } Peter Makroczy } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Peter- I have used Micro-90 for the past couple years with good results. manufactured by: International Products Corp. PO Box 70, Bulington, NJ 08016-0070, USA phone: 609-386-8770 UK Branch: 1 Church row, Chislehurst, Kent BR7 5PG, United Kingdom Phone: 0181-467-8944 -Mike
On Fri, 23 Apr 1999, Peter Makroczy wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear microscopists, } } I have question concerning cleaning of white ceramic insulator inside } electron gun of TEM JEOL 2000FX. As a consequence of electric charges } thereare brown traces on the surface of insulator. Does anybody has } experience how to remove these traces and what cleaning solution (e.g. } aceton, methanol or something else) to use. Thank you very much. } } } Yours sincerely } } } Peter Makroczy } } Dept. of Mat.Science } } Technical University of Kosice } } Slovak Republic } } makroczy-at-tuke.sk } } -- } Peter Makroczy } Technical University of Kosice } Department of Materials Science } Park Komenskeho 11 } 042 00 Kosice } Slovak Republic } E-mail: makroczy-at-tuke.sk } Tel.: +421 95 602 25 40 } Fax.: +421 95 633 27 23 } } } }
The consensus is correct. Duniway is a good place for pump repair and exchange. They charge $495 for an exchange and it is essentially next day if a like model is in stock. Otherwise they repair the unit they receive. They have the same Varian 30 L/s pump I have in stock so I should receive the exchange unit on Monday or Tuesday.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Jon:
We use an H2/O2 torch for blowing quartz fibers to make biprisms. I believe most other practitioners in the field do the same.
Check out "Practical aspects of electron holography", D. C. Joy et al, Ultramicroscopy 51(1993) 1-14 for further tips. It is a good idea to examine the fiber in the SEM after it is mounted and coated (we do a simple Au-Pd sputter coating), to assure that it is smooth, uniform, and the correct diameter. It takes practice, and there are many techniques for blowing and collecting fibers. Finally, our health and safety guys made us build a special hood with proper ventilation before we were allowed to perform this operation. You may want to check things out in this area at your place...
Best wishes,
Larry PS I'll send you a reprint...
} } } Has anyone made a biprism for a TEM using a drawn SiO_2 coated with Au? } } I am looking for ideal fibre thickness, heat source that was used (which } gases & flame temperature) and for any tips on how to make them. } The current suggestions that I have heard are an H_2 and O_2 torch to } get the temperature up to aroung 2000 K, but these gases can be quite } hazardous to work with. } } I would really appreciate any advice. } } Thanks, Jon } } -- } ***************************************** } Jonathan Barnard } } Microstructural Physics, } H.H.Wills Physics Laboratory, } University of Bristol, } Tyndall Avenue, } Bristol BS8 1TL. } } 0117 928 9000 ext 8750 } } *****************************************
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
I have 2 suggestions that may be of interest to you:
1) MicroDrill The MicroDrill is a small drill that fits into the objective port of your=
microscope and uses the rack system on your microscope to turn it into a micro drill press. You can either "drill" holes (60u) or "scribe" circle= s of diameters from 500 to 1500 microns. The drill is battery operated. T= he drill mounts into a 36 TPI and 0.8" diameter opening. If you have anothe= r size, we can work out an adapter.
2) MicroMarker The MicroMarker is a small inked marker that will fit in place of your standard objectives in the light microscope and comes in a kit with 3 colors. There is one kit for 33mm long objectives and one kit for 45mm long objectives. They will make a circle with a 0.07" ID. =
If you would like more inforamtion, please contact me off-line.
Best regards-
David Henriks South Bay Technology, Inc. 1120 Via callejon San Clemente, CA 92673 USA
Hello everyone. I'm looking for a diamond scribing attachment for old Zeiss compound =
scopes that was used to score areas on slides. It attached to the =
objective ring like a lens, and when you found the object of interest, =
you swung this scribe around and turned it to etch a circle at that =
spot. The one I have used was in Mel Fuller's lab at UGA, but it =
disappeared when he retired (sound familiar?). It could be adjusted for =
any diameter circle. It may not necessarily be specific for a Zeiss: =
that was the scope I used it on. I can't remember the labels or =
markings on it now. It was extremely useful for flat-embedded material =
that was to be cut out and glued to blank BEEMs for sectioning.
If you know where I can find one, let me know how to purchase it.
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu ********************************************
Try the great folks at Duniway http://www.duniway.com
I don't have experience with a 30 l/s pump, but the pumps with smaller flanges have to be cut apart and then rewelded, which limits the number of times they can be rebuilt.
best regards mark
Mark W. Lund, PhD VP Engineering } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"This is a YOUNG business...How can I tell you what YOUR job is when I don't know what MINE is?" --Pogo
The M & M '99 Golf Tournament is scheduled for July 31, 1999 in the Columbia River Gorge outside Portland. Space is limited to the first 50 players. Please send you golf reservation forms in as soon as possible. If you do not have a registration form you may call 877-MSA-MAS-1 (877-672-6271)and ask for one, or you may e-mail me at Ladd Research (jarnott-at-ladd.cc) with a fax number and I can fax you a registration form. This is going to be our best tournament yet, so hurry to get your slot.
Thanks, John Arnott Chairman -- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
I was asked to post this for a friend.=A0 Please reply directly to him = at the e-mail below as he is not part of this listserver.=A0 Thank-you!
Marisa
---------------
I am looking for information on any training program that would include = TEM sample preparation. I am mostly interested in the wedge polishing = technique.
I have recently acquired an old PGT IMIX EDS system. It appears the hard drive in the Sparc 1 system is dead. I am told a replacement is nowhere to be found. Is there anyone out there who could help me locate a replacement drive? If not a drive, does anyone have a computer and analyzer for sale which is compatible with a PGT Prism digital spectrometer? Michael Ingram Rodel, Inc Polishing Lab 451 Bellevue Rd Newark, DE 19713 (302) 366-0500, ext.: 2545
Can someone recommend a good instructional video for light microscopy suitable for a graduate laboratory?
Thanks,
Susan A. Fugett
Department of Chemical Engineering and Materials Science Phone: 612-625-8803 University of Minnesota 612-625-0808 421 Washington Ave SE Fax: 612-626-7246 Minneapolis, MN 55455 Email: fugett-at-cems.umn.edu
Peter: My Hitachi engineer taught me to use acetone to wipe this ceramic insulator. I have never had very stubborn deposits, so I do not know what additional cleanig would be suitable in that case.
Maureen Petersen
} Dear microscopists, } } I have question concerning cleaning of white ceramic insulator inside } electron gun of TEM JEOL 2000FX. As a consequence of electric charges } thereare brown traces on the surface of insulator. Does anybody has } experience how to remove these traces and what cleaning solution (e.g. } aceton, methanol or something else) to use. Thank you very much. } } } Yours sincerely } } } Peter Makroczy } } Dept. of Mat.Science } } Technical University of Kosice } } Slovak Republic } } makroczy-at-tuke.sk } } -- } Peter Makroczy } Technical University of Kosice } Department of Materials Science } Park Komenskeho 11 } 042 00 Kosice } Slovak Republic } E-mail: makroczy-at-tuke.sk } Tel.: +421 95 602 25 40 } Fax.: +421 95 633 27 23
************************************************************************ Maureen Petersen Department of Plant Pathology 1453 Fifield Hall University of Florida
LATE BREAKING POSTER SESSION AT MICROSCOPY AND MICROANALYSIS '99
Microscopy and Microanalysis '99 will feature a poster session composed of presentations of newly acquired data or analyses which were unavailable for submission by the February 15 deadline. A short, half page abstract describing the studies is required. The abstract should include: Title, Authors, Authors affiliation, and a Brief Description of the studies. The description should include the Aim of the studies, a short characterization of the Methods, and a brief account of the Results and their Importance. Abstracts should be e-mailed or faxed to the program chair, Jay Jerome, at jjerome-at-wfubmc (email) or 336-716-6174 (fax). Abstracts may be submitted immediately but must be received by June 25, 1999. Abstracts will be reviewed by members of the program committee. A limited number of poster boards are available and preference will be given to early submissions. Abstract authors will be notified of acceptance of their abstracts no later than July 1 (earlier for early submissions).
Additional Information on M&M'99 can be found on the meeting WWW Site
---------------------------------------------- - AKA: W. Gray Jerome, Ph.D. - - Department of Pathology - - Wake Forest University School of Medicine - - Winston-Salem, NC 27157-1092 - - Ph: 336-716-4972, 336-716-2675 - - Fax: 336-716-6174 - - E-mail: jjerome-at-wfubmc.edu - ----------------------------------------------
John, The item is called " Object marker with diamond" and is a Zeiss item # 462960 and was $960 in the 1991 catalog.I find it very useful. Russ, Xerox
-----Original Message----- } From: John Shields [mailto:jpshield-at-arches.uga.edu] Sent: Friday, April 23, 1999 9:08 AM To: msa listserver
Hello everyone. I'm looking for a diamond scribing attachment for old Zeiss compound scopes that was used to score areas on slides. It attached to the objective ring like a lens, and when you found the object of interest, you swung this scribe around and turned it to etch a circle at that spot. The one I have used was in Mel Fuller's lab at UGA, but it disappeared when he retired (sound familiar?). It could be adjusted for any diameter circle. It may not necessarily be specific for a Zeiss: that was the scope I used it on. I can't remember the labels or markings on it now. It was extremely useful for flat-embedded material that was to be cut out and glued to blank BEEMs for sectioning.
If you know where I can find one, let me know how to purchase it.
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu ********************************************
As the manufacturer of the "MicroDrill" a microscope marking tool, I feel that I should point out one of its limitiations that was not mentioned in David Henriks reply to John Sheilds.
Unlike the earlier Zeiss product, scribes made with the "MicroDrill" are not continuously adjustable for size. Individual bits of the desired scribe diameter must be substituted into its chuck to achieve various scribe sizes.
SUMMER I 1999 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-Section BA)
NASSAU COMMUNITY COLLEGE, Garden City, Long Island, New York
A five week, Summer Session I 1999 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered four days per week (Monday through Thursday) between the hours of 8:00 am and NOON. Classes will begin on May 24 and end on June 24, 1999.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $86 per credit (for Nassau County residents or New York State residents with a certificate of residency).
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
Interested individuals should register as soon as possible since the course is limited to a total enrollment of ten (10) students.
If you have further questions, you should e-mail me directly at the address below.
For information about mail or telephone registration (Dial-a-Course) point your browser to http://www.sunynassau.edu/courses/sum99/sum99.htm. Telephone registration is only available until April 29, 1999 (between the hours of 2:30 pm to 7:00 pm) The phone numbers are (516) 572-7131 or 7372 or 7425.
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
Leitz also made the same item you are talking about that would screw into a standard nosepiece. This may still be available. if you are interested in something to do the same thing that leave a ink circle around the area then I would check with Nikon rep. this does the same thing at a fraction of the cost, Maybe $200 or so opposed to $500 or $600. I may be wrong on my estimates but it is a siginificant difference. Diamond scriciber opposed to ink???
This item is used by cytologist to mark areas of interest on slides typically typically with say 10x objective then move up to 40x high dry...
Good luck
C. Passione -----Original Message----- } From: Gillmeister, Russ {RGillmeister-at-sdms.usa.xerox.com} To: 'John Shields' {jpshield-at-arches.uga.edu} Cc: 'MSA' {Microscopy-at-sparc5.microscopy.com}
} Can someone recommend a good instructional video for light microscopy } suitable for a graduate laboratory? } You'll find a good CD-ROM reviewed in the Project MICRO bibliography; URL below. It's Pagliaro et al., "Microscopy Tutor" It's definitely university level & is described in MICRO to discourage precollege teachers from ordering it.
Thjere's a recent 15 minute video on Koehler illumination: Matsudaira, "Getting Started", from the Using Microscopy series. It's from Cogito, www.cogitomedia.com & is distributed by Academic Press.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
If indeed the "Ink" Objective Marker is suitable for the application. Nikon still has them... #79032 and the List Price is $137.00. They are RMS thread and refillable with various ink colors.
Regards,
Larry Kordon Nikon, Inc. Columbia, Maryland nikon-at-jagunet.com
Cono Passione wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } John, } } Leitz also made the same item you are talking about that would screw into a } standard nosepiece. } This may still be available. if you are interested in something to do the } same thing that leave a ink circle around the area then I would check with } Nikon rep. this does the same thing at a fraction of the } cost, Maybe $200 or so opposed to $500 or $600. I may be wrong on my } estimates but it is a } siginificant difference. Diamond scriciber opposed to ink??? } } This item is used by cytologist to mark areas of interest on slides } typically typically with say } 10x objective then move up to 40x high dry... } } Good luck } } C. Passione } -----Original Message----- } } From: Gillmeister, Russ {RGillmeister-at-sdms.usa.xerox.com} } To: 'John Shields' {jpshield-at-arches.uga.edu} } Cc: 'MSA' {Microscopy-at-sparc5.microscopy.com} } Date: Friday, April 23, 1999 9:14 PM } Subject: RE: searching for special lens } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } John, The item is called " Object marker with diamond" and is a Zeiss item } # } } 462960 and was $960 in the 1991 catalog.I find it very useful. Russ, Xerox } } } } -----Original Message----- } } } From: John Shields [mailto:jpshield-at-arches.uga.edu] } } Sent: Friday, April 23, 1999 9:08 AM } } To: msa listserver } } Subject: searching for special lens } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hello everyone. } } I'm looking for a diamond scribing attachment for old Zeiss compound } } scopes that was used to score areas on slides. It attached to the } } objective ring like a lens, and when you found the object of interest, } } you swung this scribe around and turned it to etch a circle at that } } spot. The one I have used was in Mel Fuller's lab at UGA, but it } } disappeared when he retired (sound familiar?). } } It could be adjusted for } } any diameter circle. It may not necessarily be specific for a Zeiss: } } that was the scope I used it on. I can't remember the labels or } } markings on it now. It was extremely useful for flat-embedded material } } that was to be cut out and glued to blank BEEMs for sectioning. } } } } If you know where I can find one, let me know how to purchase it. } } } } ******************************************** } } John P. Shields } } Center for Ultrastructural Research } } 151 Barrow Hall } } University of Georgia } } Athens, GA 30602-2403 } } (706)542-4080 } } jpshield-at-arches.uga.edu } } ******************************************** } } } } } }
We have to reluctantly part with our Zeiss (Leo) 902 TEM with Integrated Electron Energy Spectrometer, to make way for a new TEM. Any one interested in acquiring it can get in touch with me directly through E-mail (mvp2-at-cornell.edu). Information on the 902 is provided below:
ZEISS (LEO) 902 TEM. The unit is in excellent condition and is equipped with: integral EELS prism and spectrometer and recorder, for both one and two dimensional (quantitative) EELS imaging, a low light level (SIT) video camera and monitor, a motorized goniometer stage capable of specimen tilt through 120 degrees, and full rotation; motorized specimen movement, and a plate camera. Date acquired 10/01/89 Original cost of the unit $235,750 The unit has been under service contract since it was installed.
Dear nestor please forward this message to microscopy :
Dear netter,
All my post to microscopy was rejected and I and/or my provider (what is very small company) have been unjustify accused as a source of spam, please be so kind and send to me directly if you have similar problem.
Is there any chance to punish individuals for this accusation?. Is it custom in america that you can be taken into jail for pederesty or muder only that you have similar (not same) name or address to some criminal?
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptiste (surname) of Bejsak (Bayshark)-Collorado-Mansfeld
http://www.coleoptera.org Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., P.O.Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Your accuser was text in your header. No individual per say. Your Email contained information which matched to the automatic filter in the listserver. This is something that everyone has to put up with to protect the greater good of the ListServer community. There are a number of checks and balances that have been put into the system over the years to minimize SPAM/JUNK Mail and Viruses. Your message got flag as a POTENTIAL source, it was stopped after you replied as per the instructions it subsequently was passed through to the list after HUMAN review. Yes this caused a delay in your posting, however, the number of Undesireable messages stopped by the filter is far greater than the few that are delayed. If you do not understand or appreciate this I am sorry, but you need to realize that thousands of people can be affected by inappropriate information in the Email. Maintaining and monitoring postings even if it is only by a software program is the appropriate thing and prudent job to do in today's environment.
If you wish to submit anyone's name to your lawyers it should be me as the ListServer SysOp. I am solely responsible for the installing and maintaining the filter as well as operating the server. I have setup the key word lists and I edit and maintain them. There are no other persons involved.
I accidentally ran into part of a Nova show on a PBS station showing some very interesting micro videos of cells, micro animals, and parasites, colorized SEMs, and fiber optic close-ups of insects and caterpillars. The series of 3 shows is called Odyssey of Life. I don't know if it is current issue Nova, but I think the pictures would interest many on the list. I missed the 1st show, saw part of the second. The 3d show is an interview with the maker, the person who made some fascinating human embryo pictures some years back.
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
} } It is all attitude, } } } This is an actual job application someone submitted at a McDonald's } fast-food establishment AND THEY HIRED HIM! } (editor's note: If they hire this guy, I am SURE they would hire any of } us!!) } } } } NAME: Greg Bulmash } } DESIRED POSITION: Reclining. HA But seriously, whatever's available. If I } was in a position to be picky, I wouldn't be applying here in the first } place. } } DESIRED SALARY: $185,000 a year plus stock options and a Michael Ovitz } style severance package. If that's not possible make an offer and we can } haggle. } } EDUCATION: Yes. } } LAST POSITION HELD: Target for middle management hostility. } } SALARY: Less than I'm worth. } } MOST NOTABLE ACHIEVEMENT: My incredible collection of stolen pens and } post-it notes. } } REASON FOR LEAVING: It sucked. } } HOURS AVAILABLE TO WORK: Any. } } PREFERRED HOURS: 1:30-3:30 p.m., Monday, Tuesday, and Thursday. } } DO YOU HAVE ANY SPECIAL SKILLS?: Yes, but they're better suited to a more } intimate environment. } } MAY WE CONTACT YOUR CURRENT EMPLOYER?: If I had one, would I be here? } } DO YOU HAVE ANY PHYSICAL CONDITIONS THAT WOULD PROHIBIT YOU FROM LIFTING UP } TO 50 LBS?: Of what? } } DO YOU HAVE A CAR?: I think the more appropriate question here would be "Do } you have a car that runs?" } } HAVE YOU RECEIVED ANY SPECIAL AWARDS OR RECOGNITION?: I may already be a } winner of the Publishers Clearinghouse Sweepstakes. } } DO YOU SMOKE?: Only when set on fire. } } WHAT WOULD YOU LIKE TO BE DOING IN FIVE YEARS?: Living in the Bahamas with } a fabulously wealthy super model who thinks I'm the greatest thing since } sliced bread. Actually, I'd like to be doing that now. } } DO YOU CERTIFY THAT THE ABOVE IS TRUE AND COMPLETE TO THE BEST OF YOUR } KNOWLEDGE?: No, but I dare you to prove otherwise. } } SIGN HERE: Scorpio with Libra rising.
-- End original message --
best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**Suppose you were an idiot... And suppose you were a member of Congress ... But I repeat myself.-Mark Twain**
Thank you for many hints and formulations on Epon embedding. Now I know that I have to try out a few paths. However , meanwhile I=B4m aware of that the method of trial and error is possibly the most important basic technique in embedding business. In this case, I think I will use the formula that is traded from generation to generation in our lab (see my first mail ref. to Epon ..). Bye, Michael Reiner
To all compmed and histonet and microscopy members:
I goofed big time and I'm sorry about sending the non-related humor e-mail it was NOT intentional but I hit the wrong send key. please forgive...... now I gotta tell my boss to ignore my e-mail...
best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**Suppose you were an idiot... And suppose you were a member of Congress .. But I repeat myself.-Mark Twain**
As a subscriber to the ListServer, I would like to express my appreciationn and support for all your efforts to minimize the SPAM/JUNK mail and viruses we receive.
Gill Bond New Mexico Tech
On Mon, 26 Apr 1999, Nestor J. Zaluzec wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Ricardo } } Your accuser was text in your header. No individual per say. } Your Email contained information which matched to the automatic filter in the } listserver. This is something that everyone has to put up with } to protect the greater good of the ListServer community. There } are a number of checks and balances that have been put into the } system over the years to minimize SPAM/JUNK Mail and Viruses. } Your message got flag as a POTENTIAL source, it was stopped } after you replied as per the instructions it subsequently was passed } through to the list after HUMAN review. Yes this caused a delay } in your posting, however, the number of Undesireable messages } stopped by the filter is far greater than the few that are delayed. } If you do not understand or appreciate this I am sorry, but you need } to realize that thousands of people can be affected by inappropriate } information in the Email. Maintaining and monitoring postings } even if it is only by a software program is the appropriate thing } and prudent job to do in today's environment. } } If you wish to submit anyone's name to your lawyers it should be } me as the ListServer SysOp. I am solely responsible for the installing } and maintaining the filter as well as operating the server. I have setup the } key word lists and I edit and maintain them. There are no other } persons involved. } } } Nestor } } } } } }
Dear Mr. Martin, S.E.E. manufactures turn-key microspectrophotometers for the UV, visible, and NIR regions. Our website is located at http://www.see-incorp.com . Please feel free to contact me if you have questions. Sincerely, Paul
-- Dr. Paul Martin S.E.E. Incorporated 801 Mahler Road Suite G Burlingame, CA 94010 USA
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Dear Nestor, } } As a subscriber to the ListServer, I would like to express my } appreciationn and support for all your efforts to minimize the SPAM/JUNK } mail and viruses we receive. } } Gill Bond } New Mexico Tech } } On Mon, 26 Apr 1999, Nestor J. Zaluzec wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Ricardo } } } } Your accuser was text in your header. No individual per say. } } Your Email contained information which matched to the automatic filter in the } } listserver. This is something that everyone has to put up with } } to protect the greater good of the ListServer community. There } } are a number of checks and balances that have been put into the } } system over the years to minimize SPAM/JUNK Mail and Viruses. } } Your message got flag as a POTENTIAL source, it was stopped } } after you replied as per the instructions it subsequently was passed } } through to the list after HUMAN review. Yes this caused a delay } } in your posting, however, the number of Undesireable messages } } stopped by the filter is far greater than the few that are delayed. } } If you do not understand or appreciate this I am sorry, but you need } } to realize that thousands of people can be affected by inappropriate } } information in the Email. Maintaining and monitoring postings } } even if it is only by a software program is the appropriate thing } } and prudent job to do in today's environment. } } } } If you wish to submit anyone's name to your lawyers it should be } } me as the ListServer SysOp. I am solely responsible for the installing } } and maintaining the filter as well as operating the server. I have setup the } } key word lists and I edit and maintain them. There are no other } } persons involved. } } } } } } Nestor } } } } } } } } } } } } } } } Hi,
I am amazed and grateful for the filter that Nestor manages. About every day I swear at AOL when I find my personal computer full of obnoxious messages. Why can't AOL be as good as Nestor in firewalling rot? Sincerely, Hildy Crowley
We are in the process of sifting through information on digital TEM = cameras and have been concentrating on systems from Gatan, Advanced = Microscopy Techniques (AMT) and Soft Imaging Systems. I would = appreciate feedback from experienced users on your degree of = satisfaction with any of the camera systems from these companies (i.e. = resolution, acquisition software, customer support, etc.). Thanks for = your help.
Steve
Stephen Fields, Ph.D. Oklahoma Medical Research Foundation 825 N.E. 13th Street Oklahoma City, OK 73104 Office Phone: (405) 271-7245 Fax: (405) 271-3153 steve-fields-at-omrf.ouhsc.edu
} I accidentally ran into part of a Nova show on a PBS station showing } some very interesting micro videos of cells, micro animals, and } parasites, colorized SEMs, and fiber optic close-ups of insects and } caterpillars. The series of 3 shows is called Odyssey of Life. I don't } know if it is current issue Nova, but I think the pictures would } interest many on the list. I missed the 1st show, saw part of the } second. The 3d show is an interview with the maker, the person who } made some fascinating human embryo pictures some years back. } } } Leonard Corwin } Research Chemist } Fort Dodge Animal Health } Princeton, NJ 08543-0400
Leonard -
Here'sthe description of the third tape that appears in the Project MICRO bibliography (URL below):
Nilssen, L. 1996 The Photographer's Secrets 1 hour, $19.95 from WGBH Boston Video, P.O.Box 2284, South Burlington, VT, 05407, 800-255-9424. Lennart Nilssen is famous for his beautiful images of human development. This is part three of the Nova series Odyssey of Life (or part 4 of the series The Wonder of Life) It explains his use of SEM and other imaging tools that blur the line between microscopy and macrophotography; it will be particularly interesting for budding microscopists. All ages.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
The Quebec Chapter of the Microscopical Society of Canada is proud t= o announce:
3-Day Workshop covering all aspects of VARIABLE PRESSURE Scanning Electron Microscopy. The guest speaker is Dr. David Joy and he will cover the following topics:
Introduction to Variable Pressure S= EM Beam Interaction with Solids SE electron Imaging BSED Imaging VPSEM Imaging X-ray work in Variable Pressure Mode Lab. Sessions ... Since the number of participants is very limited we strongly advise interested people to act quickly.
DATE: August 24-26 WHERE: Varennes just south-east of Montr=E9al, QC., Canada
WORKSHOP on VARIABLE PRESSURE SEM Guest Speaker : Dr. David C. Joy
For more Information please contact : Dr. Pierre Hovington, (IREQ) ovington-at-ireq.ca , = 450 652-8125 fax: 450 652-8424
Hi there, Nigel Unwin did this in the '70s using a strand of spider silk. He picked up a spider on an aperture (don't know what species, but it was in Cambridge) and dangled it so a strand fell across the centre of the aperture. Then he coated it with gold.
The original reference is in Zeitschrift fur Naturforschung Vol 29 A (1974) pp 158-163 and was also printed as a Philips Applications bulletin EM 95 Dec 1974.
***************************************************** Mel Dickson, Deputy Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Ricardo } } Your accuser was text in your header. No individual per say. } Your Email contained information which matched to the automatic filter in the } listserver. This is something that everyone has to put up with } to protect the greater good of the ListServer community. There } are a number of checks and balances that have been put into the } system over the years to minimize SPAM/JUNK Mail and Viruses. } Your message got flag as a POTENTIAL source, it was stopped } after you replied as per the instructions it subsequently was passed } through to the list after HUMAN review. Yes this caused a delay } in your posting, however, the number of Undesireable messages } stopped by the filter is far greater than the few that are delayed. } If you do not understand or appreciate this I am sorry, but you need } to realize that thousands of people can be affected by inappropriate } information in the Email. Maintaining and monitoring postings } even if it is only by a software program is the appropriate thing } and prudent job to do in today's environment. } } If you wish to submit anyone's name to your lawyers it should be } me as the ListServer SysOp. I am solely responsible for the installing } and maintaining the filter as well as operating the server. I have setup the } key word lists and I edit and maintain them. There are no other } persons involved. } } } Nestor
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
In the last 2 1/2 years, I have been working with muscle tissue and blood buffy coats from dogs infected with Hepatozoonosis. Part of my PhD study is to morphologically detail the asexual stages of the parasite, Hepatozoon americanum which is responsible for canine hepatozoonosis in North America. Based on the literature, other researchers believe the parasite lives in neutrophils; however based on preliminary TEM work, I am certain these parasite dwell in a "transformed" or " altered" macrophage. I am aware that normal macrophages and neutrophils can be easily differentiated based on there ultrastructural characteristics; however, since these cells are transformed due to the parasite's existence in the cytoplasm, the basic characteristics of the cell have changed; therefore, I am interested in finding a method which will positively identify the cell in question. Currently with light microscopy , I have tried with no success immunohistochemistry and enzyme histochemistry. Thus, my question is: Does anyone have a working TEM marker or TEM method for positively differentiating dog macrophages from neutrophils other than by their basis ultrastructural characteristics? I would appreciate any comments or suggestions.
Dear Listservers, Does anyone have a reference or simple method for quantifying viruses in solution? A colleague has a suspension which needs to be quantified. One assay technique detects a concentration of 10 (E10) per ml while a second assay technique detects a concentration of 10 (E11) per ml. I wonder if the error involved with quantification by EM would be too great to shed any light on the problem. I expect the best option would involve negative staining. Thankyou.
John Brealey Queen Elizabeth Hospital EM Unit Adelaide South Australia }
We have just installed a CM200 FEG TEM/STEM and EDS.
Does anyone know what are the best conditions (Gun lens/extraction voltage/spot sizes/C1 and C2 aperture sizes etc etc) for performing high spatial resolution microanalysis?
We are trying to look at chemically heterogenous particles which are only a few nanometres in size {smaller} . {/smaller}
I am looking for an inexpensive microscope 5X to 100 X and would probably settle for a monocular. Changing oculars is not a problem.
It is for photography. I have a 4X5 Leintz camera and a 35mm Ziess with a front reflex attachment so a vertical ocular would be ideal.
It is for personal use so the cosmetics are not very important. The optics are important. I have a pretty good range of lighting equipment so lights are not necessary.
If you have any thing that might be of interest to me please email me. I am looking to spend less than $250.00.
I realize that I probably won't be able to get everything I want in one scope.
thanks Gordon
Gordon Couger gcouger-at-couger.com www.couger.com/gcouger Stillwater, OK 405 624-2855 GMT -6:00
-----Original Message----- } From: Stan, Pat, Robert, Peter, and Angus Hansen [mailto:sehansen-at-bellatlantic.net] Sent: Monday, April 26, 1999 8:10 PM To: rschoonh-at-sph.unc.edu Cc: COMPMED-at-LISTSERV.AALAS.ORG; histonet; Microscopy ListServ
Anyone have a surplus Reichert/Leica FC4E or parts? ..or know of one available? I would like to find some back up parts and equipment and sources. Also are there othere cryo-ultramicrotomes for TEM uses other than RMC, Microstar and Leica? Also accessories? Also are tere any ultra-cryo type list servers? Jeff Day/ 'JD'
___________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com/getjuno.html or call Juno at (800) 654-JUNO [654-5866]
John: Mix 1 drop of virus solution and 1 drop of a suitable size latex solution of known and reasonably similar concentration as the unknown. The latex spheres concentrations are fairly accurately known in terms of percent w/v. This, for a certain size latex spheres can be calculated and diluted to achieve the required particle number concentration.
If greater accuracy is required the concentration of the latex stock solution can be checked in one of the electronic particle sizing/ counting instruments (eg. Coulter counter). Coated grids are applied to the mixed drop and in the case of small particles, this is negatively stained.
Disclaimer: ProSciTech provides the largest range of latex particles in small volumes (mostly 5ml) from 0.1um to 20um in 12 sizes. Applications and concentration versus size conversions and other data is online, linked to page S2, which is accessed from "Contents". Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
} } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } Quantification of viruses in solution. } } Dear Listservers, Does anyone have a reference or simple } method for } quantifying viruses in solution? A colleague has a } suspension which needs } to be quantified. One assay technique detects a } concentration of 10 } (E10) per ml while a second assay technique detects a } concentration of 10 } (E11) per ml. I wonder if the error involved with } quantification by } EM would be too great to shed any light on the problem. I } expect the best } option would involve negative staining. Thankyou. } } John Brealey } Queen Elizabeth Hospital } EM Unit Adelaide South Australia } } } }
Hi, There were a debate on the list a few month ago about anti-vibration table (home made systems and commercial ones). If anybody has collected the emails related to that, could he forward them directly to me. Thanks for your help! Cecile Dr. Cecile Prouteau School of Metallurgy & Materials The University of Birmingham Edgbaston, Birmingham B15 2TT UK E-mail : c.prouteau-at-bham.ac.uk tel : (UK=44)- (0)121 414 5170 fax : (UK=44)- (0)121 414 5232
Question: I am a relative novice at microscopy and have the task of trying to freeze-substitute samples containing large amounts of fat in the form of fat droplets (size range between sub-micron to approx. 10 microns) for TEM. Although I can produce reasonable results for semi-thin sections for LM, the fixatives (combinations of glutaraldehyde, OsO4, RuO4, and UA) and solvents (acetone and methanol) I have used do not fix the fat sufficiently for ultra-thin sectioning. (I have also varied the substitution and fixation periods from two days to over a week with little success). Can anybody suggest a fixative/solvent regime which will fix and dehydrate, but not mobilize, the fat? I have good indications that methanol and ethanol are redistributing the fat considerably at -90C, even after three days osmication. Sample preparation also requires the temperature to remain below -25C until polymerization. Any suggestions would be welcome! Regards, Nick Johnson
Nick.B.Johnson-at-Unilever.com
Dr. N. Johnson, Structural Analysis, Unilever Research, Colworth House, Beds, U.K.
We have a Hitachi S-520 SEM installed in 1983. Lately, we have experienced instability problems in the SEM and I'm not sure what to do. The Hitachi S-520 SEM has a single rotary pump, single diffusion pump and Pirani gauge. On initial pumping and warm-up, the SEM gets down to high vacuum (reading 10uA) quite normally in about 20min. On applying HT (20kV), everything appears normal for approximately 80min, then, suddenly the HT switches itself off and the vacuum gauge rises to 15uA in 30sec and then falls back to10uA in 2min. If the slightest gun emission current is applied when the SEM says it's ready, the HT instantly switches itself off. I have cleaned the filament assembly, anode and gun chamber around the anode. However, I have not cleaned the insulator at the top of the gun chamber. The Pirani gauge is clean and the diffusion pump is not too hot. Any advice would be much appreciated and gratefully received. Thankyou.
John Brealey Queen Elizabeth Hospital EM Unit Adelaide South Australia
Steve, We have two AMT systems with 2K x 2K Kodak Megaplus cameras coupled to a Kodak XLS8600 printer and a Codonics (Kodak) NP1600 printer. We just upgraded to Windows NT with a few minor glitches which for the most part have been resolved. We have been very satisifed with the results as have our customers. The support is very good as is the reliability. I have no experience with the Gatan or SIS cameras. Russ, Xerox
-----Original Message----- } From: Steve Fields [mailto:steve-fields-at-omrf.ouhsc.edu] Sent: Monday, April 26, 1999 3:42 PM To: 'Microscopy-at-MSA.Microscopy.Com'
We are in the process of sifting through information on digital TEM cameras and have been concentrating on systems from Gatan, Advanced Microscopy Techniques (AMT) and Soft Imaging Systems. I would appreciate feedback from experienced users on your degree of satisfaction with any of the camera systems from these companies (i.e. resolution, acquisition software, customer support, etc.). Thanks for your help.
Steve
Stephen Fields, Ph.D. Oklahoma Medical Research Foundation 825 N.E. 13th Street Oklahoma City, OK 73104 Office Phone: (405) 271-7245 Fax: (405) 271-3153 steve-fields-at-omrf.ouhsc.edu
Try the May '97 Microscopy Today, pg. 20 (Microscopy 101):
Quantitation of Virus Particles by Negative Staining for TEM by Robert Alain, Institut Armand-Frapplier
Phil
} } Quantification of viruses in solution. } } Dear Listservers, Does anyone have a reference or simple method for } quantifying viruses in solution? A colleague has a suspension which needs } to be quantified. One assay technique detects a concentration of 10 } (E10) per ml while a second assay technique detects a concentration of 10 } (E11) per ml. I wonder if the error involved with quantification by } EM would be too great to shed any light on the problem. I expect the best } option would involve negative staining. Thankyou. } } John Brealey } Queen Elizabeth Hospital } EM Unit Adelaide South Australia } }
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 Voice: (608) 833-2885 Fax: (608) 836-1969 (please make sure my name is on any fax) oshel-at-terracom.net
I am submitting this for the person above. Please reply to her.
} I have a question concerning the measurement of the hyphe of the fungi } Aspergillus oryzae in a agar matrix. } We have grown the fungi in a agarmatrix, then we make thin coupes and put them } under the microscope. then we want to measure the amount of fungi in a certain } volume of agarmatrix. We make pictures of the coupes and try to work on it } with the computer programe 'Adobe Photoshop'. The problem we encounter is that } the colour inside the hyphe is the same as the colour of the agarmatrix, the } only thing we do see is the wall of the hyphe. So we would like to colour the } inside of the hyphe, so we can differentiate between inside and outside the } hyphe. Using fluorchromen to colour the inside gives us the problem that it } only colours certain parts (for instance living or dead) of the hyphe. } -So i am looking for a dye that can colour all inside of the hyphe if it is } possible. } -Or a dye that colours the cell wall very clearly } -Or a stain that colours the agarmatrix without colouring the fungi. } } Thank you very much for any tips you might have } } greetings Marjolein } } } } Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
This is a good job for a grad student / post doc. You should run through the calculations of probe size vs Cs & Cc. Then do the physical measurments of probe size vs beam current and create yourself a operational / working curves. The expt. measurements are always slightly different than the theoretical, but the calculations will put you in the right general area.
We did this for the VG here at ANL, you can see the results in the AAEM Electronic Notebook, and/or look at the Proc. of Microscopy & Microanalysis '98 Atlanta pge 386.
If you go to the notebook (http://tpm.amc.anl.gov/NJZ/AAEMNoteBook.html) do a search for the key words: FEG, brightness, VOA Current
Nestor
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sorry for the flood but you asked for it. The followin e-mails contain the most recent discussions via e-mail as I am a year behind in the Tips & Tricks archives. There is also another discussion archived at the Tips & Tricks site you may be interested in. Point your browser to
http://www.biotech.ufl.edu/~emcl
follow the Tips & Tricks link and look in the Light Microscopy section.
Our whole site is under renovation so please bear with me for the next month or so while I get it finished. Comments, suggestions, and contributions are always welcome.
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
The New England Society for Microscopy will hold its Sixteenth Annual Spring Symposium at Marine Biological Laboratories, Woods Hole, Massachusetts on May 7 & 8, 1999. In addition to our scientific presentations, this year's Symposium=
will feature PROEJCT MICRO, the new Great Explorations in Math and Scienc= e Program designed to introduce Microscopy to elementary- and middle school-aged students. If you are interested in participating in this exciting new program, be sure to register and join us for dinner on Frida= y evening. =
PRIZES for Best Poster, Best Student Poster, and Best Photo-As-Art will also be awarded. If you're interested in submitting your poster for judging, please contact Dr. Changmo Sung at Changmo_Sung-at-uml.edu. =
PROGRAM Friday, May 7th
12:00 pm Registration: Swope Center
1:00 pm Welcome: Lillie Auditorium
Session I Chairperson: Doug Taatjes, UVM
1:05 pm Laser Scanning Cytometry as an Imaging System Ed Luther, Senior Scientist - Biology, CompuCyte Corporation, Cambridge, MA
1:45 pm Focused Ion Beam Microscopy: The New Kid on the Block Dr. David Casey, Senior Scientist, Micrion Corp, Peabody, MA
2:25 pm Membrane Fusion Machinery in Cells Dr. Bhanu Jena, Dept. of Surgery and Biomedical Eng., Yale University, New Haven, CT
3:05 pm Afternoon Break Coffee served in Lillie 103
Session II Chairperson: Tony Garratt-Reed, M.I.T.
3:20 pm Multi-technique Characterization of Emissive Coatings on Electrod= es Dr. Chris Peters, Senior Project Engineer R & D, Osram Sylvania, Beverly, MA
4:00 pm Microscopic Approach to the Study of Pancreatic Islets Dr. Thomas Jetton, Ergo Scientific, Inc., Cambridge, MA
4:40 pm Advances in Microanalysis: A Look at the Past, Present and Future=
Dr. William Hardy, CEO, Princeton Gamma Tech, Princeton, NJ
5:30 pm Cocktails and Dinner: Swope Center
7:30 pm Project Micro "Festival" Meigs Room, Swope Center Dr. Burton E. Goodrich, Jr., Executive Director, South Coast Educational Collaborative Janet E. Goodrich, Principal, Miscoe Hill Elementary School, Mendon, MA
Saturday, May 8th
7 to 8:00 am Breakfast: Swope Center
Session III Chairperson: Eben Oldmixon, HSPH =
8:30 am The Use of Electron Microscopy in the Characterization of Carbon Deposits Paul Anderson, Chemistry Department, Northeastern University, Boston, MA
9:10 am Confocal Microscopy of Nuclear Envelope Breakdown Dr. Mark Terasaki, Asst. Prof., Dept. of Physiology, UCONN Health=
Ctr., Farmington, CT
10:00 am Commercial Exhibits and Posters: Swope Center Coffee and doughnuts will be served
12:30 pm Presentation of Poster and Photos-As-Art Awards and Door Prizes: =
Poster Area, Swope Center
1:00 pm Lunch with Short Tour of MBL: Swope Center =
2:00 pm 2-hour Discovery Cruise aboard the R/V Patriot II Tickets must be reserved in advance
Register Today! Contact L. Kirstein at NESM-at-compuserve.com. Registratio= n Deadline will be extended to April 30th. =
SILENT AUCTION, OCEANOGRAPHY CRUISE, POSTER AWARDS, DOOR PRIZES =
I am looking for a CCD camera and image capture card for use with a standard Zeiss light microscope. I've been looking all over and I can't find any cameras designed for this use. Can anyone help me out with the name of a vendor or a website?
I recall reading somewhere a long time ago about special fixation procedures for non-conductive samples that are hard to adequately coat. I seem to remember tannic acid being involved. Does this ring a bell with anybody out there?
Specifically, we are processing pig intestine samples for SEM and are having problems with charging on the microvilli. We have repeatedly coated the samples with Au/Pd, but the problems persist. Perhaps there's a specialized processing technique that can help?
Thanks in advance. Randy
Randy Tindall Electron Microscope Core College of Veterinary Medicine University of Missouri - Columbia Phone: 573-882-8304
Dear Mel, } } Does anyone know what are the best conditions (Gun lens/extraction } voltage/spot sizes/C1 and C2 aperture sizes etc etc) for performing high } spatial resolution microanalysis? } You want to minimize both x-rays generated up-column (primarily at the condenser apertures) and those generated by scattered, possibly multiply scattered, electrons interacting in the specimen and detector areas. The former will depend on the the composition of the apertures, and is lessened by the use of low-z materials. We installed an aluminum C1 aperture for this reason, and we obtained some ~1 mm thick Be disks with 100 um holes for use as C2 apertures. The aluminum C1 works very well, but the Be C2 has charging problems from the surface oxide. Our best results for spatial resolution were not spectacular, but since the high-voltage scope is used for thick specimens where the beam will spread within the specimen, we were not too concerned. You want to have a high enough voltage that beam spread and multiple scattering within the spe- cimen are minimized, but you need to consider the efficiency of your shielding as a function of voltage. I suggest using calculations to arrive at a proposed optimum and experimenting with the relevant parameters to find the true optimum. } } We are trying to look at chemically heterogenous particles which are only } a few nanometres in size. } Will you be able to get sufficient signal from one of these so that the background produced by excitation of the rest of the specimen by stray radiation is small in comparison? Do you want to characterize the hetero- geneity within a single particle? Good luck. Yours, Bill Tivol
Thanks to you all, It's been great help Cecile Dr. Cecile Prouteau School of Metallurgy & Materials The University of Birmingham Edgbaston, Birmingham B15 2TT UK E-mail : c.prouteau-at-bham.ac.uk tel : (UK=44)- (0)121 414 5170 fax : (UK=44)- (0)121 414 5232
Dear Listpeople: Is it possible to cut reasonable quality longitudinal sections of rat molar undecalcified tooth between 100 nm to 0.1 mm with a diamond or sapphire knife? Does it perhaps require special circumstances and equipment? They will be used in immunocytochemical labeling for TEM. Thank you, Lauri
Dr. Lauri J. Pelliniemi, M.D. Telephone +358-2-3337312 University of Turku Kiinamyllynkatu 10 Internet e-mail lauri.pelliniemi-at-utu.fi FIN-20520 Turku FINLAND. Europe http://www.utu.fi/med/em/personne.html
Dear Mel, When you do EDS on STEM, you use the same sort of conditions as an SEM. Small aperature and higher condensor lens setting will give you better spacial resolution, but at the cost of x-ray count rate. On a thin sample you may only have a hundred counts per second or less, so count times may get long You wrote: } } We have just installed a CM200 FEG TEM/STEM and EDS. } } Does anyone know what are the best conditions (Gun lens/extraction voltage/spot sizes/C1 and C2 aperture sizes etc etc) for performing high spatial resolution microanalysis? } } We are trying to look at chemically heterogenous particles which are only a few nanometres in size. } ***************************************************** } Mel Dickson, } Deputy Director. } Electron Microscope Unit, } University of New South Wales. } Sydney NSW 2052 Australia } } Phone (+612) 9385-6383 } Fax (+612) 9385-6400 } Website {http://srv.emunit.unsw.edu.au} } ***************************************************** Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
} } } "Bartlett, Jeanine" {jqb7-at-cdc.gov} 04/27 6:41 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
As did I!
-----Original Message----- } From: Stan, Pat, Robert, Peter, and Angus Hansen [mailto:sehansen-at-bellatlantic.net] Sent: Monday, April 26, 1999 8:10 PM To: rschoonh-at-sph.unc.edu Cc: COMPMED-at-LISTSERV.AALAS.ORG; histonet; Microscopy ListServ
On Tue, 27 Apr 1999, Chris Bradley wrote:
} I am looking for a CCD camera and image capture card for use with a standard Zeiss light } microscope. I've been looking all over and I can't find any cameras designed for this } use. Can anyone help me out with the name of a vendor or a website?
???? Pretty much SOP today and all over the Web.*. Check Edmunds or McCrone. Also, any C-mount camera can easily be adapted to your microscope tube. The grabber cards are ubiquitous as well.
However, let me suggest you start the other way 'round. Define your application needs and decide on the software first. Then, choose grabber and camera from the supported list. Otherwise, you may buy something which won't give you what you need.
Method 1: To determine virus in solution is to do a plaque assay. Keep = diluting viral stock until there is approxamitely 1 virus per ml and = infect cell cultures with it. This determine infective particles.
Method 2: Ref. is Sharp, D.G., 1974. Proceedings of 32nd Annual = Meeting of EMSA. Clayton's Publishing Division, p. 264-265.=20
Method 3: McCombs, R.M., Benyesh-Melnick, M. and Brunschwig, J.P. 1966 = Biophysical studies of vesicular stomatitis virus. J. Bacteriol. = 91:803-812
The suggestion of using latex spheres is ok but has one major flaw in the = technique, Do the sphere's and viral particles stick to the grid in equal = proportions????? probably not.
In my experence, I am able to dectect down to 1 x 10 e6 particles per ml. = using method 2 above.
Best of luck,
Ed
Edward P. Calomeni Dept Pathology - EM Lab 3000 Arlington Ave. Toledo, OH 43614
} } } "Nestor J. Zaluzec" {zaluzec-at-Sparc5.Microscopy.Com} 04/26 9:38 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html=20=
Dear Listservers, Does anyone have a reference or simple method for quantifying viruses in solution? A colleague has a suspension which needs to be quantified. One assay technique detects a concentration of 10 (E10) per ml while a second assay technique detects a concentration of 10 (E11) per ml. I wonder if the error involved with quantification by EM would be too great to shed any light on the problem. I expect the best option would involve negative staining. Thankyou.
John Brealey Queen Elizabeth Hospital EM Unit Adelaide South Australia }
Randy, check this reference: Murakami, T (1973) A revised tanin-osmium = method for non coated sem specimens. Arch. Hist. Jap 36(3) 189-. Or = what I think is superior, the OTOTO method of Robert Kelley which uses = thiocarbohydrazide as a ligand to bind more osmium to the surface. = Hayat has this technique in one of his SEM method volumes and Kelley's = original citation is in Ultrastructure Research 45 (1973) 254-258.
Hank Adams Integrated Microscopy Core Cell Biology Baylor College of Medicine
Nick: I am more of a novice than you, and would GREATLY appreciate any tips you get on this. ( In fact would any *experienced* lab like to do this work for me for a fee?) I am doing something similar, a suspension of 15 micron liposome aggregates containing vegetable oils.
I don't have the answer. I presume osmication or rutheniation should be done extensively at room temp or preferably *above* (37C), before dehydration. A method using imidazole was highly recommended to me, as better than extended osmication, see reference below. I do expect that freeze-substitution is the way to go for dehydration. Acetone is known to extract fewer phospholipids than do the alcohols; it may be better for nonpolar lipids as well. Just keep it well-chilled.
Angermuller S, Fahimi HD, Histochem J 1982 Sep;14(5):823-35, Imidazole-buffered osmium tetroxide: an excellent stain for visualization of lipids in transmission electron microscopy. (nb: in PubMed just now I hit the "related articles" button and found a few interesting articles http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?db=m&form=6&uid=6182131&dopt=m&dispmax=100 )
I hired a local EM tech to work on this and was disappointed with the results, so hope you do find a method that works, and that you can let me know. For your smaller particles, presumably you can use cryo TEM with an energy loss filter and avoid fixing entirely, but I can't with my large aggregates
Good luck Richard
} } } Nick B Johnson {Nick.B.Johnson-at-unilever.com} 04/27/99 05:13am } } } Question: I am a relative novice at microscopy and have the task of trying to freeze-substitute samples containing large amounts of fat in the form of fat droplets (size range between sub-micron to approx. 10 microns) for TEM. Although I can produce reasonable results for semi-thin sections for LM, the fixatives (combinations of glutaraldehyde, OsO4, RuO4, and UA) and solvents (acetone and methanol) I have used do not fix the fat sufficiently for ultra-thin sectioning. (I have also varied the substitution and fixation periods from two days to over a week with little success). Can anybody suggest a fixative/solvent regime which will fix and dehydrate, but not mobilize, the fat? I have good indications that methanol and ethanol are redistributing the fat considerably at -90C, even after three days osmication. Sample preparation also requires the temperature to remain below -25C until polymerization. Any suggestions would be welcome! Regards, Nick Johnson
Nick.B.Johnson-at-Unilever.com
Dr. N. Johnson, Structural Analysis, Unilever Research, Colworth House, Beds, U.K.
We had AMT install their 12-bit system that uses a Hammamatsu camera. I'm using a Lexmark laser printer for everyday prints. but have access to a multitude of other printers on our network. Though we have no dye-sub printer, excellent results have been achieved with premium inkjet photo paper and inkjet printers.
AMT's reputation among vendors and users, performance of instrumentation, and price were all major considerations. Subsequent interaction after installation has been very good, too. One other reason for going to AMT was the non-interest by GATAN.
Overall, the system has more than lived up to expectations. I don't miss the darkroom one bit.
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Steve, We have two AMT systems with 2K x 2K Kodak Megaplus cameras coupled to a Kodak XLS8600 printer and a Codonics (Kodak) NP1600 printer. We just upgraded to Windows NT with a few minor glitches which for the most part have been resolved. We have been very satisifed with the results as have our customers. The support is very good as is the reliability. I have no experience with the Gatan or SIS cameras. Russ, Xerox
-----Original Message----- } From: Steve Fields [mailto:steve-fields-at-omrf.ouhsc.edu] Sent: Monday, April 26, 1999 3:42 PM To: 'Microscopy-at-MSA.Microscopy.Com'
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We are in the process of sifting through information on digital TEM cameras and have been concentrating on systems from Gatan, Advanced Microscopy Techniques (AMT) and Soft Imaging Systems. I would appreciate feedback from experienced users on your degree of satisfaction with any of the camera systems from these companies (i.e. resolution, acquisition software, customer support, etc.). Thanks for your help.
Steve
Stephen Fields, Ph.D. Oklahoma Medical Research Foundation 825 N.E. 13th Street Oklahoma City, OK 73104 Office Phone: (405) 271-7245 Fax: (405) 271-3153 steve-fields-at-omrf.ouhsc.edu
id PAA24629; Tue, 27 Apr 1999 15:19:40 -0500 (EST) Message-Id: {3.0.3.32.19990427162046.006fc9bc-at-biotech} X-Sender: gwe-at-biotech X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.3 (32)
Regarding TEM of fat, I might suggest two possible approaches. One would be to use a hydrophobic resin like Lowicryl HM-20 or HM-11. It might even serve as the substitution media. Another thought is to freeze dry the samples and embedd them in resin directly . One could also expose the sample, after drying, to osmium vapors from the crystals in order to keep things anhydrous. I would recommend a resin that can tolerate a little water, like Epon or its equivalent, or a methacrylate based resin.
Would using freeze-fracture EM give the results you are after??
Fats and lipids are a nightmare for EM.
Greg Erdos Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
Has anyone out there managed to add transmitted-light illumination to the optical microscope of a JEOL 840? I'd appreciate hearing from you.
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
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I have located two good prospects for CCD cameras which do not need to be tethered to a computer. They are:
Olympus DP10 - uses a smart card to record images. Has a built in LCD panel for formatting your image and a keypad for interacting with the programming. You can print directly from the camera if desired. Resolution: 1280x1024. Price ~$3000
Fugi HC-300Z: does not have built in LCD panel so it is helpful to attach a small monitor for setting up your formatting, etc. Downloads images directly to a ZIP disk. Has small keypad for interfacing with software. Resolution: 1280 x 1000. Price ~$4000
Both mount using standard C-mounts. They may need a relay lens. They can be used on a copy stand with a standard camera lens.
Kodak also makes a microscope tube adapter for use with one of their consumer cameras. Does not appear to give the quality that the above ones do and is tethered to a computer, as are SPOT cameras, etc. Also, most of the other available cameras are substantially more expensive ( most are "cooled") although they may perform better under low light.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
Chris Bradley wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi everyone, I am looking for information about selected area electron channelling technique which is used for strain measurement of crystalline materials. I have read few articles but I do not have any idea about what size sample should be prepared, what kind of backscatter dedector do i need etc. If anybody who has experience with that technique please inform me. Thanks
} Olympus DP10 - uses a smart card to record images. Has a built in LCD } panel for formatting your image and a keypad for interacting with the } programming. You can print directly from the camera if desired. Resolution: } 1280x1024. Price ~$3000
If it records on SmartMedia, one does not need a frame grabber with it. I use a Lexar SmartMedia (parallel-port) reader. It functions like a removeable disc drive permitting one to read/write the SM. It cost me $37.
} Fugi HC-300Z: does not have built in LCD panel so it is helpful to attach } a small monitor for setting up your formatting, etc. Downloads images } directly to a ZIP disk. Has small keypad for interfacing with software. } Resolution: 1280 x 1000. Price ~$4000
Again. No grabber.
} Kodak also makes a microscope tube adapter for use with one of their } consumer cameras.
Also, it (DC-120) retains its non-removeable lens and has poorer resolution.
Consider also the Pixera and the MicroVision in this class of camera.
What all of these do NOT permit is the use of real-time image processing software. All processing is done post hoc. If this is OK in one's application, fine. Otherwise, one must consider a separate camera + grabber.
How are you coating your specimens? The common approach is to place the specimen in the chamber and to let the coater get on with the task? Microvilli are difficult to coat if you use this method and if this is followed by the common observation at 20kV then I can understand why the samples charge.
Try this coating method.
1) Set the sample at approx 45deg in the coater with a working distance of 5cms 2) With a coater that indicates kV on its variable control set this = at the lowest level at which a plasma will strike (probably ~800 volts) and then tune for 20mA. If the coater has a deposition control set it at 10m= A. 3) Coat for one minute 4) Tilt the sample in exactly the opposite direction and repeat.
Try it out at 10kV
5) If not quite good enough - run the coater to obtain the best vacu= um that you can and then try to strike a plasma without gas being introduced= , or at worst the very minimum of gas - coat for one minute with the sample=
flat.
To improve your microscopes performance at the lower kV be sure to have a= t least 100uA emission current with a Japanese instrument (W) with Philips=
50uA(W) and a WD of less than 10mm. If you can not obtain these curren= t levels you need to place the filament nearer the cap.
Hope this helps?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Sorry to hear you are having problems but I am afraid high voltage is wha= t we call in the trade a "standard fault" on the otherwise superb S520.
Not being on the spot it is of course very difficult. Any fall off in vacuum level is likely to trip the high voltage, however I am inclined to=
tell you to look at the gun area. Inside the gun casing are the connections between the gun components and the high voltage cable. These=
connections in time break down and give a wide variety of problems; this = is 90% likely to be the problem! I do not recall many vacuum funnies on 520= s.
Just as a test, run the instrument up at a lower kV (say 2 or 5kV) and s= ee if the fault is identical. If the fault changes we prove that the fault = is high voltage related, then you go to the area I have outlined.
This area of the gun is sealed, but if that is where the problem is you have three routes (1) have a go yourself when we can give you step by ste= p instructions. (2) Get the local agent to sort it out. (3) Look around t= o see if there is a company nearby that specialises in high voltage repairs=
Well, I hope we have helped, remember a motto "if a man built it a man ca= n fix it", no offence to the ladies as man is used in the generic sense.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
We routinely use a variant of Mallick & Wilsons ( IITRI/SEM 1975; 259-264) OTOTO (osmium -thiocarbohydrazide) technique for cochlea specimens (organ of Corti - inner ear) which have a very convoluted shape and are extremely difficult to sputter coat. The wash must be thorough, including the neck, rim and cap of the specimen vial; any trace of TCH or OsO4 remaining will form a precipitate when the other reagent is added. We have also successfully used the osmium-tannic acid method but prefer the former for very convoluted specimens. One advantage of this method over sputter coating is that it permits subsequent redissection and SEM observation without recoating and/or embedding for TEM examination (e.g., Hunter-Duvar IM, Mount RJ (1978). The organ of Corti following ototoxic antibiotic treatment. Scanning Electron Micros/1978/II., 423-430).
O-T-O-T-O SUMMARY
- fix, postfix OsO4 - 1% aq TCH 20 min. with agitation - wash well - 1% aq OsO4 2 hr. with agitation - wash well - 1% aq TCH 20 min. with agitation - wash well - 1% aq OsO4 2 hr. with agitation - wash well - dehydrate - CPD
"Tindall, Randy D." wrote:
} I recall reading somewhere a long time ago about special fixation procedures } for non-conductive samples that are hard to adequately coat. I seem to } remember tannic acid being involved. Does this ring a bell with anybody out } there? } } Specifically, we are processing pig intestine samples for SEM and are having } problems with charging on the microvilli. We have repeatedly coated the } samples with Au/Pd, but the problems persist. Perhaps there's a specialized } processing technique that can help?
--
Richard J. Mount Auditory Science Laboratory, Department of Otolaryngology & Brain and Behaviour Division/Research Institute The Hospital for Sick Children Toronto, Ontario, Canada (416) 813-6551; Fax (416) 813-8456 http://www.sickkids.on.ca/HSCWeb/Otolaryngology/Otoalias/Earhome1.htm
Hello John: } "The suggestion of using latex spheres is ok but has one } major flaw in the technique, Do the sphere's and viral } particles stick to the grid in equal proportions????? } probably not."
Maybe you are right, but I doubt it, especially for small particles like virus. When touching a coated grid repeatedly against a drop of virus solution and part blotting it, the aim is to retain a thin film of the aqueous solution. This dries down and the particles adhere. This is my "model" for just applying an aqueous solution. I would expect that the loss of particles, latex or virus would be small and similar.
When negatively staining particles, those particles are mixed in a metallic solution. After that solution has dried down the particles are actually embedded in the metallic film. Differential retention of particles? . . . . I doubt it very much. It should also be noted that latex spheres have been used for decades to estimate/count particles in EM. Maybe all that data was wrong, but it seems rather more likely that some of those authors did their home work.
I have no hard data on this, but intuitively, with many years of negative staining experience I would be most surprised if this was otherwise. Particle retention during negative staining seems such an obvious problem, somebody is sure to have studied it. Please tell me if I am wrong. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Wednesday, April 28, 1999 4:11 AM, Ed Calomeni [SMTP:ecalomeni-at-mco.edu] wrote:
} } Hi John, } } Method 1: To determine virus in solution is to do a plaque } assay. Keep diluting viral stock until there is } approxamitely 1 virus per ml and infect cell cultures } with it. This determine infective particles. } } Method 2: Ref. is Sharp, D.G., 1974. Proceedings of } 32nd Annual Meeting of EMSA. Clayton's Publishing } Division, p. 264-265. } } Method 3: McCombs, R.M., Benyesh-Melnick, M. and } Brunschwig, J.P. 1966 Biophysical studies of vesicular } stomatitis virus. J. Bacteriol. 91:803-812 } } The suggestion of using latex spheres is ok but has one } major flaw in the technique, Do the sphere's and viral } particles stick to the grid in equal proportions????? } probably not. } } In my experence, I am able to dectect down to 1 x 10 e6 } particles per ml. using method 2 above. } } Best of luck, } } Ed } } Edward P. Calomeni } Dept Pathology - EM Lab } 3000 Arlington Ave. } Toledo, OH 43614 } } 419-383-3484 } 419-383-3066 (fax) } ecalomeni-at-mco.edu } } Edward P. Calomeni } Dept Pathology - EM Lab } 3000 Arlington Ave. } Toledo, OH 43614 } } 419-383-3484 } 419-383-3066 (fax) } ecalomeni-at-mco.edu } } } } } "Nestor J. Zaluzec" {zaluzec-at-Sparc5.Microscopy.Com} } } } } 04/26 9:38 PM } } } } } Quantification of viruses in solution. } } Dear Listservers, Does anyone have a reference or simple } method for } quantifying viruses in solution? A colleague has a } suspension which needs } to be quantified. One assay technique detects a } concentration of 10 } (E10) per ml while a second assay technique detects a } concentration of 10 } (E11) per ml. I wonder if the error involved with } quantification by } EM would be too great to shed any light on the problem. I } expect the best } option would involve negative staining. Thankyou. } } John Brealey } Queen Elizabeth Hospital } EM Unit Adelaide South Australia } } } } } }
Hello Ritchie: Some years ago I too was pushed about by some geologists who could not adapt to a perfectly good atomic number (backscattered) image. I know that compromise transmitted light scopes exist on some WD probes. The 840 was not designed for it. To be useful you require double polarisers; there is insufficient room near the centre of the stage, you require interlocks (no light with SE detector on), the carbon coating kills polarised images. The more you look at the possibility the more problems you will find. If you persist you eventually obtain crummy light microscope images. Non microprobe specialist geologist users often mistake the provided scope (on WD systems) as a weak attempt to provide a light microscope to view the specimen - not so. Its real function is to locate and focus the beam spot using a fluorescent mineral and to calibrate working distance through the low depths of field of the light scope. Use BS to find your way around the specimen. At times that is difficult, but marking slides with India ink and some rough drawings can overcome that. Better still, if available make digital images of your areas of interest and display those on a monitor when locating spots for analyses. There are several ways of "killing that particular cat", installing a transmitted light scope is rather grizzly. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Wednesday, April 28, 1999 6:18 PM, Ritchie Sims [SMTP:r.sims-at-auckland.ac.nz] wrote: } } } Hello, All } } Has anyone out there managed to add transmitted-light } illumination to } the optical microscope of a JEOL 840? } I'd appreciate hearing from you. } } thanks } } Ritchie } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : } r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand }
For an exhaustive review, visit "Curtin's Short Courses in Digital Photography": http://www.shortcourses.com/index.htm
In particular, view Curtin's "Book1: A Short Course in Digital Photography", with emphasis on Chapter 2: The Foundations of Digital Imaging (http://www.shortcourses.com/chapter02.htm) and Chapter 3: Digital Cameras (http://www.shortcourses.com/chapter03.htm)
Another excellent review/tutorial can be found at Sound Vision Incorporated's, "How Digital Cameras Work" (http://www.soundvisioninc.com/howdcw.htm) which deals with the technical nitty gritty and recommendations of camera types based on application needs.
Happy reading and hope this helps.
Best regards,
Walt Bobrowski Subcellular Pathology Parke-Davis Research 2800 Plymouth Road Ann Arbor, MI 48105
The important issue for any microscope is the availability of a phototube with C-mount from Zeiss which is used to attach the camera to the microscope. Any CCD with a C-mount should be OK.
As for image capture cards, there are a number of frame grabbers on the market. One decision you need to make here is whether to go with a digital camera which has a frame grabber on-board or to go to an analog camera with the frame grabber mounted in your computer. I'd suggest that you talk to a local system integrator (if you send me specifics on your location, I may be able to recommend a contact or two).
Two quick suggestions: Media Cybernetics has a complete system available (from camera to board to cables to software ... sort of "imaging in a box "). Just remember, you still need the Zeiss coupler or something similar (Diagnostic Instruments has great C-mounts with zoom focus).
Details are available the websites of these respective companies. Call/email me if you have any trouble finding them.
CAVEAT: MME has no commercial interest in any of these companies.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 10:12 AM 4/27/99 -0500, Chris Bradley wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The New England Society for Microscopy will hold its Sixteenth Annual Spring Symposium at Marine Biological Laboratories, Woods Hole, Massachusetts on May 7 & 8,1999. In addition to our scientific presentations, this year's Symposium will feature PROEJCT MICRO, the new Great Explorations in Math and Science Program designed to introduce Microscopy to elementary- and middle school-aged students. If you are interested in participating in this exciting new program, be sure to register and join us for dinner on Friday evening. =
PRIZES for Best Poster, Best Student Poster, and Best Photo-As-Art will also be awarded. If you're interested in submitting your poster for judging, please contact Dr. Changmo Sung at Changmo_Sung-at-uml.edu.
A COMMERCIAL TABLE-TOP EXHIBIT is scheduled for Saturday morning from 10a= m to 12:30pm. =
The deadline for Exhibitor Registration has been extended to April 30th. =
Reserve your table now. The Spring Symposium always has a strong membership turnout and the Saturday Exhibition promises to be packed. =
Please double check to make sure you have registered so you do not miss this opportunity to visit old friends and make important new contacts. =
=
PROGRAM Friday, May 7th
12:00 pm Registration: Swope Center
1:00 pm Welcome: Lillie Auditorium
Session I Chairperson: Doug Taatjes, UVM
1:05 pm Laser Scanning Cytometry as an Imaging System Ed Luther, Senior Scientist - Biology, CompuCyte Corporation, Cambridge, MA
1:45 pm Focused Ion Beam Microscopy: The New Kid on the Block Dr. David Casey, Senior Scientist, Micrion Corp, Peabody, MA
2:25 pm Membrane Fusion Machinery in Cells Dr. Bhanu Jena, Dept. of Surgery and Biomedical Eng., Yale University, New Haven, CT
3:05 pm Afternoon Break Coffee served in Lillie 103
Session II Chairperson: Tony Garratt-Reed, M.I.T.
3:20 pm Multi-technique Characterization of Emissive Coatings on Electrod= es Dr. Chris Peters, Senior Project Engineer R & D, Osram Sylvania, Beverly, MA
4:00 pm Microscopic Approach to the Study of Pancreatic Islets Dr. Thomas Jetton, Ergo Scientific, Inc., Cambridge, MA
4:40 pm Advances in Microanalysis: A Look at the Past, Present and Future=
Dr. William Hardy, CEO, Princeton Gamma Tech, Princeton, NJ
5:30 pm Cocktails and Dinner: Swope Center
7:30 pm Project Micro "Festival" Meigs Room, Swope Center Dr. Burton E. Goodrich, Jr., Executive Director, South Coast Educational Collaborative Janet E. Goodrich, Principal, Miscoe Hill Elementary School, Mendon, MA
Saturday, May 8th
7 to 8:00 am Breakfast: Swope Center
Session III Chairperson: Eben Oldmixon, HSPH =
8:30 am The Use of Electron Microscopy in the Characterization of Carbon Deposits Paul Anderson, Chemistry Department, Northeastern University, Boston, MA
9:10 am Confocal Microscopy of Nuclear Envelope Breakdown Dr. Mark Terasaki, Asst. Prof., Dept. of Physiology, UCONN Health=
Ctr., Farmington, CT
10:00 am Commercial Exhibits and Posters: Swope Center Coffee and doughnuts will be served
12:30 pm Presentation of Poster and Photos-As-Art Awards and Door Prizes: =
Poster Area, Swope Center
1:00 pm Lunch with Short Tour of MBL: Swope Center =
2:00 pm 2-hour Discovery Cruise aboard the R/V Patriot II Tickets must be reserved in advance
Register Today! Contact L. Kirstein at NESM-at-compuserve.com. Registratio= n Deadline will be extended to April 30th. =
SILENT AUCTION, OCEANOGRAPHY CRUISE, POSTER AWARDS, DOOR PRIZES =
I have used this tissue processor for several years with good results, only draw back seem to do with resin polymerization in the vials if the protocol is for an extended period of time. -Mike
On Tue, 27 Apr 1999 msteglic-at-notes.mdacc.tmc.edu-at-Sparc5.Microscopy.Com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } I am currently looking at the EMS Lynx tissue processor and would like any } feedback from anyone currently using one } } (Do you like it?, Does it function as it should? Are there any drawbacks to } the unit? etc..). } } Does anyone know if there is a lab in the Houston area using one? } } Also does anyone know of any other tissue processor available for EM } processing? } } Mannie Steglich } U. T. M. D. Anderson Cancer Center } Houston, TX. } } } }
Hi guys, I need recommendation for 3D/deconvolution software, which will be able to run on NT. Thanks, --Ciprian ________________________________________________________________ Ciprian A. Almonte Phone: (412) 648-9796 Center for Biologic Imaging FAX: (412) 648-8330 University of Pittsburgh URL:http://sbic6.sbic.pitt.edu Pittsburgh, PA 15261 USA mailto:calmonte-at-pitt.edu ________________________________________________________________
We are consolidating our EM unit and must relutantly part ways with an exceptional H-600 Hitachi EM scope.
You may contact me directly through the e-mail address: rmonahan-at-caregroup.harvard.edu
or by phone at: 617-667-5777
Basic information on the instrument follows: Hitachi H-600 TEM The instrument is in excellent condition. It has been under service agreement with Hitachi.It has been used by a single experienced electron micriscopist to do strictly biological work.
Rita Monahan-Earley Senior Electron Microscopist Beth Israel Deaconess Medical Center Harvard Medical School 330 Brookline Ave Boston,Mass. 02215 USA E-mail:rmonahan-at-caregroup.harvard.edu Phone:617-667-5777
} Well, I hope we have helped, remember a motto "if a man built it a man can } fix it", no offence to the ladies as man is used in the generic sense. }
And keep in mind Mott's Law of Recursive Repair, which states that "In order to fix anything, you always have to fix something else first."
I have found this to be widely applicable, from software to old houses. Especially old houses...
} Dear All, } } We are in the market for a new sputter coater. We need a robust, } simple to use (i.e. student-proof) instrument for routine gold or Au/Pd } coating of biological samples. I would prefer to avoid micro-processor } controlled units (who's going to repair the circuit boards in 10 years?). } Does anyone have any advice, etc., etc.? } } Bob } } } Dr. Robert R. Wise } Department of Biology and Microbiology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu } www.uwosh.edu/departments/biology/wise/wise.html
Randy: I agree with Hank Adams on the OTOTO method, and have used Bob Kelley's method with great success for many years. The first critical part of the method is using a specific thiocarbohydrazide (not all suppliers/lots are equal), and doing the "wash" process for purifying the TCH just prior to use. The second critical step for success is being compulsive about the wash/time cycle in the published method. Any slighting of either of these steps can result in most unsatisfactory results. The conditions in the microscope are also critical, and I have obtained excellent results with the OTOTO method in both W filament SEMs at 10 to 30 kV as well as LaB6 and FESEMs at 1 to 2 kV. Hope this helps.
Roger Moretz Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. Ridgefield, CT
The comments above reflect my own experience and opinions.
} -----Original Message----- } From: Tindall, Randy D. [SMTP:TindallR-at-missouri.edu] } Sent: Tuesday, April 27, 1999 12:26 PM } To: 'microscopy-at-sparc5.microscopy.com' } Subject: SEM: Fixation for non-conductive samples } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } I recall reading somewhere a long time ago about special fixation } procedures } for non-conductive samples that are hard to adequately coat. I seem to } remember tannic acid being involved. Does this ring a bell with anybody } out } there? } } Specifically, we are processing pig intestine samples for SEM and are } having } problems with charging on the microvilli. We have repeatedly coated the } samples with Au/Pd, but the problems persist. Perhaps there's a } specialized } processing technique that can help? } } Thanks in advance. } Randy } } } Randy Tindall } Electron Microscope Core } College of Veterinary Medicine } University of Missouri - Columbia } Phone: 573-882-8304 }
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Steve, We have a Gatan 791 MSC for our CM120. It takes lovely pictures. It did take me some time to master the art of acquiring the images. I took the Gatan class they offer in the spring, and it is definitely worth taking. Dr. Barbara Armbruster works in the California office and she took the time to help me work out my initial problems. She is a wonderful teacher. Let me know if you would like to see some images (pathology lab). Peggy Harger-Allen EM lab - VA Medical Center 317-554-0000x2392 harger-allen-at-indianapolis.va.gov
via smtpd (for sparc5.microscopy.com [206.69.208.10]) with SMTP; 28 Apr 1999 21:09:56 UT Received: from ptag2 (ptag2.pt.cyanamid.com [141.173.51.2]) by igate.cyanamid.com (8.9.3/8.9.3) with ESMTP id OAA02519 for {Microscopy-at-MSA.Microscopy.com} ; Wed, 28 Apr 1999 14:55:10 -0400 (EDT) Received: from ccmail.pt.Cyanamid.COM by pt.Cyanamid.COM (PMDF V5.1-9 #28913) id {01JAKJV6624W8X67N9-at-pt.Cyanamid.COM} for Microscopy-at-MSA.Microscopy.com; Wed, 28 Apr 1999 14:55:05 EDT
I need to examine some dissolution-recrystallization processes at temperatures in the 100-200 deg C range by transmitted light. I am looking for a way of making a cell, perhaps by gluing a metal washer with a hole of 3-4 mm onto a slide 26 x 26 mm to fit in a hot stage. I don't know if epoxy will withstand both heat and solvent. For another hotstage application, I once tried water glass for glass-to-glass, but it outgassed badly above 100 deg C.
I would appreciate any suggestions for other cements or from vendors with an all-glass cell, total height { 2 mm, cell ID 1-4 mm (round or rectangular), fittable on to or cut-downable to 26 x 26 mm.
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
At 12:03 PM 4/27/99 GMT+1200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi, I am trying to find a frame grabber that is able to grab images from a CCD camera and SEM(Amray 1830). The CCD camera is used for process monitoring through an optical microscope(Questar QM-100). A VCR is used to record the process on tape. After the process, sample is transfered into SEM chamber and I need to grab a few static images where particular features are found. Anyone has experience in this?
I need a copy of an old file called MSmouse.COM for an application that emulates this driver to reverse a mouse cursor on a DOS environment. The hard disk that had it crashed and the new computer does not have a 5 inch drive. Any idea of where I can get a copy will be appreciated.
*Disclaimer: Whatever... is not Tulane opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web:[ http://www.tmc.tulane.edu/ferminlab/] or [http://www.tmc.tulane.edu/imaging/] Internet: [cfermin-at-mailhost.tcs.tulane.edu] 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
There are many different systems on the market today, and they differ tremendously. You can purchase cameras that transfer the images serially, through a frame grabber, or by use of a different medium. Resolution and dynamic range of the cameras is an issue. What kind of software you need and the kind of support you expect.
For example: if you need high resolution images, you may have to consider a 3-chip TV camera or a digital color camera. For the latter you need to decide, if you can live with a low refresh rate or "offline" use (Images transferred to computer serially or by "sneaker net") or if you need a live image. If the cameras are too expensive, you may be able to acquire neighboring, overlapping images and do a multiple image montage with a less expensive camera, but that requires the appropriate software. Is it important to have automatically calibrated images? Do you need stage control, now or perhaps later? It is probably best to talk to other people who have image capture systems. Of course, as we sell those, we are happy to talk to you, too.
I would start, as Kalman Rubinson suggests, by defining the parameters for the system. Once you have done that, call a few vendors and discuss it with them. Depending on what they sell, they will suggest one or the other solution. That will help you narrow down the specs. I am sure, most vendors are happy to discuss this with you, I know, we would. Once you have narrowed down your choice, you can ask for quotes for similar systems and see, what the prices are.
If you need more help, give me a call or send me an email.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
} -----Original Message----- } From: Exchange Administrator } Sent: Wednesday, April 28, 1999 12:30 PM } To: Michael Bode } Subject: FW: CCD Camera and Image capture card } } } } ---------- } From: Kalman Rubinson[SMTP:kr4-at-is2.nyu.edu] } Sent: Tuesday, April 27, 1999 12:24 PM } To: Chris Bradley } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: CCD Camera and Image capture card } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } On Tue, 27 Apr 1999, Chris Bradley wrote: } } } I am looking for a CCD camera and image capture card for use with a } standard Zeiss light } } microscope. I've been looking all over and I can't find any cameras } designed for this } } use. Can anyone help me out with the name of a vendor or a website? } } ???? Pretty much SOP today and all over the Web.*. Check } Edmunds or McCrone. Also, any C-mount camera can easily be } adapted to your microscope tube. The grabber cards are } ubiquitous as well. } } However, let me suggest you start the other way 'round. } Define your application needs and decide on the software } first. Then, choose grabber and camera from the supported } list. Otherwise, you may buy something which won't give } you what you need. } } Kal } }
Dear Microscopists I have a client who wants to use SEM to image the surface of corneas looking at scars after laser treatment. Any ideas on preparation, etc?? TIA Alan N Hall Laboratory for Microscopy and Micro-Analysis NWII Building University of Pretoria Pretoria 0002 Republic of South Africa Tel: +27-12-420 3896(Office) +27-12-420 2075(Laboratory) Fax: +27-12-362 5150
We are seeking to replace our stock of these, and e-mail enquiries to the company themselves have got nowhere. So (a) does anyone know a supplier, and (b) is there an alternative? We have used this type because it is the only organic material for filter membranes that can stand up to Xylene at 120^C, and the anodisc alumina filters in our catalogues are all too small in pore size.
Thanks in advance,
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Can anyone tell me if there is a Canadian equivalent to the US. Project Micro and if so where I can find information or contact person? Thanks
Debbie Lietz Electron Microscopy Suite Department of Biology Trent University Peterborough, Ontario K9J 7B8 Telephone: (705)748-1486 Fax: (705) 748-1205 email: dlietz-at-trentu.ca
Have you tried a silver metal membrane? I believe these would be a suitable substitute. The company I am familiar with is Osmonics. A silver metal membrane with your required pore size can be found on their website at:
http://www.osmonics.com/products/Page2.htm
Hope this is helpful,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
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Kal, Thanks for your comments. I did not originate this thread but have been looking for a general purpose digital camera for our multi-user facility for a few months. We have good high resolution video cameras and associated computer-enhancement capabilities and s-VHS recorders for live cell use so do not need video cameras. I find that fluorescence use is down as users turn to more confocal use. Thus our need for low light cameras is decreasing.
What I see is users who want to take a few images from a stereo microscope or light microscope and do not want to shoot a whole roll. So they do not document as much as they should. Addition of a relatively inexpensive digital camera may fill the need when the $10,000 for a more versatile camera + framegrabber+computer is not available. If funds are not a concern, the tendancy is often to go with the most versatile instruments one can find. However sometimes this leads to excess features, many of which are not needed and never used, and a waste of precious $$.
Also, recently there has been a call for cameras for student use in classes. For this purpose, we need multiple cameras but certainly do not want to designate a computer for each one and, again, cost is very important. Hopefully the two non-tethered camera I mentioned are just the beginnings of new offerings for this type of instrument.
I would appreciate hearing further comments from users (not venders) of these specific cameras and also would like to hear about other potential cameras to serve these needs (again from users rather than venders....I would appreciate the real on-the-job plus and minuses, not just the specifications).
Debby
--------------------------------------
} Olympus DP10 - uses a smart card to record images. Has a built in LCD } panel for formatting your image and a keypad for interacting with the } programming. You can print directly from the camera if desired. Resolution: } 1280x1024. Price ~$3000
If it records on SmartMedia, one does not need a frame grabber with it. I use a Lexar SmartMedia (parallel-port) reader. It functions like a removeable disc drive permitting one to read/write the SM. It cost me $37.
} Fugi HC-300Z: does not have built in LCD panel so it is helpful to attach } a small monitor for setting up your formatting, etc. Downloads images } directly to a ZIP disk. Has small keypad for interfacing with software. } Resolution: 1280 x 1000. Price ~$4000
Again. No grabber.
} Kodak also makes a microscope tube adapter for use with one of their } consumer cameras.
Also, it (DC-120) retains its non-removeable lens and has poorer resolution.
Consider also the Pixera and the MicroVision in this class of camera.
What all of these do NOT permit is the use of real-time image processing software. All processing is done post hoc. If this is OK in one's application, fine. Otherwise, one must consider a separate camera + grabber.
Kal
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RECEIVED: from SF_Database by POP_Mailbox_-1286883292 ; 27 APR 99 18:53:24 UT Received: from IS2.NYU.EDU by mailcenter.btny.purdue.edu with SMTP (QuickMail Pro Server for MacOS 1.1.2); 27-Apr-1999 18:53:20 -0500 Received: from localhost (kr4-at-localhost) by is2.nyu.edu (8.8.8/8.8.7) with SMTP id TAA09208; Tue, 27 Apr 1999 19:53:41 -0400 (EDT) Date: Tue, 27 Apr 1999 19:53:41 -0400 (EDT) From: Kalman Rubinson {kr4-at-is2.nyu.edu} To: Debby Sherman {sherman-at-btny.purdue.edu} cc: "message to: MSA list" {microscopy-at-sparc5.microscopy.com} Subject: RE: CCD camera and capture card In-Reply-To: {iss.2c27.37262849.ad7f2.1-at-scribe.cc.purdue.edu} Message-ID: {Pine.OSF.3.95.990427194804.7537A-100000-at-is2.nyu.edu} MIME-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII
Are you looking to acquire images from a CCD camera AND an SEM, or is it one or the other?
} From your description ("a VCR is used to record...") I assume, that you are using an analog CCD camera. There are many frame grabbers on the market that will digitize these signals. Just look for "frame grabber" on the net, or give me a call.
If you wish to acquire images digitally from the SEM, the choice narrows down considerably, unless you only wish to use the SEM TV signal. In this case the same as for the TV camera applies. If you wish to acquire high resolution images from the SEM, you need some form of "active" or "passive" SEM interface. The reason is, that it is impossible to use TV standards (for example NTSC or PAL) for high resolution, slow scan images. So you need different hardware. As far as I know (perhaps I am wrong, in this case I apologize) we are the only ones that have "active" and "passive" acquisition boards that work as add-ons to our frame grabber. In other words, you can, with the appropriate software and hardware, use our frame grabber to acquire both TV signals AND control or synchronize to an SEM. Please give me a call if you neeed more information, or send me an email. If you want to check out our web site, look for "Grabbit" (our frame grabber) and "ADDA" (our SEM interface).
Michael Bode
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com web: www.soft-imaging.com
} ---------- } From: Tong Wang[SMTP:tong-at-jlab.org] } Sent: Wednesday, April 28, 1999 4:39 PM } To: microscopy-at-sparc5.microscopy.com } Subject: frame grabber for SEM } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Hi, I am trying to find a frame grabber that is able to grab images } from a } CCD camera and SEM(Amray 1830). The CCD camera is used for process } monitoring through an optical microscope(Questar QM-100). } A VCR is used to record the process on tape. After the process, sample } is } transfered into SEM chamber and I need to grab a few static images } where } particular features are found. Anyone has experience in this? } } }
} I need a copy of an old file called MSmouse.COM for an application that } emulates this driver to reverse a mouse cursor on a DOS environment. The } hard disk that had it crashed and the new computer does not have a 5 inch } drive. Any idea of where I can get a copy will be appreciated.
Unfortunately, I deleted your earlier message but from Jim's response, it looks like your geologists need a combination of two techniques which are less than compatible. Not being very experienced with EM, I can't comment on the stages available, but in other applications (i.e., going from a fluorescence microscope to an FT-IR microscope), we have used finder stages which gave us the ability to exactly locate a specific feature then move it to the other microscope and find that feature again. If this were the case, your geologists could do their conventional polarized light analysis then move the system to the electron microscope for other imaging and analysis.
Hope this is helpful. Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** To find out what was new in microscopy at Pittcon, see "Focus on Microscopy", American Lab, May 1999
At 10:47 PM 4/28/99 +1000, Jim J Darley wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The best way to keep an equal proportion of latex beads and virus particles is to centrifuge the mixing (well mixed) directly on a grid. For this, I use a Beckman Airfuge Ultracentrifuge with a A-100 rotor. The technique consist to mix a part of virus particles and a part of latex bead with a known concentration. Place in an Airfuge 200 µL-tube with a Formvar-carbon coated grid in the bottom. Centrifuge at 120 000 g (20 psi) during 5 min. Recuperate the grid, dry it, stain and finally count virus and beads in a TEM and compare the concentration of beads with concentration of virus particles see: - Alain, R et al., J. Virol. Meths, 16 (1987), p. 209-216 - Alain, R, Microscopy today, may, issue #97-4, D.Grimes Ed., (1997) p. 20
Robert Alain ********************************************************** Robert Alain, M.Sc. Microscopie électronique INRS-Institut Armand-Frappier-Microbiologie et Biotechnologie 531 boul. des Prairies Laval, Québec CANADA H7N 4Z3 Tel: (450)687-5010 ext#4388 Fax: (450)686-5626 e-mail: Robert.alain-at-INRS-iaf.uquebec.ca Http://www.iaf.uquebec.ca/iaf/recherche/viro/me.html **********************************************************
We are seeking vendors to provide some ocular micrometers (Olympus and Lietz eyepieces) as follows:
(1) grid configuration, divided into 400 squares (i.e., 20 x 20) (2) grid configuration, divided into 100 squares (10 x 10)
It is highly desirable to have the divisions numbered in the x-y directions.
Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Dear Bob, I recently acquired a Denton Desk II coater and etcher as part of a larger, used equiment purchase and I'm very pleased with it. It is much simpler to use than my very old Hummer was, very simple to operate and automatic enough for students. I just post a few lines of instructions on the wall behind it and everyone has had a good coat in 30 sec. so far. I like the simple target, which a flat gold sheet, which means that you don't have to get ripped off for a special target when it is consumed. I have been pleasantly surprised. I don't see any micro-processor control, just solenoid and relay. The system leaks the pump back to vacuum when you turn it off, so you don't suck oil vapours back into the chamber, something I could never teach students to do. You wrote: } } } Dear All, } } } } We are in the market for a new sputter coater. We need a robust, } } simple to use (i.e. student-proof) instrument for routine gold or Au/Pd } } coating of biological samples. I would prefer to avoid micro-processor } } controlled units (who's going to repair the circuit boards in 10 years?). } } Does anyone have any advice, etc., etc.? } } } } Bob } } } } } } Dr. Robert R. Wise } } Department of Biology and Microbiology } } University of Wisconsin-Oshkosh } } Oshkosh, WI 54901
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
I am an experienced electronics design engineer who wants to take on building something a bit more challenging in his garage: A scanning Electron Microscope.
I already have information on building diffusion pumps ( although more information is always welcome ), and I have info. on construction and testing of high vacuum apparatus.
The electronics will be straightforward enough.
What I need is help on electron optics. Are there any "beginners guide to designing electron optics" or "A home made SEM: how I did it" books out there? Can anyone suggest how I get from undergraduate physics to SEM design ( I learn well from books )?
Dear John, We've been using eyepiece reticules from Klarman Rulings, POB 4795, Manchester, NH 03108, (800)252-2401. They are not expensive, available in any diameter with a wide range of styles, and accurate.
Glen
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
On Thu, 29 Apr 1999, John J. Bozzola wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We are seeking vendors to provide some ocular micrometers (Olympus and } Lietz eyepieces) as follows: } } (1) grid configuration, divided into 400 squares (i.e., 20 x 20) } (2) grid configuration, divided into 100 squares (10 x 10) } } It is highly desirable to have the divisions numbered in the x-y directions. } } Thanks. } } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } #################################################################### } } } }
Thanks, Greg, I'll consider these lines of thinking. We have tried going from acetone into HM-20 (I think that's what was used; it was over a year ago), not sure how it would work directly, have to look into that. I'm pessimistic about the membrane connections holding up after freeze-drying and then re-wetting with resin. Will see if I can read up on what's been done.
We have done freeze-fracture-etch but the details I want to see (connections between membranes) do not show up for various reasons with that technique. I guess something to look into is the possibility of cryo TEM with energy filtering and 3-d reconstruction, but my specimens are so thick (~15 micron diameter) that, as I understand it, it's too thick for the usual technique so maybe I'd have to see if this could be done with HVEM. Thanks again for your suggestions Richard p.s. Fats and lipids are a nightmare for lots of things, but that's my job. I vividly remember a biochem professor trashing one of my class proposals because they're hard to work with. That's what makes it interesting!
} } } Greg Erdos {gwe-at-biotech.ufl.edu} 04/27/99 12:20pm } } } Regarding TEM of fat, I might suggest two possible approaches. One would be to use a hydrophobic resin like Lowicryl HM-20 or HM-11. It might even serve as the substitution media. Another thought is to freeze dry the samples and embedd them in resin directly . One could also expose the sample, after drying, to osmium vapors from the crystals in order to keep things anhydrous. I would recommend a resin that can tolerate a little water, like Epon or its equivalent, or a methacrylate based resin.
Would using freeze-fracture EM give the results you are after??
Fats and lipids are a nightmare for EM.
Greg Erdos Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
I have just been asked to submit budget numbers for a replacement SEM. We have a DS-130 with tungsten filament. I have not been keeping up on the latest and greatest and I would like to hear from end users who have recently purchase a SEM. Our needs are almost all related to inorganic and metallurgical samples. Thanks for your time.
Bruce Kaye
Dockyard Laboratory Pacific (DL(P)) Building 199 Dockyard PO Box 17000 Stn Forces Victoria BC V9A 7N2 CANADA (250) 363-2514 Fax: (250) 363-2856 Email: bruce.kaye-at-edrd.dnd.ca
I did a TEM in high school....many years ago. It was based on an RCA EMU. The cost today of a fine SEM platform is about $5K. With that, you get console, column, turbo, roughing pump, etc., etc. I would not imagine that any current efforts at trying to achieve what has already been achieve would be worth more than self gratification. If that is your goal, go for it. If you really want to see what a SEM can do and want to improve on it, get a good column and start working on digital controls and capture. That avoids reinventing the wheel and truly is where the future is as I see it.
} } } The last few times I"ve made up a section-staining solution of uranyl } acetate, (J.E.M.Techniques 16,81(1990)), an abundant brown precipitate } forms within the syringe in a day or two. It usually stays fine for at least a month or even more. } I've tried both a glass syringe and a polypro syringe with similar }
I want to buy an used SEM. I'm looking now on a Hitachi S-530 model. Is there someone that can give me their experience with that SEM model ? What I should look to make the best deal ? Thank you.
I want to buy a DIAMS to grabbe image from a SEM, from a optical microscope, and from a stereoscope. I'm evaluating now the Quartz PCI system from Quartz Imaging Corporation, and the Clemex Acquisition and R'Kive System from Clemex Company. Is there someone that can give me their experience with those DIAMS model ? Thank you.
I remember that someone asked about slide markers recently. If you look on EBAY the internet auction site there is one for sale. It is a Leitz diamond marker and the item# is 95912618. The price so far is $50 but it has an unknown reserve.
It is difficult to add something to Steve has told. But I have pricked my ears after your words that the diffusion pump is not too hot. The point is that diffusion pump works only, when its bottom is too hot, and top is too cold. If the diffusion pump does not pump, any HV turning on can led to glow discharge and to overloading of HV supply unit. Pirani gauge is too rough meter and requires often adjustment to estimate vacuum.
Regards. Victor Sidorenko, ANTRON, Moscow, Russia.
Renata Korzyniewski wrote:
} Dear Listservers, } } We have a Hitachi S-520 SEM installed in 1983. Lately, we have experienced } instability problems in the SEM and I'm not sure what to do. The Hitachi } S-520 SEM has a single rotary pump, single diffusion pump and Pirani } gauge. On initial pumping and warm-up, the SEM gets down to high vacuum } (reading 10uA) quite normally in about 20min. On applying HT (20kV), } everything appears normal for approximately 80min, then, suddenly the HT } switches itself off and the vacuum gauge rises to 15uA in 30sec and then } falls back to10uA in 2min. If the slightest gun emission current is } applied when the SEM says it's ready, the HT instantly switches itself } off. I have cleaned the filament assembly, anode and gun chamber around } the anode. However, I have not cleaned the insulator at the top of } the gun chamber. The Pirani gauge is clean and the diffusion pump is } not too hot. Any advice would be much appreciated and gratefully } received. Thankyou. } } John Brealey Queen Elizabeth Hospital EM Unit Adelaide South Australia }
EuroFE '99 is a forum to bring together interested parties in the field of Field Emission Technologies. There is also a strong emphasis on links with other display technologies such as LCD's, Light Emitting Polymers and Phosphors. The aims of this Workshop are to:
Present the activity of each group in Europe working in Field Emission (FE) research and related areas. Present the EuroFE network to the Scientific Community and Brussels and enhance its activity. Initiate, through this meeting, groups with several partners in order to present projects to Brussels. Enhance the participation of companies in the EuroFE Network and EU programs. Exchange knowledge & initiate collaboration between groups either Institutes or Companies (manufacturing joint ventures, collaborative research, development activities, etc.). Allow important student participation.
The scientific programme will cover the whole spectrum of Field Emission research and related areas including keynote lectures, oral presentations, posters and a product/instrument exhibition. Topics of interest will be for example:
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For further information please see http://www.cmp-cientifica.com/EuroFE/eurofe99.htm Or contact me direct.
Renata Korzyniewski wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Listservers, } } We have a Hitachi S-520 SEM installed in 1983. Lately, we have experienced } instability problems in the SEM and I'm not sure what to do. The Hitachi } S-520 SEM has a single rotary pump, single diffusion pump and Pirani } gauge. On initial pumping and warm-up, the SEM gets down to high vacuum } (reading 10uA) quite normally in about 20min. On applying HT (20kV), } everything appears normal for approximately 80min, then, suddenly the HT } switches itself off and the vacuum gauge rises to 15uA in 30sec and then } falls back to10uA in 2min. If the slightest gun emission current is } applied when the SEM says it's ready, the HT instantly switches itself } off. I have cleaned the filament assembly, anode and gun chamber around } the anode. However, I have not cleaned the insulator at the top of } the gun chamber. The Pirani gauge is clean and the diffusion pump is } not too hot. Any advice would be much appreciated and gratefully } received. Thankyou. } } John Brealey Queen Elizabeth Hospital EM Unit Adelaide South Australia
Hi Renata or John, Do you know if your diffusion pump has a full charge of oil? I've seen this type of vacuum behavior when the charge gets low. Things start out pumping fine, but then the system gets all of the oil up in the pump and on the side-walls. It stops pumping until some of that oil makes it back to the sump where it is re-boiled. A short burst of pumping occurs.
Might be an easier thing to check than tearing apart potted HV components for starters.
Ken Converse owner Quality Images third party SEM service Delta, PA
EMBO Practical Course on Biophysical and Mathematical Approaches to Cell Biology
4 July - 17 July 1999 -at- EMBL, Heidelberg, Germany
ORGANIZERS: Ernst H.K. Stelzer, Michael Way, J.K. Heinrich H=F6rber
The links between physics and cell biology are becoming more and more important. In many instances it is no longer sufficient to test a model quantitatively. Many processes such as microtubule dynamics (i.e. the microtubule assembly and disassembly) have only been understood since a) = the in vitro system was sufficiently well developed, b) a mathematical model that describes the events became available, and c) the events could be recorded, analyzed and compared with the model. This course is aimed fir= st of all at researchers working in the life sciences who wish to improve th= eir mathematical background and move into a more direct use of physics for quantitative biology. They wish to acquire a mathematical/physical framework for their biological system of interest and need to know how to verify it in a quantitative manner. However, another equally important group of attendees has a background in mathematics and/or physics and wis= hes to understand the problems of working with biological systems in a quantitative manner. One of the main objectives of this EMBO course is t= o improve the communication between biologists and physicists.
Speakers from the Americas and Europe including the staff at EMBL (Boulin= , Dotti, Eaton, Florin, Gonz=E1lez, Griffiths, H=F6rber, Hyman, Karsenti, N= ilsson, Pepperkok, Simons, Stelzer, Vernos, Way, Zerial) will introduce students = to: microtubule dynamics, determining growth shrinkage/rates, generating microtubule patterns, molecular motors, mechanisms of neuronal plasma membrane asymmetry, microtubule organization during development, complex membrane organelles, chromosome movement at mitosis, mitotic spindle assembly, distributions of proteins along exocytic pathway, surface polar= ity in epithelial cells, actin cytoskeleton and cell motility, membrane traff= ic in eukaryotic cells, cell polarity in Drosophila, single molecule force measurements, molecular mechanics. Further topics may be added.
The deadline for applications is 15 May 1999. The organizers will select the students. Their decision is final. Industry applications will be considered. The course participants will come from all over Europe. The selected students will be informed by 21 May 1999. The main selection criteria are: degree in natural sciences, exposure to modern cell biology for at least one year, research is related to progress in biology, and th= e ability to convince the organizers of an interest in this course by CV an= d covering letter. Students will not be exclusively post-doctoral.
Applications should be sent in printed or electronic form to: Dr. Ernst H= .K. Stelzer, European Molecular Biology Laboratory (EMBL), Cell Biology and Biophysics Programme, Meyerhofstrasse 1, D-69117 Heidelberg, Germany. stelzer-at-embl-heidelberg.de, fax +49 (6221) 387306. http://www.embl-heidelberg.de/ExternalInfo/stelzer/Courses/CellBiologyBio= phy sics
Sincerely Yours E. Stelzer
Dr. Ernst H.K. Stelzer EMBL-Heidelberg Light Microscopy Group Cell Biophysics Programme Meyerhofstrasse 1 D-69117 Heidelberg Postfach 10.2209 D-69012 Heidelberg Germany Phone +49-6221-387 354 Fax +49-6221-387 306
I'd like to find a used SEM. Since I currently have an ISI, an old SX-30 or 40 would be perfect, as I have quite a bit of ISI apparatus. I'm certainly willing to switch manufacturers if I can find a good small to medium size scope.
If any of you have a SEM in your garage that you hoped to bring back online someday, but are being nagged to get rid of it, this is your chance to make a big dent in the spring cleaning! Please make a proposal to jbest-at-elmdas.com, and not the list server. Thank You.
Jianli, Try cooling the sample slowly in liquid N2 and then lightly scribing the surface under the liquid N2 with a diamond scribe. This may flake off the film in small areas providing an edge for scanning. Make sure the adjacent film is not delaminated as this would give a false reading. Also the surface should be cleaned to remove lose particles e.g. CO2 snow cleaning works great for this. Good luck. Russ Xerox
-----Original Message----- } From: Jianli Wang [mailto:jiwang4-at-mail.vt.edu] Sent: Thursday, April 29, 1999 5:02 PM To: spm-digest-anal-at-bbfm.di.com
************************************************************** THE UNIVERSITY OF LEEDS
DEPARTMENT OF MATERIALS, SCHOOL OF PROCESS, ENVIRONMENTAL AND MATERIALS ENGINEERING.
RESEARCH OPPORTUNITY IN HIGH SPATIAL RESOLUTION MATERIALS CHARACTERISATION
RESEARCH FELLOW
The above HEFCE-funded post is available from October 1 1999 for a fixed period of three years to commission, support and apply an optimized, analytical field emission transmission electron microscope for materials analysis to a wide range of materials projects based at Leeds and in regional institutions.
Applicants should have a PhD in physical/engineering sciences and research experience in Materials, Physics or Chemistry involving particularly Transmission Electron Microscopy and Microanalysis.
Salary: Research Staff Grade 1A (stlg15,735 - stlg23,651 p.a.) according to qualifications and relevant experience.
Application forms and further particulars may be obtained from:
Ms Joy Bielby, Department of Materials, School of Process, Environmental and Materials Engineering, University of Leeds, Leeds, LS2 9JT, United Kingdom.
In all enquiries please quote the reference number 062-069-004-027
Informal enquiries on the post should be directed to Dr R. Brydson (email: mtlrmdb-at-leeds.ac.uk)
Closing date for applications 24 July 1999
Towards Equal Opportunities. **************************************************************
_____________________________ Dr. Rik Brydson, University Research Fellow, Electron Optical Unit, Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
We recently acquired a digital camera from Advanced Microscopy Techniques (AMT) and are extremely pleased with it. AMTs model 12-HR uses a Hammamatsu camera, custom lens and AMTs own software. The camera has an electronic shutter and manages better than 10 frames per sec at lower illumination than most users would use by eye on the fluorescent screen. Sharpness is excellent, and indeed publication quality. Support and vendor interaction are simply outstanding - it is hard to imagine it being any better. This is in stark contrast to the treatment our institution has received from Gatan - who says that since they pitch in a few hundred dollars to support our local microscopy group that it should be sufficient support for the local users!
Software: Admittedly, Gatans software is a mature package, whereas the AMT software is undergoing rapid revisions. However, the AMT software works perfectly and meets all of our needs - as biologists in a busy core facility. If we had chosen Gatan, each of our users would need to spend nearly $600 apiece just to view the extra image data. With AMT they can see their image as well as all recorded data (voltage, mag, comments, etc) with any tiff file reader.
There are several other reasons why I would buy AMT over Gatan any day, but it would be best if you contacted me off-line. I am sure just what I have said here is going to ruffle a few Gatan feathers.
Regards, Jim
-------------------------------------------------- James F. Sanzo, Ph.D. Codirector, Biomedical Imaging Core Laboratory B-110 Richards Building University of Pennsylvania 36th and Hamilton Walk Philadelphia, PA 19104-6085 Voice: (215) 898-6730 Fax: (215) 573-2259
You don't say where you are located, but a few hours on the phone should be able to locate a free or cheap microscope sitting on somebody's loading dock. Repairing somebody else's design might not be what you are interested in, but in this case you at least don't need to build your own pump, chamber, high voltage power supply, etc.
But, if you want to go the other way and see what can be done from scratch that is fun too. Most of the literature on the technology you are trying to duplicate is pre-1970, so the on-line literature searches won't be much help. There is a lot of practical knowledge of electron optics in the Review of Scientific Instruments and the old Journal of Scientific Instruments. The old electron optics books such as Klemperer can be useful if you don't get bogged down in the details.
As for a gun, cutting off the neck of a crt, and using the video scanning coils would be a good place to start.
Or maybe better, get a vidicon based tv camera and take off the window.
best regards mark
Mark W. Lund, PhD VP Engineering } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"This is a YOUNG business...How can I tell you what YOUR job is when I don't know what MINE is?" --Pogo
} Hi guys, } I need recommendation for 3D/deconvolution software, which will be able to run on NT.
Ciprian A. Almonte Center for Biologic Imaging
Mr. Almonte,
The appropriate product we would suggest you use would be VayTek. Our suggestion is for you to check our web site www.i-cubeinc.com Then we can provide you with a specific recommendation.
Regards
Barry Dudley
-- ************************************************************ B.T. DUDLEY I-CUBE www.i-cubeinc.com Ph 1-888-77-I-CUBE 301-858-0505 301-858-0615 (Fax) I-CUBE is a Systems Integrator and Value Added Reseller of image analysis and image processing products for scientific and industrial applications. We provide a single source for imaging products, using a consultative selling approach I-CUBE 2411 Crofton Lane; Suite 14A; Crofton; Maryland; 21114 ************************************************************
The Minnesota Microscopy Society will hold its annual Spring Symposium on May 13th in the Sheraton Midway Hotel (midway between St. Paul and Minneapolis). Full details of the meeting can be found on our website http://resolution.umn,edu/MMS/ but the highlights are as follows:
MMS Spring Symposium: Latest Trends in Microscopy Thursday, May 13, 1999
Location: Sheraton Midway Hotel, located at the intersection of I94 and Hamline Avenue.
Program
8:00 - 9:00 Registration with refreshments 9:00 - 9:45 Microcalorimeter EDS with 3 eV Energy Resolution David A. Wollman, NIST 9:45 - 10:30 Innovations in Energy-Dispersive X-ray Microanalysis John J. Friel, PGT 10:30 - 11:15 Break and Vendor Displays 11:15 - 12:00 Recent Advances in Microwave Assisted Specimen Processing: Rapid Preparation of Biological Specimens for Correlative Confocal and Electron Microscopy. Mark A. Sanders, U of M 12:00 - 1:00 Lunch Provided 1:00 - 1:30 MMS Business Meeting 1:30 - 2:15 Application of SEM Diffraction to Phase Identification Pat Camus, NORAN 2:15 - 3:00 Break and Vendor Displays 3:00 - 3:45 Imaging Mechanisms in Dynamic Force Microscopy of Polymers Greg D. Haugstad, U of M
Reservation/ Cost: Registration is $50 for members, $60 for non-member (this includes a one year membership fee). To make reservations, please contact Mike Coscio at 612-514-1331, or by e-mail at mike.coscio-at-medtronic.com.
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist Office:(612) 626-7594 CIE Characterization Facility, University of Minnesota Desk: (612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
I would be interested in hearing about the value of service contracts. Our SEM (Hitachi S-3100H) is about 2.5 years old and has been operating without problems. If I operated without a service contract, handling all preventitive maintenance myself, am I begging for trouble? Those of you who don't operate with a service contract have you found the cost of emergency services so expensive that a service contract would have been preferable? Do you find the service difficult to get without a contract? Are the parts that would need replacing terribly expensive (more than the cost of the service contract)? As the instrument gets older is there a greater concern? Is my concern about the cost of the service contract unwarranted?
Thanks.
David Rose W.L. Gore & Associates 297 Blue Ball Road Elkton, MD 21921
Dear Marc, I have used the Quartz PCI system for some years and it should be able to do everything you want to do. I only have two SEM's and a TWAIN flat-bed scanner on mine, but the icons and tools for video cameras, digital cameras and other devices are all there and active all the time. I know others who use SEM and video acquisition iinto the same system and it works perfectly. The database allows you to organize all the images and search for them by job, session, job or any search string you like. I have over 2800 images in my database from two SEM's and the scanner. You wrote:
} } I want to buy a DIAMS to grabbe image from a SEM, from a optical } microscope, and from a stereoscope. I'm evaluating now the Quartz PCI } system from Quartz Imaging Corporation, and the Clemex Acquisition and } R'Kive System from Clemex Company. Is there someone that can give me } their experience with those DIAMS model ? Thank you. } } Marc Montreuil } Lachine (Qu=E9bec) Canada } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
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Dmitri.
} Hi all, some toughts: } } a database containing user manuals for a range of older popular microscopes } would be a very good idea, especially since the purchase of a second hand } microscope is usualy the only way for an amateur microscopist to get hold of } a decent microscope at a (more or less) reasonable price. That's why I have } put some Reichert Zetopan manuals on my website. } } In the best case scenario this service should be easy accesible and free if } at all possible. If that isn't possible a small fee could be asked to cover } the costs. } } But it isn't simple: } } I don't think that the large manufacturers would like to support this idea, } as it isn't in their short-term intrests to support users of second-hand } equipment, they rather like to sell new gear (one example: I know from a } very reliable German source, that Zeiss has, after the "wende", destroyed } large stocks of spare parts for older "Eastern-German Zeiss } manufactureres"). I think they see it the wrong way: I can't imagine much } amateurs who spend the exorbitant prices the large manufactureres ask for } their products, at least I wouldn't... } } And, unfortunally, as far as I know, the manufacturers are the owners of the } copyrights of their manuals, so we're stuck here, that would be the first } problem to be solved... } } After that: finding someone to coordinate the project, finding the manuals } and scan those (can't be much of a problem I suppose), finding a server to } host the documents and finding some people to do investigations regarding } brands, models, serial numbers... to match manuals and models/versions... } } I would like to volunteer for such a project... } } Hello Royal microscopical society (England), Microscopy Society of America, } German microscopy clubs, Micscape Magazine, manufacturerers...? } } Yvan Lindekens.
__________________________________________ Dmitri V. Sokolov, Doctor Course student Research Center for Interface Quantum Electronics, Hokkaido University, North 13, West 8, Kitaku, Sapporo 060, Hokkaido, Japan Phone 81-11-706-7174 Fax 81-11-716-6004 http://www.geocities.com/SiliconValley/Campus/1314 AOL Instant Messenger FalconDot ICQ 9418072 __________________________________________
I' m thinking to do double staining with peroxidase-DAB AND gold particles with silver enhancement in cultured cells (pre-embbeding treatment and I will use Epon). Do you know if it is possible? Do you have any reference about this technique? I will be very grateful for any information
Sincerely
Francisco Javier Hernandez-Blazquez IARC-WHO Unit of Multistage Carcinogenesis 150 Cours Albert Thomas 69372 - Lyon France fhernandez-at-iarc.fr Telephone: (33) 472738536 Fax: (33)472738442
is there any nice SEM pictures of beetles on WEB? I would like to create link from www.coleoptera.org to some nice image sites...
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptiste (surname) of Bejsak (Bayshark)-Collorado-Mansfeld
http://www.coleoptera.org Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., P.O.Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.37 / Virus Database: 30 - Release Date: 6.4.1999
Two major international databases of entomological and agricultural literature are now available for free on WWW---
you can find both on www.coleoptera.org in directory {Coleoptera Bibliography} and in first subdirectory {Search}
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptiste (surname) of Bejsak (Bayshark)-Collorado-Mansfeld
http://www.coleoptera.org Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., P.O.Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.37 / Virus Database: 30 - Release Date: 6.4.1999
At 12:18 PM 4/30/99 , you wrote: } } Dear List, } } I would be interested in hearing about the value of service contracts. Our SEM } (Hitachi S-3100H) is about 2.5 years old and has been operating without } problems. If I operated without a service contract, handling all preventitive } maintenance myself, am I begging for trouble? Those of you who don't operate } with a service contract have you found the cost of emergency services so } expensive that a service contract would have been preferable? Do you find the } service difficult to get without a contract? Are the parts that would need } replacing terribly expensive (more than the cost of the service contract)? As } the instrument gets older is there a greater concern? Is my concern about the } cost of the service contract unwarranted? } } Thanks. } } David Rose } W.L. Gore & Associates } 297 Blue Ball Road } Elkton, MD 21921 } }
A good question and one that is difficult to answer. There are many possibilities.
It is true that as scopes age, as with any instrument, something can go terribly wrong. A pump can fail, lens coil can short out, driver blows up, CRT dies, etc. Depending on the particular scope and what type of source it uses, there is probably some crossover point where it makes sense to have the contract. For example, if you have a system running field emission, then the cost to replace the emitter is about $3500 just for the emitter. A LaB6 would run about $550 and W about $20. The cost of the annual contract would include the cost of replacing the emitter whether it actually needed replacement or not. If the emitter did not fail, you lost money. If it did, maybe you broke even. You probably just did not lose as much.
Suppose an ion pump fails or needs overhaul. This can cost between $500 and $1000. A mechanical pump rebuild/repair is about $350. Apertures are cheap and are simply consumables. The pumps will eventually need repair and overhaul. With normal use and reasonalble care, that is probably at the 4-6 year time point.
What I found works for me is to set aside about 1/4 of the annual maintenance contract cost in a separate account. I perform the PM myself. Then, if I need factory or independent service help, I have a pool to pay for it. And it does not hurt to have a factory or independent expert come in once in awhile just to check the system over and do some of the more exotic cleaning and alignment chores. These people generally charge straight time plus travel. some independents only charge for on-the-job time. The going rates seem to be about $125-$175 per hour. Most jobs tend to run around 4 hours.
If your system is really unreliable or you are not able to perform most of the PM functions yourself, then I would think that the maintenance contract is a good idea...either with the factory or an independent.
parts don't seem to be a problem. It is true that as systems get older it can be more difficult to obtain parts. However, this depends on the brand and what it is that needs replaced. The major items that need replacement and more or less industry standards. Like Edwards or Alcatel mechanical pumps. Varian ion pumps, etc. O-rings are widely available. It would seem that an item must be very scope specific to become an issue of availability. Within 5-7 years of a scope coming out, parts don't seem to be a problem. My early Amray is still supported by Amray.
We are seeking to replace our stock of these, and e-mail enquiries to the company themselves have got nowhere. So (a) does anyone know a supplier, and (b) is there an alternative? We have used this type because it is the only organic material for filter membranes that can stand up to Xylene at 120^C, and the anodisc alumina filters in our catalogues are all too small in pore size. =================================================== We (e.g. SPI Supplies) have offered the SPI Silver Membrane filters since 1976. They might be an acceptable alternative to the other mentioned choices. They are available in a number of different pore sizes including your desired 0.8 um. They are inert to xylene or just about any other organic solvent for that matter. You can get full information on our website given below. They handle more or less like other membrane filters including the fact they can be critical point dried.
While the expense per membrane is higher than most polymer membranes, depending on what you are collecting, if strictly organic, exposure to an oxygen plasma etcher can usually regenerate them for further use with other samples, something you could not do with polymeric membranes.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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we have to analyse by TEM magnetic samples coming from steel industry. The first tries we made in the TEM (LEO 912 Omega) were really not successfull, and we have no experience with this kind of samples.
Does anyone know how to handle magnetic samples in a TEM?
Many thanks in advance, Nathalie _________________________________________________
Dr. Nathalie Bozzolo Laboratoire d'Analyse des Materiaux Centre de Recherche Public - Centre Universitaire 162a, avenue de la Faiencerie L-1511 Luxembourg tel : (352)46 66 44 402 fax : (352)46 66 44 400 e-mail : Nathalie.Bozzolo-at-crpcu.lu _________________________________________________
I truly appreciate all of you MSA list members taking the time to respond to my query about the old version of MSmouse.com driver. I was able to resurrect the old computer and discovered a bad RAM socket as main culprit. I will be able to copy the file from there to a new machine this week. Much appreciated and gracias!
*Disclaimer: Whatever... is not Tulane opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web:[ http://www.tmc.tulane.edu/ferminlab/] or [http://www.tmc.tulane.edu/imaging/] Internet: [cfermin-at-mailhost.tcs.tulane.edu] 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
have fun... sigh... I certainly have a lot more respect for the pioneers of EM who got all those nice photos of steel in the 50's.
Since the sample is likely to be a low carbon steel, you will probably find that the surface is covered with an oxide film which obscures any structure inside the sample. Even storing the samples in a vacuum desiccator did not prevent the formation of the oxide layers. Immediately before putting the sample in the scope, I have used a Fischione plasma cleaner running pure argon to clean this oxide layer off the surface. You may be able to briefly sputter in an ion mill to do the same.
One of the more frustrating aspects of doing EM on magnetic samples is that every time you move the sample (especially tilt), you will find it necessary to reset your current centering. I will often define one of the DF channels to be my bright-field condition so that I can easily reset my illumination tilt. (If readjusting the BF tilt is easy on your scope, this may not be an issue for you.) Otherwise, just remember that patience is a virtue.
Good luck, Henk
At 10:05 AM 5/3/99 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
I'm not quite sure if you got problems in sample prep or analysis 'cause magnetic material samples usually are not only tough to prepare but also tough to be analyzed. The following is just two tips:
1. Demagnetization This may work well only for hard magnets. Soft magnetic materials are easily magnetized anyway and therefore demagnetization wont help much. 2. ALWAYS turn off the objective lens while you are loading your specimen into the TEM.
As for analysis, that's at least a 3 credit 600 level course. You may want to see cells, walls, precipitates in hard magnets, or you may want to see anti-phase-boundaries in soft magnets and therefore superlattices, or most probably you may want to see magnetic domains and therefore you may even want to modify your TEM with special pole pieces, etc. etc. For functional multilayers, there are a bunch of other stories.
Good luck.
-cy
On Mon, 3 May 1999, Nathalie Bozzolo wrote:
} Date: Mon, 3 May 1999 10:05:53 +0200 (MET DST) } From: Nathalie Bozzolo {bozzolo-at-crpcu.lu} } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM on magnetic samples } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear colleagues, } } we have to analyse by TEM magnetic samples coming from steel industry. } The first tries we made in the TEM (LEO 912 Omega) were really not successfull, } and we have no experience with this kind of samples. } } Does anyone know how to handle magnetic samples in a TEM? } } Many thanks in advance, Nathalie } _________________________________________________ } } Dr. Nathalie Bozzolo } Laboratoire d'Analyse des Materiaux } Centre de Recherche Public - Centre Universitaire } 162a, avenue de la Faiencerie } L-1511 Luxembourg } tel : (352)46 66 44 402 } fax : (352)46 66 44 400 } e-mail : Nathalie.Bozzolo-at-crpcu.lu } _________________________________________________ } } }
I work for a NASA contractor whose job it is to design and construct experimental packages for use in space. Currently we are designing a package which will make use of a Leica microscope to be flown aboard the International Space Station.
My question regards the use of immersion oil in conjunction with an objective. We have purchased an objective designed to be used as such, and it's operation is understandable on ground. What I need to know is if there is any one who has done any rersearch into the wetting properties of these types of oils, such that when we try to deploy them in space we can wet the surfaces of interest, i.e. the sample slide, and the objective? The necessity for this type of information becomes evident when you realize that there will be no gravity to assist in the deployment of the oil droplet.
Any assistance in the this matter would be greatly appreciated. Thank you in advance
Our lab owns a tem (philips 420, 15 years old) and a sem (nanolab le21000 = 12-15 years old) and we never had a service contract on any of them...raiso= n it is too expensive...
I believe that if you do basic maintenance such as oil change, filament = change...etc.... and you train and watch carefully people that are using = the instruments you should not run into deep trouble... I have compagny maintenance when I encounter a problem that I am unable to = solve and some of them are simply handle by the phone,=20 and since I have been working here (for the past 10 years), it cost me a = fraction of the cost of a contract service.... we now own a new SEM and intend to do exactly the same thing.
Hope this will help,=20
Diane Montpetit Electron microscopy lab Agriculture Canada Food research center St-Hyacinthe, Qu=E9bec Canada
Dear Aley, Because it is easy to slightly over-saturate the filament, I always run at just under the saturation point. Also, the saturation point actually creeps down as the filament ages, so check it, reset it and recentre it every hour for the first five hours or so of the new filament's life. The vacuum condition is also important. My filaments last an average of one month (100 hrs.) At 02:38 PM 02/05/99 +0300, you wrote: } Hello everyone, } } What is the average lifetime of a hair-pin tungsten filament? } We have a JEOL-840A SEM which we operate at 20 KV, and which we run at } asturation (on average) for ~ 7 hours everyday 5 days a week. For the } past several years, filaments used to last 6 weeks or longer, but } recently, the filament is burning out every 5 - 7 days! What could be } the reasons for this, and what are the solutions? } Many thanks. } } Aley El-Shazly } Department of Earth Sciences } College of Science } Sultan Qaboos University } POBox 36, Al-Khod PC 123 } Oman } e-mail: aley-at-squ.edu.om } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
} My question regards the use of immersion oil in conjunction with an } objective. We have purchased an objective designed to be used as such, and } it's operation is understandable on ground. What I need to know is if there } is any one who has done any rersearch into the wetting properties of these } types of oils, such that when we try to deploy them in space we can wet the } surfaces of interest, i.e. the sample slide, and the objective? The } necessity for this type of information becomes evident when you realize } that there will be no gravity to assist in the deployment of the oil droplet. }
Dear John, I have not studied the properties of immersion oil, but from what I've seen, the oil will wet both the glass of the slide and the parts of the lens. This property is a function of the composition of the oil and the microscope parts and will be independent of gravitation. That is, once the oil is in contact with the slide and the scope, it will form much the same contact angles as it does in an earth- bound lab. I suggest filling a syringe with the oil. On the space station, when examining a specimen on a slide place the lens near the slide. The distance should be somewhat smaller than the size of a droplet being expelled from the syringe. Place the syringe near the end of the lens and slowly express a drop of oil. When the drop makes contact with the slide and lens, it should wet both. Continue until the appropriate amount of oil is on the lens and slide. Good luck. Yours, Bill Tivol
Sounds interesting. I have no real experience in this matter but just a few things I thought about: Oil should work regarding stickiness but the surface tension may be a problem. Have you considered using the 63X/1.2 water immersion objective from Leica. I've heard it's really good and it will be less greasy:-) The surface tension of water will make it easy to control the droplet. If one equip the scope with both objective one can always see. I think there is a good 100X water objective from Leica as well.
Regards
Lars
____________________________________________________________ Lars Bjork,PhD
Dept of Immunology Dept. of Biology Wenner-Gren Institute Sect. for Cell & Mol. Biology Stockholm University Pharmacia & Upjohn S-106 91 Stockholm S-112 87 Stockholm SWEDEN SWEDEN
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Gentlemen, } } I work for a NASA contractor whose job it is to design and construct } experimental packages for use in space. Currently we are designing a } package which will make use of a Leica microscope to be flown aboard the } International Space Station. } } My question regards the use of immersion oil in conjunction with an } objective. We have purchased an objective designed to be used as such, and } it's operation is understandable on ground. What I need to know is if there } is any one who has done any rersearch into the wetting properties of these } types of oils, such that when we try to deploy them in space we can wet the } surfaces of interest, i.e. the sample slide, and the objective? The } necessity for this type of information becomes evident when you realize } that there will be no gravity to assist in the deployment of the oil droplet. } } Any assistance in the this matter would be greatly appreciated. } Thank you in advance } } John } John Eustace } Optical Physicist } Dynacs Engineering } 2001 Aerospace Parkway } Brookpark, Ohio 44142 } Ph (216) 977 - 1244 } Fax (216) 977 - 1269 } }
} } What is the average lifetime of a hair-pin tungsten filament? } } We have a JEOL-840A SEM which we operate at 20 KV, and which we run at } } asturation (on average) for ~ 7 hours everyday 5 days a week. For the } } past several years, filaments used to last 6 weeks or longer, but } } recently, the filament is burning out every 5 - 7 days! What could be } } the reasons for this, and what are the solutions? } } Many thanks.
Check the vacuum first. We had a long term problem with ~30 hours filament lifetime on a XL30/tmp. Tried all sorts of other fixes first. Replacing the gun o-ring fixed the problem and we are now up to ~160 hours. The gun o-ring is the cheapest fix, easy to do. Should have tried that first.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
We work on magnetic samples here and one of the primary methods we use to handle them is to reduce the volume of material as much as possible. This includes making the disks smaller than 3 mm (2.3 mm is another standard) and keeping them thin. I believe one technique has been to punch out a 1mm disk of the steel, and insert it into a 3mm disk of another material with a 1mm hole punched out. If you have questions about that technique, please contact me "offline" and I'll put you in touch with one of my colleagues. A more difficult sample prep method that has been around for some time is the "window" electropolishing technique. This method, outlined in many microscopy texts (Williams and Carter, for example), results in a small sample volume. It takes practice but can yield good results.
Additionally, we have ordered our microscopes with "extra strength" objective stigmator coils. If you will do many of these samples, perhaps they can be retrofitted (is that a word?) into your LEO.
Best of luck.
Cheers, JSV
*************************** John S. Vetrano Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 USA Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
---------- } From: bozzolo-at-crpcu.lu Sent: Monday, May 3, 1999 12:05 AM To: Microscopy-at-Sparc5.Microscopy.Com
Dear colleagues,
we have to analyse by TEM magnetic samples coming from steel industry. The first tries we made in the TEM (LEO 912 Omega) were really not successfull, and we have no experience with this kind of samples.
Does anyone know how to handle magnetic samples in a TEM?
Many thanks in advance, Nathalie _________________________________________________
Dr. Nathalie Bozzolo Laboratoire d'Analyse des Materiaux Centre de Recherche Public - Centre Universitaire 162a, avenue de la Faiencerie L-1511 Luxembourg tel : (352)46 66 44 402 fax : (352)46 66 44 400 e-mail : Nathalie.Bozzolo-at-crpcu.lu _________________________________________________
Couldn't you duck the issue and use high/dry objectives instead? These offer VERY good optics without the necessity for immersion oil.
Ann Lehman Trinity College Hartford CT
-----Original Message----- } From: John Eustace [mailto:John.Eustace-at-lerc.nasa.gov] Sent: Monday, May 03, 1999 9:15 AM To: Microscopy-at-Sparc5.Microscopy.Com
Gentlemen,
I work for a NASA contractor whose job it is to design and construct experimental packages for use in space. Currently we are designing a package which will make use of a Leica microscope to be flown aboard the International Space Station.
My question regards the use of immersion oil in conjunction with an objective. We have purchased an objective designed to be used as such, and it's operation is understandable on ground. What I need to know is if there is any one who has done any rersearch into the wetting properties of these types of oils, such that when we try to deploy them in space we can wet the surfaces of interest, i.e. the sample slide, and the objective? The necessity for this type of information becomes evident when you realize that there will be no gravity to assist in the deployment of the oil droplet.
Any assistance in the this matter would be greatly appreciated. Thank you in advance
The usual problem with the kind of samples you might get from a steel = company is that normal TEM-size disk samples made by jet electropolishing interact = so strongly with the objective lens (OL) magnetic field that they grossly = deflect the electron beam off axis and cause high image astigmatism. Moreover, = the beam axial alignment and astigmatism change each time the sample is tilted = (or sometimes even translated). In addition, thin foil areas tend strongly = to align normal to the lens axis, and will bend to maintain this alignment whan = you attempt to tilt the sample. What usually happens during tilting is, = just as you're approaching the orientation or diffraction condition you wanted, = the sample area reorients itself. This is a semi-reversable process, but = after you tilt back and forth a few times, the sample tends to become deformed = and worthless. In really severe cases of sample-OL field interaction, the = sample can be ripped out of the holder or will reorient the holder tilt = mechanism so the holder cannot be withdrawn without scratching the OL polepieces. = If this happens, don't try to remove the holder. Just turn off the OL, open = the OL chamber, and carefully free the stuck parts. In any case, you'll want = to make sure your sample is firmly held before inserting the sample holder. = The infamous 'j-ring' holder clips (c-shaped spring clips) in certain = holders are prone to losing magnetic samples and should be avoided. Screw-down = holder mechanisms are best. =20
OK. So what to do. Use the least amount of magnetic material you can = in the sample. Make the samples as thin as possible by reducing the sheet = thickness before punching disks for jet thinning. Disk samples 30-50 =B5m thick = can be used in most microscopes with foolproof sample mounting if you realign beam = tilts and correct OL astigmatism each time the sample is tilted. Many = microscopes increase the beam deflection range in the darkfield mode, so I = typically use one of the darkfield channels for magnetic sample work. Also, it's a good = idea to switch off the OL before inserting the sample holder. On some = microscopes, you can also increase the range of stigmator strength, but that's extreme. = Just remember to align the microscope beforehand with a nonmagnetic sample. = Also, your co-workers will appreciate it if you restore normal settings when = you finish. =20
In some cases, you might want to take the idea of minimizing magnetic = sample sizes further. A colleague working with radioactive samples has = developed the technique of punching out a 1 mm diameter disk sample and fastening it = into an annular disk of nonmagnetic material (usually 316 stainless steel) = before jet electropolishing the center. (I'll send the reference to you if I can = find it.) For much greater sample reduction, I have had success window-thinning = sheet samples (see standard references on TEM sample preparation for details) = and cutting off thin shards sandwich between two grids. In this case, = sample/OL interaction was almost nil. Good luck
Larry Thomas Mechanical and Materials Engineering Washington State University Pullman, WA 99352 USA
thomas-at-mme.wsu.edu tel: 509 372-0793 ---------- From: bozzolo-at-crpcu.lu Sent: Monday, May 3, 1999 1:05 AM To: Microscopy-at-Sparc5.Microscopy.Com Subject: TEM on magnetic samples
The Microscopy ListServer -- Sponsor: The Microscopy Society of = America=20 To Subscribe/Unsubscribe -- Send Email to = ListServer-at-MSA.Microscopy.Com On-Line Help = http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html = -----------------------------------------------------------------------.=
Dear colleagues,
we have to analyse by TEM magnetic samples coming from steel industry. =
The first tries we made in the TEM (LEO 912 Omega) were really not successfull,=20 and we have no experience with this kind of samples.
Does anyone know how to handle magnetic samples in a TEM?=20
Many thanks in advance, Nathalie _________________________________________________
Dr. Nathalie Bozzolo Laboratoire d'Analyse des Materiaux Centre de Recherche Public - Centre Universitaire 162a, avenue de la Faiencerie L-1511 Luxembourg tel : (352)46 66 44 402 fax : (352)46 66 44 400 e-mail : Nathalie.Bozzolo-at-crpcu.lu _________________________________________________
Dear Sir/Madam, I am conducting experimental studies on surfactants. I would like to know if the temperature and concentration phase diagram of teepol is available. If it is please tell me where. Thanking You, Yours Sincerely, Geetha Basappa.
****************************************** Dr Geetha Basappa, Project Associate, Physics Department, Indian Institute of Science, Bangalore, India.
Aley, Check your upper & lower gun cylinder walls area for a faint bluish-gray tint(tungsten oxide). You could have a slight vacuum leak. You would probably never see the leak, it being so far away from any vacuum gauge. Good luck.
Gary M. Easton, Pres. Scanners Corporation ----- Original Message ----- } From: Mary Mager {mager-at-interchange.ubc.ca} To: aley {aley-at-squ.edu.om} Cc: {Microscopy-at-Sparc5.Microscopy.Com} Sent: Monday, May 03, 1999 11:39 AM
Mary Mager wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Aley, } Because it is easy to slightly over-saturate the filament, I always run at } just under the saturation point. Also, the saturation point actually creeps } down as the filament ages, so check it, reset it and recentre it every hour } for the first five hours or so of the new filament's life. The vacuum } condition is also important. My filaments last an average of one month (100 } hrs.) } At 02:38 PM 02/05/99 +0300, you wrote: } } Hello everyone, } } } } What is the average lifetime of a hair-pin tungsten filament? } } We have a JEOL-840A SEM which we operate at 20 KV, and which we run at } } asturation (on average) for ~ 7 hours everyday 5 days a week. For the } } past several years, filaments used to last 6 weeks or longer, but } } recently, the filament is burning out every 5 - 7 days! What could be } } the reasons for this, and what are the solutions? } } Many thanks. } } } } Aley El-Shazly } } Department of Earth Sciences } } College of Science } } Sultan Qaboos University } } POBox 36, Al-Khod PC 123 } } Oman } } e-mail: aley-at-squ.edu.om } } } Regards, } Mary } } Mary Mager } Electron Microscopist } Metals and Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } e-mail: mager-at-interchg.ubc.ca
Aley, With a sudden drop in filament life, as you describe, and assuming that you are treating your microscope the same as always, I would look for a vacuum leak in the immediate area of the gun. It could be a single lint fiber causing enough degradation of your vacuum to shorten your filament life as you descibe. Have you opened anything either on the back side of the gun (around thge stanpipe to the DP) or down on the column (fimal apertures, beam current probe, isolation valve)? If you have, that is the first place I would look.
Ken Converse owner Quality Images third party SEM service Delta, PA
Diane Montpetit wrote ... } } Our lab owns a tem (philips 420, 15 years old) and a sem } (nanolab le21000 12-15 years old) and we never had a service } contract on any of them...raison it is too expensive... } } ...
This is my viewpoint as well ... HOWEVER, I am a firm believer in finding the money (by whatever means possible) to cover a service contract for the two years after the initial warranty period. If anything is going to go wrong it will generally happen during this time period. This also gives the facility manager and technicians the time to become acquainted with the little idiosyncracies of the intrument with the help of those who are most aware.
... my $0.02 :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Dear Listers, I am searching for an old JEOL 100SX TEM which could be used for parts. I would appreciate any leads. Thank you and please contact me at my address rather than on the list. Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB - 499 1807 West Slaughter Lane #200 Austin, Texas 78748-6200 Phone: 512/282-5507 Fax: 512/280-0702
At a recent MSA meeting (within the last 5 years or so) there was a session on microscopes in the classroom that featured something affectionately referred to as a 'Scope on a Rope'.
It was a video camera tethered to a TV with a built in light source and medium mag. All you did was hold it up to something and a magnified picture appeared on the TV.
Of course, now that I need to know more about it, I can't find any references. Anyone remember it or have an idea of where to start looking?
As always, thanks a million.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Has anyone had any experience with phycoerythrin-conjugated antibodies and fluorescence microscopy? I'm specifically interested in hearing whether there are any "anti-fade" reagents that work well with PE. Although I've heard that PE antibodies don't work well with fluorescence microscopy, I don't have a choice of conjugates, unfortunately! Any help would be much appreciated!
Rizwan Haq Ontario Cancer Institute Toronto, Ontario Canada haqr-at-oci.utoronto.ca
Using Ultramicrotomy in Materials Science - September 21-24, 1999
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Lodging, tuition, meals, and materials are all covered in the fee of $1950 (only transportation excluded). Lectures and lodging are at the beautiful resort, Lodge on the Desert, below the Santa Catalina mountains. Lab sessions are at the RMC/Ventana's Tucson laboratory. For more information please contact Steve Miller, tel: 520-903-9366, SMiller-at-Ventanamed.com or see our web site at RMC-Scientific.com/microtomes/
} John Eustace wrote: } } } My question regards the use of immersion oil in conjunction with an } } objective. We have purchased an objective designed to be used as such, and } } it's operation is understandable on ground. What I need to know is if there } } is any one who has done any rersearch into the wetting properties of these } } types of oils, such that when we try to deploy them in space we can wet the } } surfaces of interest, i.e. the sample slide, and the objective? The } } necessity for this type of information becomes evident when you realize } } that there will be no gravity to assist in the deployment of the oil droplet. } } } John,
There is also a water immersion lens. It does not have as large an aperture as a oil immersion lens. But the water is a lot less noxious stuff to be floating around in the air than oil.
It won/t wet out as well as oil but spilled water is of little consequence in a weightless environment.
Water immersion should work for most experiments and you can always put the oil lens in if it is needed.
Gordon
Gordon Couger gcouger-at-couger.com www.couger.com/gcouger Stillwater, OK 405 624-2855 GMT -6:00
At 02:38 PM 02/05/99 +0300, you wrote: } Hello everyone, } } What is the average lifetime of a hair-pin tungsten filament? } We have a JEOL-840A SEM which we operate at 20 KV, and which we run at } asturation (on average) for ~ 7 hours everyday 5 days a week. For the } past several years, filaments used to last 6 weeks or longer, but } recently, the filament is burning out every 5 - 7 days! What could be } the reasons for this, and what are the solutions? } Many thanks. } } Aley El-Shazly } Department of Earth Sciences } College of Science } Sultan Qaboos University } POBox 36, Al-Khod PC 123 } Oman } e-mail: aley-at-squ.edu.om
I get about 50 hours of life from a standard W filament. I can get over 225 hours life from an Energy Beam Sciences SG filament. A good vacuum is very important no matter which filament make you use. I use an ion pump routinely and switch back and forth between W and LaB6.
I'm not sure which type filament your Jeol uses but here are the two types that are available for Jeol:
I did double labelling of whole retinal tissue years ago. The method came from the refs below. It was fairly easy, though it was necessary to be rigorous with controls (as in any double labelling). I didn't persevere as fixation wasn't great and labelling intensity was low. I could probably have improved it, but didn't feel it was worth it. The papers below had (from memory; I no longer have the originals) good results.
Sako H, et al (1986) Simultaneous detection of B-cells and T-cells by a double immunohistochemical technique using immunogold-silver staining and the avidin-biotin-peroxidase complex method. Histochemistry 86:1-4
van den Pol AN (1985) Silver-intensified gold and peroxidase as dual ultrastructural immunolabels for pre- and postsynaptic neurotransmitters. Science 228: 332-5
van den Pol AN (1986) Tyrosine hydroxylase immunoreactive neurons throughout the hypothalamus receive glutamate decarboxylase immunoreactive synapses: a double pre-embedding immunocytochemical study with particulate silver and HRP. J Neurosci 6:877-91
van den Pol AN, Smith AD and Powell JF (1985) GABA axons in synaptic contact with dopamine neurons in the substantia nigra: double immunocytochemistry with biotin-peroxidase and protein A-colloidal gold. Brain Res 348:146-54
Chan J, Aoki C and Pickel VM (1990) Optimization of differential immunogold-silver and peroxidase labelling with maintenance of ultrastructure in brain sections before plastic embedding. J Neurosci Methods 33:113-27
Demonstration of two antigens using a novel combination of immunogld-silver staining and immunoenzymatic labelling. J Histochem. Cytochem. 38:307-13
Krenacs T, Laszik Z and Dobo E. (1989) Application of immunogold-silver staining and immunoenzymatic methods in multiple labelling of human pancreatic Langerhans islet cells. Acta Histochem. 85:79-85
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
Does anyone out there in cyber space have a control board (90014438-611) = for an LKB 2188 NOVA Ultratome. We are quite willing to pay for it as = our ultratome is now off the air. Any suggestions would be gratefully = received.
Terry A Robertson
=20 Dr Terry Robertson (PhD) Senior Research Fellow Department of Pathology University of Western Australia Nedlands 6907
Hi All, I have two boxes of brand new JEOL E-type filaments here. I have no idea what they are doing in the lab: there has never been an instrument that uses these in the vicinity. Anybody have a use for them? Cheers, Malc. Dr MP Roberts Department of Geology Rhodes University Grahamstown 6140 South Africa Tel: +27 46 6038316 Fax: +27 46 6229715 ******************************* "If God had meant birds to fly, he would have given them engines" Anon.
I guess you will have a mass of replies on this one?
It is my experience, working around the world, that filament life almost=
seems more important in many laboratories than the quality of the final image ;-).
What sets the filament life?
1. The magnification level and the kV you wish to use - as you go higher in magnification (} 20,000X) you need to work with a better formed=
probe with as many electrons as possible. Therefore high magnification means on a JSM 840 100 to 120 uA of current, more current means less life= ? =
If you are running to attain true surface images you will find the gun is=
far less efficient at {10kV so the filament life will fall as you drive t= he filament even harder to get the emission current up. Low magnification( {5,000X), when the instrument is not really being used very=
hard at all, the filaments should go on for ever (see later).
2. The way you saturate and align - a wave form is the best method a= s a wide variety of people all come to the same conclusion with this method= . =
However as the filament thins with use, requiring less current, you shoul= d be rechecking your version of saturation and alignment each time you swit= ch the kV on and after each kV change. Then we come on to under saturating!= =
Fine if you run at very low magnifications and do basic x-ray analysis, b= ut do not complain when your images are not so good, poor source production equals poor image production as the final beam spot mimics the source!.
3. The vacuum level - you may fudge your saturation or find excuses for running at a low emission current but the vacuum system cannot be fudged. The life of a filament is directly related to the vacuum level i= n the gun chamber, the better the vacuum the better the filament life! A dirt gun chamber will also "spoil" the gun vacuum so a clean chamber is also important. Remember vacuum gauges tend to be a long way from the gu= n and do not represent its vacuum! JSM840 has the advantage of using an exchange airlock which should give a very good column vacuum.
4. The quality of the filaments - having worked for a number of manufacturers we found from time to time we were supplied from the factor= y with poor quality filaments. They seemed to be made up of wire plus rubbish and they only lasted about 10 hours making a mess of the cathode and gun chamber. We heard about some problems but you will be surprised how few boxes came back. Standard practice should be NEVER use a complet= e box of filaments if they are working well, always save two. When you hav= e a new box of filaments, where the first one gives a poor filament life, u= se one more and work with particular care. If this life too is poor fit one=
of your "good box" filaments and try again. "Good box" filament gives go= od life - problem =3D new box of filaments. "Good box" filament gives poor = life - problem =3D gun vacuum.
5. How you measure it - if you run an instrument that has a meter fitted to the filament on switch you soon learn that even if the instrume= nt is "on" for seven hours a day the actual filament time is far short of that! Many checks have shown that in a seven hour day the filament is unlikely to be on for more than half the time with a single specimen exchange system. With a multi user facility, with operator changes, the filament time is reduced even further. X-ray analysis is the biggest ti= me consumer as people leave the filament on when playing with their spectrum= . =
HT on makes sense filament on costs money!
So to answer the question what is a good filament life? Well it depends
SEM } 50,000X a pointed filament will last about 10-15 hours a V about 20-= 30 hours ~20,000X a V about 30-40 hours {5,000X a V about 40-60 hours EDX a V in excess of 60 hours
TEM it is a different game as we do not drive the gun hard under many applications and to make cross over easy to recognise we run slightly und= er saturated. So TEM depending on application and instrument age 70 hours plus.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
I'm afraid that I didn't see the original posting on this, so I am relying to this reply.
I find it interesting that this thread should run concurrent to a thread regarding the value of service contracts. You often don't kow what you may be missing...
A simple tungsten filament should work for around 80 hours of use. If you are getting less than this, you are either operating the filament at too high a temperature or there are vacuum leaks in your system that permit air at a location that reduces the vacuum in the gun. The major gun seal is opened when replacing filaments and should be checked. A typical SEM vacuum system puts the pumps at one end of the volume being evacuated and the electron gun at the other end. Small leaks in the gun, or in the standpipe that evacuates the gun, can have enhanced effects on the filament life. The standpipe is often the culprit. This is the vacuum connection between the specimen chamber and the electron gun that is provided to enhance the pumpdown of the gun and prevent the gun being pumped down through the column and its apetures.
The standpipe o-ring seals at the specimen chamber and the gun are often a weak point in design. Manufacturers have used a variety of seal designs at both the chamber and gun that often are marginal at best. Another, insiduous source of problems are the seals at the secondary electron detector, EDS detector or other chamber ports. Since they are often near the standpipe opening to the specimen chamber, they to can lead to poor gun vacuum.
If your instrument has a gun translation system, this should also be checked. The gun translation provides for the mechanical adjustment of the position of the electron gun, usually through opposing set screw adjustments at the top of the electron column. Many, if not most, recent model instruments have electronic controls, rather than mechanical controls, that don't present this problem.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com
-----Original Message----- } From: Dr. Gary Gaugler {gary-at-gaugler.com} To: MSA listserver {Microscopy-at-sparc5.microscopy.com}
John - your concerns are unfounded. Modern immersion fluids are essentially non-toxic and they adhere because of extreme surface tension makes them quite sticky and that would not change outside gravity. They are also essentially non-drying and would last in a reduced atmosphere with no appreciable drying. In a clean environment the medium need only be wiped off occasionally. In gravity or with g-forces applied, a hanging drop could move, but that is unlikely to apply in space and during blast-off presumably no oil would be on the objective. If there was concern about the drop of immersion oil being dislodged, I would recommend Cargille's Type NVH with high viscosity of 21,000cST , this type is used for inclined and inverted optics. The drop will never fall off, but it is harder to wipe off. Sure you can do away with oil immersion, but in light microscopy this will lower top resolution attainable.
Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Monday, May 03, 1999 11:15 PM, John Eustace [SMTP:John.Eustace-at-lerc.nasa.gov] wrote: } } Gentlemen, } } I work for a NASA contractor whose job it is to design and } construct } experimental packages for use in space. Currently we are } designing a } package which will make use of a Leica microscope to be } flown aboard the } International Space Station. } } My question regards the use of immersion oil in } conjunction with an } objective. We have purchased an objective designed to be } used as such, and } it's operation is understandable on ground. What I need to } know is if there } is any one who has done any rersearch into the wetting } properties of these } types of oils, such that when we try to deploy them in } space we can wet the } surfaces of interest, i.e. the sample slide, and the } objective? The } necessity for this type of information becomes evident } when you realize } that there will be no gravity to assist in the deployment } of the oil droplet. } } Any assistance in the this matter would be greatly } appreciated. } Thank you in advance } } John } John Eustace } Optical Physicist } Dynacs Engineering } 2001 Aerospace Parkway } Brookpark, Ohio 44142 } Ph (216) 977 - 1244 } Fax (216) 977 - 1269
I'm doing strange observations on a group of tiny mites and I have found that they are able to produce fine drops of idrophobic substance ("saliva" ?) when they are immersed in oil. I have tried some types of oil but "immersion oil for microscopy (ordinary use) nd =3D 1.516 at 23=B0C produced= by Olympus optical Co., LTD" is the only one in which it happens.
My knowledge about the composition of this oil was that it is produced from cedar. So, I tested condensate cedar oil but the mite died soon (after 30 minutes) without any dropplets visible.
My question is:
What is the exact composition of the immersion oil?
Thanks for all help.
dr Enrico de Lillo Istituto di Entomologia agraria - Universit=E0 Bari - Italy tel. +39 080 5443105 fax +39 080 5442876 email: delillo-at-agr.uniba.it http://193.204.185.103/de_lillo.htm
Ann Lehman says: } Couldn't you duck the issue and use high/dry objectives instead? These } offer VERY good optics without the necessity for immersion oil.
Hmm, could you explain how to duck the issue of theoretical impossibility to achieve N.A. greater than 1.0 in the air, with the consequent effects on resolution and meaningful magnification? Also, all other conditions equal, immersion objectives tend to beat dry ones when resolution and magnification are important - so "VERY good" becomes "MUCH better" with oil... High/dry is likely to be 40x to 63x... Oil is likely to be 90x to 100x...
Or have you seen a "very good" high/dry 100x na=1.0 objective?
John, you may need oil-on-oil, i.e. placing the oil-drop on the top condenser lens to oil the specimen slide to the condenser, and oil-drop on the cover-slip to oil the front lens of the objective to the cover-slip. Otherwise your resolution will suffer.
In zero-G your oil droplet will probably just stay a ball so you'd have to "squash" it over the surface to be oiled... This is my rough guess... I'd really like to hear the "actual" solution you eemploy. -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
Have solid-state annular BSE detectors for 15kv+ SEM applications attained true TV-rate performance in recent times? If so, could a manufacturer be mentioned, please?
I can't speak for the 'Scope on a Rope, but I saw an interesting all-in-one device from Cole Parmer at PITTCON. It is a digital microscope system which has a 4 MB smart card for collecting images and a 2"liquid crystal screen. It has its own set of lenses (30x comes standard; 1x, 50x, 100x, and 200x also available) so it can act as a traveling microscope for field use but also connects directly to a microscope via a C mount. It also has an adapter kit for interface with a PC or MAC. While not inexpensive, it is a neat package.
Cole-Parmer can b reached at 800-323-4340 or www.coleparmer.com.
I look forward to hearing more about 'Scope on a Rope.
CAVEAT: MME has no financial interest in this product.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 03:39 PM 5/3/99 -0700, Jon Krupp wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} At a recent MSA meeting (within the last 5 years or so) there was a session } on microscopes in the classroom that featured something affectionately } referred to as a 'Scope on a Rope'. } } It was a video camera tethered to a TV with a built in light source and } medium mag. All you did was hold it up to something and a magnified picture } appeared on the TV. } } Of course, now that I need to know more about it, I can't find any } references. Anyone remember it or have an idea of where to start looking? } } As always, thanks a million.
Jon Krupp
Hi, Jon. I believe Carolina Biological sells it. You can get info from Bill & Cindy Henk of Louisiana (check the MSA membership list), who presented it at MSA. It's quite expensive, so the Ken-A-Vision version (Insights, listed on the MICRO page under "microscope suppliers" is one source) is more popular for precollege use. But it must be mounted on a microscope. } Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Check out the following web site. Richard Sherman is the patent holder, sells the guns, and can give you reprints of the articles. (Look in JVST circa 1992 for them.) His Email address is co2clean-at-aol.com
A colleague of mine works on primary cortical cultures. When she does immunocytochemistry on them she usually uses indirect (immunofluorescence) method, which works well. When she uses fluorescence labeled avidin-biotin method and she gets extremely high background. I have looked at the controls. The non specific binding is as bright as the specific one. I have never worked with cultures. Could there be endogenous biotin in them? Or would you have any other suggestions on what could be the cause and what can one do to prevent this? Thanking you in advance for your input, Lilith -------------------------------------------------------- Lilith Ohannessian-Barry National Research Council Institute of Biological Sciences CANADA Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
Is there a blocking step used in either procedure. When I was doing IHC on TC we used to routinely block, whether it was with a serum or a detergent such as Tween 20. We had success with blocking and felt the background was just a cell culture thing, but were thankful that it was virtually eliminated.
Mary Sr. Lab Tech Bone Marrow Transplant Medical College of Wisconsin Mary_M-at-bmt.mcw.edu
On Tue, 4 May 1999, Barry, Lilith wrote:
} Date: Tue, 4 May 1999 14:48:00 -0400 } From: "Barry, Lilith" {Lilith.Barry-at-nrc.ca} } To: Histonet {histonet-at-Pathology.swmed.edu} , } microscopy {microscopy-at-Sparc5.Microscopy.Com} } Subject: Tissue culture background } } A colleague of mine works on primary cortical cultures. When she does } immunocytochemistry on them she usually uses indirect (immunofluorescence) } method, which works well. When she uses fluorescence labeled } avidin-biotin method and she gets extremely high background. I have looked } at the controls. The non specific binding is as bright as the specific one. } I have never worked with cultures. Could there be endogenous biotin in them? } Or would you have any other suggestions on what could be the cause and what } can one do to prevent this? } Thanking you in advance for your input, } Lilith } -------------------------------------------------------- } Lilith Ohannessian-Barry } National Research Council } Institute of Biological Sciences } CANADA } Tel;613-993-6460 } Fax;613-941-4475 } e-mail; lilith.barry-at-nrc.ca }
I am to prepare samples for TEM from Au 38at% Ni 100um. thick foil.
I will be using a Fischione polisher.
Does anyone have a polishing solution for this alloy?
Thanks in advance
Fred Pearson
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
I have a paper from Bernie Kestel at Argonne National Lab which describes=
the use of a non-acid electrolyte for thinning Au with the South Bay Technology Model 550 Jet Polisher. The recipe is for his BK-2 solution a= s follows:
The sample polished was annealed, polycrystalline gold and it was thinned=
at -55 degrees C with a potential at 150V at one half the maximum electrolyte flow rate. I believe the Fischione unit only goes up to 120V= , so you may need to adjust the recipe to account for the lower voltage. =
I have been presented with a new challenge and I need to be brought up to speed on a few things.
I need to help some biochemists look at something they call liposomes. Actually they call them SR vesicles and BR vesicles, sensory and bacterial rhodopsin vesicles. This is what I know: The liposomes are in 4M salt, they are supposed to be something like 15 nm in dia. and all they want to know is whether they are intact spheres or broken fragments. Previously they had someone look at them using freeze fracture, but that person is no longer available and we do not have freeze fracture available here.
They suggested negative staining, but they are short on references and advice. I was just going to dive in and do a simple negative stain with UA, but thought I better look for help wherever I can get it.
I am not sure about a couple of things. Does anyone have advice about how to apply the liposomes to the grid, I thought I would just drop some onto a formvar coated grid to start. The 15 nm size bothers me too. Seems small for our TEM and telling one little tiny blob from another little tiny blob is no fun for me.
Does anyone have advice about how the 4M salt will work out. I have visions of looking at salt boulders rather than liposomes. I asked about rinsing out the salt, but they say that will burst the liposomes.
Yes, I do remember a thread about liposomes here recently. Of course, as usual, I did not give it the attention it deserved given my new assignment. Perhaps someone could refresh my memory and clue me in.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
My company is considering to send out samples on a regular basis for EDS analysis with high spatial resolution (in the nm range). I would appreciate to be contacted direcly by email with referrals of analytical labs with FE-TEM or FE-STEM capabilities willing to offer this service.
Adsorption of Poly(2-vinylpyridine) Wormlike Polyelectrolyte Brushes on Mica Studied in-situ by MAC Mode(tm) AFM
U. Schmidt~, S. Prokhorova+, S.S. Sheiko+, M. Möller+, P. Dziezok*, M. Schmidt* ~Molecular Imaging / Roper Scientific, Sollner Str 61, D-81497 Munchen, Germany +Dept. of Organic Chemistry III, Univ. of Ulm, Albert Einstein Allee 11, 89069 Ulm, Germany. * Inst. of Physical Chemistry, Univ. of Mainz, Jakob-Wedler Weg 11, 55128 Mainz, Germany.
Bart Cannon wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } } Have solid-state annular BSE detectors for 15kv+ SEM applications } attained true TV-rate performance in recent times? If so, could a } manufacturer be mentioned, please? } } Thanks. } } Bart Cannon
Bart, Either a Robinson Detector or a GW Electronics detector should work at TV rates. Anybody else have any candidates?
Ken Converse owner Quality Images third party SEM service Delta, PA
While on the filament "thread" (oops, a pun slipped in), I am wondering if someone can make recommendations for suppliers of LaB6 filaments for use in a Hitachi H-7100FA (hi res, small probe). Hitachi makes a wonderful assembly but it is getting pricey ($3,500) and we would like to explore other possibilities. The purse keeps getting smaller .....
Thanks, John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I do agree with several of the respondents that, though filament life can be influenced by a number of factors, the most common cause of an abruptly decreasing life is likely to be poor vacuum. However, I do not recall anyone mentioning one of the simplest ways to check for this condition: by inspecting the tip of the expired filament under a low-power microscope (or use your SEM if you like to indulge in overkill).
Under optimal conditions, a filament's life is determined exclusively by the evaporation of the tungsten. Under optimal conditions (i.e., good vacuum and the filament drive current is not so high that the wire melts), the filament wire will gradually thin as the tungsten evaporates away. But because the thickness of the filament wire is not perfectly uniform and the temperature distribution is also not perfectly uniform, some parts of the filament wire will evaporate faster than others. Eventually, a localized region of the filament wire will have thinned enough so that it will become appreciably hotter than the rest of the filament, this causes the local rate of evaporation to accelerate even more -- the local thin spot has a higher resistance which makes it hotter, which makes it evaporate faster, which makes it thinner, which makes it hotter, etc. -- a regenerative process which quickly results in enough of a local "hot spot" to melt the tungsten and the filament breaks open at this point. The dynamics of filament construction are such that the regions just to either side of the apex of the "V" tip are naturally going to be a little thinner than the tip itself. Thus, when a filament expires "normally" (i.e., solely by evaporation) you will see that the break is on one of these legs -- the wire adjacent to the break will be noticeably thinned to a taper and there will likely be a small "ball" on either side of the break where the molten tungsten solidified. (A pair of large globs of tungsten with negligible thinning indicates that the filament was blown by a high current surge -- just the way a common fuse wire blows.)
Bad vacuum always produces a different kind of filament failure. This is because gas molecules in the gun will be ionized by collisions with the electron beam. The positive ions are accelerated towards the cathode and are focused into the filament tip. This impact sputters the tungsten tip and creates a crater. A filament which has failed because of poor vacuum, when inspected under a microscope, will look like someone took a bite out of the very tip of the "V". There is negligible thinning of the wire along the legs.
These effects are very easy to recognize with a small amount of practice and practice in doing so should really, at least in my opinion, be part of the training for anyone who operates a tungsten filament microscope (similar effects can also be observed for LaB6 filaments). ------ I do want to take a small exception to several of the statements that were made regarding altering the filament life by varying filament height, emission, magnification, kilovoltage, and the like. My objection is not that these statements are not in some sense true, but rather that they confuse cause and effect. Saying that dropping the filament height in the wehnelt will lengthen its life is sort of like stating that putting out-of-balance wheels on your car will result in better gas mileage -- it will -- because you will need to drive slower.
With a properly constructed and operated tungsten filament (i.e., assuming negligible gas sputtering), the rate of evaporation (and thus the life of the filament) are dictated solely by two factors: (1) the temperature of the filament; and (2) the thickness of the wire -- PERIOD. There are no other operational factors.
If you want long filament life, use filaments which have a somewhat thicker wire (many microprobes use this strategy) or simply run the filament at a lower temperature. So why doesn't everyone simply use thick filament wire and/or low temperature? Because these both result in sub-optimal optical performance. In other words, you have a choice: (a) operating your filament for high brightness and best resolution and accepting short filament life; or (b) operating at a lowered temperature for long filament life and accepting reduced source brightness and poorer imaging.
So why do so many microscopists state that they can alter their filament life by changing the height of the filament, changing the emission, altering the kV, etc.? Because these changes can shift the operating point at which "saturation" of the filament is achieved -- thus when the operator subsequently saturates the filament by adjusting the filament drive, he/she is actually changing the filament temperature. Thus, changing one of these parameters may in fact be a practical means of achieving longer filament life, but it is strictly a secondary effect. If you doubt this, I propose a simple experiment: instead of adjusting the filament drive so as to achieve saturation, instead maintain constant filament drive. If you do this, you will find that your filament lifetime is unaffected by any of these other operating conditions (though the quality of your imaging may well deteriorate unacceptably).
There is nothing magic about saturation. It was shown by Haine back in the '30s that the effect which we refer to as "saturation" is solely a function of the way in which the bias resistor circuit regulates the filament's emission. By varying the bias resistor, you can make the filament saturate at any temperature you please. Of course, you are still limited by the fundamental fact that a long-lived filament is a not-so-bright filament (no social commentary intended). But if a long filament life is more important than maximum image quality, then this is a good strategy.
I know these things have been discussed on this listserver before, but they seem to bear repeating in the context of the question which was asked and the discussion which has followed.
Many of my friends/clients have been waiting for me to say this so here goes -
"Just what do people get from an SEM which gives 100 hours filament life?= "
When I visit labs who need to get more from their SEM the problem always = is that they get 100+ hours life (by their means of measurement) and the images die at } 3,000X. The first thing we have to do is give them more current and throw away the filament life. As I have said so many times before "in some labs filament life is more important than image quality!"=
When I am abroad the fuel consumption of my car is amazing, it sits in th= e garage at home and does nothing, should I boast?
Come on guys lets start putting realistic figures on filament life rather=
than hours in a day and lets relate this to the maximum magnification we use and the lowest kV we use. Then and only then will we all have a base=
to work from!
Try this equation - multiply the actual filament on time by the thousand digits of the maximum magnification used and the emission current , then divide by the average kV you use. My typical customer would have a 6.7K value.
I think that would be interesting, anyone with other ideas?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain
I want to thank all of you who responded (on- and off-line) to my question about magnetic samples in the TEM. I got many usefull advises. I will keep all these messages, if anybody is interesting in getting a summary, just let me know!
Dr. Nathalie Bozzolo Laboratoire d'Analyse des Materiaux Centre de Recherche Public - Centre Universitaire 162a, avenue de la Faiencerie L-1511 Luxembourg tel : (352)46 66 44 402 fax : (352)46 66 44 400 e-mail : Nathalie.Bozzolo-at-crpcu.lu _________________________________________________
} From: "drose-at-wlgore.com"-at-Sparc5.Microscopy.Com To: Microscopy-at-Sparc5.Microscopy.Com Date sent: Tue, 4 May 1999 09:43:03 -0400
Greetings. This may be a bit out of the main-stream of our usual concerns but I have a question that many of us may share. In my microscopy lab we have an older model Sorvall Superspeed centrifuge, useful for all sorts of things. I have funds to purchase a new fixed angle rotor but cannot decide on a standard aluminum rotor or one of the new carbon fiber composite rotors. Does anyone have experience with the carbon fiber? No brand names need be involved, but do they have any hidden drawbacks compared to the standard? Anecdotes are welcome. Thank you in advance. Bill P.
I maintain the Tips & Tricks site, biologic archives of this listserver. You are correct there was a thread or two but I don't have them posted to web yet so here are the raw messages. If there is ever anything you need and it is not on the web site below, simply ask and I will search the unposted material. There are roughly 300 discussions not yet posted which exceeds what is posted. I will be addressing this in the next few months as the site gets a facelift. Sorry for the flood, but you asked for it
At 04:25 PM 5/4/1999 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
Hello dear all! I am looking for any information on the microscope, called Grumhauser and on the person who made it. The spelling I have might be incorrect. Best regards, Dima My email address is la_dima-at-hotmail.com
_______________________________________________________________ Get Free Email and Do More On The Web. Visit http://www.msn.com
BSE at TV bandwidths depends heavily on the performance of preamplifier following the detector. We've marketed a black-box add-on for a little over a decade, through one of the mainstream SEM manufacturers. The preamp offers full TV bandwidth at moderate gain, and 0.6 MHz (fuzzy TV) bandwidth at full gain, to allow real time viewing for positioning the sample.
Still wider band performance is possible, but it's been unclear whether there's a need for it - from here it looked like BSE was being supplanted by environmental detectors.
Jon - Liposomes will be invisible among all the salt crystals. I used to wash such isolates in an ammonium acetate solution. This is a volatile buffer and would sublime after application to the grid. However, you may get sufficiently clean (salt free) preparation by: 1 Apply and blot substrated grid repeatedly to liposome solution. 2 Blot 3 Apply to a series of drops of negative stain (try a couple different ones PTA, UA, ammonium molybdate) and blot in between. Its generally a good idea to use this simple method to obtain good distribution of particles. In this case, you need to eliminate the salt and that requires numerous apply and blot cycles. You could also let the grid float for a while on a drop of stain to allow dilution of the salt. Negative staining solutions do not cause osmotic shock - except when dealing with extreme halophiles.
When you finally see the image, don't be disappointed. Anything prepared by biochemists appears to microscopists as if prepared from a festering cadaver. Remaining salt is easily distinguished: its cubic. But how to judge those liposomes: are they uniform and intact? Its always a matter of degree and never a pretty sight. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Wednesday, May 05, 1999 9:26 AM, Jon Krupp [SMTP:jmkrupp-at-cats.ucsc.edu] wrote: } } I have been presented with a new challenge and I need to } be brought up to } speed on a few things. } } I need to help some biochemists look at something they } call liposomes. } Actually they call them SR vesicles and BR vesicles, } sensory and bacterial } rhodopsin vesicles. This is what I know: The liposomes are } in 4M salt, they } are supposed to be something like 15 nm in dia. and all } they want to know } is whether they are intact spheres or broken fragments. } Previously they had } someone look at them using freeze fracture, but that } person is no longer } available and we do not have freeze fracture available } here. } } They suggested negative staining, but they are short on } references and } advice. I was just going to dive in and do a simple } negative stain with UA, } but thought I better look for help wherever I can get it. } } I am not sure about a couple of things. Does anyone have } advice about how } to apply the liposomes to the grid, I thought I would just } drop some onto a } formvar coated grid to start. The 15 nm size bothers me } too. Seems small } for our TEM and telling one little tiny blob from another } little tiny blob } is no fun for me. } } Does anyone have advice about how the 4M salt will work } out. I have visions } of looking at salt boulders rather than liposomes. I asked } about rinsing } out the salt, but they say that will burst the liposomes. } } Yes, I do remember a thread about liposomes here recently. } Of course, as } usual, I did not give it the attention it deserved given } my new assignment. } Perhaps someone could refresh my memory and clue me in. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } }
John - Don't know about that model Hitachi. We supply "normal" Lab6 cathodes (Kimball Physics), which I understand are as good as any and they are arguably the most robust. I and no doubt any supplier of LaB6 cathodes would love to sell those to Hitachi (or anybody) for less than 20% of your quoted price.
The difference is so large that I wonder. Do those cathodes have the same base as do Hitachi tungsten filaments? Do you know the size of the microflat atop the LaB6 cone? Normal are 15 square micrometers, I assume that for a high resolution TEM you probably would want that small flat. The larger flats, say 40 sq. micrometers are more suited to microprobes where long term stabillity is more important than is greatest brightness. The larger flats are more expensive. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Wednesday, May 05, 1999 12:01 PM, John J. Bozzola [SMTP:bozzola-at-siu.edu] wrote: } } } While on the filament "thread" (oops, a pun slipped in), I } am wondering if } someone can make recommendations for suppliers of LaB6 } filaments for use in } a Hitachi H-7100FA (hi res, small probe). Hitachi makes a } wonderful } assembly but it is getting pricey ($3,500) and we would } like to explore } other possibilities. The purse keeps getting smaller .... } } } Thanks, } John } } ########################################################## } ########## } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ###################################################### #### } ########## } }
Thanks for to those who reponded. Below is information on the CO2 Snow Gun.
--------------------------------- Applied Surface Technologies manufactures CO2 Snow Gun. They can be reached at 908-464-6675, email at co2clean-at-aol.com or on the www at www.co2clean.com.
There are many microscopy examples on the www site by using AFM or SEM. Please look at the site to look for similar applications.
If you need more information, just call or email.
--------------------------------- Check out the following web site. Richard Sherman is the patent holder, sells the guns, and can give you reprints of the articles. (Look in JVST circa 1992 for them.) His Email address is co2clean-at-aol.com
Jon, Freeze Fracture is the best answer considering the salt problem. The other alternative is cryo-TEM, best done with Leo/Zeiss energy filtering imaging. We neagtive stain liposomes just as we would any other sample for conventional TEM. What you see are little bubbles. We have also freeze-dried and shadowed or freeze dried and replicated. You may be able to get around the salt problem if you osmicate first. Just a guess. At 04:25 PM 5/4/1999 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
(a) told me about silver membrane filters, which I had not heard of before (b) reminded me of Gelman, who recommended their PTFE membranes.
As to which I use for which application, it's horses for courses!
And now the question: one has a scanned or digitally acquired micrograph. The background is a gently varying grey, with rather low contrast local features (blobs, bacteria, or whatever). The task is to Fourier transform, then filter out the lowest frequencies with a high-pass filter, leaving one with the features on a uniform grey background. One can then increase the contrast to make the features more prominent for printed reproduction. Is there software (Win 95) that can do this? (preferably inexpensive, though we could make arrangements to use plugins for Adobe Photoshop elsewhere, if necessary).
You can see an example of what I'm trying to do on:
http://www.reading.ac.uk/~spsolley/fourier.htm
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I only worry about filament life if I have to change a filament too frequently. Normal operation would have the filament saturated fully. = If the image quality is unimportant and analysis is the main focus then the current is reduced slightly. Image quality is more important than a =A3= 10 filament which only takes a few minutes to swap ( We keep a spare assembl= y ready ).
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Steve Chapman {PROTRAIN-at-CompuServe.COM} To: American {microscopy-at-sparc5.microscopy.com} is } that they get 100+ hours life (by their means of measurement) and the } images die at } 3,000X. The first thing we have to do is give them more } current and throw away the filament life. As I have said so many times } before "in some labs filament life is more important than image quality!= " } } When I am abroad the fuel consumption of my car is amazing, it sits in t= he } garage at home and does nothing, should I boast? } } Come on guys lets start putting realistic figures on filament life rathe= r } than hours in a day and lets relate this to the maximum magnification we } use and the lowest kV we use. Then and only then will we all have a bas= e } to work from! } } Try this equation - multiply the actual filament on time by the thousand } digits of the maximum magnification used and the emission current , then } divide by the average kV you use. My typical customer would have a 6.7K } value. } } I think that would be interesting, anyone with other ideas? } } Steve Chapman } } Senior Consultant E.M. } Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. } Tel & Fax 44 (0)1844 353161 } Web Site - http://ourworld.compuserve.com/homepages/protrain } }
} ... } } } While on the filament "thread" (oops, a pun slipped in), I am } wondering if } someone can make recommendations for suppliers of LaB6 } filaments for use in } a Hitachi H-7100FA (hi res, small probe). Hitachi makes a wonderful } assembly but it is getting pricey ...
I don't know about your SEM but we've had excellent performance and lifetimes using FEI cathodes, LaB6 and CeB6 ... see:
http://www.feibeamtech.com/lab6/lab6page.htm
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Is she using Thermanox coverslips to grow the cells on? If so, they exhibit bright autofluorescence. Don't know what the coating is. Ronnie Houston Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children Dallas, TX
Barry, Lilith wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } A colleague of mine works on primary cortical cultures. When she does } immunocytochemistry on them she usually uses indirect (immunofluorescence) } method, which works well. When she uses fluorescence labeled } avidin-biotin method and she gets extremely high background. I have looked } at the controls. The non specific binding is as bright as the specific one. } I have never worked with cultures. Could there be endogenous biotin in them? } Or would you have any other suggestions on what could be the cause and what } can one do to prevent this? } Thanking you in advance for your input, } Lilith } -------------------------------------------------------- } Lilith Ohannessian-Barry } National Research Council } Institute of Biological Sciences } CANADA } Tel;613-993-6460 } Fax;613-941-4475 } e-mail; lilith.barry-at-nrc.ca
I looked in some FIM books for polishing solutions. I found one for gold that might work on your alloy.
for gold: equal volumes of conc. HNO3 and conc HCl, 1-10Vac
Note 10%(5-10Vdc)HCl will work for Ni (I also found 40% solution at 1-2Vdc) so this might work for your Au-Ni alloy.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Fred Pearson To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hello Microscopists:
I am to prepare samples for TEM from Au 38at% Ni 100um. thick foil.
I will be using a Fischione polisher.
Does anyone have a polishing solution for this alloy?
Thanks in advance
Fred Pearson
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
There is an old analog trick of unsharp masking which might give you a similar result. You make an out-of-focus contact print of your original negative, then sandwich the two together in your enlarger to make your final print. This effectively subtracts the low frequency background from the original image and adjusts the contrast of fine details in high-contrast images. My reference was Kodak Techbits 1990, issue 1 (pub # P-#-90-1).
You can mimic this analog technique by using a very large blurring kernal on the original and subtracting the blurred image from the original image. If necessary, you can multiply the blurred image by a constant to adjust the final intensity. You can also define a custom kernel to get something large enough to do what you want. This should help even out the background in your image.
Henk
At 03:17 PM 5/5/99 +0100, you wrote: } } } And now the question: one has a scanned or digitally acquired micrograph. } The background is a gently varying grey, with rather low contrast local } features (blobs, bacteria, or whatever). The task is to Fourier } transform, then filter out the lowest frequencies with a high-pass filter, } leaving one with the features on a uniform grey background. One can then } increase the contrast to make the features more prominent for printed } reproduction. Is there software (Win 95) that can do this? (preferably } inexpensive, though we could make arrangements to use plugins for Adobe } Photoshop elsewhere, if necessary). } } You can see an example of what I'm trying to do on: } } http://www.reading.ac.uk/~spsolley/fourier.htm } } +------------------------------------------------------------------------+ } | Robert H.Olley Phone: | } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } | University of Reading {University internal extension 7867 | } | Whiteknights Fax +44 (0) 118 9750203 | } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } | England URL: http://www.reading.ac.uk/~spsolley | } +------------------------------------------------------------------------+ }
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu "Progress does not consist in replacing a wrong theory with a right one. It consists of replacing a wrong theory with one that is more subtly wrong."
Looking for an old JEOL TEM 2000FX. Please let me know if someone has a clue of a place who want to give away their old TEM for free to a University (we will pay the frieght). A JEOL 2000FX TEM even if it is not in working order would be o.k. as we just need it for parts as a back-up for other TEM (same model) that we have.
Please reply directly on my e-mail: zur-at-mmae.engr.ucf.edu
Thank you.
Zia ur Rahman Electron Microscope Engineer University of Central Florida, Orlando, Florida
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Reply to: RE: Tissue culture background Dear Barry, This sounds like a case of the wrong blocking agent. From what you = describe, my guess is your colleague treats the cells with dilute serum (= BSA or FCS) prior to addition of the fluorescent marker. It also looks as = if she does not dilute the fluorescent avidin in blocking agent either. The reason for the strange result is that serum contains biotin-like = molecules. When applied to the cells it will stick all over and the = fluorescent avidin will bind to it giving high background). Try the = labeling protocol without blocking agents and see if there is a difference.= Alternative strategies include substituting a non-serum blocker (such as = cold-water fish skin gelatin) for the serum, or even using antibodies to = biotin as a replacement for the avidin. Biotin-like molecules are also = present in mitochondria. If the cells have an unusually large number of = mitochondria in them, then this too could give the "high background" = described. Regards, Paul Webster
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Barry, Lilith wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
id {2LL9JGKA} ; Wed, 5 May 1999 13:39:29 -0500 Message-ID: {D8F9EBE8536ED111920F00005A422A315A474A-at-commserver.srrc.usda.gov}
Fred makes a good point about examining the tip end of an "expired" tungsten filament. Besides indicating possible vacuum problems, the size of the metal ball at the tip of the filament (usually on the shorter tip) can also indicate user error during filament saturation.
} ---------- } From: Fred Schamber (personal)[SMTP:fhscham-at-sgi.net] } Sent: Tuesday, May 04, 1999 10:27 PM } To: Listserver, Microscopy } Subject: Re: filament life } } } I do agree with several of the respondents that, though filament life } can be influenced by a number of factors, the most } common cause of an abruptly decreasing life is likely to be poor } vacuum. However, I do not recall anyone mentioning one } of the simplest ways to check for this condition: by inspecting the tip } of the expired filament under a low-power } microscope (or use your SEM if you like to indulge in overkill). } } The dynamics of filament construction are such that the } regions just to either side of the apex of the "V" tip } are naturally going to be a little thinner than the tip itself. Thus, } when a filament expires "normally" (i.e., solely by } evaporation) you will see that the break is on one of these legs -- the } wire adjacent to the break will be noticeably thinned } to a taper and there will likely be a small "ball" on either side of the } break where the molten tungsten solidified. (A pair of } large globs of tungsten with negligible thinning indicates that the } filament was blown by a high current surge -- just the way } a common fuse wire blows.) } This "high current surge" can often be found with SEM's which have multiple users. Especially with older and less expansive instruments, the tip failure shape will indicate oversaturation by less experienced users. This is less of a problem with those instruments with "lockout" features where the primary user can set the filament saturation limits on the SEM.
} Bad vacuum always produces a different kind of filament failure. This } is because gas molecules in the gun will be ionized } by collisions with the electron beam. The positive ions are accelerated } towards the cathode and are focused into the } filament tip. This impact sputters the tungsten tip and creates a } crater. A filament which has failed because of poor } vacuum, when inspected under a microscope, will look like someone took a } bite out of the very tip of the "V". There is } negligible thinning of the wire along the legs. } } These effects are very easy to recognize with a small amount of practice } and practice in doing so should really, at least in } my opinion, be part of the training for anyone who operates a tungsten } filament microscope (similar effects can also be } observed for LaB6 filaments). } If the instrument does not have multiple users or new users, I would check for vacuum leaks as a possible cause of repeated filament failure.
Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124-4305
Hello!=A0=A0 We are looking for some help in choosing and/or finding an objective for our Nikon Diaphot scope.=A0 We use the scope (setup for epifluorescence microscopy) to measure the response of neurons to mechanical deformation as a model for traumatic brain injury.=A0 We currently use either Fura-2 (calcium) or Di-8-ANNEPS (membrane potential) to measure the cellular response while the mechanical injury is applied to the cells by a device that is attached to the microscope.=A0 The trouble we're having is that the cells are attached to an elastic membrane and they need to be in saline during the experiment.=A0 Since the microscope is inverted (it's a long story), we think the best way to image the cells is with a water immersion objective.=A0 However, our current objective is a 40X oil immersion and we are using it in liquid (not the ideal situation).=A0 We have funds for a new objective and we'd like your input as to what would be the best objective given our current hardware (old Nikon with 160 mm tube length) and device (cells must be immersed in saline and there is no coverslip) limitations. Another requirement is a long working distance since we need to be able to change the medium between the objective and the cells during an experiment.
What we think we need is a high magnification (40X or greater) fluor water immersion objective with a long working distance on the order of millimeters. We know that newer objectives are made to meet these needs but they do not fit our old scope. Do you have any ideas or recommendations for objectives?
Thanks in advance,
Donna M. Geddes GT/Emory Department of Bioengineering Georgia Institute of Technology Atlanta Georgia, 30332
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I agree Steve.
I only worry about filament life if I have to change a filament too frequently. Normal operation would have the filament saturated fully. = If the image quality is unimportant and analysis is the main focus then the current is reduced slightly. Image quality is more important than a =A3= 10 filament which only takes a few minutes to swap ( We keep a spare assembl= y ready ).
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Steve Chapman {PROTRAIN-at-CompuServe.COM} To: American {microscopy-at-sparc5.microscopy.com} is } that they get 100+ hours life (by their means of measurement) and the } images die at } 3,000X. The first thing we have to do is give them more } current and throw away the filament life. As I have said so many times } before "in some labs filament life is more important than image quality!= " } } When I am abroad the fuel consumption of my car is amazing, it sits in t= he } garage at home and does nothing, should I boast? } } Come on guys lets start putting realistic figures on filament life rathe= r } than hours in a day and lets relate this to the maximum magnification we } use and the lowest kV we use. Then and only then will we all have a bas= e } to work from! } } Try this equation - multiply the actual filament on time by the thousand } digits of the maximum magnification used and the emission current , then } divide by the average kV you use. My typical customer would have a 6.7K } value. } } I think that would be interesting, anyone with other ideas? } } Steve Chapman } } Senior Consultant E.M. } Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. } Tel & Fax 44 (0)1844 353161 } Web Site - http://ourworld.compuserve.com/homepages/protrain } }
Looking for an old JEOL TEM 2000FX. Please let me know if someone has a clue of a place who want to give away their old TEM for free to a University (we will pay the frieght). A JEOL 2000FX TEM even if it is not in working order would be o.k. as we just need it for parts as a back-up for other TEM (same model) that we have.
Please reply directly on my e-mail: zur-at-mmae.engr.ucf.edu
Thank you.
Zia ur Rahman Electron Microscope Engineer University of Central Florida, Orlando, Florida
} We have a Sony DCR-TRV9 digital video camera recording. We would like to } be able to take the video stream, send it to a pc based computer, edit the } file, and grab individual frames and save them as separate files. Has } anyone out there got any suggestions? We'll also need a way to get the } video to the computer. We've tried frame grabbers but because there are } so many frames per seconds, it locks it up. The video is stored on an } Mini DV Digital Video Cassette. } Thanks in advance for any ideas. } } Robin Griffin } UAB } }
Here is the recipe for better performance, a 15 year old SEM is just a youngster!.
1. Use the wave form and the main peak (the one when heating the filament does not increase the signal level) do not undersaturate and ali= gn as accurately as you can
2. You do not say which SEM you have but if it has a bias control adjust the emission current to nearer 100uA. If you do not have a bias control half the distance between your filament and the front face of the=
cathode aperture. If the current is then 70 to 100 at saturation that is=
much better if not shorten the distance again.
3. Work at less than 15mm working distance.
4. Do not worry about how noisy the screen image is, concentrate on getting the best photograph. To do this reduce the spot size (screen goe= s dimmer) until the image when focussed and stigmated looks good on a mediu= m slow scan rate. With a noisy image focus and stigmate for maximum contrast, never stigmate on a directional image.
All this said simply increasing the emission current and shortening the W= D should be more than enough. If you let me know exactly which instrument you use I would be pleased to give specific instructions for that instrument.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
We also have a Hitachi 7100FA, running with a LaB6 . We bought it with two Hitachi cathodes. When these were used we put in a Denka that we had sitting in a drawer - just the straight (M3?) hunk of crystal on a tungsten loop design - and it has worked very well indeed. In spite of being a completely different shape to the Hitachi crystal - short and fat rather than long and thin. We dont use the machine much for really high res work, so I couldnt say if the ultimate performance is different, but we routinely adjust the alignment using a TV camera and screen giving an 8 million X mag, and havent noticed a difference, and the stability has been very good. We always use the auto filament run up and are very conservative - 10 min in the morning, 5 min subsequently, the filament goes off at every specimen and camera change. The Denka performance surprised us, because on an SEM we have had consisitently much better experiences with the Kimball Physics cathode.. .. We cleaned and readjusted the height at about 600 hours, which is similar to what we did with the Hitachis. I dont know if we just dropped lucky with this Denka or what . Is the slow run-up what makes the difference? - so I'm still uncertain what to get when it gives up the ghost! I'd also appreciate hearing what other people's experience has been with LaB6s on the 7100.
regards Sally Stowe
Dr Sally Stowe Facility Coordinator Australian National University EM Unit Research School of Biological Sciences Box 475, ACT 2601, Canberra, Australia FAX 06 (0)2 6279 8525
} } } John J. Bozzola {bozzola-at-siu.edu} 5/05/99 12:00 } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
While on the filament "thread" (oops, a pun slipped in), I am wondering if someone can make recommendations for suppliers of LaB6 filaments for use in a Hitachi H-7100FA (hi res, small probe). Hitachi makes a wonderful assembly but it is getting pricey ($3,500) and we would like to explore other possibilities. The purse keeps getting smaller .....
Thanks, John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
John J. Bozzola wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } While on the filament "thread" (oops, a pun slipped in), I am wondering if } someone can make recommendations for suppliers of LaB6 filaments for use in } a Hitachi H-7100FA (hi res, small probe). Hitachi makes a wonderful } assembly but it is getting pricey ($3,500) and we would like to explore } other possibilities. The purse keeps getting smaller ..... } } Thanks, } John } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ####################################################################
John, I have a couple of customers who have had good luck with the Kimbal Physics LaB6 tips. They run in the vicinity of $500, last I knew, and they last a long time. They also are not nearly so sensitive to thermal stress as the competing mounting systems, so failure tends to be due to evaporation of the Lab6, not mount failure.
Barry Scientific in Massachusetts may be a little closer to deal with than Jim (sorry Jim!). Kimball no longer retails, but they are in New Hampshire.
Ken Converse owner Quality Images third party SEM service Delta, PA
My company deals in Refurbished Lab Equipment, however we do not do a large business in EM and I admit that I am not well versed in this field. I do from time to time come across used EMs of different brands & styles that I broker.
I do have a few related items in stock, i.e.: a Leica (Reichert) Super Nova Ultra Microtome.
If anyone is interested, please contact me for details.
Ford M. Royer Analytical Instruments, Ltd. 9921 13th Ave. N. Minneapolis, MN 55441 phone: (800) 565-1895, ext. 17 FAX: (612) 929-1895 email: froyer-at-bitstream.net Web Site: www.aibltd.com
Shalvoy, Richard B CHES wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I'm looking for contact information for vendors of used equipment (ie my old } SEM). } } I'm sure they're out there but being pretty new to this area I'm not sure } where to find them. } } Richard Shalvoy } Arch Chemicals (formerly Olin Corporation) } Cheshire, CT
Are there any non volatile fixatives for animal and vegetable material for LM? I am an amateur with very serious asthma. Formaldehyde and formic acid both cause very serious reactions. I don't think it is formaldehyde but some of the less soluble stuff that is formed causing the problem. All my research shows that formaldehyde is too soluble to make it to the lungs. It reacts with the sinuses and throat before it gets to the lungs.
I can't really visualize something that isn't pretty volatile and reactive that would fix tissue. I see references to HgCl, picric acid, acetic acid. potassium permanganate and silver nitrate being used in fixing solutions. I am comfortable handling any of these and none of them cause me a big problem. But I only see them used in conjunction with an aldehyde. Can any one give me some alternatives to the ayldhydes?
While I had several hours of microbiology as an undergraduate the microscope was just one of the tools. With digital image capture and unexpected retirement it opens a whole new world to LM.
I find this list to be an outstanding resource. Nestor and the group do an outstanding job keeping on topic without being heavy handed. The group also seems to be spending a lot less of their employer's time on the net than in most groups. There will be no post during working hours and a flock of messages an evening. I guess that it being an industry mail list I would think twice before posting in working hours knowing that my next resume would probably go to some one on the list.
Thanks Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
A discussion I was engaged in today relative to my last-night's posting suggests to me that there is a bit more to be said regarding the role of emission relative to filament life. There seems to be a common assumption that emission current plays a role in filament life. This is not true.
The typical heating power applied to a tungsten filament in an electron microscope is on the order of 6 watts (product of heater current times heater voltage). Since the filament is operating in a vacuum, this power can be dissipated via three routes: (1) heat conducted through the posts of the filament; (2) photons (light) radiated by the incandescent wire; and (3) energy carried off by emitted electrons (emission current). How large is the latter effect? A WRONG way to estimate this is to take the product of the emission current and the beam voltage. For example, when operating at 30 keV with 100 microamps of emission (a higher emission than most modern scopes use), you would come up with a dissipated power of 3 watts, which is appreciable. This is in fact the power which the gun assembly as a whole imparts to the beam, but it is irrelevant to the question of power dissipation by the filament. This is because that 30 keV of kinetic energy is imparted AFTER the electron leaves the filament.
In fact, an electron is "boiled off" the heated filament with a very low energy. There is actually a distribution of energies, depending on the temperature, but the average is about 1 eV. If one uses 1 eV as the electron energy, then at 100 microamps of emission the dissipated power being carried off through the emitted electons is only 100 microwatts. This is obviously insignificant relative to the five watts of heating power being applied to the filament. Thus, we can conclude that emission plays no role in the filament heat balance and thus does not affect filament life in any practical sense. Furthermore, since the energy at which the electrons leave the filament is independent of accelerating voltage, we also see clearly that the beam voltage plays no direct role in filament life.
So where does the heat go? It has been some years since I did this calculation, but I remember the conclusion that I obtained for one geometry -- heat conducted through the posts is about half of the heat radiated as light. This will, of course, depend considerably on the geometry specific to an instrument, and since I ignored a lot of the complexities of reflected light, etc., my computation was only an approximation, but I think it is safe to say that the radiated and conducted heat components are comparable in magnitude.
I think the reason that so many people believe that emission current is related to filament life is because if they "undersaturate" the filament they observe both a lower emission current and a longer filament life. But both of these are consequences of the fact that they have actually reduced the filament temperature, which is the controlling factor on filament evaporation rate.
We have a well-used but perfectly functional Hitachi S-450 SEM available to anyone who may have a use for it as spares or a fully functional instrument. Cost highly negotiable, transport your problem.
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Tel +27 (0)331 260 5155 Fax +27 (0)331 260 5776 website:http:www.nu.ac.za (departments} units) Email:bruton-at-emu.unp.ac.za postal address; Private Bag X01, Scottsville, 3209 KwaZulu-Natal South Africa
Synoptics were selling something called `Massram' literally that - a mass of RAM (memory) to allow continuous video to be captured to memory without the problems of access time to any disc storage device. The problem is that storing continuous video really eats up the memory (250Kb? per frame x 25 or 30 frames/sec x time of video sequence). It will be too expensive to buy for a one off but there are some out there somewhere. From memory Bath University in the UK had a system and offered a service but I don't know the cost.
Try Synoptics in the UK for contacts +44 (0) 1223 727100 or fax +44 (0) 1223 727101.
Good luck, Ron
On Wed, 5 May 1999 15:07:33 -0500 "rgriffin-at-eng.uab.edu"-at-Sparc5.Microscopy.Com wrote:
} } We have a Sony DCR-TRV9 digital video camera recording. We would like to } } be able to take the video stream, send it to a pc based computer, edit the } } file, and grab individual frames and save them as separate files. Has } } anyone out there got any suggestions? We'll also need a way to get the } } video to the computer. We've tried frame grabbers but because there are } } so many frames per seconds, it locks it up. The video is stored on an } } Mini DV Digital Video Cassette. } } Thanks in advance for any ideas. } } } } Robin Griffin } } UAB } } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Does anybody out there know of a spare HT cable for a Siemens Elmiskop 102 TEM? I can't afford to buy a custom built new one, so my only real alternative is to try and track down a second hand one. Can anybody help?
I've used Denka, FEI and Kimball LaB6 emitters in my Jeol 2000. The Denka gave the best image quality but required to be run up very carefully. Out of 4 only one died of old age; the rest were destroyed by careless users, in spite of death threats from me. The FEI and Kimball emitters do not give quite such a good image, but they are really quite robust. I think the Kimball is slightly tougher than the FEI. I've caught people treating them worse than I would treat a tungsten filament, without any ill effects.
I have no financial interst in any of these suppliers.
Regards, Eric
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
I am looking for informations about nitrogen quantitative analyses by WDX system and specially in biological samples. I am interested also to know the sensitivities (minimum detection limits) and what is the result with biological sample (matrix effect such absorption with about 40% of c and 15% of O). Many thanks in advances.
Didier
-------------------------------------------- Didier Le Thiec I.N.R.A. Centre de Recherches Forestieres Unite d'Ecophysiologie Forestiere Laboratoire de Pollution Atmospherique 54280 Champenoux - France
Thank you for responding to my question on tissue culture background. I don't think the background is because the fixation or the immuno method because I routinely use the same method on brain sections and do not get the same background. It isn't the coverslip autofluorescing either because one can see beautiful labeled structures in the cells except on the controls. So it has to be something specific about cortical cultures and the strept or neutravidin-biotin method. Do you have any other thoughts? Lilith ------------------------------------------------------------------------ Lilith Ohannessian-Barry National Research Council Institute of Biological Sciences CANADA Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
Ron: As I recall, Texas Memory offers something similar. See http://www.texmemsys.com/ -Mike
Ron Doole wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Robin, } } Synoptics were selling something called `Massram' } literally that - a mass of RAM (memory) to allow continuous } video to be captured to memory without the problems of } access time to any disc storage device. The problem is that } storing continuous video really eats up the memory (250Kb? } per frame x 25 or 30 frames/sec x time of video sequence). } It will be too expensive to buy for a one off but there are } some out there somewhere. From memory Bath University in } the UK had a system and offered a service but I don't know } the cost. } } Try Synoptics in the UK for contacts +44 (0) 1223 727100 or } fax +44 (0) 1223 727101. } } Good luck, } Ron } } On Wed, 5 May 1999 15:07:33 -0500 } "rgriffin-at-eng.uab.edu"-at-Sparc5.Microscopy.Com wrote: } } } } We have a Sony DCR-TRV9 digital video camera recording. We would like to } } } be able to take the video stream, send it to a pc based computer, edit the } } } file, and grab individual frames and save them as separate files. Has } } } anyone out there got any suggestions? We'll also need a way to get the } } } video to the computer. We've tried frame grabbers but because there are } } } so many frames per seconds, it locks it up. The video is stored on an } } } Mini DV Digital Video Cassette. } } } Thanks in advance for any ideas. } } } } } } Robin Griffin } } } UAB } } } } } } } } } } ---------------------- } Mr. R.C. Doole } Department of Materials, } University of Oxford. } Parks Road, Oxford. OX1 3PH. UK. } Phone +44 (0) 1865 273701 } Fax +44 (0) 1865 283333 } ron.doole-at-materials.ox.ac.uk
Dear listers, Can anyone help me? I am trying to put a grant application together at very very short notice, and I need to find out the cost of a GOOD TEM. Not one that is the BMW of TEMs, more like an Alfa Romeo - if you know what I mean, and preferably one that has some sort of computerised image assistance with it. Thanks for your help in advance. Please email me as soon as humanly possible at
alex.black-at-nuigalway.ie
Thanks again.
Alex
__________________________________ Alexander Black Department of Anatomy National University of Ireland, Galway Republic of Ireland
via smtpd (for sparc5.microscopy.com [206.69.208.10]) with SMTP; 6 May 1999 20:19:17 UT Received: FROM toto.research.aa.wl.com BY akita.research.aa.wl.com ; Thu May 06 16:12:42 1999 Received: by toto.research.aa.wl.com with Internet Mail Service (5.5.2232.9) id {JNYX3544} ; Thu, 6 May 1999 16:12:41 -0400 Message-ID: {602B9620D104D2118E0A00805FBBC43602826DE0-at-redfox.research.aa.wl.com} {MICROSCOPY-at-Sparc5.Microscopy.Com}
I recommend you contact (if they don't contact you from monitoring this List) a few of the major EM suppliers, especially those from your area. An internet search for Philips, Hitachi, Zeiss, Jeol or any other brand (sorry to those I left out) should get you in contact with a willing sales representative who can work with your urgent time schedule. It will not be easy, since the features are many, and very specific to your lab research focus. Your general query to this list will only get you personal biases, and will likely mislead you more than help you.
Better yet, these companies can fax or express-mail you an official quote to include in your proposal.
Gregg Sobocinski Parke-Davis Pharmaceuticals Ann Arbor, Michigan USA Gregg.Sobocinski-at-wl.com
-----Original Message----- } From: ALEX BLACK [mailto:ALEXANDER.BLACK-at-NUIGALWAY.IE] Sent: Thursday, May 06, 1999 2:22 PM To: MICROSCOPY-at-Sparc5.Microscopy.Com
Dear listers, Can anyone help me? I am trying to put a grant application together at very very short notice, and I need to find out the cost of a GOOD TEM. Not one that is the BMW of TEMs, more like an Alfa Romeo - if you know what I mean, and preferably one that has some sort of computerised image assistance with it.
Thanks for your help in advance. Please email me as soon as humanly possible at
alex.black-at-nuigalway.ie
Thanks again.
Alex
__________________________________ Alexander Black Department of Anatomy National University of Ireland, Galway Republic of Ireland
I was a happy user of a plain and standard SEM Zeiss DSM 940, which gave = me nice shots. I work with systematics of sphecid wasps and used to photogra= ph uncoated specimens. I got my best pictures using BSE detection, although = our microscope has no detector for backscatering electrons. Now we have a bra= nd new machine, a LEO 440, with almost all acessories, including a BSE detector. However, I cannot acheive the same results that I got with the Zeiss SEM. As the LEO 440 is wholly automatized, its software do not allo= w me to make the same adjustments that I used in the Zeiss machine. I can u= se only SE detection, that gives me pictures of bugs with a methalic aspect, which is pretty but not usefull. So, let post you two questions. Why a go= t nice pictures in a SEM using a dection method without the proper detector= ? How I can get similar results in a fully coputer controled machine?
S=E9rvio T=FAlio Pires Amarante
serviopa-at-usp.br
Museu de Zoologia da Universidade de S=E3o Paulo Caixa Postal 42694-970 04299-970 S=E3o Paulo BRASIL
Steve Chapman responded to my recent posting with the suggestion that I am missing the point of filament "saturation" and included some further thoughts regarding his theory of how this relates to filament lifetime (quote appended). He concludes with an observation which suggests that "getting it right" in terms of understanding the mechanism is important to operting a SEM effectively. Consistent with that sentiment, I am going to launch into a fairly lengthy discussion of the phenomenon that we know as "saturation", since it is my experience that it is widely misunderstood. I'm also going to touch on some related subjects like the role of "brightness". But please be warned, this is NOT a brief posting. OK?
} I read with interest Fred's note but fear he has missed one important } feature of saturation? } } Saturation occurs when the aperture in the front of the cathode is filled } with electrons, generate more and they are unable to pass through therefore } no change in emission current. } } What affects this aperture? The physical size of the aperture and the bias } field. Apply more bias and the effective aperture size is reduced, } therefore the number of electrons needed to fill the aperture is reduced, } therefore the amount of heat required to produce the required number of } electrons for saturation is also reduced. Hence apply more bias you need } less heat you get a longer filament life as heat (oxidation and } evaporation) is the determining factor. } Steve's analysis is based on the perception that "saturation" is a fundamental electron-optical effect which limits the emission directly. For an electron microscope operated in the normal way, this is simply not the case.
The "kernel of truth" in Steve's argument is that there is indeed an effect of the kind he describes in what are referred to as "high perveance" guns. The effect is due to "space charge" effects encountered when there is a high density of electrons in front of the cathode. Since electrons repell each other, a sufficiently high density of electrons will effectively screen the cathode from the accelerating field, and indeed there will be the kind of effect he describes where you simply cannot add more electrons to the beam.
Unfortunately for his argument, that is not the regime in which electron microscopes operate by design. A microscope designer tries hard to stay away from this condition because the screening action defocuses the beam and increases the energy spread (the Boersch effect ) long before it appreciably limits the emission current. This effect becomes more important at low accelerating voltages since the electrons, being accelerated more slowly, can hang around in front of the filament tip longer. This is why you want to move the anode closer to the filament under low voltage imaging -- you want to increase the field gradient and get those electrons out of there as rapidly as possible. So it is in fact possible to operate an electron microscope under high-perveance conditions where there is a true space-charge "saturation" effect (set the filament way back, the anode at its furthest spacing, and crank the beam voltage way down). But this seriously degrades the imaging and, given any choice in the matter, you want to stay as far away from this operating condition as you can; this kind of aberrant situation is definitely not what people normally mean when they talk about a "saturated filament". So I will stand by my original assertion that the effect that we call "saturation" is simply a consequence of the common bias-resistor circuit, nothing more.
In the early days of electron microscopy, it was generally assumed that the "saturation" condition was something like what Steve describes -- you were "saturating" the phase space available for electrons. Haine soundly disproved this in a 1952 paper (in an earlier posting I erroneously referred to this as being published in 1938) [Haine & Einstein, Characteristics of the hot cathode electron microscope gun, British Journal of Applied Physics, Volume 3, p40, 1952]. On page 45 of this paper he states: "There has been some confusion in discussions on electron guns as to the effect of self-biasing (automatic biasing), the bias being obtained from the voltage drop across a resistance connected in series with the h.v. supply. With this type of biasing, an electron gun exhibits the property, frequently termed 'saturation,' whereby the current rises with temperature to a limiting value beyond which increase in temperature produces little increase in beam current. This has been attributed to the gun running into the space charge limited condition. That this is not so is readily seen when this type of bias is replaced by a variable independent bias; no such effect then takes place." He then goes on to prove his point with graphs. I personally have had extensive experience in working with an electron microscope gun equipped with a separate bias supply and can verify what he states -- there is no "saturation" effect without the bias resistor..
Let's look at the situation in detail (darn, I wish I had a blackboard!). Any electron optics reference will show you a figure of a self biased triode gun. The negative high voltage is connected directly to the wehnelt (grid) and thence to the filament (cathode) via the bias resistor. All current flowing from the filament must pass through the bias resistor, resulting in a voltage drop which makes the cathode more positive than the wehnelt. Thus, interposed between the cathode and the anode is the wehnelt which is at a somewhat more negative potential than the cathode, thus inhibiting the emission of the cathode's electrons. This creates a self regulating situation: any increase in emission current causes the bias to increase, thus choking off emission; a decrease in emission causes the bias to decrease, thus permitting greater emission. The effect is that a constant emission current is maintained, despite minor variations in filament temperature, cathode-tip geometry, and the like. This is a marvelous circuit -- both simple and highly effective with a near-instantaneous response. However, it is not unique to electron microscopy. Anyone old enough to have worked with vacuum tubes has seen it used for a constant current source and any idea about fancy optical effects is clearly not applicable there. To drive the point home: the effect which microscopists call "saturation" is nothing more than reaching the operating point in the emission current stabilization circuit established by the bias resistor -- there is no profound electron optical effect involved. Certainly it is not a matter of the electrons having "filled" a virtual "aperture" established by the bias field.
This may have some readers perplexed at this point. If the "knee" in the saturation curve is caused only by reaching the operating point in a simple emission stabilization circuit, why is this the best point to operate the gun? The most basic answer is: "because you would have to be an idiot to design a gun any other way"! Assuming you know the best conditions for imaging, why would you stabilize your emission current anywhere else?
Let's consider the way the wehnelt does its job of restricting cathode emission. The only reason that electrons can get past the negatively biased wehnelt at all is because the orifice in the wehnelt allows some of the anode's positive field to penetrate to the tip of the filament (the "zero equipotential"). When properly adjusted, there is a small circular region at the tip where electrons can escape to the anode -- everywhere else on the filament, the low energy electrons which are boiled off of it are repelled back into the filament. As the relative negative bias on the wehnelt increases, the anode field doesn't penetrate as far and the size of the emitting region on the filament tip shrinks so that fewer electrons can escape. As the bias decreases, the anode field penetrates further and the size of the emitting region increases, resulting in more emission. The objective of gun biasing is thus to select an emission region right at the tip of the filament. This is common knowledge covered in any optics text.
If you are controlling the bias with an independent voltage supply, you can completely cut off emission by applying a very high bias, or you can create a very high emission (up to the current limit of your supply) by setting the bias at a very low value. What you actually want to do, of course, is set the bias so that just the tip of the filament is able to emit. In this "independent bias" configuration, changing the filament temperature has no effect on the bias, and thus no effect on the position of the emitting region. Increasing the filament temperature causes the emission to increase without limit (and there is no "saturation" rolloff). In this kind of arrangement, it is really easy to understand the relationship of filament life and emission since both are direct consequences of filament temperature.
When you introduce the "self-biasing" resistor circuit, it gets a little more confusing since the effects are coupled. For one thing, you can't set the gun into a "cutoff" condition, because that would require an infinitely large bias resistance. For another thing, the relationship between filament temperature and emission current is different above and below the "saturation" knee. But the fact remains that you do establish a particular bias voltage value via this circuit and for the same bias value, you will get exactly the same emission as with "independent biasing". However, now in order to change the bias you either need to change the bias resistor (R) or to change the filament heating current so as to change the emission current (I) so as to effect the desired I*R drop (bias). But, if you have any sense at all, you select the bias resistor value such that the "saturation" knee occurs at a filament temperature which gives both a nice compact emission pattern AND an acceptable filament lfe. But this is a product of good design practice, not any kind of optical effect. Fortunately, this target is rather easy to hit.
As Steve has pointed out, the effect on the emission pattern of changing the bias can be readily observed on a TEM. This can also be seen on an SEM which is equipped with scanning coils in the gun. We provide this kind of "source imaging" feature on our Personal SEM(TM) and we train our users to use it to adjust the filament drive so as to achieve the most compact source emission pattern. With this tool, it is easy to demonstrate that if you mis-adjust the bias resistor, you can observe an emission "saturation" effect (emission current stops increasing as the filament temperature is increased) when the emission pattern is actually far from optimal. Typically, this occurs when the bias resistor is set too small and thus one has selected too large a region of the filament tip for emission.
So I hope that I have now clearly made my point that "saturating the gun" is just a technique we use to achieve an otherwise desireable result -- putting the microscope into a stable current configuration which we have set up to also provide the best imaging conditions. The notion of "saturation" per se is really without meaning for electron microscope optics. It's rather unfortunate that this term entered our vocabulary since it is quite misleading (maybe we should call it "stabilization"). But that is the term we are stuck with, it seems (kind of like my other pet peeve: Energy "Dispersive" Spectroscopy). =================
Steve's comments about the role of filament depth, kilovoltage, and anode spacing are generally sound advice, but when he speaks about "efficiency", he starts to blur the issue. As already noted, one wants to get the electrons away from the cathode as rapidly as possible so that there is a minimal space-charge effect. This is accomplished by maintaining a high field gradient in the gun and this can be accomplished by upping the acclerating voltage, moving the anode closer to the cathode, or moving the cathode closer to the wehnelt (you could also insert a separate "extraction" electrode in the gun as has been done on occassion [Yamazaki et al, 1984] and is effectively what is done in field emission guns). Note, however, that one's objective in doing this is not to generate more emission as such, but rather to improve the quality of the beam by overcoming the space charge "Boersch effect". In other words, you don't necessarily need more electrons, but more electrons going in the right direction.
When Steve speaks about the "efficiency of the gun", the implication is that the goal is to produce lots of electrons (emission). I doubt that this was his actual intent, since that would badly miss the point. When we speak of the "efficiency" of a gun, we are properly speaking about the ratio of "brightness" to total emission. The differentiation between emission and brightness seems to confuse a lot of people. I like to illustrate the difference by posing the following problem: suppose I take two people and offer a prize to the individual who can use a water hose to spray the maximum amount of water through a knothole 100 feet away. I then tell each contestant that they have a choice of two hoses, one with a discharge rate of 10 gallons per minute and the other with a discharge rate of 1 gallon per minute. The "foolish" contestant immediately opts for the higher flow rate, and quickly reliazes his error when I then hand him a hose which has a nozzle producing a fan-shaped stream -- although the flow rate is high, only a small fraction of it can be directed through the knothole. The more thoughtful contestant asks to see the shapes of the streams produced by the two hoses and chooses the lower-flow hose which happens to have a narrowly collimated stream -- and wins the contest. The point, of course, is that it is not how many electrons the gun produces, but rather how many can be directed through the electron optical system. We call this quality "brightness" -- it is the combined spatial and angular density of the beam. An ideally "bright" beam would be a non-divergent "pencil" of electrons which is very dense (the water hose analogy breaks down here since water isn't very compressible). Brightness is the quality which we should worry about (rather than emission current) because it dictates the quality of the image spot which we can form on the sample.
There is a well-known theory due to Langmuir in which he establishes the maximum brightness value for a thermionic (e.g., tungsten or LaB6) cathode. The variables are the emissivity of the cathode material, the accelerating voltage, and the temperature of the filament. So how does one design a gun to achieve this maximum brightness? Haine answered this question in that same paper referenced above. The answer is that over quite a wide range, you can achieve the Langmuir limit with a variety of gun designs. In particular, you can vary the size of the wehnelt opening and the spacing of the filament behind the wehnelt over wide ranges and still approach the Langmuir limit so long as you adjust the biasing appropriately. What does change, however, is the "efficiency" of the gun. By efficiency, he means the ratio of brightness to total emission. Returning to the "water through a knothole" example, you can imagine that the shapes of the two hose streams are such that either hose can deliver the absolutely maximum volume of water through the distant knothole at the operating pressure of the supply line (like achieving the Langmuir limit for a given beam energy with two different gun designs). But in this case, the lower volume hose does the job with much higher efficiency, and normally would be the preferred solution (unless we enjoy wasting water).
If the wehnelt opening is large and/or the filament set well back, then you will be able to achieve the Langmuir limit only by operating with a rather high emission current. Since there is really no advantage to having all of that wasted emission current (and some practical disadvantages) it is preferable to design the gun for high efficiency. SEMs today are generally designed for more efficient cathode operation than was the case in the past. By way of comparison, a 1970's vintage ETEC operated with a 200 microamp emission current, whereas most modern SEMs operate at 50 microamps or less. The highest efficiency is obtained by placing the filament tip close behind a small wehnelt aperture. But the closer the spacing, the higher the required bias and this can lead to unstable operation if pressed too far. A very small aperture makes precise alignment of the gun particularly critical. Thus, there are tradeoffs and the gun designer will try to weigh a whole lot of factors. The end user doesn't need to worry about these things. If the design and setup is proper, setting the filament to "saturation" will approximate the Langmuir brightness limit. =====================
I do have to take strong exception to Steve's statement that: "if you do not have enough [electrons] to start with you soon run out, hence poor signal levels and low magnification microscopy!" I'm sure that this isn't precisely what Steve meant to say, but I'm going to pick on the statement anyway, since a lot of people seem to think it is true. The important question isn't how MANY electrons you have to start with, but rather, how they are DISTRIBUTED in the beam. Lose sight of that fact and you can end up drawing some rather odd conclusions. For example, would you conclude that you are going to get better imaging with an old Etec with its 200 microamp emission current, or a modern cold field emitter where the emission is orders of magnitude smaller? Clearly, emission QUALITY is vastly more important than emission QUANTITY. I make this point emphatically because so many people seem to feel that anything they do to their microscope which increases the emission will somehow increase the image quality. Sometimes it is just the opposite. To make an extreme example, if you want tons of emission current, just drill out the hole in the end of your wehnelt to a half inch diameter -- LOTS of emission, but TERRIBLE imaging. =====================
This has gotten a bit off topic from the simple question of "filament life", and I hope that anyone who has stayed with me till here can pardon me for such a lengthy reply (but I did warn you). I do hope this lengthy diatribe helps to illustrate that though filament spacing and the like do have a relationship to gun performance and ultimately to filament life, these are actually secondary effects which represent design and/or operational decisions rather than fundamental optical considerations.
} } } I read with interest Fred's note but fear he has missed one important } feature of saturation? } } Saturation occurs when the aperture in the front of the cathode is filled } with electrons, generate more and they are unable to pass through therefore } no change in emission current. } } What affects this aperture? The physical size of the aperture and the bias } field. Apply more bias and the effective aperture size is reduced, } therefore the number of electrons needed to fill the aperture is reduced, } therefore the amount of heat required to produce the required number of } electrons for saturation is also reduced. Hence apply more bias you need } less heat you get a longer filament life as heat (oxidation and } evaporation) is the determining factor. } } With regard to the position of the filament the further it is away from the } cathode aperture the greater the affect of the bias field. Hence a long } filament to aperture distance is effectively increasing the bias field } requiring less electrons for saturation therefore less heat and a longer } filament life. } } The best instrument to use to see this in action is the TEM where we may } visualise the virtual source and may see the effects of bias on saturation. } } When we lower the kV the gun becomes less efficient due to the anode to } cathode distance being too great for the applied voltage and the resulting } field being less efficient in helping to create the virtual source. The } source is also affected by the higher residual bias field in most SEM which } throttles the cathode aperture still further. If you lower the kV and want } maximum efficiency you should move the filament forward to overcome the } bias field and raise the anode to increase the efficiency of the } anode-cathode field. } } Hope this helps, the truth is that even with field emission if you do not } start with a suitable emission current the potential for good quality } images is drastically reduced. Remember everything we do once the beam has } left the electron gun throws electrons away, if you do not have enough to } start with you soon run out, hence poor signal levels and low magnification } microscopy! } } The SEM is an exciting SCIENTIFIC INSTRUMENT, understand it and it becomes } an amazing tool, mis understand and it becomes a super light microscope; } what a waste? } } Steve Chapman } } Senior Consultant E.M. } Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. } Tel & Fax 44 (0)1844 353161 } Web Site - http://ourworld.compuserve.com/homepages/protrain } For Consultancy and Courses in Electron Microscopy World Wide }
The problem that you outline is very common when people change instrument= s. Zeiss using a SE detector gives satisfactory BSE images, LEO using a BSE=
detector you do not like the image as much! =
Using the so called SE detector for BSE collection you may achieve a good=
signal but only with a large surface area detector. This signal also contains a shadow effect due to the specimen-detector geometry. Trying t= o use the LEO in the same way as the Zeiss will not work as the LEO SE detector has too small a surface area for this style of operation.
The LEO BSE detector being directly above the specimen will be free from shadows, unless you set it up in the TOPO mode, or move the BSE detector off axis sufficient to clear the beam path. Detector variations will be available to you in the DETECTOR menu under SELECT SIGNAL, either do not switch all the segments on or switch to the TOPO mode. Try operating at different WD to optimise the LEO specimen-detector geometry?
Another problem, as you work at very low magnifications, may be you do no= t have enough signal, out up the probe current? Also the bigger the spot t= he better under low mag BSE circumstances. Do not tell anyone I said this b= ut you may find you get a better image by running at the false peak often called the first peak of saturation. This gives you a bigger source and hence a larger final probe size.
Good luck
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Regarding improving the performance of your Amray SEM the same rules appl= y as I passed on to Paula. (Sorry I do not have specific instructions for a=
1600 only an 1800 and I do not know how they compare?)
1. You need current - through higher emission levels
2. You need to saturate and align correctly to aid the above
3. You need to run in a minimum aberration condition - short working=
distance
4. Plus the following which are Basic SEM Adjustment Procedures
Image is Too Noisy?
1 Spot size is too small? 2 Emission current is too low? Variable aperture is too small? 4 Specimen position is not ideal - too far from the detector or tilted away from the =
detector? 5 High voltage is too low?
Resolution is Poor?
1 Spot size too large? 2 Working distance too long? 3 Final Aperture too large? 4 Emission current too low or saturation and alignment poor? 5 High voltage too low? 6 Image does not contain high resolution information? 7 Too much backscatter in the image (kV too high or specimen tilted=
towards the =
detector)?
Presentation of the Image is Poor?
1 Spot size incorrect, too small or too large? 2 Would tilt help present the information? 3 Have you the ideal WD? 4 Have you the correct kV for the job? 5 Have you the correct aperture (small) to obtain a good depth of field, do you need to =
move the specimen further away from the final lens? 6 Are you seeing a mixed SE/BSE image when you need a BSE or SE image. Lower =
the kV for the latter raise the kV for the former.
Hope this helps?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
----------------------------------------------------------------- PHILADELPHIA SOCIETY FOR MICROSCOPY ----------------------------------------------------------------- AN AFFILIATE OF THE MICROSCOPY SOCIETY OF AMERICA and THE MICROBEAM ANALYSIS SOCIETY -----------------------------------------------------------------
MEETING NOTICE - SPECIAL EVENT: Thursday, May 13, 1999
METALLOGRAPHY AND THE STUDY OF THE JAPANESE SWORD
Michael R.Notis, Lehigh University, Bethlehem, PA 18015
To be presented at The Egyptian Gallery, University of Pennsylvania Museum of Archaeology and Anthropology Reception at 5:30 followed by Dinner at 6:30 and the talk at 7:30
This meeting is intended to be of interest to both microscopists and a more general audience. We encourage you to bring guests and spouses. Cosponsor: the Museum Applied Science Center for Archaeology (MASCA), University of Pennsylvania Museum of Archaeology and Anthropology. ----------------------------------------------------------------- Meeting Details
Location: The Egyptian Gallery, University of Pennsylvania Museum of Archaeology and Anthropology (map and directions enclosed). Parking is available behind the LRSM after 5:00 PM and in the parking lots indicated on the attached map.Cost of Dinner: Members and members spouses $20.00, Students $15,Non-Members $30. Schedule: 5:30 Social hour hosted by our meeting sponsor. Beer, wine, soda and munchies 6:30 Dinner Salad- Watercress, Belgian endive, apples, alfalfa sprouts and raspberry vinaigrette. Entree- Seared breast of chicken served with porcini and shiitake mushrooms, onions, scallions, ginger and garlic topped with a black bean sauce. Vegetarian entrees available upon request. Dessert- Tiramisu 7:30 TALK ----------------------------------------------------------------- Reservations
By E-Mail (preferred): Send your name and affiliation to PSM-RESERVATIONS-at-INAME.COM By Phone: Call Dr. John Reffner, 215-619-5283 I will reply to confirm all email reservations received. DEADLINE for RESERVATIONS is Noon, Monday May 10th. ----------------------------------------------------------------- Abstract
The ancient Japanese sword is a marvel of empirical technology and has become a well known example of the beauty of metals as an art form. The manufacturing process (religious ritual) through which the sword is made, the heat treatment it receives and the microstructure that is developed will all be described. By the late 19th and into the early 20th century, the first Japanese metallurgists were studying modern advances in metallographic techniques. Among them were K.Tawara and his student H.Tanimura. This connection between sword study and metallography led to interactions between Tanimura and C.S.Smith, whose own metallographic studies of ancient metal objects, including the Japanese sword, are well known. A group or important swords originally studied by Cyril Stanley Smith using light optical microscopy have been reexamined using scanning electron microscopy and energy dispersive x-ray spectroscopy in order to better examine the microstructure and to perform microchemical analysis on the slag inclusions. The use of these newer methods for analysis demonstrates the power of microchemical analysis in combination with metallographic examination to extend our knowledge concerning these ancient swords and their fabrication. A VIDEO OF THE SWORD MAKING PROCESS WILL BE SHOWN
1. Take I-76 (Schuylkill Expressway) to the South Street; exit (left lane exit). From the west, it's the exit after 30th Street; from the east, the exit after University Avenue. Turn west (right from the west; left from the east) at the light at the top of the exit ramp. The Museum is on theleft past the next light. 2. From the PA Turnpike: Valley Forge Exit to I-76 (Schuylkill Expressway, east). Then follow directions in #1. 3. From the New Jersey Turnpike: Take Exit 3 to Walt Whitman Bridge. Once across, follow signs to I-76 (Schuylkill Expressway, west). Then follow directions in #1. 4. From I-95 North: Exit at Vine Street. Take Vine Street west to I-76. Stay in left lane. Take exit marked "To Airport" (a left U-turn). Take second exit (South Street). Then follow directions in #1. 5. From I-95 South: Just past Philadelphia Airport, follow signs toI-76. Once on I-76 follow directions in #1.
Parking lots (pay) are available behind the Museum, one block over the bridge off Convention Blvd (turn left). The Museum is on South Street, with its main entrance at the corner of 33rd and Spruce Streets. In addition, parking will be available at the usual location, and free parking for about 15 cars is available at the Sharpe Circle. Information on the Egyptian Gallery is available at
DENTON VACUUM- Three decades of manufacturing vacuum equipment for the preparation of specimens for electron microscopy.
PHILLIPS ELECTRON OPTICS, Phillips Electron Optics, part of FEI Company,is celebrating the 50th anniversary of the first commercial EM-100 TEM in 1949. We are setting the pace in electron microscopy for the new century with our latest TECNAI family of TEM's along with the XL Series SEM's and focused ion beam (FIB) systems.
EXCEL TECHNOLOGIES, INC. is a premier distributor of supplies andinstruments for industrial applications. Products include: EXTEC metallographic equipment and supplies, video imaging microscopes and products, a complete line of optical microscope products and micro-hardness testers. EXCEL maintains an extensive and computerized inventory that enables us to process and ship most orders within 24 hours. We strive to satisfy our customers with fast, efficient service as well as excellent technical assistance. With these qualities in mind, we would greatly appreciate the opportunity to fulfill your requirements. Please contact John Long at (610) 688-4440 and visit our website at www.extec.com
The Microscopy Society of America is also providing financial support for this meeting.
} You can mimic this analog technique by using a very large blurring kernal } on the original and subtracting the blurred image from the original image. } If necessary, you can multiply the blurred image by a constant to adjust } the final intensity. You can also define a custom kernel to get something } large enough to do what you want. This should help even out the background } in your image. } } Henk } } At 03:17 PM 5/5/99 +0100, you wrote: } } } } } } And now the question: one has a scanned or digitally acquired micrograph. } } The background is a gently varying grey, with rather low contrast local } } features (blobs, bacteria, or whatever). The task is to Fourier } } transform, then filter out the lowest frequencies with a high-pass filter, } } leaving one with the features on a uniform grey background. One can then } } increase the contrast to make the features more prominent for printed } } reproduction. Is there software (Win 95) that can do this? (preferably } } inexpensive, though we could make arrangements to use plugins for Adobe } } Photoshop elsewhere, if necessary).
You could look for the key-words 'median filter', 'blurring', 'non-linear filter'. NIH-Image might well have such filters. If not there are other free programs, such as Osiris: http://www.expasy.ch/www/UIN/html1/projects/osiris/osiris.html Image Tool: http://ddsdx.uthscsa.edu/dig/itdesc.html
You can find more software at the shareware-site: http://www.nsctoronto.com/nissei-sangyo/ftpsw.html
To use the median-filter do as Hendrik Colijn describes above. If the filter kernel is twice as big as the largest object in your image the filtered image will just show the smooth background. Subtract the filtered image (maybe scaled) from the original to get the objets on a flat background.
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
There is one ethanol/ethylenglycol-based fixative on the market=20 (Kryofix/Merck No. 5211) which was developped and used as a substitut= e of =20 formaldehyde ("formalin") for routine preparation in a pathology/hist= ology lab=20 of M.E. Boon in Leyden (Belgium). Best results were obtained in=20 combination with microwave irradiation and in immunostaining. See= =20 references in the "Microwave cookbook of pathology. the art of micros= copic=20 visualization" from M.E. Boon and L.P. Kok, Coulomb Press Leyden. 3rd= =20 edition 1992. Best regards Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=84tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
Please unsubscribe the following id: srinivasan-at-alpha.caer.uky.edu Sincerely Ram Srinivasan Center for Applied Energy Research 2540 Research Park Drive Lexington, KY 40511 (606) 257 - 9695 (Off) (606) 223 - 7378 (Home)
There are several solutions out there for capture of video from DV format cameras. They include the following:
Bravado DV2000 DPS Spark Miro DV300 Fast DV Master Canopus DVRex-M1 Radius EditDV
All of these are video capture cards that usually include some video editing software designed to work with the card. The last suggestion from Radius has included within the package a program called MotoDV for video capture and I believe it will capture still frames. A lot depends upon how much you want to spend, what computer platform you are using, and the type of hard drive it writes to. For a 2:1 compression ratio (good quality video) you need a drive with a transfer rate of at least 10 Megabytes/sec. Your frame size will be about 333 kilobytes and you will need 1.67 Gigabytes of space for each minute of video. Not sure what your application requires for the still frames but there quality (such as for printing) may not be great.
http://www.videoguys.com/jump.htm
http://www.videotexsystems.com/
Try these WEB sites for more information. Hope this is helpful to you.
Regards
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
Do beware that the typical IDE HDD with an advertized data transfer rate of 33 Meg/sec cannot usually sustain that rate. For example, My WD 8G (UDMA) drive benchmarks in the area of 4 Meg/sec sustained. Depends on the drive, interface type, etc.
There are several solutions out there for capture of video from DV format cameras. They include the following:
Bravado DV2000 DPS Spark Miro DV300 Fast DV Master Canopus DVRex-M1 Radius EditDV
All of these are video capture cards that usually include some video editing software designed to work with the card. The last suggestion from Radius has included within the package a program called MotoDV for video capture and I believe it will capture still frames. A lot depends upon how much you want to spend, what computer platform you are using, and the type of hard drive it writes to. For a 2:1 compression ratio (good quality video) you need a drive with a transfer rate of at least 10 Megabytes/sec. Your frame size will be about 333 kilobytes and you will need 1.67 Gigabytes of space for each minute of video. Not sure what your application requires for the still frames but there quality (such as for printing) may not be great.
http://www.videoguys.com/jump.htm
http://www.videotexsystems.com/
Try these WEB sites for more information. Hope this is helpful to you.
Regards
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
[frequent skips; } delineates Fred's post, and } } delineates Steve's]
} Consistent with that sentiment, I am } going to launch into a fairly lengthy discussion of the phenomenon that } we know as "saturation", since it is my experience that it is widely } misunderstood.
} } Saturation occurs when the aperture in the front of the cathode is filled } } with electrons, generate more and they are unable to pass through } } therefore no change in emission current. } } } So I will stand by my original assertion that the } effect that we call "saturation" is simply a consequence of the common } bias-resistor circuit, nothing more. } } Haine in a 1952 paper on page 45 states: "...With this type of biasing, } an electron gun exhibits the property, frequently termed 'saturation,' } whereby the current rises with temperature to a limiting value beyond } which increase in temperature produces little increase in beam current." } } To drive the point home: the effect which } microscopists call "saturation" is nothing more than reaching the } operating point in the emission current stabilization circuit } established by the bias resistor -- there is no profound electron } optical effect involved. Certainly it is not a matter of the electrons } having "filled" a virtual "aperture" established by the bias field. } } What you actually want to do, of } course, is set the bias so that just the tip of the filament is able to } emit. In this "independent bias" configuration, changing the filament } temperature has no effect on the bias, and thus no effect on the } position of the emitting region.
} For another thing, the relationship } between filament temperature and emission current is different above and } below the "saturation" knee.
} But, if you have any sense at all, you select the bias } resistor value such that the "saturation" knee occurs at a filament } temperature which gives both a nice compact emission pattern AND an } acceptable filament lfe.
} As Steve has pointed out, the effect on the emission pattern of changing } the bias can be readily observed on a TEM. } } So I hope that I have now clearly made my point that "saturating the } gun" is just a technique we use to achieve an otherwise desireable } result -- putting the microscope into a stable current configuration } which we have set up to also provide the best imaging conditions. The } notion of "saturation" per se is really without meaning for electron } microscope optics.
Dear Fred, I realize that the situation is considerably different for the TEM from that for the SEM; however, what I have always heard and observed is that for low filament currents electron emission occurs only from some regions of the filament tip. This can be seen by imaging the filament at crossover, where it appears as an annulus. As the filament current is raised, additional regions of the tip begin to emit electrons, and the image of the filament becomes more uniform and smaller. As the filament current reaches a value such that all regions of the tip emit, the image has no dark regions. This is what I have called "saturation"; it makes sense from the standpoint that emission cannot occur from more than the complete tip area and, therefore, cannot increase with further increases in filament current. AFAIK, this has nothing to do with space charge, current stabilization, virtual apertures, or bias resistors, but with the temperature, geometry and work function of the tip. Are there two uses of "saturation"? Yours, Bill Tivol
First I would like to thank those who responeded to my earlier inquiry about the Lynx tissue processor. I have since received info on the RMC EMP 5160 tissue processor and would like any info anyone can give me on this processor, be it good or bad.
Instead of writing directly to disk, you can always write to a RAM buffer first. For (uncompressed) video you have 30fps and each frame is about 1/3 MB (640x480), so that's only 10MB/s. You can buy RAM boxes with multiple-GByte capacity -- Texas memory has such a box with 12GB of RAM and a 1.2GB/s bandwidth that would buffer about 20 minutes worth of (uncompressed) video -- you could then "drain" this memory to your IDE HDD at 4MB/s. At the high end, you could buy a box with 128GB to hold over 3 hours of video!
See: http://www.texmemsys.com/samsys.htm
-Mike O'Keefe
"White, Woody N" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Do beware that the typical IDE HDD with an advertized data transfer rate of } 33 } Meg/sec cannot usually sustain that rate. For example, My WD 8G (UDMA) } drive } benchmarks in the area of 4 Meg/sec sustained. Depends on the drive, } interface } type, etc. } } Woody White
I have been working for a long time in the field of inorganic materials. Now I am getting into biomaterials and my first task is to determine porosity of a bone, I think I need some help. I guess that if I record images of the surface of the bone and apply sequential sectioning and image recording, I will be able to reconstruct the 3d characteristics of the object. I wonder if there is any software available to do the reconstruction or maybe someone can suggest some references where I could get started.
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I have been off-line for some time and may have missed relevant postings.
I have been asked to enquire about collecting digitised TEM and STEM images from the above microscope in order to improve the facility. The instrument dates from 1981. We are a little behind the times in this area so any information would be appreciated. Maybe pointers to the archives? Clues as to when this may have already been discussed !
Thanks in advance
Keith Ryan Marine Biological Association of the UK Citadel Hill Plymouth Devon PL9 9DR England
Firstly I have to say that when talking about improving performance I tak= e it for granted that people clean the column and gun components. My objec= t is always to move on from this point.
Now when we use a SEM at low magnifications ( {3,000X) the size of the pro= be encourages BSE to totally dominate the image particularly when using in excess of 10kV. This means that the SE detector collects a signal made u= p of direct SE, plus SE which are converted from BSE that hit the lens etc,=
plus line of site BSE (well documented in the literature).
When we view an image which contains high levels of SE it tends to be rather flat and devoid of high contrasts, the best sign of a BSE contribution is the formation of shadows. Lowering the kV we obtain an image that displays more of its SE content and less of the BSE content so=
this is the best way to analise the imaged information. =
To fully understand an unknown specimen use this procedure
1. Start at low kV ( {2kV) when you see the true specimen surface 2. Increase the kV until you see an change in the image form, now yo= u have increased the reaction volume to such an extent that the BSE start t= o dominate and you are now bringing in sub surface detail. 3. The higher the kV you use the greater the sub surface (BSE) contribution. 4. If as you go up in kV the image becomes more exciting (more detail?) this means that there is more sub surface data coming out by way=
of BSE. 5. If you go up in kV and the image becomes flat this means that the=
sub surface zones do not have much detail and the result will is a softening of the image.
(The ideal test specimen to demonstrate the above in action is a TEM specimen. I use a SPI carbon grating replica which I tack to a stub with=
the carbon side up. Take a look at different kV and blow your mind as yo= u see so much information goes missing when the kV goes up. Sure it is a very thin specimen but it is a great demonstration.)
It is for the above reasons that I frown upon those who insist on using 20kV plus for imaging, because most of the time their information is not = of the surface but of the sub surface. This is fine if you know what is happening, but most people do not understand the specimen-beam reactions and the way they relate to specimen-detector geometry.
Hope this helps?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Whilst I think people should change their scintillator every few years (y= ou clean your glasses don't you?) I have to say that, when experimenting wit= h signal and different types of scintillator material many years ago, the results did not push me too hard in this direction.
We found that a new scintillator gave a reading which very quickly fell a= nd then took an age (years) to drop to a level that one would consider was unacceptable.
Another area where we have had fun is that of photomultipliers, when the BSE signal from its detector is far stronger than the Everhart-Thornley detector its about time you changed it!
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain
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Bill Tivol makes a good point when he asks whether there are two different meanings attached to the term "saturation" (see appended quote at end).
As Bill correctly observes, there is a real physical effect where the beam emission pattern can be observed to transition from a diffuse annulus (what we used to call a "smoke ring") into a nice compact disk. This can easily be observed on a TEM (or so I'm told -- I'm not a TEM guy and have only seen this illustrated in publications). But a phenomenon which looks very much the same can be observed on a SEM equipped with gun scanning coils -- I happen to like this feature a lot and have designed it into our SEM since I find it the most reliable way of optimizing the gun. (By the way, I am unaware of any practical differences between SEM guns and TEM guns other than is related to the differences required by the TEM's higher operating voltage -- the basic optical principles are the same.)
When you do look at the emission pattern in this way, as Bill has noted, you can clearly see what it is you are trying to achieve. When "undersaturated", the beam profile is "empty" in the center. If you look at a "line scan" through the pattern, the profile looks a bit like the letter "M" -- this is clearly not what you want. As the biasing is increased (accomplished in the self-biased gun by either increasing the filament temperature or by increasing the bias resistor) the pattern collapses into a disk. In cross section, this distribution is high in the center and falls off steeply to the edges -- looking very much like a gaussian distribution (though I have no proof that is actually its mathematical shape). This is the condition you are trying to achieve, and calling this "saturation" makes a lot of sense. So Bill is perfectly correct in noting that there is a second meaning to the term "saturation" which has real validity for tuning of the gun.
My problem with the term "saturation" is that it has been used to refer to at least three different phenomena: 1. The idea of saturating the emission "phase space" with electrons such that no more can be crammed into the beam (the "high perveance" condition I referred to yesterday) -- as near as I can figure out, this is what people thought was happening when they originally used the term "saturation" in electron microscopy since it does occur with other types of electron guns. 2. The coalescing of the emission into a compact pattern as the emission becomes restricted to the tip of the filament. 3. The "knee" in the emission curve in an auto-biased gun -- the point where measured emission current stops increasing as filament temperature is increased
The first effect just doesn't apply to an electron microscope, and the third effect is simply the manifestation of the auto-bias circuit (these are the two points I argued 'ad nauseum' in yesterday's posting). The second effect is both real, and desirable -- this is what we are trying to achieve when a microscopist "saturates" the gun. I don't have any trouble with calling this "saturation" (and usually do so myself) since it is a pretty good description of what one is visually observing. The problem is, unless you have a TEM, or a SEM equipped with "source imaging" (which is apparently not all that common), people tend to think that the third effect is due to one of the first two -- and this is just not so. This mistaken notion is what I was laboring to correct. But it was never my intention to dispute the fact that something very useful happens to the beam shape when you correctly adjust the biasing (effect #2). If your microscope is setup correctly, the auto-bias "knee" (effect #3) will occur in the vicinity of the compact source distribution (#2) and using the "knee" to saturate works just fine. But if one starts arbitrarily tinkering with filament spacing and the like so as to increase emission, extend filament life, etc., without understanding what is happening, effect #3 may not match effect #2 anymore and when the operator thinks that he/she has optimized the gun by "saturating the filament" via the emission curve knee, the emission pattern may actually be non-optimal.
To address Bill's remarks about the mechanism which results in this localization of the beam in the emission image, I'm now going to restrict myself to discussing effect #2 only -- and I'll call it 'saturation" since that's what we're used to.
What causes this 'saturation' of the emission distribution to occur? Or equivalently, why does one get the hollow-centered "smoke ring" type of pattern when the filament is "undersaturated"? Bill postulates that this is because electrons are only emitted from the periphery of the circular emitting region on the filament tip (i.e., the region selected by the biasing). I don't think this is quite correct since it is energetically possible for electrons to be emitted from anywhere within the "zero equipotential" region established by the wehnelt bias. Instead, the answer is related to the fact that biasing the wehnelt not only establishes the emitting region on the filament tip, but also creates an electrostatic lens which focuses the beam (I've ignored the focusing aspect thus far in the discussion since things were complicated enough without getting into this -- and my posts were already plenty long!).
But focusing does need to be introduced if we are going to understand the emission pattern since we need to be aware that what we are seeing in the "filament image" is not an image of the filament per se, but rather, an image of the "crossover" distribution, which is the point where the focused electrons from the filament converge to their smallest cross-section. (Someone may want to argue with me about this, since if you get into really undersaturated conditions, you can actually see structure on the filament tip. However, this is because the "crossover" is in effect a spatial image of the filament tip under these conditions. In the optical sense, the "object" you are looking at in a "filament image" is this crossover image, not the physical tip as such.) So just because we don't see any electrons in the center of this emission pattern does not mean that the filament is not emitting from its center -- it means that the focusing action is such that the center of the crossover pattern isn't getting electrons directed into it.
I again like to use a garden hose analogy to describe what is happening here. Imagine how you adjust the nozzle on your garden hose to move from a fan-shaped (open-centered) distribution to a compact stream. I visualize the bias on the wehnelt shaping the beam in a rather analogous fashion. The emission distribution image you observe is thus like observing the cross-section of the water stream at some distance from the nozzle. If you think about trying to direct the tightest possible stream of electrons down the optics of the column, you can then see why "saturating" the beam (in sense #2, of course) gives you the best imaging conditions. (I hasten to add the disclaimer that I NEVER want to be accused of claiming that a garden hose is a perfectly accurate model of gun dynamics -- just a useful visual metaphor for some phenomena.)
Two other related topics suggest themselves. One is the way that this "garden hose" model explains the phenomenon of "false peaks" which you obtain with a misaligned gun when you are trying to "saturate" by watching the specimen current. The second relates to the question of why the "compact distribution" condition is optimized when the emitting region is restricted to the filament's tip. However, I don't want to engage in another marathon post like yesterday's (the Bandwidth Conservation League will probably censure me as it is) and I also have no idea whether anyone but myself is interested in such arcane topics. So I will (for once) refrain.
Fred Schamber RJ Lee Instruments
====================== Bill Tivol wrote:
} Dear Fred, } I realize that the situation is considerably different for the TEM } from that for the SEM; however, what I have always heard and observed is } that for low filament currents electron emission occurs only from some } regions of the filament tip. This can be seen by imaging the filament at } crossover, where it appears as an annulus. As the filament current is } raised, additional regions of the tip begin to emit electrons, and the } image of the filament becomes more uniform and smaller. As the filament } current reaches a value such that all regions of the tip emit, the image } has no dark regions. This is what I have called "saturation"; it makes } sense from the standpoint that emission cannot occur from more than the } complete tip area and, therefore, cannot increase with further increases } in filament current. AFAIK, this has nothing to do with space charge, } current stabilization, virtual apertures, or bias resistors, but with } the temperature, geometry and work function of the tip. Are there two } uses of "saturation"? } Yours, } Bill Tivol }
Barry, Lilith wrote: } } Thank you for responding to my question on tissue culture background. I } don't think the background is because the fixation or the immuno method } because I routinely use the same method on brain sections and do not get the } same background. It isn't the coverslip autofluorescing either because one } can see beautiful labeled structures in the cells except on the controls. So } it has to be something specific about cortical cultures and the strept or } neutravidin-biotin method. Do you have any other thoughts? } Lilith } ------------------------------------------------------------------------ } Lilith Ohannessian-Barry } National Research Council } Institute of Biological Sciences } CANADA } Tel;613-993-6460 } Fax;613-941-4475 } e-mail; lilith.barry-at-nrc.ca
Hi Lilith, Have you ruled out the endogenous biotin? Either try a biotin block prior to your staining or try another detection method, which does not utilize biotin. Katri
Is there any significant differences in SEM filament saturation respective to W, LaB6 or field emission cathodes? If so, what are they and how should one treat these different electron sources?
TONS, yes the difference between W, LaB6 and an FEG "saturation" (lets s= ay source set up) is LARGE.
For your application LaB6 saturation should be carried out when the virtu= al source is being visualised as there are a number of peaks that you may us= e depending upon the work you wish to carry out, high brightness, medium resolution and high resolution. =
May I suggest you contact the supplier of your LaB6 cathodes and they are=
sure to be able to give you a sheet relating to saturation and good housekeeping for LaB6, you just cannot treat these sources like W, they a= re very sensitive! =
A LaB6 tip will become damaged by discharge (a W will usually not unless=
at TEM voltages) such that it may emit from more than one area. In order=
to prevent this you should run up the kV and filament very very slowly. =
This allows the gas emitted through these actions to be removed before th= e gun environment becomes suitable for discharge. W filaments will take al= l that you throw at them LaB6 I say again are very very sensitive but boy d= o you get some gains when they are up and running? Higher brightness less aberration therefore higher resolution and particularly at low kV.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Scanners Corporation has an opening for a Senior SEM Field Service Engineer. This position is for the Eastern U.S. region.
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I think a good starting-point could be the Reconstruction-Homepage with several references, software descriptions and links: http://biocomp.stanford.edu/3dreconstruction/index.html For basic information, maybe the following article is of help: http://www.videomicroscopy.com/Tutorials/3D%20Reconstruction/October%20hows% 20that%20work.htm I guess that for your task volume-renderer or ray-tracer software will be a better choice than surface-renderer. Some of the volume based programs are free, e.g. NIH-image for Mac (with integrated 3D functions) http://rsb.info.nih.gov/nih-image/ There is also a Windows version, called Scion-Image http://www.scioncorp.com/frames/fr_download_now.htm However, with such a complex structure like bone I would think about a way to use a confocal laser scanning microscope. This would avoid the problem of image alignement and you can use the integrated 3D-functions of the cLSM software. Maybe someone on the list does know a protocol for thick sectioning of bones and visualization with the LSM?
Best regards
Jens
} Hi everyone: } } I have been working for a long time in the field of inorganic materials. Now } I am getting into biomaterials and my first task is to determine porosity of } a bone, I think I need some help. I guess that if I record images of the } surface of the bone and apply sequential sectioning and image recording, I } will be able to reconstruct the 3d characteristics of the object. I wonder } if there is any software available to do the reconstruction or maybe someone } can suggest some references where I could get started. } } Thanks in advance } } } Hector Calderon
------------------------------------------------------------------- Dr. Jens Buecking Tel. +49-(0)421-218 3745 University of Bremen Fax. +49-(0)421-218 4620 Dep. of Biology Email jbueck-at-biologie.uni-bremen.de Leobener Str. - NW2 URL http://www1.uni-bremen.de/~jbueck/ D- 28359 Bremen (AG Prof. Dr. H. Witte) GERMANY
2tes Acarologisches Kolloquium in Bremen, 14.-16.10.99 http://www-user.uni-bremen.de/~acari/ -------------------------------------------------------------------
I've received a few replies to my inquiry about TV rate performance of solid-state vs. scintillator BSE detectors. Thank you.
It seems that the p-n type solid state BSE detectors are not quite up to the performance at TV rates of the scintillator types, but solid-state systems such as those provided by GW Electronics do provide good TV rate performance.
Bruce E. Brinson has informed me that micro-channel plate type BSE detectors also perform adequately at TV rate and much better at that rate than some p-n solid state type detectors he has used.
In my case, I was attempting to use an "in-my-lab" modified Robinson BSE detector system. My TV-rate performance was not good enough. The light pipe shape that I prepared for my ARL SEMQ's BSE system is far from optimum because of little available space for an "overhead" scintillator and a slightly remote location needed for the PMT.
Another modification performed a few days ago has now given good TV rate performance. A simple idea. Tricky to execute. I cored out the lens bore area of the plastic scintillator of the original and replaced that volume with a cerium doped YAG crystal. A much, much brighter scintillator material than the plastic. Better performance yet will be achieved with fiber optics instead of the solid plastic light pipe.
We use John Russ's Image Processing Tool Kit (Reindeer Games Inc.) that works as Photoshop plug-ins to do this sort of thing with collagen banding. I just work on the FT image as a greyscale using Photoshop tools to select or reject the parts of interest then do a inverse filtered FT. Very easy to work with. You can remove bands or enhance bands either way.
Bob Derm Imaging Center
On Wed, 5 May 1999, Robert H. Olley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } First, thanks to all of you who either: } } (a) told me about silver membrane filters, which I had not heard of before } (b) reminded me of Gelman, who recommended their PTFE membranes. } } As to which I use for which application, it's horses for courses! } } And now the question: one has a scanned or digitally acquired micrograph. } The background is a gently varying grey, with rather low contrast local } features (blobs, bacteria, or whatever). The task is to Fourier } transform, then filter out the lowest frequencies with a high-pass filter, } leaving one with the features on a uniform grey background. One can then } increase the contrast to make the features more prominent for printed } reproduction. Is there software (Win 95) that can do this? (preferably } inexpensive, though we could make arrangements to use plugins for Adobe } Photoshop elsewhere, if necessary). } } You can see an example of what I'm trying to do on: } } http://www.reading.ac.uk/~spsolley/fourier.htm } } +------------------------------------------------------------------------+ } | Robert H.Olley Phone: | } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } | University of Reading {University internal extension 7867 | } | Whiteknights Fax +44 (0) 118 9750203 | } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } | England URL: http://www.reading.ac.uk/~spsolley | } +------------------------------------------------------------------------+ } } }
There is a software package called EIKONA3D, developed by a company called AlphaTec Ltd., which is a 3D image processing and analysis package that provides special features and tools for working with image sequences/serial sections originated from microscopy (e.g. 3D image processing, image alignment, 3D reconstruction, 3D visualization, volume rendering, surface rendering, 3D registration,= etc.).
Further information and a free demo version of EIKONA3D are available through AlphaTec's web site: http://www.alphatecltd.com
Regards, Nikos Nikopoulos
At 02:04 =EC=EC 7/5/1999 -0500, you wrote: } } Hi everyone: } } I have been working for a long time in the field of inorganic materials.= Now } I am getting into biomaterials and my first task is to determine porosity= of } a bone, I think I need some help. I guess that if I record images of the } surface of the bone and apply sequential sectioning and image recording, I } will be able to reconstruct the 3d characteristics of the object. I wonder } if there is any software available to do the reconstruction or maybe= someone } can suggest some references where I could get started.=20 } } Thanks in advance } } } Hector Calderon }
I inquired about a classified section and was given the following response:
"There isn't a "classified" section, but you can post such a notice on the listserver. I hope that's helpful to you.
Cindy Clark MSA Administrator"
The message I want posted is as follows:
We are selling a Cambridge S100 SEM with an Energy Dispersive X-ray analysis attachment. We are looking for $5000 to $10,000 exclusive of shipping costs. Anyone interested please send e-mail to tpizzey-at-canspec.com.
We do not need to routinely capture TEM images with a digital camera, as is being discussed. It would be nice though to be able to scan negatives that are to be used for say publication or other presentation.
Is there a scanner available than can produce a high resolution image, i.e. suitable for publication?
Also, thanks to those who provided responses on basement membrane labeling.
After using our SEM's cold stage for the first time last week we have a few observations. The phenomenon which were demonstrated is with regards to cathodo-luminescence and backscatter emission, so I'll send this query to the list in two parts. I hope that someone will comment on our observations as to verify what is mysterious, and possibly explain what we are seeing.
We are examining the CL emission of quartz so as to study and possibly characterize features and textures with regards to source and history (e.g., if we were to examine a sandstone, we might be able to say the source of the qtz was metamorphic or igneous). This particular hydrothermal specimen we thought would be ideal for cold stage introduction because it had demonstrated marked areas of absolute absence of CL as well as otherwise. What we saw at low temperatures was a bit disappointing. Whereas the increase in CL intensity was very dramatic for temperatures {-100C, all of the features disappeared!!! It would be interesting to examine this phenemenon in color so as to evaluate whether the color becomes homogenous as well.
Lastly ... this particular qtz sample demonstrates a phenomenon of retaining an overprint of e-beam bombardment. That is, if we were to zoom in for a focus, a subsequent image capture would show us a small bright rectangle in the middle. This phenomenon is common, BUT it isn't demonstrated by every type of qtz. Has anyone correlated this "overprinting" with a type of qtz or possibly specimen preparation??
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
We are looking for an old Philips 201. We currently have one we are using and since parts are a little hard to come by, we would like to find another one that could be purchased for parts. If you have one you would like to sell or donate, please contact me off list.
Thanks,
Michael D. Standing Microscopy Lab Brigham Young University P.O. Box 25181 Provo, UT 84602-5181
After using our SEM's cold stage for the first time last week we have a few observations. The phenomenon which were demonstrated is with regards to ... backscatter emission .. I hope that someone will comment on our observations as to verify what is mysterious, and possibly explain what we are seeing.
We are examining the CL emission of quartz and whereas BSE imaging is of little use, we do like to document our CL imagery with an accompanying BSE image. We never expected BSE imaging at cold temperatures to be any different, so we were surprised at what was demonstrated ... which was a marked increase in the topographic contribution to the BS emission. Our BS detector is the twin solid-state type where the BS signal acquired by both halves shows atomic number contrast and the difference signal shows topography. We were using a 25mm working distance so it is difficult to believe the cold stage cooled the pole-piece mounted detector. At temperatures less than -100C we couldn't see any Z contrast at all, even for MoS inclusions in the qtz vein!!! Even at temps near zero C the polishing defects were easily seen and didn't disappear 'til ambients temps were again achieved. I am not sure the Z information actually disappears, but the topo component definitely floods the signal and is impossible to remove withour sacrificing general contrast. It would first be interesting to know if someone else has seen this with both types of BS detectors ... i.e., the scintilator type (e.g., Robinson) as well.
I've never heard of anyone speaking of this phenomenon ... does anyone have any ideas or similar observations??
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Maybe your topographic information isn't changing (use SE at ambient and cold temps to check). But there could be a surface layer of ice which masks the atomic number contrast. You could also monitor x-ray counts at ambient and cold to see if there is a change.
: cooled the pole-piece mounted detector. At temperatures less : than -100C we couldn't see any Z contrast at all, even for MoS : inclusions in the qtz vein!!! Even at temps near zero C the : polishing defects were easily seen and didn't disappear 'til : ambients temps were again achieved. I am not sure the Z : information actually disappears, but the topo component definitely : floods the signal and is impossible to remove withour sacrificing : general contrast. It would first be interesting to know if someone : else has seen this with both types of BS detectors ... i.e., the : scintilator type (e.g., Robinson) as well. :
Carl : :
Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA -------------------------------- Voice: (734) 936-1550 FAX: (734) 763-4690 E-mail: chender-at-umich.edu --------------------------------
Some where I seem to remember that phosphorus has enough of a radioactivity isotope to measure. Okstate built a scintillation counter that would hold a cow. You wouldn't need one nearly so bit but if phosphorus is correlated to density it would be an easy test.
While on the subject of bones I took some pictures of some rat bones being broken. It was the usual dietary calcium trial but some of the rats had been fed small amounts of vadium(sp). The rats fed vadium(sp) did not have any stronger bones than the corresponding rats in the test the only difference is while the bones broke at the same load they did not snap. The went through a very noticeable plastic deformation before separating.
I never saw any more about the research.
Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
-----Original Message----- } From: info {info-at-zeus.csd.auth.gr} To: microscopy-at-Sparc5.Microscopy.Com {microscopy-at-Sparc5.Microscopy.Com}
Dear Hector,
There is a software package called EIKONA3D, developed by a company called AlphaTec Ltd., which is a 3D image processing and analysis package that provides special features and tools for working with image sequences/serial sections originated from microscopy (e.g. 3D image processing, image alignment, 3D reconstruction, 3D visualization, volume rendering, surface rendering, 3D registration, etc.).
Further information and a free demo version of EIKONA3D are available through AlphaTec's web site: http://www.alphatecltd.com
Regards, Nikos Nikopoulos
At 02:04 7/5/1999 -0500, you wrote: } } Hi everyone: } } I have been working for a long time in the field of inorganic materials. Now } I am getting into biomaterials and my first task is to determine porosity of } a bone, I think I need some help. I guess that if I record images of the } surface of the bone and apply sequential sectioning and image recording, I } will be able to reconstruct the 3d characteristics of the object. I wonder } if there is any software available to do the reconstruction or maybe someone } can suggest some references where I could get started. } } Thanks in advance } } } Hector Calderon }
I would very much appreciate the complete name of the journal Boll. Zool. Please, answer me not to the list. Thanks in advance.
Gabriel Adriano Rosa Area Microscopia Electronica, Depto. Cs. Biologicas Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA TE -(54-11)-4576-3349 e-mail micros-at-bg.fcen.uba.ar FAX (54-11)-4576-3384
I thought it was a nice touch when Nestor added the MSA plug to each of the posts and included the instructions for (un)subscribing. However, there has been a recent rash of unsubscription requests coming to the list by mistake. Apparently ones are not reading the instructions. The header plainly states that the requests are to be sent to
ListServer-at-MSA.Microscopy.Com
not to the list itself. They should have the subscription action included as the body in the form
By following these instructions, you can make the changes yourself. The only time Nestor (or the list) should have to be bothered by subscription requests is if a subscriber can no longer mail from the same address they subscribed under. In those cases, this list server (and most others) will require intervention to make sure the request is authentic. But for the majority of cases, I encourage those that wish to unsubscribe to read the header more closely. Nestor does enough for this list already.
Gatan DigitalMicrograph (DM) software can do precisely what you want to do with your images. They are all the standard features in DM. There are a large number of life science microscopists using DM. For further info, check out www.gatan.com, or send me an email.
Good luck!
Ming Pan Gatan, Inc.
} } } } And now the question: one has a scanned or digitally acquired micrograph. } } The background is a gently varying grey, with rather low contrast local } } features (blobs, bacteria, or whatever). The task is to Fourier } } transform, then filter out the lowest frequencies with a high-pass filter, } } leaving one with the features on a uniform grey background. One can then } } increase the contrast to make the features more prominent for printed } } reproduction. Is there software (Win 95) that can do this? (preferably } } inexpensive, though we could make arrangements to use plugins for Adobe } } Photoshop elsewhere, if necessary). } } } } You can see an example of what I'm trying to do on: } } } } http://www.reading.ac.uk/~spsolley/fourier.htm } } } } +------------------------------------------------------------------------+ } } | Robert H.Olley Phone: | } } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } } | University of Reading {University internal extension 7867 | } } | Whiteknights Fax +44 (0) 118 9750203 | } } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } } | England URL: http://www.reading.ac.uk/~spsolley | } } +------------------------------------------------------------------------+ } } } } } }
I am not an expert in this area but guess the procedure is as follows.
1. Use a fine metal polish to remove, by polishing, the aluminium a= nd phosphor from the old scintillator.
2. Clean with solvent the scintillator glass.
3. Mix a solution of phosphor in a solvent (?) with one or two drops=
of a plastic solution like formvar (to give strength). Ultrasonic this solution.
4. Place the scintillator in a beaker and pour onto it the phosphor solution. Cover but with a space for ventilation.
5. After some hours the solution will settle, decant off the excess and wait several days for the coating to dry.
6. Remove scintillator and if no defects are visible coat in a high vacuum, coating unit with a very thin layer of aluminium.
7. Check in microscope for efficiency.
8. If not good experiment with phosphor coating thickness and/or wit= h type of phosphor, P54 comes to mind(?)
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Dear Gabriel and others, I have sent this to the list as some of this info is quite generally useful and not just for Gabriel's question.
Chemical abstracts service provides a list of the approved abbreviations for all the journals abstracted in "Chemical Abstracts" at http://info.cas.org/sent.html There is a list of biological journal abbreviations at: http://arachne.prl.msu.edu/journams/ and a list of journals in the ISI at: http://library.caltech.edu/admin/abbreviations/ although none of these have Boll. Zool.
Hope this helps
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren Department of Experimental Physics Chalmers University of Technology S-412 96 G=F6teborg Sweden Tel: +46 31 772 36 33 FAX: +46 31 772 32 24 email: maclaren-at-fy.chalmers.se or: ianmaclaren-at-hotmail.com Research Group Homepage: http://fy.chalmers.se/microscopy/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Gabriel Adriano Rosa wrote: } I would very much appreciate the complete name of the journal Boll. Zool. } Please, answer me not to the list. } Thanks in advance.
I am not in the biology sector, but I guess it should be Bull. Zool. - there is at least a journal named Raffles Bulletin of Zoology.
Joergen.
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
I am trying to visualize by negative staining the attachment of phages to bacteria. However the bacteria shrink under the EM vacum, leaving an artefactual space between the negative stain and the cell wall which do not allow to take good pictures of the attached phages.
Here is the protocol I am using: I am using 2% uranyl acetate as the negative stain The cells are adsorbed to carbon coated formvar grids for 1 minute Excess is drained with filter paper The grids are floated in the negative stain for 1 minute Excess is drained with filter paper The grid goes into the TEM
I have tryed a few modifications such as fixing the material and mixing the stain with the bacteria suspension, but it did not work.
can anyone send further suggestions???
Sincerelly
Dr. A.P. Alves de Matos EM Unit, Pathology Department Curry Cabral Hospital Lisbon
Does anyone know the formula/ chemical name for an etchant called glyceregia? It is used to show carbonitrides in stainless steels for optical microscopy and I'ld like to know how to prepare some.
Im forwarding this message to the list. Hopefully someone in the states will be able to help.
Please email him direct at pwilsonarzu-at-Hotmail.com
} Dear Sir: } } I would like some information on buying a used electron microscope in the } United States. } } I would really appreciate any information you may have available. } } My e-mail: pwilsonarzu-at-Hotmail.com } } Thank you in advance, } } Philip E. Wilson
---------------------- Kevin Mackenzie Electron Microscope unit Dept Zoology University of Aberdeen Tillydrone avenue Aberdeen AB24 2TZ ----------------- Tel 01224 272847 Fax 01224 272396 email k.s.mackenzie-at-abdn.ac.uk
Hi everyone, I have heard of radiation damage to the specimens, and I'm curious how much beam current and current densities are there on the specimen in HREM imaging and nano-diffraction. Your replies are highly appreciated.
A search of the library here turned up "Bollettino di zoologia" published as vols. 1-62, 1930-1995, and since continued as the Italian Journal of Zoology.
Best regards,
Dan Luchtel, Ph.D. Professor University of Washington Environmental Health Box 357234 Seattle, WA 98195-7234
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Good day all, Does anyone have a schematic for a Coolwell chiller? Model # SE-075W C2?? I need some help to find out what the electronics are suppose to be? not what they are. Any help would be nice, since what I have heard they where bought out, and the holding company only wanted the name and management, so no tech help. Sincerly,
************************************************ Robert J. Derby New Mexico Institute of Technology Socorro, N.M. Phone - 505-835-5866 E-mail - rjderby-at-excite.com derby-at-nmt.edu ************************************************
_______________________________________________________ Get your free, private email at http://mail.excite.com/
} Some where I seem to remember that phosphorus has enough of a } radioactivity isotope to measure. Okstate built a scintillation counter } that would hold a cow. You wouldn't need one nearly so bit but if } phosphorus is correlated to density it would be an easy test. } Dear Gordon, Natural phosphorus is 100% P-31. The two radioactive isotopes are P-32 & P-33 with half-lives of 14 & 25 days, respectively. They both emit relatively high-energy beta-. Even so, the beta's would not make it to a scintillation counter if they originated in cow bones (assuming that the rest of the cow was still attached :-)). You are probably thinking of potassium, which has a significant amount of K-40. It is this isotope which gives a measurably larger dose of radiation from being inside a concrete building, as opposed to a wood building. Yours, Bill Tivol
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} I am trying to visualize by negative staining the attachment of phages to } bacteria. } However the bacteria shrink under the EM vacum, leaving an artefactual } space between the negative stain and the cell wall which do not allow to } take good pictures of the attached phages. } } Here is the protocol I am using: } I am using 2% uranyl acetate as the negative stain } The cells are adsorbed to carbon coated formvar grids for 1 minute } Excess is drained with filter paper } The grids are floated in the negative stain for 1 minute } Excess is drained with filter paper } The grid goes into the TEM } } I have tryed a few modifications such as fixing the material and mixing the } stain with the bacteria suspension, but it did not work. } } can anyone send further suggestions??? } Dear A.P., I am most definitely not an expert on this. The protocol you're using is the same as I use in my work. Since the problem is with shrink- age, perhaps placing a few ul of bacterial suspension on the grid, air drying and placing it in a vacuum, then applying the UAc will prevent the problem. Of course, such treatment could induce artifacts--it is a pretty severe process--but it might still tell you what you want to know. Freeze-drying could be another way to go, but this can also pro- duce artifacts; I've seen the results when this was done to kidney, and it's not pretty. Good luck. Yours, Bill Tivol
Hello! I need a heater to keep mammalian cells warm while viewing them with a light microscope. I know there are various types avaiable but I'm not sure where to find them. Can anyone recommend a company that sells this sort of thing? Cheers, Jennifer
-- Jennifer Shuler, Ph.D. Director of Imaging Facility Adjunct Assistant Professor Biology Department, Box 7325 Wake Forest University Winston-Salem, NC 27109
Adam wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello, } } Does anyone know the formula/ chemical name for an etchant called } glyceregia? It is used to show carbonitrides in stainless steels for } optical microscopy and I'ld like to know how to prepare some. } } Thank you, } } Adam Scott } } ascott1-at-engfac.uct.ac.za
Adam,
Please go to the http://www.kaker.com/mvd/vendors.html, there is a link to my free etchant's database. Bellow is search result:
Material: Wrought stainless steels (Fe) Type: Microetchant Method: Chemical etching Etchant (Electrolyte): 10 ml HNO3, 20 to 50 ml HCl, 30 ml glycerol (Glyceregia). Procedure: Mix HCl and glycerol thoroughly before adding HNO3. Discard before solution attains a dark orangr color. Immerse or swab specimen for a few seconds to a few minutes. Higher percentage of HCl minimizes pitting. Remarks: General structure. Reference: Metallography, Structures and Phase Diagrams, Metals Handbook, 8th Edition, Vol. 8, ASM (American Society for Metals), Metals Park, Ohio 44079, USA, 1973, p. 98.
Henrik
-- Henrik Kaker, Ph.D. SEM-EDS Laboratory Metal Ravne, Koroska cesta 14 Ravne, Slovenia, Phone: +386 602 21 131 Fax: +386 602 20 436 Mailto:Henrik.Kaker-at-guest.arnes.si http://www2.arnes.si/guest/sgszmera1/index.html
} -----Original Message----- } } Hello, } } Does anyone know the formula/ chemical name for an etchant called } glyceregia? It is used to show carbonitrides in stainless steels for } optical microscopy and I'ld like to know how to prepare some. } } Thank you, } } Adam Scott } } ascott1-at-engfac.uct.ac.za ------------------------------------------------------
Adam,
According to the ASM Metals Handbook, 8th Edition, Volume 8, page 98
"Glyceregia: 10ml HNO3, 20 to 50 ml HCL, 30 ml glycerol."
"Procedure: Mix HCL and glycerol thoroughly before adding HN03. Discard before solution attains a dark orange color. Immerse or swab specimen for a few seconds to a few minutes. Higher percentage of HCL minimizes pitting"
Hope this helps.
Neither TIMET nor William Giles assume any responsibility for the above quoted formula.
This is a follow-up on my query about fixing isolated mitochondria for TEM. First, thanks to all who responded. A summary of replies is below. We solved our immediate problem by fixing in 4% glutaraldehyde in 0.1M sodium cacodylate, r.t. for ca 30 min, and then into the fridge until the next morning. I followed with a fairly routine postfixation, dehydration, and embedding. We got good enough results to answer the first questions. However, there will be many more!
I would like to mine the collective expertise further. For the next step the client would like to localize calcium and, if possible, do some crude quantitation. From my background in neurophys I know this is tricky. Do any of you have a current favorite localization technique? In the (far distant) past I have used K-pyroantimonate to precipitate Ca with mixed results, and I can't find that particular recipe. New ideas would be enthusiastically received.
Soon I hope to have a better instrument to help in this and many other quests. We just installed a LEO (Zeiss) 912 AB TEM with integrated omega energy filter. What a beautiful instrument! What potential! How exciting! I wish I knew how to use it... I've only had it a couple of weeks and, while I can make the basic instrument work well, I still haven't had my "applications training" to learn how to use the energy filter for ESI, EELS, and all the really useful and fun stuff. I've tried to bull my way through the manuals and run on intuition, but that approach isn't working...
However, I am confident it will be useful for this kind of problem. I would be interested in striking up an e-mail penpal relationship with anyone else with this instrument.
We do not yet have a cryostage for it, and so looking at ultrarapidly cryofixed, unprecipitated ions is not yet a possibility.
Secondly, I would like to hear from anyone who might have an explanation and/or citations for the appearance of these isolated mitochondria that have been subjected to high Ca concentrations. We are seeing cup-shaped mitochondria (which I see in my inverts all the time), but the outer membrane on the side of the concavity is NOT concave, and is swollen outward. This wouldn't bother us so much, except it nags at me that there is something asymetrical about the mitochondria membranes that causes the inner and outer membranes to stay attached on one side and not on the other. Plus, sometimes the cristae are swollen and sometimes not. I would appreciate being pointed to appropriate literature, if any. My searches have not been very fruitful at this point. Control mitochondria processed at the same time do not exhibit this morphology.
Here is the summary of responses I received a few weeks ago:
Have you tried/considered fixing them in the last buffer they see before fixation? I always worry about isolation protocols....the mitos may look blah because of something they've been in during the isolation protocol, not because of your solutions. Now I try to at least match the final solution in the prep. As an example - organelles that have been purified through sucrose gradients are going to look like crap if they are taken from their sucrose and fixed in 0.1M any buffer! (Been there, done that...). And mitos are so fussy, anyway.......Investigators can be reticent about their preps. I'm sure you've had the, "Oh, didn't I tell you? These fractions are in 5M Tris" experience. Homicide doesn't seem to be a viable response :) ********************************************************** Yes I did, thanks. They looked really terrible! ********************************************************** A long time ago back in the mists of time I used to fix isolated cell fractions, mitochondrial, lysosomal, nuclear, gap junctions, etc.
We used a double fixative of cold Glutaraldehyde and Osmium Tetroxide mixed.
Reference as follows:-
Ultrastructure of Human Leukocytes after simultaneous fixation with Ultrastructure of Human Leukocytes after simultaneous fixation with Glutaraldehyde and Osmium Tetroxide and "post-fixation" in Uranyl Acetate. Hirsch, J.G. & Fedorko, M.E. (1968) J.Cell. Biol. 38, 615.
This excellent for cell suspensions and fractions. The fixative is mixed on ice and used immediately after mixing. Fixation also on ice. Fixation time for a small pellet about 30 minutes.
I used this method a lot at one time and can thoroughly recommend it. But, only for suspensions and cell pellets. Not for solid tissue.
Membranes are well defined, swelling/shrinkage artefacts are not a problem. ******************************************************************* We have had good luck with high-pressure freezing. We have examined unstained, frozen-hydrated mitos on the 400 kV instrument here. Although this procedure requires instruments not available at all facilities, I expect to get minimal artefact. ******************************************************************
Mahalo to you all- Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
You better check on the polishing step with someone who knows for sure. If there is a conductive coating on the glass under the phosphor (and I'm sure there is), such as tin oxide or indium tin oxide, you'll probably take that off too. The thin aluminum on top of the phosphor protects the phosphor and is not used as the applied voltage contact. Hard rubbing without the abrasive will not take the coating off the glass. A product like Soft Scrub might do the trick for you, but again, I would double check before I would risk it.
You have to be careful in Steve's step 5. If you cause the water solution to move too much, you can get a wavy coating. One trick that I used when making transparent screens, is to run a small diameter tube into the container that the plate being coated was place down to the bottom of the bottom. I would then tape the tube to the outside and attach a syringe to the bottom. The syringe was below the bottom of the container with the tube and syringe full. When it was time to decant the liquid, I pulled the plunger and started a siphon into a beaker. It took a while because the tubing was so small, but it worked fine.
Another addition that you can put in the solution is potassium silicate. (Sometimes this is called liquid glass.) It helps to bond the phosphor to the glass. Sorry, but I can't remember how much to put in. It was only a few percent.
-Scott
---------- } From: Steve Chapman To: de Lillo Enrico; American -----------------------------------------------------------------------.
I am not an expert in this area but guess the procedure is as follows.
1. Use a fine metal polish to remove, by polishing, the aluminium and phosphor from the old scintillator.
2. Clean with solvent the scintillator glass.
3. Mix a solution of phosphor in a solvent (?) with one or two drops of a plastic solution like formvar (to give strength). Ultrasonic this solution.
4. Place the scintillator in a beaker and pour onto it the phosphor solution. Cover but with a space for ventilation.
5. After some hours the solution will settle, decant off the excess and wait several days for the coating to dry.
6. Remove scintillator and if no defects are visible coat in a high vacuum, coating unit with a very thin layer of aluminium.
7. Check in microscope for efficiency.
8. If not good experiment with phosphor coating thickness and/or with type of phosphor, P54 comes to mind(?)
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
First, try the company from which you purchased your microscope. Secondly, BioOptechs has great stages for live cell work. Finally, there are several manufacturers for warming stages. Visit either www.mwrn.com or the microscopy society site (www.msa.microscopy.com) for vendors. Email me if you have trouble finding contact information.
Best regards Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 02:34 PM 5/11/99 -0400, Jennifer Waters wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} I am trying to visualize by negative staining the attachment of phages to } bacteria. } However the bacteria shrink under the EM vacum, leaving an artefactual } space between the negative stain and the cell wall which do not allow to } take good pictures of the attached phages. } } Here is the protocol I am using: } I am using 2% uranyl acetate as the negative stain } The cells are adsorbed to carbon coated formvar grids for 1 minute } Excess is drained with filter paper } The grids are floated in the negative stain for 1 minute } Excess is drained with filter paper } The grid goes into the TEM } } I have tryed a few modifications such as fixing the material and mixing the } stain with the bacteria suspension, but it did not work. } } can anyone send further suggestions??? }
You can freeze-dry the negative stains. Look for the papers of N.V. Nermut e.g. in the Proc 5th European Congress on Electron Microscopy 1972 page 236. ***************************************************** Mel Dickson, Deputy Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Has anyone had success in staining wax in a rubber matrix? Either optical or electron contrast methods would be of interest, particularly those for SEM. Thanks for your thoughts.
Hi all, Boll.Zool. stands for Bollettino di Zoologia, now known as "The Italian Journal of Zoology". Here's their website: http://www.uniroma1.it/bau/uzi/itjz.htm
Glyceregia formula =3D 10 ml HNO3 + 20 to 50 ml HCl + 30 ml glycerine (ref.: ASM Metal Handbook, Desk Edition, page 35.35)
Marc Montreuil Rolls-Royce Canada
Adam a =E9crit:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------= =2E } } Hello, } } Does anyone know the formula/ chemical name for an etchant called } glyceregia? It is used to show carbonitrides in stainless steels for } optical microscopy and I'ld like to know how to prepare some. } } Thank you, } } Adam Scott } } ascott1-at-engfac.uct.ac.za
-----Original Message----- } From: William Tivol {tivol-at-wadsworth.org}
} Dear Gordon, } Natural phosphorus is 100% P-31. The two radioactive isotopes } are P-32 & P-33 with half-lives of 14 & 25 days, respectively. They both } emit relatively high-energy beta-. Even so, the beta's would not make } it to a scintillation counter if they originated in cow bones (assuming } that the rest of the cow was still attached :-)). } You are probably thinking of potassium, which has a significant } amount of K-40. It is this isotope which gives a measurably larger dose } of radiation from being inside a concrete building, as opposed to a wood } building.
Bill,
I always did have trouble mixing up P & K when it was listed in any format but N-P-K in fertilizer.
If we wanted to measure P-32 or 33 we would have to drop the cow in the whole cow grinder. They have one that will grind up a cow hide, hair, guts and all for finding out what actually makes up a cow. It wouldn't be very useful for estimating bone destiny but the K-40 might if it is correlated to bone density. It would probably give skewed results for a lot of the world due to Chernoble and nuclear testing.
Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
Bacteria are a bit thick for negative staining. Changing the stain concentration helps a little. The rule is that thick specimens appear better with a lower concentration of stain, say 2%. Thin specimens are better with a lower concentration, say 0.5%. You may get better appearance trying different staining solutions, PTA or Ammonium molybdate are good candidates. Don't grow cultures on a shaker, many flagella drop off and even fimbria may suffer.
This is a difficult subject for negative staining because you require both, bacteria and flagella in one picture. Much prettier images can be obtained with shadow casting. 1 Apply bacteria solution to the face of substrated grid. (apply and blot lightly several times to obtain better distribution) Drying would leave too much debris and salts, so use a volatile buffer to wash and maintain molarity. 2 Apply and blot bacteria coated grid repeatedly to a 0.1 M solution of ammonium acetate. 3 After air drying, angle (or rotary) shadow) grid in a vacuum evaporator using fine grain evaporating material. (simultaneous C/Pt or if available high resolution Cr coater as used for FESEM)
Another alternative: If the equipment is available would be FESEM of bacteria and flagella and this could be supplemented with an image of negatively stained flagella only, taken in TEM.
Its a little challenging project, but some nice images could result. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
} I am trying to visualize by negative staining the attachment of phages to } bacteria. } However the bacteria shrink under the EM vacum, leaving an artefactual } space between the negative stain and the cell wall which do not allow to } take good pictures of the attached phages. } } Here is the protocol I am using: } I am using 2% uranyl acetate as the negative stain } The cells are adsorbed to carbon coated formvar grids for 1 minute } Excess is drained with filter paper } The grids are floated in the negative stain for 1 minute } Excess is drained with filter paper } The grid goes into the TEM } } I have tryed a few modifications such as fixing the material and mixing the } stain with the bacteria suspension, but it did not work. } } can anyone send further suggestions???
you can try to adsorb your bacteria/phage mixture onto carbon film, freshly prepared onto cleaved mica. Just cut out a small piece of the carbon film, a little bit smaller than your grid diameter. Put it in the sample solution under a 45=B0 angle and the carbon film will float from the mica. Take care that not all the carbon film floats of the mica! Let the samples adsorb for about 30 sec or 1 min depending on the optical density of your sample. Take the piece of mica back from the solution; the carbon film will reattach to the mica. Rinse in TE-buffer (Tris-EDTA buffer, 20 mM Tris, 1 mM EDTA,. pH 7.0)several times. Subsequentely, float the entire carbon film on 4% aqueous uranyl acetate, leave it for 10 sec and then pick up the carbon film with your grid. Soak access staining solution to such an extent that the surface of the grids is just wet; than immediately dry the grid under a lamp. You can also try to vary the amount of staining solution residing on the grid, i.e. to perform a "deep" stain and a "shallow" stain. Good luck. Manfred
At 12:50 11.05.99 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20 the } stain with the bacteria suspension, but it did not work. } } can anyone send further suggestions??? } } } Sincerelly } } Dr. A.P. Alves de Matos } EM Unit, Pathology Department } Curry Cabral Hospital } Lisbon } } } } }
Although the formulation for GLYCEREGIA is not the same as that for for making Nitroglycerine, nevertheless mixtures of alcohols (and other organics) with nitric acid need to be handled with care.
In the present case, I would not expect an explosion, but as in Henrik Kaker's reply, these mixtures can turn a dark orange colour. This is a warning that there may be a runaway reaction impending - generally the stuff boils over with copious quantities of brown nitric oxide gas being given off. The quantity of HCl in the mixture should prevent that happening too suddenly, but DO work in a fume cupboard and DON'T leave such a mixture around unattended.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I obtained quite good results on E.coli with T4 phage by fixing in 2.5% glutaraldehyde, washing and then staining with about 1% sodium silicotungstate. There were no gaps and staining was good enough to visualise bacteria and fine structure of phage. I used sodium silicotungstate because it seems OK for both bacteria and viruses and seems better behaved than PTA or molybdate and I usually find that uranyl acetate can be very unpredictable in terms of quality and consistency and doesn't keep well.The reason I fixed wasn't to improve preservation - it was just so I could do a time-scale for the infection cycle to produce sections and negative stains
It was over 10 years ago and it's just my opinion, anyway.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
---------- } From: jim-at-proscitech.com.au To: A.P. Alves de Matos Cc: 'microscopy-at-sparc5.microscopy.c
teria are a bit thick for negative staining. Changing the stain concentration helps a little. The rule is that thick specimens appear better with a lower concentration of stain, say 2%. Thin specimens are better with a lower concentration, say 0.5%. You may get better appearance trying different staining solutions, PTA or Ammonium molybdate are good candidates. Don't grow cultures on a shaker, many flagella drop off and even fimbria may suffer.
This is a difficult subject for negative staining because you require both, bacteria and flagella in one picture. Much prettier images can be obtained with shadow casting. 1 Apply bacteria solution to the face of substrated grid. (apply and blot lightly several times to obtain better distribution) Drying would leave too much debris and salts, so use a volatile buffer to wash and maintain molarity. 2 Apply and blot bacteria coated grid repeatedly to a 0.1 M solution of ammonium acetate. 3 After air drying, angle (or rotary) shadow) grid in a vacuum evaporator using fine grain evaporating material. (simultaneous C/Pt or if available high resolution Cr coater as used for FESEM)
Another alternative: If the equipment is available would be FESEM of bacteria and flagella and this could be supplemented with an image of negatively stained flagella only, taken in TEM.
Its a little challenging project, but some nice images could result. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
} I am trying to visualize by negative staining the attachment of phages to bacteria. However the bacteria shrink under the EM } vacum, leaving an artefactual space between the negative stain and the cell wall which do not allow to } take good pictures of the attached phages. } } Here is the protocol I am using: } I am using 2% uranyl acetate as the negative stain The cells are adsorbed to carbon coated formvar grids for 1 minute } Excess is drained with filter paper The grids are floated in the negative stain for 1 minute Excess is drained with filter paper } The grid goes into the TEM } } I have tryed a few modifications such as fixing the material and mixing the stain with the bacteria suspension, but it did not work. } } can anyone send further suggestions???
I have had good results by mixing a drop of1% phosphotungsic acid (PTA) with an equal amount of my liquid bacterial culture on a glass slide, and drying about 0.5 mucroliters of this mixture on a formvar coated grid. You can eliminate distracting salt crystals by first gently centrifuging the bacteria (4 minutes at 5K rpm in an epindorf microcentrifuge) and resuspending the pellet in distilled water. You may not get away with this and still have phages adhering to the cells, but you could try before or after adding the phages. Good luck.
I have always made my lead citrate according to Reynold's original protocol starting with lead nitrate and sodium citrate. Somebody borrowed my bottle of lead nitrate and never returned it but I have a fresh bottle of granular lead citrate. I know there is a modification of Reynold's that starts with lead citrate. Does anyone have the formula and/or opinion on whether it is any different than the original. Thanks, Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
The protocol I have used in the past is to hold the grid in forceps, apply about 10-20 ul of bacterial suspension for 1 min, and then draw off with filter paper. I then apply 20 ul of 2% aqueous phosphotungstic acid, leave on about 10 sec. and draw off and air-dry. You may have to fool around with the dilution of the bacterial suspension, so it's not too thick. Good luck.
Mary McKee
On Tue, 11 May 1999, A.P. Alves de Matos wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear collegues } } I am trying to visualize by negative staining the attachment of phages to } bacteria. } However the bacteria shrink under the EM vacum, leaving an artefactual } space between the negative stain and the cell wall which do not allow to } take good pictures of the attached phages. } } Here is the protocol I am using: } I am using 2% uranyl acetate as the negative stain } The cells are adsorbed to carbon coated formvar grids for 1 minute } Excess is drained with filter paper } The grids are floated in the negative stain for 1 minute } Excess is drained with filter paper } The grid goes into the TEM } } I have tryed a few modifications such as fixing the material and mixing the } stain with the bacteria suspension, but it did not work. } } can anyone send further suggestions??? } } } Sincerelly } } Dr. A.P. Alves de Matos } EM Unit, Pathology Department } Curry Cabral Hospital } Lisbon } } } }
HREM data is fairly easy to work out. We would generally use a 2 second exposure at 300k to 500k times mag. For our film (at 400kV) 20pA/cm2 gives the correct exposure. Ignoring the beam absorbed by the specimen, which should be thin for HREM, this works out at around 2x10^7 to 5x10^7 A/m2 on the specimen (assuming my maths is OK). This should be a reasonable estimate of the current density on the specimen. The beam area would typically be of the order of 0.3um to 1um diameter (approx. 10^-12 to 10^-13 m2).
Nano diffraction is much more variable - it depends on the type of electron gun, probe size, probe defocus, energy spread, etc. However, as a guide we can get a current of 1nA into a spot of 1nm for a FEG which is claimed to be 100 times brighter than a LaB6.
I hope this helps.
Regards, Ron
On Tue, 11 May 1999 09:35:09 -0500 (CDT) WENTAO QIN {s987041-at-jinx.umsl.edu} wrote: } } Hi everyone, } I have heard of radiation damage to the specimens, and I'm curious } how much beam current and current densities are there on the specimen in } HREM imaging and nano-diffraction. Your replies are highly appreciated. } } Wentao } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Call Frank Haze, 800-367-5665. He can furnish you with whatever information is available for your Coolwell chiller. He also has coolant fluid still available. Good luck!
I have been using an HP Scanjet 4c which is about 4 years old and has a resolution of 300dpi. I find that this gives excellent results when printed on my Codonics 1600 dye sub printer. In fact I have found that scanning in at 150 to 200 dpi is sufficent in most cases and results in files of 1/2 the size of a 300dpi file. I may be mistaken but I believe the resolution of most prints in journals is 75 to 125 dpi, so if you scan in at 200 and send them an electronic version, that should suffice.
If you need photographic quality prints, there are numerous printers (ie Epson, HP, etc) that rivel photographs. My only complaint is the paper they use is far from being similar to actual photographic paper.
I am working on making AFM topography and phase images of a series of polymer samples which are made up of varying amounts of polycarbonate, polypropylene and mostly (} 80%) ABS. I have some nice images already (soon to be on our web site) but want to improve the quality. I also want to look at the bulk regions by using cryo-microtomy to create a nice flat face for imaging. Can someone share their experiences with these types of samples and any tips on the microtomy technique (best temperatures, etc.)? I also plan to use plasma etching to see if that will clean up the surface.
Thanks in advance.
Robert J. Plano Staff Analyst, SPM Services Surface Science Labs (650)962-8767, ext. 742
Dear Tom, The formula you are looking for is that of Venable, J. M., and Coggeshall, R. (1965). A simplified lead citrate stain for use in electron microscopy. J. Cell Biol. 25, 407. Add 0.01-0.04 g of lead citrate and 0.1 ml of 10 N NaOH to 10 ml distilled water to a screw-topped vial. Use distilled water that has been freshly boiled for at least 5 min. to ensure solutions are CO2-free. The mixture should be sonicated or vigorously shaken until the lead citrate is dissolved. Centrifuge before use. Use stain immediately. Usual staining time is 1-5 min. My experience is that this formula is much more intense than Reynolds (1963) lead citrate formula. This is helpful for getting more contrast from Spurrs resin sections for which I use 0.035g of lead citrate and stain for 5 -7 min. Word of caution: this stain is very sensitive to CO2. Thus to avoid precipitation make fresh each time of use and keep staining times short, and rinse well in CO2-free distilled water.
} I know there is a modification of Reynold's that starts with } lead citrate. Does anyone have the formula and/or opinion on whether it is } any different than the original.
I think the one you are looking for is the formulation described by Venable and Coggeshall (1965) (J.Cell.Biol. 25, 407-408). I haven't checked the original reference but in our notes the instructions are: weigh out 0.02g lead citrate, add 10ml distilled water followed by 0.1ml 10M NaOH. Shake vigorously until the solution is clear. Allow to stand for at least 30 mins and do not agitate the bottle.
It is also important not to use any of the staining solution from close to the bottom of the bottle.
We used this stain for a while many years ago but reverted to Reynolds which we have found to be less prone to contamination.
I seem to remember that there is another formulation using lead citrate as the starting material, by Sato, I think, but I cannot find it right now!
Regards
Robin
} } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax) }
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za (or eurc-at-giraffe.ru.ac.za) tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
This has possibly been covered so I'll apologise in advance.
Does anyone out there know of a method to accurately determine the thickness of carbon films used on TEM grids? EELS is a possibility but I was looking for a more direct measurement eg using an SPM.
Thanks very much.
Colin Veitch
Instrumentation Scientist Electron Microscopy Textile and Material Technology Group CSIRO Textile and Fibre Technology PO Box 21, BELMONT, Vic. 3216. Australia.
Hello Malcolm and other involved in this discussion
} I obtained quite good results on E.coli with T4 phage
It will always be difficult to optimize negative stain for both viruses and bacteria because they are so different in size and density. For those who would like to see what I mean have a look at some results from one of our recent Microbiology undergrad practicals (http://www.ru.ac.za/emu/im4301.htm) - low res pictures, I'm afraid (can't afford a slow scan CCD camera!) but they do illustrate how the staining conditions vary for the different structures involved.
Regards
Robin
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za (or eurc-at-giraffe.ru.ac.za) tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
I have been looking at some digital cameras for optical microscopy that "integrate". I would have called it "frame averaging" or "pixel averaging", but it appears to be the same image pre-processing technique. Specifically, I saw an "Optronics" camera and it seemed to be able to process low light levels as low as many dark field situations without the memory affects seen on low lux type cameras. Has anyone had any experience with this particular trade name? I am looking for service and dependability technical issues. Also any similar cameras? Please respond to my email. I don't want to flood the server with this issue,especially if any comments are slightly critical or complimentary and appear commercially biased. I don't mind commercial responses. I believe they are a real necessary technical component of this and all issues. Vendors are some of the best and most motivated technical resources we have available to us.(..but that's another subject...)
(I have no commercially beneficial interest in any cameras except the benefits of my laboratory applications, that is I do not sell them. "I am not spam."....did I cover this?)
jeffrey/ 'JD'
___________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com/getjuno.html or call Juno at (800) 654-JUNO [654-5866]
I have been looking at some digital cameras for optical microscopy that "integrate". I would have called it "frame averaging" or "pixel averaging", but it appears to be the same image pre-processing technique. Specifically, I saw an "Optronics" camera and it seemed to be able to process low light levels as low as many dark field situations without the memory affects seen on low lux type cameras. Has anyone had any experience with this particular trade name? I am looking for service and dependability technical issues. Also any similar cameras? Please respond to my email. I don't want to flood the server with this issue,especially if any comments are slightly critical or complimentary and appear commercially biased. I don't mind commercial responses. I believe they are a real necessary technical component of this and all issues. Vendors are some of the best and most motivated technical resources we have available to us.(..but that's another subject...)
(I have no commercially beneficial interest in any cameras except the benefits of my laboratory applications, that is I do not sell them. "I am not spam."....did I cover this?)
jeffrey/ 'JD' Email: WA5EKH-at-juno.com
___________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com/getjuno.html or call Juno at (800) 654-JUNO [654-5866]
Dear Microcopists: I am doing HREM image simulation to compare my HREM image obtained by Philips EM 300 FEG. The parameters needed for calculation are: spherical aberration coeffienct (Cs), Gaussin defocus ("DELTA" for the Defocus spread ), beam convergent semiangle. I appreciate it if any one can give me the numbers of these three parameters for EM 300 FEG microscope. Thank you in advance.
You really need to measure these yourself as they do vary from 'scope to 'scope. I would first get hold of an amorphous Ge sample and do a tilted series of high resolution images for equal and opposite beam tilts (make sure the tilts are calibrated). If you take four tilts +/- X and +/- Y directions you can extract C_s. Using a higher number of tilts you can extract the third order aberration coefficient of your objective lens as well. Here are some references that may help you:
1: Measurement of the spherical aberration coefficient of TEMs by beam tilt induced image displacement: A.J.Koster & A.F. deJong: Ultramicroscopy 38 (1991) pp235-240
2:Improved methods for the determination of the spherical aberration coefficient in HREM from micrographs of an amorphous object: W.M.J.Coene & T.J.J.Denteneer: Ultramicroscopy 38 (1991) pp 225-233
3:Three fold astigmatism in HREM: O Krivanek Ultramicroscopy 55 (1994) pp419-433
4:A spherical aberration corrected 200 keV TEM M Haider, H Rose, S Uhlemann, E Scwann, B Kabius & K Urban Ultramicroscopy 75 (1998) pp53-60
Number 4 is definitely worth a look since it has some nice pictures to demonstrate.
-- ***************************************** Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
Tom, Oops. I gave you the wrong amount for lead. I should proof my own messages more carefully. It's 0.04 gms of lead citrate , not .4 like I previously said. Also if you place sodium hydroxide pellets around the perimeter of the staining dish it will absord the CO2.
I only dream about 300 keV FEG TEMs, so I don't have the numbers right handy. If you don't get them from the list, you might want to get a hold of J. C.H. Spence's book (Experimental High Resolution Electron Microscopy( Amazon says it's out of print)). As I recall there are step by step procedures for determining at least some of these values. A test sample like a holey gird that has seen a few seconds in an Au sputter-coater will help too.
cheers, John MSU
} Dear Microcopists: } I am doing HREM image simulation to compare my HREM image obtained by } Philips EM 300 FEG. The parameters needed for calculation are: } spherical aberration coeffienct (Cs), Gaussin defocus ("DELTA" for the } Defocus spread ), beam convergent semiangle. } I appreciate it if any one can give me the numbers of these three parameters } for EM 300 FEG microscope. Thank you in advance. } } DAI Jiyan } IMRE
I would like to thank those who send any information about the complete name of the journal Boll. Zool. I would also like to tell that it was not a missprint and that the journal=B4s name is Bolletino di Zoologia now known as The Italian Journal o= f Zoology (Ital. J. Zool.) since 1996. Bye.
Gabriel Adriano Rosa Area Microscopia Electronica, Depto. Cs. Biologicas Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA TE -(54-11)-4576-3349 e-mail micros-at-bg.fcen.uba.ar FAX (54-11)-4576-3384
Hi All, We have a Zeiss TEM. Normally when the filament is first switched on with both current and heating knobs are turned to mininimum the voltage indicator shoots up to 2V then immeadiatly starts going down to 1.3-1.4V within no more than 2 minutes, then we start raising the voltage by turning the heating knob until the voltage meter reads between 1.5-1.6V (the recomended voltage stated in the manual). Recently though we have been noticing that the voltage shoots up to 2.2V and stays there for several minutes then starts coming down very slowly and settles at more than 1.6V!!... Needless to say that we have also been burning too many filaments too quickly!! The filament lives have been no more than 10-15hours. We would appreciate any hints or suggestions on why would this happen. We are suspecting an electric problem in the filament control circuitry. Thank you.
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NORAN is actively seeking qualified software engineers or senior software engineers to support existing products (which are both UNIX and NT-based), and to assume a role in new product development. Mostly C and C++ programming at present. Familiarity with scientific programming in the areas of microscopy, imaging, microanalysis, or spectroscopy would be a definite plus.
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-------------------------------------------------- Timothy G. Moeller | NORAN Instruments Inc., Sr. Software Engineer | a ThermoSpectra company {tmoeller-at-noran.com} | {www.noran.com} -------------------------------------------------- DISCLAIMER: The statements and opinions expressed here are my own, and may not represent those of my employer, NORAN, nor of its parent, ThermoSpectra.
We routinely use lead citrate as described in a method by Venable and Coggeshall (1965) in J.Cell Biol.,25: 407 . It is quick to make up and always clean as long as you only use it on the day of making. We keep pre weighed portions of lead citrate ( 0.1 to 0.4 gm) in 10 ml specimen tubes When we need the stain we add 1 ml of N1 sodium hydroxide , wait a few minutes for the lead citrate to dissolve and add 9 ml of distilled water. Use only carbonate free NaOH and fresh distilled water.
Regards
John Manston John Manston Electron Microscope Unit Division of Biomedical Sciences Queen Mary and Westfield College University of London Mile End Road London E1 4NS Tel +171 982 6961 Fax +181 983 0613
We have a Aeiss 10A for sale. Duke University Equipment Appraisal Office and appraised it at $10K. Address at bottom. S Miller
On Thu, 15 Apr 1999, Peter Jordan wrote:
} Date: Thu, 15 Apr 1999 22:26:29 -0700 } From: Peter Jordan {emsi-at-pe.net} } To: EM Listserver {Microscopy-at-sparc5.microscopy.com} } Subject: Used Zeiss 10C } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All: } I am still looking to buy a used Zeiss 10 TEM. There must be one out } there for sale. Please let me know. Thanks. } Peter Jordan, EMSI } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
since you requested information through email, we will send you information about our high resolution digital light microscope cameras that way. If anybody else is interested in getting this information, please give us a call or contact us otherwise.
A technical note: Integrating is not necessarily the same as frame or pixel averaging. Frame or Pixel averaging means, that the camera acquires several frames and they are averaged on the frame grabber or the computer. Integration means, that the camera integrates the light on the chip by allowing longer exposure times.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
} ---------- } From: c j day[SMTP:wa5ekh-at-juno.com] } Sent: Tuesday, May 11, 1999 3:37 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Integrating Digital Cameras? } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } I have been looking at some digital cameras for optical } microscopy that "integrate". I would have called it "frame averaging" } or } "pixel averaging", but it appears to be the same image pre-processing } technique. Specifically, I saw an "Optronics" camera and it seemed } to } be able to process low light levels as low as many dark field } situations } without the memory affects seen on low lux type cameras. Has anyone } had } any experience with this particular trade name? I am looking for } service } and dependability technical issues. Also any similar cameras? Please } respond to my email. I don't want to flood the server with this } issue,especially if any comments are slightly critical or } complimentary } and appear commercially biased. I don't mind commercial responses. I } believe they are a real necessary technical component of this and all } issues. Vendors are some of the best and most motivated technical } resources we have available to us.(..but that's another subject...) } } (I have no commercially beneficial interest in any cameras except the } benefits of my laboratory applications, that is I do not sell them. "I } am } not spam."....did I cover this?) } } jeffrey/ 'JD' } } ___________________________________________________________________ } You don't need to buy Internet access to use free Internet e-mail. } Get completely free e-mail from Juno at } http://www.juno.com/getjuno.html } or call Juno at (800) 654-JUNO [654-5866] }
There are a few companies working on making CMOS (rather than = CCD-based) digital cameras which are perfect for the type of integrated solutions = that you describe.=A0 Unfortunately, the technology will only be ready for = the end of this year/beginning of 2000.=A0 If I were you, I would wait before investing, as it is predicted that digital cameras will take a big leap forward.=A0 Our company, Symagery Microsystems, is working on a chip = called the VCA1280.=A0 You can see the specs on our website at = www.symagery.com.=A0 We do not make the cameras themselves, but the following companies are = working on making CMOS cameras using our chip (I hope): Optronics (www.optronics.se), Wintriss, Xillix, SMD, Costar/JAI.
Hope this helps with your research.=A0 If you have any further = questions please do not hesitate to contact me.
-----Original Message----- } From: c j day [ mailto:wa5ekh-at-juno.com {mailto:wa5ekh-at-juno.com} ] Sent: May 11, 1999 5:59 PM To: microscopy-at-Sparc5.Microscopy.Com
Dear DAI Jiyan,
You can measure all three parameters or use accepted values. Cs depends on the polepiece. Nominal values for the CM300 are 0.65mm for the UT lens, 1.2mm for the ST, and 2.0mm for the T lens. Delta depends on Cc, high-voltage ripple and beam energy spread. Nominal values for Cc are 1.5mm UT, 1.5mm ST, and 2.0 for the T lens. You can measure the effective energy spread with a GIF or PEELS. Then a good estimate of delta is given by DELTA (in Angstroms) = 10 x Cc (in mm) x EnergySpread (ppm RMS). The EnergySpread is in parts-per-million (ppm) and is the Root-Mean-Square -- not the Full-Width-at-Half-Maximum value given by the GIF or PEELS measurement. You can use EnergySpread (ppm RMS) = EnergySpread (ppm FWHM)/2.335. For example, if you measure a FWHM energy spread of 1.0eV on a 300keV TEM, it is equivalent to1.0eV/300,000keV = 3.33 ppm FWHM. Then this is equal to 1.43 ppm RMS. Then the delta is 10 x 1.5 x 1.43 = 21.4 Angstrom. Since there is a small contribution from the lens current ripple, the final figure would be about 25 Angstrom. Beam-convergence depends of condenser lens defocus and condenser aperture. Easily measured by viewing the diffraction pattern of a known test structure with the illumination set as for imaging. For our CM300FEG/UT we use Cs = 0.65mm, delta = 25 A, alpha (convergence) = 0.35 milliradian.
A table of Cs and Cc values is given in Ultramicroscopy 47 (1992) 282-297. Measurement of alpha is also explained in Acta Cryst. 31 (1975) 307-310.
A good way to learn how to use image simulation is to attend the NCEM Summer School on Computing for HREM. Go to http://ncem.lbl.gov/ and click on "Summer School".
-Mike
DAI Jiyan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Microcopists: } I am doing HREM image simulation to compare my HREM image obtained by } Philips EM 300 FEG. The parameters needed for calculation are: } spherical aberration coeffienct (Cs), Gaussin defocus ("DELTA" for the } Defocus spread ), beam convergent semiangle. } I appreciate it if any one can give me the numbers of these three parameters } for EM 300 FEG microscope. Thank you in advance. } } DAI Jiyan } IMRE
Dear colleagues, We want to hire a suitable person for the Central Facility for Electron Microscopy at the Center for Materials Research and Analysis at the University of Nebraska-Lincoln (UNL). The salary will at least likely be in the range of the mid $30k's and is dependent on experience. The medical, dental and retirement benefits package is substantial and includes subsidized tuition for employees to take UNL courses. (BTW, Lincoln is growing, has low unemployment, is great for families, has a good range of live music, dance and theater, and has very good public and other schools.) The Facility provides user-access for ~60 faculty plus their students and other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM and a VG HB501 field-emission STEM, along with a full range of accessories, specimen preparation equipment, computers and darkroom. Some development of new research instrumentation is in progress for mapping of magnetic materials and more development is planned for the Facility. You can find out more about the Facility on our web site (that does need a little work) at URL: http://www.unl.edu/CMRAcfem/ Thanks for passing on the word, Brian Robertson
Analyze/characterize materials using electron microscopy, materials preparation and computer instrumentation. Supervise/train students, faculty and visiting researchers utilizing these methods. Bachelor's with major in physical science, engineering or related field plus three years experience in the operation, repair or design of electron microscopes or other scientific instrumentation. Master's preferred. Must have excellent computer and interpersonal/communication skills. TEM, SEM, x-ray diffraction or materials sample preparation experience preferred. Excellent benefits. Submit cover letter, resume and names, addresses and telephone numbers of three professional references to Professor Brian Robertson, CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position will remain open until a suitable candidate is found. UNL is committed to AA/EEO and ADA/504. If you require accommodation, please call (402) 472-7886.
*********************************************************** Assoc. Prof. Brian W. Robertson Department of Mechanical Engineering and Center for Materials Research and Analysis University of Nebraska-Lincoln, N124 WSEC, 17th & Vine Sts., Lincoln, NE 68588-0656, USA ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 **
I don't want to start a thread of "CMOS vs. CCD" here, that probably belongs to a newsgroup or a different listserver, but I want to respond quickly to one posting regarding CMOS cameras. If you want to respond to this posting, please send me email directly and don't respond to the listserver:
CMOS technology is definitely something to watch, it promises higher integration of the sensor and the electronics, which will evetually lead to less expensive cameras. On the other hand, CCD technology has been around for more than 20 years and is a mature technology with proven quality.
CMOS will be targeted first at the consumer market with inexpensive cameras. Whether these cameras and sensors will be sufficient for scientific equipment remains to be seen.
While I think, there is a tremendous potential in CMOS cameras, I personally think that a good CCD camera is the best choice at the moment. If you can afford to wait for a year or two, the situation may change, but for now, I'd go with CCD.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
} ---------- } From: Marisa Ahmad[SMTP:mahmad-at-semiconductor.com] } Sent: Thursday, May 13, 1999 10:49 AM } To: 'MSA listserver' } Cc: Brigitte Smiley } Subject: FW: Integrating Digital Cameras? } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Jeffrey, } } There are a few companies working on making CMOS (rather than } CCD-based) } digital cameras which are perfect for the type of integrated solutions } that } you describe. Unfortunately, the technology will only be ready for } the end } of this year/beginning of 2000. If I were you, I would wait before } investing, as it is predicted that digital cameras will take a big } leap } forward. Our company, Symagery Microsystems, is working on a chip } called } the VCA1280. You can see the specs on our website at } www.symagery.com. We } do not make the cameras themselves, but the following companies are } working } on making CMOS cameras using our chip (I hope): Optronics } (www.optronics.se), Wintriss, Xillix, SMD, Costar/JAI. } } Hope this helps with your research. If you have any further questions } please do not hesitate to contact me. } } Brigitte Smiley } } brigitte-at-symagery.com {mailto:brigitte-at-symagery.com} } } } } -----Original Message----- } } From: c j day [ mailto:wa5ekh-at-juno.com {mailto:wa5ekh-at-juno.com} ] } Sent: May 11, 1999 5:59 PM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Integrating Digital Cameras? } } } From: WA5EKH-at-juno.com } } I have been looking at some digital cameras for optical } microscopy that "integrate". I would have called it "frame averaging" } or } "pixel averaging", but it appears to be the same image pre-processing } technique. Specifically, I saw an "Optronics" camera and it seemed } to } be able to process low light levels as low as many dark field } situations } without the memory affects seen on low lux type cameras. Has anyone } had } any experience with this particular trade name? I am looking for } service } and dependability technical issues. Also any similar cameras? Please } respond to my email. I don't want to flood the server with this } issue,especially if any comments are slightly critical or } complimentary } and appear commercially biased. I don't mind commercial responses. I } believe they are a real necessary technical component of this and all } issues. Vendors are some of the best and most motivated technical } resources we have available to us.(..but that's another subject...) } } (I have no commercially beneficial interest in any cameras except the } benefits of my laboratory applications, that is I do not sell them. "I } am } not spam."....did I cover this?) } } jeffrey/ 'JD' } Email: WA5EKH-at-juno.com } } } ___________________________________________________________________ } You don't need to buy Internet access to use free Internet e-mail. } Get completely free e-mail from Juno at } http://www.juno.com/getjuno.html } {http://www.juno.com/getjuno.html} } or call Juno at (800) 654-JUNO [654-5866] } } }
Mid-$30K? Is this for real? Is this the going rate for SEM specialists? That is about 1/4 of what I would consider. There must be some subtle differentiation of what is expected from such positions vs. the qualifications and experience of others in the field. Am I the only one shocked about this?
} JOB ANNOUNCEMENT BELOW -- PLEASE POST } } Dear colleagues, } We want to hire a suitable person for the Central Facility for Electron } Microscopy at the Center for Materials Research and Analysis at the } University of Nebraska-Lincoln (UNL). The salary will at least likely be in } the range of the mid $30k's and is dependent on experience. The medical, } dental and retirement benefits package is substantial and includes } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } growing, has low unemployment, is great for families, has a good range of } live music, dance and theater, and has very good public and other schools.) } The Facility provides user-access for ~60 faculty plus their students and } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM } and a VG HB501 field-emission STEM, along with a full range of accessories, } specimen preparation equipment, computers and darkroom. Some development } of new research instrumentation is in progress for mapping of magnetic } materials and more development is planned for the Facility. You can find } out more about the Facility on our web site (that does need a little work) } at URL: http://www.unl.edu/CMRAcfem/ } Thanks for passing on the word, } Brian Robertson } } ********************************************************** } Job Announcement: } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } UNL Center for Materials Research and Analysis } } Analyze/characterize materials using electron microscopy, materials } preparation and computer instrumentation. Supervise/train students, } faculty and visiting researchers utilizing these methods. Bachelor's with } major in physical science, engineering or related field plus three years } experience in the operation, repair or design of electron microscopes or } other scientific instrumentation. Master's preferred. Must have excellent } computer and interpersonal/communication skills. TEM, SEM, x-ray } diffraction or materials sample preparation experience preferred. Excellent } benefits. Submit cover letter, resume and names, addresses and telephone } numbers of three professional references to Professor Brian Robertson, } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } will remain open until a suitable candidate is found. UNL is committed to } AA/EEO and ADA/504. If you require accommodation, please call (402) 472-7886. } } } *********************************************************** } Assoc. Prof. Brian W. Robertson } Department of Mechanical Engineering } and Center for Materials Research and Analysis } University of Nebraska-Lincoln, N124 WSEC, } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 **
****************************************************************************** You have been carefully selected to receive the following as a person interested in opportunities based upon your previous Internet postings or visits to one of our affiliate web sites. If you have received this message in error, please accept our apology as a responsible e-mailer. However, this is a ONE-TIME only announcement and is not intended as a SPAM letter, therefore you need not reply. You are automatically moved to our remove list. Again, if you are not interested, we sincerely apologize for the inconvenience. Thank you. ******************************************************************************
Dear Friend,
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"AS SEEN ON NATIONAL TV"
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*** Print This Now For Future Reference ***
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INSTRUCTIONS:
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1st level--your 10 members with $5.......................................$50 2nd level--10 members from those 10 ($5 x 100)..................$500 3rd level--10 members from those 100 ($5 x 1,000)...........$5,000 4th level--10 members from those 1,000 ($5 x 10,000).....$50,000 THIS TOTALS ----------} $55,550
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AVAILABLE REPORTS
*** Order Each REPORT by NUMBER and NAME *** Notes: -- ALWAYS SEND $5 CASH (U.S. CURRENCY) FOR EACH REPORT. CHECKS NOT ACCEPTED. -- ALWAYS SEND YOUR ORDER VIA FIRST CLASS MAIL. -- Make sure the cash is concealed by wrapping it in at least two sheets of paper. On one of those sheets of paper, include: (a) the number & name of the report you are ordering, (b) your e-mail address, and (c) your name & postal address.
PLACE YOUR ORDER FOR THESE REPORTS NOW:
REPORT #1
----------------------------------------------------------------- "The Insider's Guide to Advertising for Free on the Internet" ORDER REPORT #1 FROM: J. Grainger Via del Perugino, 5 40139 Bologna, Italy ----------------------------------------------------------------- REPORT #2 "The Insider's Guide to Sending Bulk E-mail on the Internet" ORDER REPORT #2 FROM: D. Clark P.O. Box 685 Grangeville, Idaho 83530 ----------------------------------------------------------------- REPORT #3 "The Secrets to Multilevel Marketing on the Internet" ORDER REPORT #3 FROM: Peter Gooijer Galileistraat 33 5621 AE Eindhoven The Netherlands ----------------------------------------------------------------- REPORT #4 "How to become a Millionaire utilizing the Power of Multilevel Marketing and the Internet" ORDER REPORT #4 FROM: J. Russ 8034 Linda Vista Rd. # 1M San Diego, Ca 92111 ----------------------------------------------------------------
About 50,000 new people get online every month!
******* TIPS FOR SUCCESS ******* -- TREAT THIS AS YOUR BUSINESS! Be prompt, professional, and follow the directions accurately. -- Send for the four reports IMMEDIATELY so you will have them when the orders start coming in because: When you receive a $5 order, you MUST send out the requested product/report. -- ALWAYS PROVIDE SAME-DAY SERVICE ON THE ORDERS YOU RECEIVE. -- Be patient and persistent with this program. If you follow the instructions exactly, your results WILL BE SUCCESSFUL! -- ABOVE ALL, HAVE FAITH IN YOURSELF AND KNOW YOU WILL SUCCEED!
******* YOUR SUCCESS GUIDELINES ******* Follow these guidelines to guarantee your success:
If you don't receive 20 orders for REPORT #1 within two weeks, continue
advertising or sending e-mails until you do. Then, a couple of weeks later you should receive at least 100 orders for REPORT#2. If you don 't, continue advertising or sending e-mails until you do. Once you have received 100 or more orders for REPORT #2, YOU CAN RELAX, because the system is already working for you, and the cash will continue to roll in!
THIS IS IMPORTANT TO REMEMBER: Every time your name is moved down on the list, you are placed in front of a DIFFERENT report. You can KEEP TRACK of your PROGRESS by watching which report people are ordering from you. If you want to generate more income, send another batch of e-mails or continue placing ads and start the whole process again! There is no limit to the income you will generate from this business!
Before you make your decision as to whether or not you participate in this program. Please answer one question. DO YOU WANT TO CHANGE YOUR LIFE? If the answer is yes, please look at the following facts about this program:
1. You are selling a product which does not Cost anything to PRODUCE, SHIP OR ADVERTISE. 2. All of your customers pay you in CASH! 3. E-mail is without question the most powerful method of distributing information on earth. This program combines the distribution power of e-mail together with the revenue generating power of multi-level marketing. 4. Your only expense--other than your initial $20 investment--is your time! 5. Virtually all of the income you generate from this program is PURE PROFIT! 6. This program will change your LIFE FOREVER.
ACT NOW!Take your first step toward achieving financial independence. Orderthe reports and follow the program outlined above--SUCCESSwill be yourreward.
Thank you for your time and consideration.
PLEASE NOTE: If you need help with starting a business, registering a business name, learning how income tax is handled, etc., contact your localoffice of the Small Business Administration (a Federal Agency) 1-800-827-5722 for free help and answers to questions. Also, the InternalRevenue Service offers free help via telephone and free seminars aboutbusiness tax requirements. Your earnings are highly dependant on youractivities and advertising. The information contained on this site and in the report constitutes no guarantees stated nor implied. In the event that it is determined that this site or report constitutes a guarantee of any kind, that guarantee is now void. The earnings amounts listed on this site and in the report are estimates only. If you have any questions of the legality of this program, contact the Office of Associate Director for Marketing Practices, Federal Trade Commission, Bureau of Consumer Protection in Washington, DC.
///////////////////////////////////////////////////////////////// Remove at rhobbs12-at-yahoo.com /////////////////////////////////////////////////////////////////
****************************************************************************** You have been carefully selected to receive the following as a person interested in opportunities based upon your previous Internet postings or visits to one of our affiliate web sites. If you have received this message in error, please accept our apology as a responsible e-mailer. However, this is a ONE-TIME only announcement and is not intended as a SPAM letter, therefore you need not reply. You are automatically moved to our remove list. Again, if you are not interested, we sincerely apologize for the inconvenience. Thank you. ******************************************************************************
Dear Friend,
You can earn $50,000 or more in next the 90 days sending e-mail. Seem impossible? Read on for details.
"AS SEEN ON NATIONAL TV"
Thank you for your time and interest. This is the letter you've been reading about in the news lately. Due to the popularity of this letter on the Internet, a major nightly news program recently devoted an entire show to the investigation of the program described below to see if it really can make people money.
The show also investigated whether or not the program was legal. Their findings proved once and for all that there are absolutely no laws prohibiting the participation in the program. This has helped to show people that this is a simple, harmless and fun way to make some extra money at home.
The results of this show have been truly remarkable. So many people are participating that those involved are doing much better than ever before. Since everyone makes more as more people try it out, its been very exciting to be a part of lately. You will understand once you experience it.
HERE IT IS BELOW:
*** Print This Now For Future Reference ***
The following income opportunity is one you may be interested in taking a look at. It can be started with VERY LITTLE investment and the income return is TREMENDOUS!!!
$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ If you would like to make at least $50,000 in less than 90 days ! Please read the enclosed program...THEN READ IT AGAIN!!! $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$
THIS IS A LEGITIMATE, LEGAL, MONEY MAKING OPPORTUNITY.It does not require you to come into contact with people, do any hard work, and best of all, you never have to leave the house except to get the mail. If you believe that someday you'll get that big break that you 'vebeen waiting for, THIS IS IT! Simply follow the instructions, andyour dreams will come true. This multi-level e-mail order marketingprogram works perfectly...100% EVERY TIME.
E-mail is the sales tool of the future. Take advantage of this non-commercialized method of advertising NOW!!! The longer you wait, the more people will be doing business using e-mail. Get your piece of this action!!!
MULTI-LEVEL MARKETING (MLM) has finally gained respectability. It is being taught in the Harvard Business School, and both Stanford Research and the Wall Street Journal have stated that between 50% and 65% of all goods and services will be sold through multi-level methods by the mid to late 1990's. This is a Multi-Billion Dollar industry and of the 500,000 millionaires in the U.S., 20% (100,000) made their fortune in the last several years in MLM. Moreover, statistics show 45 people become millionaires everyday through Multi-Level Marketing.
You may have heard this story before, but over the summer Donald Trump made an appearance on the David Letterman show. Dave asked him what he would do if he lost everything and had to start over from scratch. Without hesitating, Trump said he would find a good network marketing company and get to work. The audience started to hoot and boo him. He looked out at the audience and dead-panned his response: "That's why I'm sitting up here and you are all sitting out there!"
The enclosed information is something I almost let slip through my fingers. Fortunately, sometime later I re-read everything and gave somethought and study to it. My name is Johnathon Rourke. Two years ago, the corporation I worked at for the past twelve years down-sized and my position was eliminated. After unproductive job interviews, I decided to open my own business. Over the past year, I incurred many unforeseen financial problems. I owed my family, friends and creditors over $35,000. The economy was taking a toll on my business and I just couldn't seem to make ends meet. I had to refinance and borrow against my home to support my family and struggling business. AT THAT MOMENT something significant happened in my life and I am writing to share the experience in hopes that this will change your life FOREVER FINANCIALLY!!!
In mid December, I received this program via e-mail. Six month's prior to receiving this program I had been sending away for information on various business opportunities. All of the programs I received, in my opinion, were not cost effective. They were either too difficult for me to comprehend or the initial investment was too much for me to risk to see if they would work or not. One claimed that I would make a million dollars in one year...it didn't tell me I'd have to write a book to make it!
But like I was saying, in December of 1997 I received this program. I didn't send for it, or ask for it, they just got my name off a mailing list.THANK GOODNESS FOR THAT!!! After reading it several times, to make sure I was reading it correctly, I couldn't believe my eyes. Here was a MONEY MAKING PHENOMENON. I could invest as much as I wanted to start, without putting me further into debt. After I got a pencil and paper and figured it out, I would at least get my money back. But like most of you I was still a little skeptical and a little worried about the legal aspects of it all. So I checked it out with the U.S. Post Office (1-800-725-2161 24-hrs) and they confirmed that it is indeed legal! After determining the program was LEGAL and NOT A CHAIN LETTER, I decided "WHY NOT."
Initially I sent out 10,000 e-mails. It cost me about $15 for my time on-line. The great thing about e-mail is that I don't need any money for printing to send out the program, and because all of my orders are fulfilled via e-mail, my only expense is my time. I am telling you like it is I hope it doesn't turn you off, but I promised myself that I would not "rip-off" anyone, no matter how much money it made me.
In less than one week, I was starting to receive orders for REPORT #1 By January 13, I had received 26 orders for REPORT #1. Your goal is to "RECEIVE at least 20 ORDERS FOR REPORT #1 WITHIN 2 WEEKS. IF YOU DON'T, SEND OUT MORE PROGRAMS UNTIL YOU DO!" My first step in making $50,000 in 90 days was done. By January 30, I had received 196 orders for REPORT #2. Your goal is to "RECEIVE AT LEAST 100+ ORDERS FOR REPORT #2 WITHIN 2 WEEKS. IF NOT, SEND OUT MORE PROGRAMS UNTIL YOU DO. ONCE YOU HAVE 100 ORDERS, THE REST IS EASY, RELAX, YOU WILL MAKE YOUR $50,000 GOAL." Well, I had 196 orders for REPORT #2, 96 more than I needed. So I sat back and relaxed. By March 1, of my e-mailing of 10,000, I received $58,000 with more coming in every day.
I paid off ALL my debts and bought a much needed new car. Please take time to read the attached program, IT WILL CHANGE YOUR LIFE FOREVER!! ! Remember, it won't work if you don't try it. This program does work , but you must follow it EXACTLY! Especially the rules of not trying to place your name in a different place. It won't work and you'll lose out on a lot of money! In order for this program to work, you must meet your goal of 20+ orders for REPORT #1, and 100+ orders for REPORT #2 and you will make $50,000 or more in 90 days. I AM LIVING PROOF THAT IT WORKS!!!
If you choose not to participate in this program, I am sorry. It really is a great opportunity with little cost or risk to you. If you choose to participate, follow the program and you will be on your way to financial security. If you are a fellow business owner and are in financial trouble like I was, or you want to start your own business, consider this a sign. I DID!
Sincerely, Johnathon Rourke
A PERSONAL NOTE FROM THE ORIGINATOR OF THIS PROGRAM:
By the time you have read the enclosed program and reports, you should have concluded that such a program, and one that is legal, could not have been created by an amateur.
Let me tell you a little about myself. I had a profitable business for 10 years. Then in 1979 my business began falling off. I was doing the same things that were previously successful for me, but it wasn't working. Finally, I figured it out. It wasn't me, it was the economy. Inflation and recession had replaced the stable economy that had been with us since 1945.I don't have to tell you what happened to the unemployment rate... because many of you know from first hand experience. There were more failures and bankruptcies than ever before.
The middle class was vanishing. Those who knew what they were doing invested wisely and moved up. Those who did not, including those who never had anything to save or invest, were moving down into the ranks of the poor. As the saying goes, "THE RICH GET RICHER AND THE POOR GET POORER." The traditional methods of making money will never allow you to "move up" or "get rich", inflation will see to that.
You have just received information that can give you financial freedom for the rest of your life, with "NO RISK" and "JUST A LITTLE BIT OF EFFORT." You can make more money in the next few months than you have ever imagined. I should also point out that I will not see a penny of this money, nor anyone else who has provided a testimonial for this program. I have already made over 4 MILLION DOLLARS!I have retired from the program after sending thousands and thousands of programs.
Follow the program EXACTLY AS INSTRUCTED. Do not change it in any way . It works exceedingly well as it is now. Remember to e-mail a copy of this exciting report to everyone you can think of. One of the people you send this to may send out 50,000...and your name will be on everyone of them!
Remember though, the more you send out the more potential customers you will reach.
So my friend, I have given you the ideas, information, materials and opportunity to become financially independent. IT IS UP TO YOU NOW!
"THINK ABOUT IT"
Before you delete this program from your mailbox, as I almost did, take a little time to read it and REALLY THINK ABOUT IT. Get a pencil and figure out what could happen when YOU participate. Figure out the worst possible response and no matter how you calculate it, you will still make a lot of money! You will definitely get back what you invested. Any doubts you have will vanish when your first orders come in. IT WORKS!
Jody Jacobs, Richmond, VA
HERE'S HOW THIS AMAZING PROGRAM WILL MAKE YOU THOUSANDS OF DOLLAR$
INSTRUCTIONS:
This method of raising capital REALLY WORKS 100% EVERY TIME. I am sure that you could use up to $50,000 or more in the next 90 days. Before you say "BULL... ", please read this program carefully.
This is not a chain letter, but a perfectly legal money making opportunity. Basically, this is what you do: As with all multi-level businesses, we build our business by recruiting new partners and selling our products. Every state in the USA allows you to recruit new multi-level business partners, and we offer a product for EVERY dollar sent. YOUR ORDERS COME BY MAIL AND ARE FILLED BY E-MAIL, so you are not involved in personal selling. You do it privately in your own home, store or office. This is the GREATEST Multi-Level Mail Order Marketing anywhere.
This is what you MUST do:
1. Order all 4 reports shown on the list below (you can't sell them if youdon't order them). -- For each report, send $5.00 CASH, the NAME & NUMBER OF THE REPORT YOU ARE ORDERING, YOUR E-MAIL ADDRESS, and YOUR NAME & RETURN ADDRESS (in case of a problem) to the person whose name appears on the list next to the report. MAKE SURE YOUR RETURN ADDRESS IS ON YOUR ENVELOPE IN CASE OF ANY MAIL PROBLEMS! -- When you place your order, make sure you order each of the four reports. You will need all four reports so that you can save them on your computer and resell them. -- Within a few days you will receive, via e-mail, each of the four reports. Save them on your computer so they will be accessible for you to send to the 1,000's of people who will order them from you.
2. IMPORTANT DO NOT alter the names of the people who are listed next to each report, or their sequence on the list, in any way other than is instructed below in steps "a" through "f" or you will lose out on the majority of your profits. Once you understand the way this works, you'll also see how it doesn't work if you change it. Remember, this method has been tested,and if you alter it, it will not work. a. Look below for the listing of available reports. b. After you've ordered the four reports, take this advertisement and remove the name and address under REPORT #4. This person has made it through the cycle and is no doubt counting their $50,000! c. Move the name and address under REPORT #3 down to REPORT #4. d. Move the name and address under REPORT #2 down to REPORT #3. e. Move the name and address under REPORT #1 down to REPORT #2. f. Insert your name/address in the REPORT #1 position.
Please make sure you COPY ALL INFORMATION, every name and address, ACCURATELY!
3. Take this entire letter, including the modified list of names, and save it to your computer. Make NO changes to the instruction portion of this letter.
Your cost to participate in this is practically nothing (surely you can afford $20). You obviously already have an Internet connection and e-mail is FREE!
There are two primary methods of building your downline:
METHOD #1: SENDING BULK E-MAIL
Let's say that you decide to start small, just to see how it goes, and we'll assume you and all those involved send out only 2,000 programs each. Let's also assume that the mailing receives a 0.5% response. Using a good list the response could be much better. Also, many people will send out hundreds of thousands of programs instead of 2,000. But continuing with this example, you send out only 2,000 programs. With a 0.5% response, that is only 10 orders for REPORT #1. Those 10 people respond by sending out 2,000 programs each for a total of 20,000. Out of those 0.5%, 100 people respond and order REPORT #2. Those 100 mail out 2,000 programs each for a total of 200,000. The 0.5% response to that is 1,000 orders for REPORT #3. Those 1,000 send out 2,000 programs each for a 2,000,000 total. The 0.5% response to that is 10,000 orders for REPORT #4. That's 10,000 $5 bills for you. CASH!!! Your total income in this example is $50 + $500 + $5,000 + $50,000 for a total of $55,550!!! REMEMBER FRIEND, THIS IS ASSUMING 1,990 OUT OF THE 2,000 PEOPLE YOU MAIL TO WILL DO ABSOLUTELY NOTHING AND TRASH THIS PROGRAM! DARE TO THINK FOR A MOMENT WHAT WOULD HAPPEN IF EVERYONE, OR HALF SENT OUT 100,000 PROGRAMS INSTEAD OF 2,000. Believe me, many people will do justthat, and more! By the way, your cost to participate in this is practically nothing. You obviously already have an Internet connection and e-mail is FREE!!! REPORT #2 will show you the best methods for bulk e-mailing, tell you where to obtain free bulk e-mail software and where to obtain e-mail lists.
METHOD #2 - PLACING FREE ADS ON THE INTERNET
Advertising on the internet is very, very inexpensive, and there are HUNDREDS of FREE places to advertise. Let's say you decide to start small just to see how well it works. Assume your goal is to get ONLY 10 people to participate on your first level. (Placing a lot of FREE ads on the Internet will EASILY get a larger response.) Also assume that everyone else in YOUR ORGANIZATION gets ONLY 10 downline members. Follow this example to achieve the STAGGERING results below:
1st level--your 10 members with $5.......................................$50 2nd level--10 members from those 10 ($5 x 100)..................$500 3rd level--10 members from those 100 ($5 x 1,000)...........$5,000 4th level--10 members from those 1,000 ($5 x 10,000).....$50,000 THIS TOTALS ----------} $55,550
Remember friends, this assumes that the people who participate only recruit 10 people each. Think for a moment what would happen if they got 20 people to participate! Most people get 100's of participants! THINK ABOUT IT! For every $5.00 you receive, all you must do is e-mail them the report they ordered. THAT'S IT! ALWAYS PROVIDE SAME-DAY SERVICE ON ALL ORDERS! This will guarantee that the e-mail THEY send out with YOUR name and address on it will be prompt because they can't advertise until they receive the report!
AVAILABLE REPORTS
*** Order Each REPORT by NUMBER and NAME *** Notes: -- ALWAYS SEND $5 CASH (U.S. CURRENCY) FOR EACH REPORT. CHECKS NOT ACCEPTED. -- ALWAYS SEND YOUR ORDER VIA FIRST CLASS MAIL. -- Make sure the cash is concealed by wrapping it in at least two sheets of paper. On one of those sheets of paper, include: (a) the number & name of the report you are ordering, (b) your e-mail address, and (c) your name & postal address.
PLACE YOUR ORDER FOR THESE REPORTS NOW:
REPORT #1
----------------------------------------------------------------- "The Insider's Guide to Advertising for Free on the Internet" ORDER REPORT #1 FROM: J. Grainger Via del Perugino, 5 40139 Bologna, Italy ----------------------------------------------------------------- REPORT #2 "The Insider's Guide to Sending Bulk E-mail on the Internet" ORDER REPORT #2 FROM: D. Clark P.O. Box 685 Grangeville, Idaho 83530 ----------------------------------------------------------------- REPORT #3 "The Secrets to Multilevel Marketing on the Internet" ORDER REPORT #3 FROM: Peter Gooijer Galileistraat 33 5621 AE Eindhoven The Netherlands ----------------------------------------------------------------- REPORT #4 "How to become a Millionaire utilizing the Power of Multilevel Marketing and the Internet" ORDER REPORT #4 FROM: J. Russ 8034 Linda Vista Rd. # 1M San Diego, Ca 92111 ----------------------------------------------------------------
About 50,000 new people get online every month!
******* TIPS FOR SUCCESS ******* -- TREAT THIS AS YOUR BUSINESS! Be prompt, professional, and follow the directions accurately. -- Send for the four reports IMMEDIATELY so you will have them when the orders start coming in because: When you receive a $5 order, you MUST send out the requested product/report. -- ALWAYS PROVIDE SAME-DAY SERVICE ON THE ORDERS YOU RECEIVE. -- Be patient and persistent with this program. If you follow the instructions exactly, your results WILL BE SUCCESSFUL! -- ABOVE ALL, HAVE FAITH IN YOURSELF AND KNOW YOU WILL SUCCEED!
******* YOUR SUCCESS GUIDELINES ******* Follow these guidelines to guarantee your success:
If you don't receive 20 orders for REPORT #1 within two weeks, continue
advertising or sending e-mails until you do. Then, a couple of weeks later you should receive at least 100 orders for REPORT#2. If you don 't, continue advertising or sending e-mails until you do. Once you have received 100 or more orders for REPORT #2, YOU CAN RELAX, because the system is already working for you, and the cash will continue to roll in!
THIS IS IMPORTANT TO REMEMBER: Every time your name is moved down on the list, you are placed in front of a DIFFERENT report. You can KEEP TRACK of your PROGRESS by watching which report people are ordering from you. If you want to generate more income, send another batch of e-mails or continue placing ads and start the whole process again! There is no limit to the income you will generate from this business!
Before you make your decision as to whether or not you participate in this program. Please answer one question. DO YOU WANT TO CHANGE YOUR LIFE? If the answer is yes, please look at the following facts about this program:
1. You are selling a product which does not Cost anything to PRODUCE, SHIP OR ADVERTISE. 2. All of your customers pay you in CASH! 3. E-mail is without question the most powerful method of distributing information on earth. This program combines the distribution power of e-mail together with the revenue generating power of multi-level marketing. 4. Your only expense--other than your initial $20 investment--is your time! 5. Virtually all of the income you generate from this program is PURE PROFIT! 6. This program will change your LIFE FOREVER.
ACT NOW!Take your first step toward achieving financial independence. Orderthe reports and follow the program outlined above--SUCCESSwill be yourreward.
Thank you for your time and consideration.
PLEASE NOTE: If you need help with starting a business, registering a business name, learning how income tax is handled, etc., contact your localoffice of the Small Business Administration (a Federal Agency) 1-800-827-5722 for free help and answers to questions. Also, the InternalRevenue Service offers free help via telephone and free seminars aboutbusiness tax requirements. Your earnings are highly dependant on youractivities and advertising. The information contained on this site and in the report constitutes no guarantees stated nor implied. In the event that it is determined that this site or report constitutes a guarantee of any kind, that guarantee is now void. The earnings amounts listed on this site and in the report are estimates only. If you have any questions of the legality of this program, contact the Office of Associate Director for Marketing Practices, Federal Trade Commission, Bureau of Consumer Protection in Washington, DC.
///////////////////////////////////////////////////////////////// Remove at rhobbs12-at-yahoo.com /////////////////////////////////////////////////////////////////
****************************************************************************** You have been carefully selected to receive the following as a person interested in opportunities based upon your previous Internet postings or visits to one of our affiliate web sites. If you have received this message in error, please accept our apology as a responsible e-mailer. However, this is a ONE-TIME only announcement and is not intended as a SPAM letter, therefore you need not reply. You are automatically moved to our remove list. Again, if you are not interested, we sincerely apologize for the inconvenience. Thank you. ******************************************************************************
Dear Friend,
You can earn $50,000 or more in next the 90 days sending e-mail. Seem impossible? Read on for details.
"AS SEEN ON NATIONAL TV"
Thank you for your time and interest. This is the letter you've been reading about in the news lately. Due to the popularity of this letter on the Internet, a major nightly news program recently devoted an entire show to the investigation of the program described below to see if it really can make people money.
The show also investigated whether or not the program was legal. Their findings proved once and for all that there are absolutely no laws prohibiting the participation in the program. This has helped to show people that this is a simple, harmless and fun way to make some extra money at home.
The results of this show have been truly remarkable. So many people are participating that those involved are doing much better than ever before. Since everyone makes more as more people try it out, its been very exciting to be a part of lately. You will understand once you experience it.
HERE IT IS BELOW:
*** Print This Now For Future Reference ***
The following income opportunity is one you may be interested in taking a look at. It can be started with VERY LITTLE investment and the income return is TREMENDOUS!!!
$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ If you would like to make at least $50,000 in less than 90 days ! Please read the enclosed program...THEN READ IT AGAIN!!! $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$
THIS IS A LEGITIMATE, LEGAL, MONEY MAKING OPPORTUNITY.It does not require you to come into contact with people, do any hard work, and best of all, you never have to leave the house except to get the mail. If you believe that someday you'll get that big break that you 'vebeen waiting for, THIS IS IT! Simply follow the instructions, andyour dreams will come true. This multi-level e-mail order marketingprogram works perfectly...100% EVERY TIME.
E-mail is the sales tool of the future. Take advantage of this non-commercialized method of advertising NOW!!! The longer you wait, the more people will be doing business using e-mail. Get your piece of this action!!!
MULTI-LEVEL MARKETING (MLM) has finally gained respectability. It is being taught in the Harvard Business School, and both Stanford Research and the Wall Street Journal have stated that between 50% and 65% of all goods and services will be sold through multi-level methods by the mid to late 1990's. This is a Multi-Billion Dollar industry and of the 500,000 millionaires in the U.S., 20% (100,000) made their fortune in the last several years in MLM. Moreover, statistics show 45 people become millionaires everyday through Multi-Level Marketing.
You may have heard this story before, but over the summer Donald Trump made an appearance on the David Letterman show. Dave asked him what he would do if he lost everything and had to start over from scratch. Without hesitating, Trump said he would find a good network marketing company and get to work. The audience started to hoot and boo him. He looked out at the audience and dead-panned his response: "That's why I'm sitting up here and you are all sitting out there!"
The enclosed information is something I almost let slip through my fingers. Fortunately, sometime later I re-read everything and gave somethought and study to it. My name is Johnathon Rourke. Two years ago, the corporation I worked at for the past twelve years down-sized and my position was eliminated. After unproductive job interviews, I decided to open my own business. Over the past year, I incurred many unforeseen financial problems. I owed my family, friends and creditors over $35,000. The economy was taking a toll on my business and I just couldn't seem to make ends meet. I had to refinance and borrow against my home to support my family and struggling business. AT THAT MOMENT something significant happened in my life and I am writing to share the experience in hopes that this will change your life FOREVER FINANCIALLY!!!
In mid December, I received this program via e-mail. Six month's prior to receiving this program I had been sending away for information on various business opportunities. All of the programs I received, in my opinion, were not cost effective. They were either too difficult for me to comprehend or the initial investment was too much for me to risk to see if they would work or not. One claimed that I would make a million dollars in one year...it didn't tell me I'd have to write a book to make it!
But like I was saying, in December of 1997 I received this program. I didn't send for it, or ask for it, they just got my name off a mailing list.THANK GOODNESS FOR THAT!!! After reading it several times, to make sure I was reading it correctly, I couldn't believe my eyes. Here was a MONEY MAKING PHENOMENON. I could invest as much as I wanted to start, without putting me further into debt. After I got a pencil and paper and figured it out, I would at least get my money back. But like most of you I was still a little skeptical and a little worried about the legal aspects of it all. So I checked it out with the U.S. Post Office (1-800-725-2161 24-hrs) and they confirmed that it is indeed legal! After determining the program was LEGAL and NOT A CHAIN LETTER, I decided "WHY NOT."
Initially I sent out 10,000 e-mails. It cost me about $15 for my time on-line. The great thing about e-mail is that I don't need any money for printing to send out the program, and because all of my orders are fulfilled via e-mail, my only expense is my time. I am telling you like it is I hope it doesn't turn you off, but I promised myself that I would not "rip-off" anyone, no matter how much money it made me.
In less than one week, I was starting to receive orders for REPORT #1 By January 13, I had received 26 orders for REPORT #1. Your goal is to "RECEIVE at least 20 ORDERS FOR REPORT #1 WITHIN 2 WEEKS. IF YOU DON'T, SEND OUT MORE PROGRAMS UNTIL YOU DO!" My first step in making $50,000 in 90 days was done. By January 30, I had received 196 orders for REPORT #2. Your goal is to "RECEIVE AT LEAST 100+ ORDERS FOR REPORT #2 WITHIN 2 WEEKS. IF NOT, SEND OUT MORE PROGRAMS UNTIL YOU DO. ONCE YOU HAVE 100 ORDERS, THE REST IS EASY, RELAX, YOU WILL MAKE YOUR $50,000 GOAL." Well, I had 196 orders for REPORT #2, 96 more than I needed. So I sat back and relaxed. By March 1, of my e-mailing of 10,000, I received $58,000 with more coming in every day.
I paid off ALL my debts and bought a much needed new car. Please take time to read the attached program, IT WILL CHANGE YOUR LIFE FOREVER!! ! Remember, it won't work if you don't try it. This program does work , but you must follow it EXACTLY! Especially the rules of not trying to place your name in a different place. It won't work and you'll lose out on a lot of money! In order for this program to work, you must meet your goal of 20+ orders for REPORT #1, and 100+ orders for REPORT #2 and you will make $50,000 or more in 90 days. I AM LIVING PROOF THAT IT WORKS!!!
If you choose not to participate in this program, I am sorry. It really is a great opportunity with little cost or risk to you. If you choose to participate, follow the program and you will be on your way to financial security. If you are a fellow business owner and are in financial trouble like I was, or you want to start your own business, consider this a sign. I DID!
Sincerely, Johnathon Rourke
A PERSONAL NOTE FROM THE ORIGINATOR OF THIS PROGRAM:
By the time you have read the enclosed program and reports, you should have concluded that such a program, and one that is legal, could not have been created by an amateur.
Let me tell you a little about myself. I had a profitable business for 10 years. Then in 1979 my business began falling off. I was doing the same things that were previously successful for me, but it wasn't working. Finally, I figured it out. It wasn't me, it was the economy. Inflation and recession had replaced the stable economy that had been with us since 1945.I don't have to tell you what happened to the unemployment rate... because many of you know from first hand experience. There were more failures and bankruptcies than ever before.
The middle class was vanishing. Those who knew what they were doing invested wisely and moved up. Those who did not, including those who never had anything to save or invest, were moving down into the ranks of the poor. As the saying goes, "THE RICH GET RICHER AND THE POOR GET POORER." The traditional methods of making money will never allow you to "move up" or "get rich", inflation will see to that.
You have just received information that can give you financial freedom for the rest of your life, with "NO RISK" and "JUST A LITTLE BIT OF EFFORT." You can make more money in the next few months than you have ever imagined. I should also point out that I will not see a penny of this money, nor anyone else who has provided a testimonial for this program. I have already made over 4 MILLION DOLLARS!I have retired from the program after sending thousands and thousands of programs.
Follow the program EXACTLY AS INSTRUCTED. Do not change it in any way . It works exceedingly well as it is now. Remember to e-mail a copy of this exciting report to everyone you can think of. One of the people you send this to may send out 50,000...and your name will be on everyone of them!
Remember though, the more you send out the more potential customers you will reach.
So my friend, I have given you the ideas, information, materials and opportunity to become financially independent. IT IS UP TO YOU NOW!
"THINK ABOUT IT"
Before you delete this program from your mailbox, as I almost did, take a little time to read it and REALLY THINK ABOUT IT. Get a pencil and figure out what could happen when YOU participate. Figure out the worst possible response and no matter how you calculate it, you will still make a lot of money! You will definitely get back what you invested. Any doubts you have will vanish when your first orders come in. IT WORKS!
Jody Jacobs, Richmond, VA
HERE'S HOW THIS AMAZING PROGRAM WILL MAKE YOU THOUSANDS OF DOLLAR$
INSTRUCTIONS:
This method of raising capital REALLY WORKS 100% EVERY TIME. I am sure that you could use up to $50,000 or more in the next 90 days. Before you say "BULL... ", please read this program carefully.
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///////////////////////////////////////////////////////////////// Remove at rhobbs12-at-yahoo.com /////////////////////////////////////////////////////////////////
-} We have a Zeiss TEM. Normally when the filament is first switched on with } both current and heating knobs are turned to mininimum the voltage indicator } shoots up to 2V then immeadiatly starts going down to 1.3-1.4V within no more } than 2 minutes, then we start raising the voltage by turning the heating knob } until the voltage meter reads between 1.5-1.6V (the recomended voltage stated in } the manual). } Recently though we have been noticing that the voltage shoots up to 2.2V and } stays there for several minutes then starts coming down very slowly and settles } at more than 1.6V!!... Needless to say that we have also been burning too many } filaments too quickly!! The filament lives have been no more than 10-15hours. } We would appreciate any hints or suggestions on why would this happen. We } are suspecting an electric problem in the filament control circuitry. Thank } you.
I have seen this happen in other filaments and I and a couple of EE's are at a loss to explain it. The solution we came up wiht was to use a current controlled power supply and slowly bring up the current insted of the voltage. You may have to switch power supplies when the heater is up to temperature.
Good luck Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
----------------------------------------------------. } } } Mid-$30K? Is this for real? Is this the going rate for SEM } specialists? That is about 1/4 of what I would consider. } There must be some subtle differentiation of what is expected } from such positions vs. the qualifications and experience of } others in the field. Am I the only one shocked about this? }
We have graduates with experience in electrical engineering technology graduating for 50k & 60 K my sun is still fishing he is probably going to take a lower paid job to get into embed systems but has passed up 70k as a system admin. He does have 10 years experience in the field. All but the 50K kid are older and have some real work behind them. Last I heard the 60K was the top at the university. It annoys some of the engineers that look at EET as where you go when you can't cut engineering.
Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
Sres MSA: Dado que mi tema de investigaci=F3n es el an=E1lisis de propiedades mec=E1nicas de pol=EDmeros semicritalinos, necesitar=EDa contactarme con alguna persona que pueda informarme sobre estudios de microestructura de los mismos.Desde ya muchas gracias.=20 Ing elena Brandaleze.
Judging from last summer's C&E News salary survey (7/27/98) $35k is about right for a new BS chemist. The average salary for 10 years experience goes up to $48k. The MS person that they prefer will cost substantially more however - $45-55k.
Now a second question is whether they'll be happy with a new BS in this position - I think not, but as in many things you do get what you pay for.
Richard Shalvoy Arch Chemicals (formerly Olin Corporation) Cheshire, CT
-----Original Message----- } From: Dr. Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Thursday, May 13, 1999 10:59 PM To: MSA listserver
Keep in mind that this is a university doing the hiring in a field where (at least in biological EM positions) people tend to enjoy their work. Also, my first job at a university only had a 7.5 hour workday. I agree that it's disappointing, but they figure someone with minimal graduate work/experience receiving full medical, tuition reimbursement, 4 weeks of vacation, and the casual atmosphere of a university, they can get away with it. Especially for someone with minimal prior experience.
I'm not trying to justify it, just trying to offer a window into how they are thinking. They will likely fill the position with some willing individual looking for that extra job experience and generous vacation time.
Gregg Sobocinski Parke-Davis Pharmaceuticals Ann Arbor, Michigan USA Gregg.Sobocinski-at-wl.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ********************************************************** } Job Announcement: } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } UNL Center for Materials Research and Analysis } } Analyze/characterize materials using electron microscopy, materials } preparation and computer instrumentation. Supervise/train students, } faculty and visiting researchers utilizing these methods. Bachelor's with } major in physical science, engineering or related field plus three years } experience in the operation, repair or design of electron microscopes or } other scientific instrumentation. Master's preferred. Must have excellent } computer and interpersonal/communication skills. TEM, SEM, x-ray } diffraction or materials sample preparation experience preferred. Excellent } benefits. Submit cover letter, resume and names, addresses and telephone } numbers of three professional references to Professor Brian Robertson, } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } will remain open until a suitable candidate is found. UNL is committed to } AA/EEO and ADA/504. If you require accommodation, please call (402) 472-7886. }
We are to begin the process of using tripod polishing for preparing TEM samples, and request vendors to contact me with regards to types of polishers available, style, cost, ease of use.
I also welcome users of this method to comment online (or email directly) as to the ease of use and technique of sample preparation, since our lab has not gone this route before.
thanks in advance
Fred
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
This is similar to what I make and I do SEM, TEM, LM and immuno. at LM and TEM levels for an animal research lab at UTSW in Dallas. I have over 15 years of experience, a MS, and the position I hold is considered a non-tenure, PhD position. This is reality for working in academia in the midwest (I should mention I started out at the U of MN, where I was paid alot less.). Why do I stay? Cost of living here is low, so you can live on it and I like the field so much I'm getting a PhD in neuroscience to continue to grow in it.
Karen Pawlowski Sr. Research Assoc. UT Southwestern, Dallas, TX PhD candidate, UT Dallas, Dallas, TX
On Thu, 13 May 1999, Dr. Gary Gaugler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Mid-$30K? Is this for real? Is this the going rate for SEM } specialists? That is about 1/4 of what I would consider. } There must be some subtle differentiation of what is expected } from such positions vs. the qualifications and experience of } others in the field. Am I the only one shocked about this? } } } } } JOB ANNOUNCEMENT BELOW -- PLEASE POST } } } } Dear colleagues, } } We want to hire a suitable person for the Central Facility for Electron } } Microscopy at the Center for Materials Research and Analysis at the } } University of Nebraska-Lincoln (UNL). The salary will at least likely be in } } the range of the mid $30k's and is dependent on experience. The medical, } } dental and retirement benefits package is substantial and includes } } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } } growing, has low unemployment, is great for families, has a good range of } } live music, dance and theater, and has very good public and other schools.) } } The Facility provides user-access for ~60 faculty plus their students and } } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM } } and a VG HB501 field-emission STEM, along with a full range of accessories, } } specimen preparation equipment, computers and darkroom. Some development } } of new research instrumentation is in progress for mapping of magnetic } } materials and more development is planned for the Facility. You can find } } out more about the Facility on our web site (that does need a little work) } } at URL: http://www.unl.edu/CMRAcfem/ } } Thanks for passing on the word, } } Brian Robertson } } } } ********************************************************** } } Job Announcement: } } } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } } } UNL Center for Materials Research and Analysis } } } } Analyze/characterize materials using electron microscopy, materials } } preparation and computer instrumentation. Supervise/train students, } } faculty and visiting researchers utilizing these methods. Bachelor's with } } major in physical science, engineering or related field plus three years } } experience in the operation, repair or design of electron microscopes or } } other scientific instrumentation. Master's preferred. Must have excellent } } computer and interpersonal/communication skills. TEM, SEM, x-ray } } diffraction or materials sample preparation experience preferred. Excellent } } benefits. Submit cover letter, resume and names, addresses and telephone } } numbers of three professional references to Professor Brian Robertson, } } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } } will remain open until a suitable candidate is found. UNL is committed to } } AA/EEO and ADA/504. If you require accommodation, please call (402) 472-7886. } } } } } } *********************************************************** } } Assoc. Prof. Brian W. Robertson } } Department of Mechanical Engineering } } and Center for Materials Research and Analysis } } University of Nebraska-Lincoln, N124 WSEC, } } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 ** } } Cheers, } Gary Gaugler, Ph.D. } } }
Sres MSA: Dado que mi tema de investigaci=F3n es el an=E1lisis de propiedades mec=E1nicas de pol=EDmeros semicritalinos, necesitar=EDa contactarme con alguna persona que pueda informarme sobre estudios de microestructura de los mismos.Desde ya muchas gracias.=20 Ing elena Brandaleze.=20
Elena Brandaleze asks whether there is anyone who can provide information on studies of the microstructure of semicrystalline polymers. She works on the mechanical properties of the same.
At 05:50 AM 5/14/99 , you wrote: } Gary: Where are all the 120K jobs. I might want one. } }
They are in Silicon Valley and all over California. These are senior positions. Junior and journey positions gross about $65K-$90K, plus benefits. A brand new person might start at $50K.
Nebraska and academia. Yep. Now I understand the low amount. Perhaps based on cost of living, it is not so bad. Cost of living here in CA is certainly higher. So hopefully, the compensation is relative and commensurate.
Just for your info, we have one of the best junior colleges in the country (and one of the few) that train and produce SEM techs. San Joaquin Delta College, Stockton CA has an excellent program and a fine reputation. With tons of biotech, microelectronics and materials companies all over the place, there are ample opportunities for people to get a start and to move up. Most companies would like people to have a BS but they really want someone who can do the job. They need results, not credentials.
Judy Murphy, who leads the Microscopy Technology Center at San Joaquin Delta College is on this list-server so if anyone has any questions about their program, I'm sure she would be glad to answer them.
My recommendation is to get someone to help you set up your lab and that can come in, spend a couple of days, and show you how to do it. I taught myself from the literature and it was a painful process. Once I could do it, then it was easy for me to show others how to do it. You should also get the literature from the MRS series of TEM sample prep books on how to do it and pay particular attention to Ron Anderson group's works. Number three has a fairly detailed outline of how to do it by John Benedict, Ron Anderson and Stanley Klepeis, MRS Vol 254, 121.
You should seriously consider the South Bay Technology course that is periodically taught. SBT has a very good relationship with Ron Anderson and since his group developed and taught the world the technique, I highly recommend that you contact them.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Fred Pearson To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hello
We are to begin the process of using tripod polishing for preparing TEM samples, and request vendors to contact me with regards to types of polishers available, style, cost, ease of use.
I also welcome users of this method to comment online (or email directly) as to the ease of use and technique of sample preparation, since our lab has not gone this route before.
thanks in advance
Fred
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
This can end up being a very lengthy discussion for the listserver and I don't know your level of expertise with ultramicrotomy so I will keep this very brief. Please feel free to contact me or our applications staff directly for more detail.
You will need to be concerned with the Tg of each phase, their size and relative volumes in the sample. If the ABS phase is 80% and the other phases are not too different in Tg then start at 10-15 degrees above the ABS Tg.
If possible trim before putting in the Ultramicrotome, section at 100nm first to see how it behaves, keep the block face small (less than 1mm). You can try large faces later.
You are looking for consistent sections (no skips), no curling, no chip or flakes. You should see highly specular finish and relatively flat sections. There are a host of remedies for each variation, I will not try to put them all here. If you can get to 70nm you should have a good SPM sample.
Curling or skipping usually means sectioning too warm or a dull knife. Chips or flakes, with or instead of sections means one or more phases are too cold (glassy). Adjust temperature only a few degrees at a time.
A point that some people miss is that the back side of the sections are usually smoother than the front ( the block face which becomes the next section face). If the best you are able to do still shows some chatter try to hold a section face down and do your SPM on the back side.
We are a commercial manufacturer of Ultramicrotomes and cryo attachments to fit nearly all ultramicrotomes. We run a Materials Science Ultramicrotomy Course each Fall which addresses SPM sample preparation. Please see our web site for details.
www.Ventanamed.com Look for RMC button Steve Miller Director of North American Sales Ventana EM Products Group Ventana Medical Systems, Inc. 3450 S. Broadmont Tucson, AZ 85713 Direct phone: 520-205-4118 Fax: 520-903-0132
-----Original Message----- } From: Robert Plano [mailto:RPLANO-at-cea.com] Sent: Wednesday, May 12, 1999 3:51 PM To: 'spm-at-di.com'; 'Microscopy-at-Sparc5.Microscopy.Com'
Greetings.
I am working on making AFM topography and phase images of a series of polymer samples which are made up of varying amounts of polycarbonate, polypropylene and mostly (} 80%) ABS. I have some nice images already (soon to be on our web site) but want to improve the quality. I also want to look at the bulk regions by using cryo-microtomy to create a nice flat face for imaging. Can someone share their experiences with these types of samples and any tips on the microtomy technique (best temperatures, etc.)? I also plan to use plasma etching to see if that will clean up the surface.
Thanks in advance.
Robert J. Plano Staff Analyst, SPM Services Surface Science Labs (650)962-8767, ext. 742
You are not the only one shocked about this. Looking at the figure as low as Mid-$30K, I would say there is something seriously not quite right. These days a simple graduate in computer programming with 2-3 years experience easily gets a starting salary of $50k+ and as they go ahead with few more years of experience, they quickly move into the scale of $70k-80k and crosses $100k within a span of well less than 10 years of career.
- Is there something going wrong with microscopists and SEM/TEM engineers? - Is Mid-$30k fair to expect for electron microscopists (3 years experienced) who maintains the equipment as well? - What if they have 12 -15 years of experience? - Is $60k max (typically at Universities) is fair that these gorgeous people are expected to get max. in their lifetime? - Is that what the people who are the backbone for running and maintaining research facilities are supposed to rewarded with?
I mean just compare it with professionals with same qualifications and experience in other fields. In my personal opinion, there is something not quite right and needs to be re-assessed.
Zia ur Rahman University of Central Florida, Orlando, Florida
At 07:59 PM 5/13/1999 -0700, you wrote:
} Mid-$30K? Is this for real? Is this the going rate for SEM } specialists? That is about 1/4 of what I would consider. } There must be some subtle differentiation of what is expected } from such positions vs. the qualifications and experience of } others in the field. Am I the only one shocked about this? } } } } } JOB ANNOUNCEMENT BELOW -- PLEASE POST } } } } Dear colleagues, } } We want to hire a suitable person for the Central Facility for Electron } } Microscopy at the Center for Materials Research and Analysis at the } } University of Nebraska-Lincoln (UNL). The salary will at least likely be in } } the range of the mid $30k's and is dependent on experience. The medical, } } dental and retirement benefits package is substantial and includes } } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } } growing, has low unemployment, is great for families, has a good range of } } live music, dance and theater, and has very good public and other schools.) } } The Facility provides user-access for ~60 faculty plus their students and } } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM } } and a VG HB501 field-emission STEM, along with a full range of accessories, } } specimen preparation equipment, computers and darkroom. Some development } } of new research instrumentation is in progress for mapping of magnetic } } materials and more development is planned for the Facility. You can find } } out more about the Facility on our web site (that does need a little work) } } at URL: http://www.unl.edu/CMRAcfem/ } } Thanks for passing on the word, } } Brian Robertson } } } } ********************************************************** } } Job Announcement: } } } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } } } UNL Center for Materials Research and Analysis } } } } Analyze/characterize materials using electron microscopy, materials } } preparation and computer instrumentation. Supervise/train students, } } faculty and visiting researchers utilizing these methods. Bachelor's with } } major in physical science, engineering or related field plus three years } } experience in the operation, repair or design of electron microscopes or } } other scientific instrumentation. Master's preferred. Must have excellent } } computer and interpersonal/communication skills. TEM, SEM, x-ray } } diffraction or materials sample preparation experience preferred. Excellent } } benefits. Submit cover letter, resume and names, addresses and telephone } } numbers of three professional references to Professor Brian Robertson, } } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } } will remain open until a suitable candidate is found. UNL is committed to } } AA/EEO and ADA/504. If you require accommodation, please call (402) 472-7886. } } } } } } *********************************************************** } } Assoc. Prof. Brian W. Robertson } } Department of Mechanical Engineering } } and Center for Materials Research and Analysis } } University of Nebraska-Lincoln, N124 WSEC, } } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 ** } } Cheers, } Gary Gaugler, Ph.D. } } } }
} -} We have a Zeiss TEM. Normally when the filament is first switched on } with } } both current and heating knobs are turned to mininimum the voltage } indicator } } shoots up to 2V then immeadiatly starts going down to 1.3-1.4V within no } more } } than 2 minutes, then we start raising the voltage by turning the heating } knob } } until the voltage meter reads between 1.5-1.6V (the recomended voltage } stated in } } the manual). } } Recently though we have been noticing that the voltage shoots up to 2.2V } and } } stays there for several minutes then starts coming down very slowly and } settles } } at more than 1.6V!!... Needless to say that we have also been burning too } many } } filaments too quickly!! The filament lives have been no more than } 10-15hours. } } We would appreciate any hints or suggestions on why would this happen. } We } } are suspecting an electric problem in the filament control circuitry. } Thank } } you. } } Gordon Cougar wrote: } I have seen this happen in other filaments and I and a couple of EE's are } at a loss to explain it. The solution we came up wiht was to use a current } controlled power supply and slowly bring up the current insted of the } voltage. } You may have to switch power supplies when the heater is up to temperature. } } Good luck } Gordon
Gordon has a good point. Also recall a recent post that suggested the use of "good" and "bad" filaments. That is, once you have a batch of filaments that are good, don't use up all of them...save a couple. Then, get a new batch, use them, and decide if they too are good. Then, when a batch performs differently than the previous good ones, you can verify that the earlier good ones are still good or that there is a vacuum leak (the post was based on filament lifetime and vacuum quality).
So, have you changed filament suppliers or gotten a new batch of filaments recently?
The nature of filaments is that their resistance is lowest when cold. As they heat, their resistance increases. Thus, for a given applied voltage, the circuit current will be highest at first, then decrease as the filament heats up. In a circuit where the current limiter is the filament (most circuits are this way), the circuit is generally constant voltage, not constant current.
What you are seeing is counter to what I would think is normal for sure. The only filament-based reason for the voltage to shoot up is if the filament's resistance shot up. That seems hard to explain. However, that the steady state voltage is higher than before could indicate a batch of filaments with higher intrinsic resistance. Without seeing the circuit diagram of the filament supply section, it is hard to make a qualified recommendation on it or even to speculate. Since the filament is at high potential, there are many novel methods that manufacturers use to achieve this and to supply an isolated filament current.
You might also check that the filament is well seated, the terminals are tight and all associated connections are tight.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
OK OK can we move this "debate" off the list? It has been pointed out that academic salaries are low compared to industry (always have been, always will be), that people in certain fields (e.g., engineers - why not use MDs for a real eye-opener?!) make more than those from other fields (e.g., biologists) at all levels of experience, that salaries in any position fluctuate with location, and that academics are generally not satisfied with their salaries (once they discover how much others are making!). I think that covers all the bases.... OOPS I forgot the gender comparision - maybe someone can throw in one final thought-provoking message along those lines.
Rob Palmer Academic
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At 09:19 AM 5/14/99 , you wrote: } Welcome to our world!!! This is pretty much the going rate...sad, isn't } it? For biological microscopists, too. Techs don't get paid very well. } } Tamara Howard } CSHL }
Yeah...very sad and perplexing. Microscopy and EM is not at all a trivial undertaking. Especially if one is looking for useful results. TEM work is even more challenging from what I have seen. I am on the SEM side.
While I admit that I am not 100% a microscopist (it is a means to another end), I am awed by the pool of expertise out there. I don't claim to be an expert by any stretch and I appreciate the free flow of information on this list-server and the one-on-one communications.
I know that there are well-paying tech jobs out there--my brother had several. But even he jumped ship for software work. I suppose that pay is geographically based and contorted by supply and demand. I can also understand low pay in an academic instutution where non-academia is viewed differently. Nevertheless, expertise is not something that just happens. Maybe "You get what you pay for" applies here? In Northern CA, and especially Silicon Valley, EM techs make good wages. And they are in demand. Unfortunately, at both U.S. Coasts, the semiconductor business seems to be imploding. In this respect, I'm like a vulture, waiting for a great deal on a recent SEM being dumped by a company. But for jobs, the only bright spot appears to be biotech.
There is also a vocational institute in Madison Wisconsin that teaches electron microscopy. The graduates of this program are well trained in SEM, TEM, AFM, FIB, materials and biological prep, etc. For more information contact Glenn Boda at (608) 246-6254 or Michael Kostrna at (608) 246-6762.
A textbook editor is looking for a micrograph. He isn't a list subscriber, so please contact him directly.
} ...the image we need as given to us from the chapter editor: } } A picture of aluminum atoms from a scanning tunneling microscope. The } individual atoms of aluminum should appear as dots. The individual atoms } in } the photograph may be accentuated by adding color so that it is easier } to see } them. This image would be used in an upcoming high school Science } textbook.
} Andy Christiansen } Photo Researcher } Holt, Rinehart and Winston } achristiansen-at-hrw.com
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I know there have been some discussions on this listserver in the past about computer sign up systems for electron microscopes. Are there any new programs out there? We are working on a web-based instrument sign up program to replace our existing sign up/microscope accounting system. The old system is not Y2K friendly. Besides the Universty of Minnesota, is there any other group which has a working web-based system?
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
Actually this salary range is not as low as you would think.. I have a M.S. degree and 5 years of EM experience and it took a while for me to find a job in EM throughout the country.. I was applying for jobs that were offering $25K to start as a salary and some were more depending on what part of the country you were looking in... I ended up in Seattle at a job for $32K with benefits to start... I am now currently in Los Angeles at UCLA Medical Center and the salary here is quite a bit more than in Seattle.. But that is outweighed due to the high cost of just living here...So the salary for this job is not as bad as one might think...
==========================
} You are not the only one shocked about this. Looking at the figure as low } as Mid-$30K, I would say there is something seriously not quite right. } These days a simple graduate in computer programming with 2-3 years } experience easily gets a starting salary of $50k+ and as they go ahead with } few more years of experience, they quickly move into the scale of $70k-80k } and crosses $100k within a span of well less than 10 years of career. } } - Is there something going wrong with microscopists and SEM/TEM engineers? } - Is Mid-$30k fair to expect for electron microscopists (3 years } experienced) who maintains the equipment as well? } - What if they have 12 -15 years of experience? } - Is $60k max (typically at Universities) is fair that these gorgeous } people are expected to get max. in their lifetime? } - Is that what the people who are the backbone for running and maintaining } research facilities are supposed to rewarded with? } } I mean just compare it with professionals with same qualifications and } experience in other fields. In my personal opinion, there is something not } quite right and needs to be re-assessed. } } Zia ur Rahman } University of Central Florida, } Orlando, Florida } } } At 07:59 PM 5/13/1999 -0700, you wrote: } } } Mid-$30K? Is this for real? Is this the going rate for SEM } } specialists? That is about 1/4 of what I would consider. } } There must be some subtle differentiation of what is expected } } from such positions vs. the qualifications and experience of } } others in the field. Am I the only one shocked about this? } } } } } } } } } JOB ANNOUNCEMENT BELOW -- PLEASE POST } } } } } } Dear colleagues, } } } We want to hire a suitable person for the Central Facility for Electron } } } Microscopy at the Center for Materials Research and Analysis at the } } } University of Nebraska-Lincoln (UNL). The salary will at least likely be in } } } the range of the mid $30k's and is dependent on experience. The medical, } } } dental and retirement benefits package is substantial and includes } } } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } } } growing, has low unemployment, is great for families, has a good range of } } } live music, dance and theater, and has very good public and other schools.) } } } The Facility provides user-access for ~60 faculty plus their students and } } } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM } } } and a VG HB501 field-emission STEM, along with a full range of accessories, } } } specimen preparation equipment, computers and darkroom. Some development } } } of new research instrumentation is in progress for mapping of magnetic } } } materials and more development is planned for the Facility. You can find } } } out more about the Facility on our web site (that does need a little work) } } } at URL: http://www.unl.edu/CMRAcfem/ } } } Thanks for passing on the word, } } } Brian Robertson } } } } } } ********************************************************** } } } Job Announcement: } } } } } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } } } } } UNL Center for Materials Research and Analysis } } } } } } Analyze/characterize materials using electron microscopy, materials } } } preparation and computer instrumentation. Supervise/train students, } } } faculty and visiting researchers utilizing these methods. Bachelor's with } } } major in physical science, engineering or related field plus three years } } } experience in the operation, repair or design of electron microscopes or } } } other scientific instrumentation. Master's preferred. Must have excellent } } } computer and interpersonal/communication skills. TEM, SEM, x-ray } } } diffraction or materials sample preparation experience preferred. Excellent } } } benefits. Submit cover letter, resume and names, addresses and telephone } } } numbers of three professional references to Professor Brian Robertson, } } } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } } } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } } } will remain open until a suitable candidate is found. UNL is committed to } } } AA/EEO and ADA/504. If you require accommodation, please call (402) } 472-7886. } } } } } } } } } *********************************************************** } } } Assoc. Prof. Brian W. Robertson } } } Department of Mechanical Engineering } } } and Center for Materials Research and Analysis } } } University of Nebraska-Lincoln, N124 WSEC, } } } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } } } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 ** } } } } Cheers, } } Gary Gaugler, Ph.D. } } } } } } } } }
We have a researcher who would like to use the technique of Green and Linstead (Protoplasma (1990) 158:33-38) for making a mold, then a cast of plant shoot apices. They reference dental impression polymers from Kerr UK Ltd., Peterborough, UK. I may have missed it but I didn't find it (or the likes) in the half-dozen mainstream microscopy suppliers' catalogs we have on hand.
Can anyone suggest a source for these products? It would probably be easiest and fastest if we could get a supplier in the USA.
Thanks in advance for any help you can give.
+++++++++++++++++ Dale A. Callaham Senior Microscopist, Stuck Jar Lid Opener, Odd Job Bear Central Microscopy Facility The University of Massachusetts Amherst, MA 01007 ------------------------- Phone 413-545-3751 FAX 413-545-3243 email dac-at-bio.umass.edu
I am looking for an objective aperture for a Hitachi H-9000 (UHV) TEM with sizes of 10,20,30 & 50 microns. Any suggestions about suppliers -- we can supply dimensions.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Hi, The subject line says all: I am using LM to look at sections embedded with Spurr's, and I would like to know its refractive index. Haven't been able to track it down on the web or through my vendor.
Thanks, Jill
------------------------------------------------------------------------------ Jill L. Schmidt Phone: (206) 685-1926 University of Washington FAX: (206) 543-0275 Oceanography Box 357940 e-mail: jills-at-ocean.washington.edu Seattle, WA 98195-7940 http://www.ocean.washington.edu/people/grads/jills/Schmidt.html ------------------------------------------------------------------------------
Hi, I would like to initiate a discussion with someone about the following specific questions, but I don't want to bog down the list. If you can help, please reply directly to me.
First of all, I'm not much of a microscopist and becoming more experienced in order to work on a particular problem: where are bacteria located on undisturbed sand grains. I embed "undisturbed" samples with Spurr's (after staining), cut (very) thick sections (0.5 mm) and look at them with fluorescence microscopy (100 x oil objective).
I would like to look at the sections without using cover slips for the following reasons:
1) Reduced sample preparation time if I am looking "into" the sample rather than right at the surface. I don't have to be as careful in preparing the surface, and can worry less about the polishing compounds becoming embedded in the resin.
2) I have actually found better results ("better images" to my eye) looking into the sample, and this confuses me. The resin itself is autofluorescent, and the DAPI-stained bacteria are brighter still. Could it be that by placing resin between the "focal plane" (quotes because I'm not sure if it's the correct term) and the objective that I am attenuating some of the light with the resin, and therefore can see the contrast between the bacteria and the resin better? I have found that stopping down the diaphragm near the light source helps too.
Now, I know that if the resin does not have the same refractive index as glass+oil, it will lead to distortion. My question is, does it matter for my purposes: I am _not_ looking at the size of the bacteria, just presence/absence. I am tracing the sediment-grain edges (I have a digital camera hooked up to the microscope). Wouldn't the distortion merely affect the apparent width of the grain broundary, rather than the shape (pattern) of the grain outline itself? The grains are 50-200 micrometer in diameter, i.e. a whole grain will not typically be within a field of view.
I am mainly after a record of where the bacteria are located on grain edges, and am not in need of an exact visual representation. I am would rather have large sample sizes (thus the desire for reduced prep time) rather than high precision because I want to be able to treat the results (bacterial position compared with pore width) statistically. I want to be able to publish these results, and don't want to proceed ahead with what just "works best" if it doesn't make any sense.
In searching the archives, I came across someone's lament about the lack of curricula for microscopy and I completely agree! I am at a major research university and there are no classes given on light microscopy.
One other question is: we have a 100x oil objective NA 1.25 (Zeiss) with a collar that adjusts from .8 - 1.25. The microscope owner _thinks_ this is to adjust for different refractive indexes of the specimen. I can see that it functions as an aperature, and because it goes up to 1.25 is seems that it would be related to NA. I can't understand why someone would want a lower NA, unless it would increase the "depth of field". Any ideas? Do you control NA and refractive index the same way?
Thanks for any help you can give me, Jill ------------------------------------------------------------------------------ Jill L. Schmidt Phone: (206) 685-1926 University of Washington FAX: (206) 543-0275 Oceanography Box 357940 e-mail: jills-at-ocean.washington.edu Seattle, WA 98195-7940 http://www.ocean.washington.edu/people/grads/jills/Schmidt.html ------------------------------------------------------------------------------
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Reply to: Re: Electron Microscopy Position Available = Immediately An interesting topic that I hope will be explored in a little more detail, = is not so much how microscopists are paid (which is usually too little) = but what is expected of them. In addition to knowing evey preparation = technique and piece of equipment, we have to know how to fix the equipment,= teach students and post-docs and figure out ways to fund the facility. = No problem.
The really dangerous aspect to this is that quite often the head of the EM = lab is expected to provide a data collection and image interpretation = service for the majority of users. Although I am sure everyone feels able = to do this, for microscopy (and in particular electron microscopy) to = continue to grow, it is important that individual researchers become = responsible for their own specimen preparation, data collection and = results interpretation. Not knowing the technology is no longer a good = excuse to avoid the work. =
On the subject of pay, I always thought that the low initial salaries = being offered were to cover the risk involved with new hires. The true = story could only be that once settled in the salary actually rose to a = comfortable level! =
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Karen S Pawlowski wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
} TEM levels for an animal research lab at UTSW in Dallas. I have over 15 } years of experience, a MS, and the position I hold is considered a } non-tenure, PhD position. This is reality for working in academia in the } midwest (I should mention I started out at the U of MN, where I was paid } alot less.). Why do I stay? Cost of living here is low, so you can live = } on it and I like the field so much I'm getting a PhD in neuroscience to } continue to grow in it. } } Karen Pawlowski } Sr. Research Assoc. UT Southwestern, Dallas, TX } PhD candidate, UT Dallas, Dallas, TX } } On Thu, 13 May 1999, Dr. Gary Gaugler wrote: } } } ------------------------------------------------------------------------=
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America = } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.= Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.= html } } -----------------------------------------------------------------------.=
} } = } } = } } Mid-$30K? Is this for real? Is this the going rate for SEM } } specialists? That is about 1/4 of what I would consider. } } There must be some subtle differentiation of what is expected } } from such positions vs. the qualifications and experience of = } } others in the field. Am I the only one shocked about this? } } = } } = } } = } } } JOB ANNOUNCEMENT BELOW -- PLEASE POST } } } } } } Dear colleagues, } } } We want to hire a suitable person for the Central Facility for = Electron } } } Microscopy at the Center for Materials Research and Analysis at the } } } University of Nebraska-Lincoln (UNL). The salary will at least likely = be in } } } the range of the mid $30k's and is dependent on experience. The = medical, } } } dental and retirement benefits package is substantial and includes } } } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } } } growing, has low unemployment, is great for families, has a good range = of } } } live music, dance and theater, and has very good public and other = schools.) = } } } The Facility provides user-access for ~60 faculty plus their students = and } } } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A = SEM } } } and a VG HB501 field-emission STEM, along with a full range of = accessories, } } } specimen preparation equipment, computers and darkroom. Some = development } } } of new research instrumentation is in progress for mapping of magnetic } } } materials and more development is planned for the Facility. You can = find } } } out more about the Facility on our web site (that does need a little = work) } } } at URL: http://www.unl.edu/CMRAcfem/ = } } } Thanks for passing on the word, } } } Brian Robertson } } } } } } ********************************************************** } } } Job Announcement: } } } } } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } } } } } UNL Center for Materials Research and Analysis } } } } } } Analyze/characterize materials using electron microscopy, materials } } } preparation and computer instrumentation. Supervise/train students, } } } faculty and visiting researchers utilizing these methods. Bachelor's = with } } } major in physical science, engineering or related field plus three = years } } } experience in the operation, repair or design of electron microscopes = or } } } other scientific instrumentation. Master's preferred. Must have = excellent } } } computer and interpersonal/communication skills. TEM, SEM, x-ray } } } diffraction or materials sample preparation experience preferred. = Excellent } } } benefits. Submit cover letter, resume and names, addresses and = telephone } } } numbers of three professional references to Professor Brian Robertson, } } } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-= 0656 or } } } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Posit= ion } } } will remain open until a suitable candidate is found. UNL is = committed to } } } AA/EEO and ADA/504. If you require accommodation, please call (402) = } 472-7886. } } } } } } } } } *********************************************************** } } } Assoc. Prof. Brian W. Robertson } } } Department of Mechanical Engineering } } } and Center for Materials Research and Analysis } } } University of Nebraska-Lincoln, N124 WSEC, } } } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } } } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 ** } } = } } Cheers, } } Gary Gaugler, Ph.D. } } = } } = } } = } } } } } RFC822 header } ----------------------------------- } } Received: from Sparc5.Microscopy.Com [206.69.208.10] by mailhouse.hei.= org } (SMTPD32-4.07) id A18E691011E; Fri, 14 May 1999 13:03:26 PST } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.= 11) = } id IAA02338 for dist-Microscopy; Fri, 14 May 1999 08:40:47 -0500 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id IAA02333 for = } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 14 May 1999 08:40:16 -= 0500 } Received: from poteidaia.utdallas.edu (poteidaia.utdallas.edu = } [129.110.10.1]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id = IAA02322 for = } {Microscopy-at-sparc5.microscopy.com} ; Fri, 14 May 1999 08:40:03 -0500 } Received: from apache.utdallas.edu (kna101-at-apache.utdallas.edu [129.110.= 16.9]) } by poteidaia.utdallas.edu (8.9.1/8.9.1/null-3.5) with ESMTP id IAA14724;=
} Fri, 14 May 1999 08:42:30 -0500 (CDT) } Received: from localhost (kna101-at-localhost) } by apache.utdallas.edu (8.9.1/8.9.1) with SMTP id IAA01748; } Fri, 14 May 1999 08:42:29 -0500 (CDT) } X-Authentication-Warning: apache.utdallas.edu: kna101 owned process = doing -bs } Date: Fri, 14 May 1999 08:42:29 -0500 (CDT) } From: Karen S Pawlowski {kna101-at-utdallas.edu} } To: "Dr. Gary Gaugler" {gary-at-gaugler.com} } cc: MSA listserver {Microscopy-at-Sparc5.Microscopy.Com} } Subject: Re: Electron Microscopy Position Available Immediately } In-Reply-To: {4.1.19990513195604.009dc490-at-pop.calweb.com} } Message-ID: {Pine.GSO.3.96.990514083332.1144A-100000-at-apache.utdallas.edu} =
Working together with process engineers and laser designers, we aim to improve the performance of current generation high power lasers and to develop the next.
If you are at, or near the end of a PhD thesis in microscopy (TEM), preferably in the semiconductor area, and are looking to widen your horizons in the world of industry, this could be for you.
A basic knowledge of german is essential.
If you are interested please contact our Human Resources Manager, Teresa Eichholzer, Binzstrasse 17, 8045 Zurich, Switzerland, or by e-mail at teresa.eichholzer-at-uniphasele.com
For further information please visit our website at http://www.uniphase.com.
-----Mensaje original-----De: Antonio Molina {ifrm111-at-if.csic.es} Para: Microscopy-at-MSA.Microscopy.Com {Microscopy-at-MSA.Microscopy.Com} Fecha: viernes 14 de mayo de 1999 17:06Asunto: LM, SEM (may be TEM or AFM) - Ice cristal size and shape: how to do it Dear microscopists, I am interested in measuring the size, shape, growth, dynamics of recrystallization, etc, of ice, contained in a variety of foods, and generated through different freezing processes (different freezing speeds, hydrostatic pressure, antifreezers). I wonder which could be the easier way to obtain quantitative data. I am considering light microscopy with a cryostage, but I don't know if I will be needing special optics (will phase contrast or polarisation help?). I am also starting to do experiments on cryo-SEM, but I am frightened of altering the ice distribution of my sample, if I cool it further. If you have come across with this problem, your comments could be very useful to me. Thanks, Antonio D. Molina-Garcia Inst. del Frio, CSIC 28040 Madrid SPAINPhone (+34) 91 5445607 (+34) 91 5492300 Fax (+34) 91 5493627E-mail : ifrm111-at-if.csic.es http://www.csic.es/ifrio/ingind.htm
} Date: Fri, 14 May 1999 10:57:51 +0000 } To: "Shalvoy, Richard B CHES" {RBShalvoy-at-archchemicals.com} } From: Lou Solebello {microls1297-at-mindspring.com} } Subject: RE: Electron Microscopy Position Available Immediately } X-Attachments: A:\loucv98.doc; } In-Reply-To: {39E38D236504D211B2BD00805FE616BF3DF9F6-at-ealt-exc-02.corp.olin.com} } } Thats about where I was in my last position. A similar, yet less comprehensive survey of microscopist salaries by Microscopy Today put the AVERAGE national salary for an MS degreed microscopist in all fields with 10+ yrs experience at $57,000 per year. One has to consider that this is an average, across all ecomomic demographics from a college par timer to a CEO. } } Would they be happy with a BS chemist out of school for $35k? probably not.....who is going to teach him, be his mentor? They are expecting too much for too little which makes an excellent recipe for disaster. The level of responsibility and knowledge they are expecting is way out of line for the compensation. } } I almost am embarrased at what I was earning in my last position, and I am considered to be "Senior level". I was earning $46,000/yr prior to benes which worked out to be about $60k/yr with benes. That however is my fault for not being savy enough business wise to research what I was worth. But, I didnt view it that way. Naive as it may sound, I too honesty in compensation for granted because I do not take advantage of others. Besides, I was doing what I love to do, which I cannot put a dollar amount on. A quick glance at my CV (attached) clearly shows that I am not average, but a good deal above average as a microanalyst. You might also be surprised to know that I earned less than that at well known prestigious microanalytical company where I had been for close to 10 yrs. That should dispel quite a few myths people have about the earnings of scientists that company....they are lower than you would expect. I have also worked with others making in the six digit range, and simply was amazed because they completely lacked the intelligence or expertise that would normally be warranted for that salary level. In fact, they shouldnt have been making a technicians salary. } } I thank you for humoring my dissertation on this subject, but I believe that everyone in the scientific community should be alarmed and discuss this topic mor openly. We wonder why there is scientific fraud, and many people have a negative opinion of scientists?, or why companies have been taking the attitudes they have? It is self promulgated by those same companies who are blind to their own errors, but have egos too big to admit it, and need to protect and justify their own high six digit figures. I personally do not like unions, but I strongly believe that if scientists do not do something about the situation, we will be our own demise by simply not acknowledging the problem. } } Lou Solebello } At 07:02 AM 5/14/99 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Not all jobs in California pay as much as Gary says. I'm at UCB and don't make anywhere near that much (and I've been doing this for 17 years). Maybe in Silicon Valley in the semiconductor business, but not in academia. I guess academic technicians are underpaid all over the states. Unfortunately, it's very expensive to live in to SF bay area unless you live east about an hour drive.
But we know what we're in for when we take the job. So I guess it's a case of let the technician beware.
Paula :-)
} At 05:50 AM 5/14/99 , you wrote: } } Gary: Where are all the 120K jobs. I might want one. } } } } } } They are in Silicon Valley and all over California. These } are senior positions. Junior and journey positions gross } about $65K-$90K, plus benefits. A brand new person might } start at $50K. }
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Friends of mine with similar degrees and experience in other fields have second cars, vaction homes and boats. I have a 13 year old Honda. Of course, if she gets accepted, my daughter will go to BU for four years, tuition free 8-)
Ron ----- Original Message ----- } From: Robert J. Palmer Jr. {rjpalmer-at-utkux.utcc.utk.edu} ,Snip} - } maybe someone can throw in one final thought-provoking message along those } lines. } } Rob Palmer } Academic } } } Gordon, } } } } You are not the only one shocked about this. Looking at the figure as low } } as Mid-$30K, I would say there is something seriously not quite right. } } These days a simple graduate in computer programming with 2-3 years } } experience easily gets a starting salary of $50k+ {snip}
Henrik Kaker wrote: ============================================= I'm looking e-mail address of Graticules Limited from UK. ============================================== Try the following:
Mr. Brian Kyte General Manager Pyser Ltd./Graticules E-mail: BrianKyte-at-debscom.demon.co.uk
Yes, this is the same Brian Kyte formerly associated with VG Microtech and Polaron and Bio-Rad.
Chuck
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Look for us! ############################ WWW: www.2spi.com ############################ =======================================================
I am editing some early papers of Sigmund Freud from the years before he turned to the mind. He published several papers in the 1870s and 1880s on his microscopical work, mainly on the nervous system of lower vertebrates.
He refers to some lenses, stains and fixation techniques I'm familiar with and some I'm not familiar with. For instance, he mentions Hartnac lenses of various numbers and serum iodide.
Does anyone know the state of microscopy during this period? Is there a person familiar with this era or book that might list the various stains, etc. in use? Does anyone know how to translate the Hartnac numbers into something meaningful to us? I suppose they refer to what we call power. Even general information such as what was visible and what not, whether artificial light or only sunlight was used, etc. would be helpful. He does not name the microscope he uses so we have to surmise what what available and standard.
Any help will be greatly appreciated and acknowledged.
- Larry Kunstadt, PhD The Psychoanalytic Institute New York University Medical Center
Kerr Specialties as a water soluble wax http://www.kerrcasting.com/Product/English/Wax/Sol-u-wax.htm It melts at 140 F and will dissolve in water. Would this have a use in embedding with out having to fully clear the subject. It will clear a lot faster after it is sliced than before. Just a thought.
Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
We are also considering developing a web-based reservation system, and I think many others would also be interested what other labs are doing in this area. I would be interested in learning of any software which is freely available or low cost and open source.
We are in the very early stages of modifying an open source (Perl) web-calender and scheduling system to suite our needs. The reference url is: http://curiosityshoppe.tierranet.com/framecal/index.shtml
I found this and a few others by searching the web with I believe was combinations of ("resource" and "scheduling" and "calenders")
If we develop a system based on FrameCal or something else we will make it freely available as licensing permits.
Of course, if someone else has a suitable system available freely or at low cost it would save us development time and costs.
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.
} } I know there have been some discussions on this listserver in the past
} about computer sign up systems for electron microscopes. Are there any new } programs out there? We are working on a web-based instrument sign up } program to replace our existing sign up/microscope accounting system. The } old system is not Y2K friendly. Besides the Universty of Minnesota, is } there any other group which has a working web-based system? } } John C. Wheatley } Lab Manager } Arizona State University } Center for Solid State Science } PSA-213 } BOX 871704 } Tempe, AZ 85287-1704 } } Phone: (602) 965-3831 } FAX: (602) 965-9004 } John.Wheatley-at-ASU.Edu
At 01:22 PM 5/14/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Try Energy Beam Sciences
http://www.ebsciences.com
800.992.9037
They offer a wide assortment of apertures and also strip apertures for TEMs custom made to order. Not sure if they support Hitachi.
I would like to invite you to opening Image gallery of cartoons on www.coleoptera.org .
I am wonder if someone would like to put SEM nice picture of beetles to Image gallery.
Keep care and be of good cheer.
Regards
Ricardo
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.37 / Virus Database: 30 - Release Date: 6.4.1999
L. D. Marks wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } I am looking for an objective aperture for a } Hitachi H-9000 (UHV) TEM with sizes of 10,20,30 & 50 } microns. Any suggestions about suppliers -- we } can supply dimensions. } } ++++++++++++++++++++++++++++++++++++++++++++++++ } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } fax: (847) 491-7820 } mailto:l-marks-at-nwu.edu } http://www.numis.nwu.edu } ++++++++++++++++++++++++++++++++++++++++++++++++
Dear Mr. Marks,
Ladd Research has been drilling apertures for EM use for the last forty plus years and we would be very happy to quote you on this. Please contact me directly and we can compare notes on sizes and give you a price. Since we drill ourselves we can give you a choice on hole sizes if you wish to vary from the norm.
Sincerely,
JD Arnott President
-- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
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Hi, I would like to initiate a discussion with someone about the following specific questions, but I don't want to bog down the list. If you can help, please reply directly to me.
First of all, I'm not much of a microscopist and becoming more experienced in order to work on a particular problem: where are bacteria located on undisturbed sand grains. I embed "undisturbed" samples with Spurr's (after staining), cut (very) thick sections (0.5 mm) and look at them with fluorescence microscopy (100 x oil objective).
I would like to look at the sections without using cover slips for the following reasons:
1) Reduced sample preparation time if I am looking "into" the sample rather than right at the surface. I don't have to be as careful in preparing the surface, and can worry less about the polishing compounds becoming embedded in the resin.
2) I have actually found better results ("better images" to my eye) looking into the sample, and this confuses me. The resin itself is autofluorescent, and the DAPI-stained bacteria are brighter still. Could it be that by placing resin between the "focal plane" (quotes because I'm not sure if it's the correct term) and the objective that I am attenuating some of the light with the resin, and therefore can see the contrast between the bacteria and the resin better? I have found that stopping down the diaphragm near the light source helps too.
Now, I know that if the resin does not have the same refractive index as glass+oil, it will lead to distortion. My question is, does it matter for my purposes: I am _not_ looking at the size of the bacteria, just presence/absence. I am tracing the sediment-grain edges (I have a digital camera hooked up to the microscope). Wouldn't the distortion merely affect the apparent width of the grain broundary, rather than the shape (pattern) of the grain outline itself? The grains are 50-200 micrometer in diameter, i.e. a whole grain will not typically be within a field of view.
I am mainly after a record of where the bacteria are located on grain edges, and am not in need of an exact visual representation. I am would rather have large sample sizes (thus the desire for reduced prep time) rather than high precision because I want to be able to treat the results (bacterial position compared with pore width) statistically. I want to be able to publish these results, and don't want to proceed ahead with what just "works best" if it doesn't make any sense.
In searching the archives, I came across someone's lament about the lack of curricula for microscopy and I completely agree! I am at a major research university and there are no classes given on light microscopy.
One other question is: we have a 100x oil objective NA 1.25 (Zeiss) with a collar that adjusts from .8 - 1.25. The microscope owner _thinks_ this is to adjust for different refractive indexes of the specimen. I can see that it functions as an aperature, and because it goes up to 1.25 is seems that it would be related to NA. I can't understand why someone would want a lower NA, unless it would increase the "depth of field". Any ideas? Do you control NA and refractive index the same way?
Thanks for any help you can give me, Jill ------------------------------------------------------------------------------ Jill L. Schmidt Phone: (206) 685-1926 University of Washington FAX: (206) 543-0275 Oceanography Box 357940 e-mail: jills-at-ocean.washington.edu Seattle, WA 98195-7940 http://www.ocean.washington.edu/people/grads/jills/Schmidt.html ------------------------------------------------------------------------------
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I am looking for an objective aperture for a Hitachi H-9000 (UHV) TEM with sizes of 10,20,30 & 50 microns. Any suggestions about suppliers -- we can supply dimensions.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
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Reply to: Re: Electron Microscopy Position Available Immediately An interesting topic that I hope will be explored in a little more detail, is not so much how microscopists are paid (which is usually too little) but what is expected of them. In addition to knowing evey preparation technique and piece of equipment, we have to know how to fix the equipment, teach students and post-docs and figure out ways to fund the facility. No problem.
The really dangerous aspect to this is that quite often the head of the EM lab is expected to provide a data collection and image interpretation service for the majority of users. Although I am sure everyone feels able to do this, for microscopy (and in particular electron microscopy) to continue to grow, it is important that individual researchers become responsible for their own specimen preparation, data collection and results interpretation. Not knowing the technology is no longer a good excuse to avoid the work. On the subject of pay, I always thought that the low initial salaries being offered were to cover the risk involved with new hires. The true story could only be that once settled in the salary actually rose to a comfortable level!
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Karen S Pawlowski wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dr. Gaugler, } } This is similar to what I make and I do SEM, TEM, LM and immuno. at LM and } TEM levels for an animal research lab at UTSW in Dallas. I have over 15 } years of experience, a MS, and the position I hold is considered a } non-tenure, PhD position. This is reality for working in academia in the } midwest (I should mention I started out at the U of MN, where I was paid } alot less.). Why do I stay? Cost of living here is low, so you can live } on it and I like the field so much I'm getting a PhD in neuroscience to } continue to grow in it. } } Karen Pawlowski } Sr. Research Assoc. UT Southwestern, Dallas, TX } PhD candidate, UT Dallas, Dallas, TX } } On Thu, 13 May 1999, Dr. Gary Gaugler wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Mid-$30K? Is this for real? Is this the going rate for SEM } } specialists? That is about 1/4 of what I would consider. } } There must be some subtle differentiation of what is expected } } from such positions vs. the qualifications and experience of } } others in the field. Am I the only one shocked about this? } } } } } } } } } JOB ANNOUNCEMENT BELOW -- PLEASE POST } } } } } } Dear colleagues, } } } We want to hire a suitable person for the Central Facility for Electron } } } Microscopy at the Center for Materials Research and Analysis at the } } } University of Nebraska-Lincoln (UNL). The salary will at least likely be in } } } the range of the mid $30k's and is dependent on experience. The medical, } } } dental and retirement benefits package is substantial and includes } } } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } } } growing, has low unemployment, is great for families, has a good range of } } } live music, dance and theater, and has very good public and other schools.) } } } The Facility provides user-access for ~60 faculty plus their students and } } } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM } } } and a VG HB501 field-emission STEM, along with a full range of accessories, } } } specimen preparation equipment, computers and darkroom. Some development } } } of new research instrumentation is in progress for mapping of magnetic } } } materials and more development is planned for the Facility. You can find } } } out more about the Facility on our web site (that does need a little work) } } } at URL: http://www.unl.edu/CMRAcfem/ } } } Thanks for passing on the word, } } } Brian Robertson } } } } } } ********************************************************** } } } Job Announcement: } } } } } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } } } } } UNL Center for Materials Research and Analysis } } } } } } Analyze/characterize materials using electron microscopy, materials } } } preparation and computer instrumentation. Supervise/train students, } } } faculty and visiting researchers utilizing these methods. Bachelor's with } } } major in physical science, engineering or related field plus three years } } } experience in the operation, repair or design of electron microscopes or } } } other scientific instrumentation. Master's preferred. Must have excellent } } } computer and interpersonal/communication skills. TEM, SEM, x-ray } } } diffraction or materials sample preparation experience preferred. Excellent } } } benefits. Submit cover letter, resume and names, addresses and telephone } } } numbers of three professional references to Professor Brian Robertson, } } } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } } } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } } } will remain open until a suitable candidate is found. UNL is committed to } } } AA/EEO and ADA/504. If you require accommodation, please call (402) } 472-7886. } } } } } } } } } *********************************************************** } } } Assoc. Prof. Brian W. Robertson } } } Department of Mechanical Engineering } } } and Center for Materials Research and Analysis } } } University of Nebraska-Lincoln, N124 WSEC, } } } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } } } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 ** } } } } Cheers, } } Gary Gaugler, Ph.D. } } } } } } } } } } } RFC822 header } ----------------------------------- } } Received: from Sparc5.Microscopy.Com [206.69.208.10] by mailhouse.hei.org } (SMTPD32-4.07) id A18E691011E; Fri, 14 May 1999 13:03:26 PST } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) } id IAA02338 for dist-Microscopy; Fri, 14 May 1999 08:40:47 -0500 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id IAA02333 for } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 14 May 1999 08:40:16 -0500 } Received: from poteidaia.utdallas.edu (poteidaia.utdallas.edu } [129.110.10.1]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id IAA02322 for } {Microscopy-at-sparc5.microscopy.com} ; Fri, 14 May 1999 08:40:03 -0500 } Received: from apache.utdallas.edu (kna101-at-apache.utdallas.edu [129.110.16.9]) } by poteidaia.utdallas.edu (8.9.1/8.9.1/null-3.5) with ESMTP id IAA14724; } Fri, 14 May 1999 08:42:30 -0500 (CDT) } Received: from localhost (kna101-at-localhost) } by apache.utdallas.edu (8.9.1/8.9.1) with SMTP id IAA01748; } Fri, 14 May 1999 08:42:29 -0500 (CDT) } X-Authentication-Warning: apache.utdallas.edu: kna101 owned process doing -bs } Date: Fri, 14 May 1999 08:42:29 -0500 (CDT) } From: Karen S Pawlowski {kna101-at-utdallas.edu} } To: "Dr. Gary Gaugler" {gary-at-gaugler.com} } cc: MSA listserver {Microscopy-at-Sparc5.Microscopy.Com} } Subject: Re: Electron Microscopy Position Available Immediately } In-Reply-To: {4.1.19990513195604.009dc490-at-pop.calweb.com} } Message-ID: {Pine.GSO.3.96.990514083332.1144A-100000-at-apache.utdallas.edu} } MIME-Version: 1.0 } Content-Type: TEXT/PLAIN; charset=US-ASCII } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-UIDL: 907884271 } Status: U }
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Actually this salary range is not as low as you would think.. I have a M.S. degree and 5 years of EM experience and it took a while for me to find a job in EM throughout the country.. I was applying for jobs that were offering $25K to start as a salary and some were more depending on what part of the country you were looking in... I ended up in Seattle at a job for $32K with benefits to start... I am now currently in Los Angeles at UCLA Medical Center and the salary here is quite a bit more than in Seattle.. But that is outweighed due to the high cost of just living here...So the salary for this job is not as bad as one might think...
==========================
} You are not the only one shocked about this. Looking at the figure as low } as Mid-$30K, I would say there is something seriously not quite right. } These days a simple graduate in computer programming with 2-3 years } experience easily gets a starting salary of $50k+ and as they go ahead with } few more years of experience, they quickly move into the scale of $70k-80k } and crosses $100k within a span of well less than 10 years of career. } } - Is there something going wrong with microscopists and SEM/TEM engineers? } - Is Mid-$30k fair to expect for electron microscopists (3 years } experienced) who maintains the equipment as well? } - What if they have 12 -15 years of experience? } - Is $60k max (typically at Universities) is fair that these gorgeous } people are expected to get max. in their lifetime? } - Is that what the people who are the backbone for running and maintaining } research facilities are supposed to rewarded with? } } I mean just compare it with professionals with same qualifications and } experience in other fields. In my personal opinion, there is something not } quite right and needs to be re-assessed. } } Zia ur Rahman } University of Central Florida, } Orlando, Florida } } } At 07:59 PM 5/13/1999 -0700, you wrote: } } } Mid-$30K? Is this for real? Is this the going rate for SEM } } specialists? That is about 1/4 of what I would consider. } } There must be some subtle differentiation of what is expected } } from such positions vs. the qualifications and experience of } } others in the field. Am I the only one shocked about this? } } } } } } } } } JOB ANNOUNCEMENT BELOW -- PLEASE POST } } } } } } Dear colleagues, } } } We want to hire a suitable person for the Central Facility for Electron } } } Microscopy at the Center for Materials Research and Analysis at the } } } University of Nebraska-Lincoln (UNL). The salary will at least likely be in } } } the range of the mid $30k's and is dependent on experience. The medical, } } } dental and retirement benefits package is substantial and includes } } } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } } } growing, has low unemployment, is great for families, has a good range of } } } live music, dance and theater, and has very good public and other schools.) } } } The Facility provides user-access for ~60 faculty plus their students and } } } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM } } } and a VG HB501 field-emission STEM, along with a full range of accessories, } } } specimen preparation equipment, computers and darkroom. Some development } } } of new research instrumentation is in progress for mapping of magnetic } } } materials and more development is planned for the Facility. You can find } } } out more about the Facility on our web site (that does need a little work) } } } at URL: http://www.unl.edu/CMRAcfem/ } } } Thanks for passing on the word, } } } Brian Robertson } } } } } } ********************************************************** } } } Job Announcement: } } } } } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } } } } } UNL Center for Materials Research and Analysis } } } } } } Analyze/characterize materials using electron microscopy, materials } } } preparation and computer instrumentation. Supervise/train students, } } } faculty and visiting researchers utilizing these methods. Bachelor's with } } } major in physical science, engineering or related field plus three years } } } experience in the operation, repair or design of electron microscopes or } } } other scientific instrumentation. Master's preferred. Must have excellent } } } computer and interpersonal/communication skills. TEM, SEM, x-ray } } } diffraction or materials sample preparation experience preferred. Excellent } } } benefits. Submit cover letter, resume and names, addresses and telephone } } } numbers of three professional references to Professor Brian Robertson, } } } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } } } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } } } will remain open until a suitable candidate is found. UNL is committed to } } } AA/EEO and ADA/504. If you require accommodation, please call (402) } 472-7886. } } } } } } } } } *********************************************************** } } } Assoc. Prof. Brian W. Robertson } } } Department of Mechanical Engineering } } } and Center for Materials Research and Analysis } } } University of Nebraska-Lincoln, N124 WSEC, } } } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } } } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 ** } } } } Cheers, } } Gary Gaugler, Ph.D. } } } } } } } } }
no creo que seamos muchos los que hablamos espanhol en esta lista, te recomendaria que escribieras en Ingles y asi ampliar tus posibilidades de obtener ayuda.
Elena,
I dont think that many of us speak spanish, I will recommend you to write in english, increasing, in this way, your possibilities of obtainig help from this list.
saludos
Gary=20 EPhD Trondheim, Norway.
On Tue, 27 Apr 1999, Elena Brandaleze wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America= =20 } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Sres MSA: } Dado que mi tema de investigaci=F3n es el an=E1lisis de propiedades } mec=E1nicas de pol=EDmeros semicritalinos, necesitar=EDa contactarme con } alguna persona que pueda informarme sobre estudios de microestructura de } los mismos.Desde ya muchas gracias.=20 } =09=09=09=09Ing elena Brandaleze. } =20
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Hi, The subject line says all: I am using LM to look at sections embedded with Spurr's, and I would like to know its refractive index. Haven't been able to track it down on the web or through my vendor.
Thanks, Jill
------------------------------------------------------------------------------ Jill L. Schmidt Phone: (206) 685-1926 University of Washington FAX: (206) 543-0275 Oceanography Box 357940 e-mail: jills-at-ocean.washington.edu Seattle, WA 98195-7940 http://www.ocean.washington.edu/people/grads/jills/Schmidt.html ------------------------------------------------------------------------------
} Date: Sun, 16 May 1999 14:42:53 +0000 } To: Paul Webster {pwebster-at-mailhouse.hei.org} } From: Lou Solebello {microls1297-at-mindspring.com} } Subject: Re: Electron Microscopy Position Available Immediately } In-Reply-To: {199905142059.PAA08933-at-Sparc5.Microscopy.Com} } } What is expected of an electron microscopist, or ANY microscopist for that matter (and I use that term loosely....there are a lot people who carry the title but are nothing more than a user), is DIRECTLY relate to what they get paid. } } The amount of responsibility that we microscopist take on relative to other professionals in other fields is a great deal under compensated. We are expected to know everything there is about our individual discipline as well as all other aspects of microscopy. We are expected to deliver THE ANSWER nearly immediately at a low cost. We are expected to know how to assemble and dissasemble our instruments and fix them at the same cost and rate. We are looked upon sourly when we say "cant be done" as if we are inferior or lying. We are expected to handle samples that potentially are extremely hazardous to our health.....the danger of many unknowns on the consulting level are unknown until we discover what we are analyzing. We are expected to write SOP's, develop QA/QC manuals, submitt contract bids, obtain grant funding, supervise people, be walking encyclopedia, be able to read client minds, maintain proficiency in round robins and regulatory complance audits, be an MBA and business wiz, and work 60 hours a week on salary if required at a moments notice. } } That is the real point that I have been trying to get across to others on this list server. We are GROSSLY underpaid considering our responsibilities, level of education and experience. The only thing that keeps most of us in these positions is the fact that we love our work. Nobody said life was fair.... } } At 02:06 PM 5/14/99 -0700, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
John writes ... } } } I know there have been some discussions on this listserver in the past } about computer sign up systems for electron microscopes. Are there any new } programs out there? We are working on a web-based instrument sign up } program to replace our existing sign up/microscope accounting system. The } old system is not Y2K friendly. Besides the Universty of Minnesota, is } there any other group which has a working web-based system?
There is an interesting wwweb based calendar for individuals and groups at
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results, and are in demand. This is a must. (What if you had products with 9 years of extensive double blind studies, worked in seconds or minutes, and affordable! What if you could add products announced on CNN and weight loss products that really work! Naw!! Really?? This is only a dream, right?!)
2) What if in addition to other products IN DEMAND, you had an absolutely DELICIOUS nutritional snack bar. (What if this nutritional bar had 3 major medical health benefits and it contained: 1) Live enzymes with digestive capabilities and studies to prove that it aids in the prevention and the reversal of any degenerative disease including cancer, 2) Micro nutrients that have been proven to prevent and reduce the risk of cancer dramatically and actually has the endorsement of The National Cancer =46oundation,
3) The only nutritional snack bar in the industry to be registered with the =46DA and have a drug code right on the label. All this and=
still be amazingly delicious and filling! This couldn't possibly be for real, could it?!?)
3) Then, you would want a company who would be willing to expose these products thru the media, at their cost: TV and radio commercials, newspapers, talk shows, etc., to help you get the word out. Offer people the chance to purchase products thru an 800 number while viewing or listening to the advertisement. (I haven't seen anything like this before! Could you just imagine!)
4) Then you would want your ideal company to let you share in the sales generated from this media exposure. Imagine what kind of volume this
national exposure would generate. (What company would do this? We had nothing to do with these sales! This would be radical!)
5) Wouldn't it be great if we were given the names and addresses of the people who ordered product this way, so that we could follow-up with the customer to either sell more product as needed or introduce him or her to our ideal business opportunity. That would take care of our distributors having to constantly be finding their own qualified leads. (Companies now let us find our own leads, they follow-up with these people and THEY pocket the proceeds!)
6) You would want your company's compensation plan structured in such a way that we wouldn't need hundreds or even thousands in our downline before we start earning a decent check. Most people cannot do this. (True, but who's going to tell you this when you start?)
7) Wouldn't it be great if we could share in the successes of other distributors, and not just those that are in our downline. We would want everybody to be successful, not just some. (Nope - all we can do is admire the others' success!)
Yes, wouldn't it be great if a company like this existed? One that confronted all the major drawbacks in this industry and actually solved them....a company that was built by distributors, for distributors... . a company that would want it's distributors to be successful....wouldn 't it be great?
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now for complete details. You now have the power to achieve your goals!
******************************************************** To be removed from our mailing list, please send an email to mailto:glarge99-at-yahoo.com?subject=3Dremove ********************************************************
Well folks.... I screwed up and during a reprogramming of the Junk Mail Filter managed to muck up the works. I did not notice that the "Listserver" Mail was off-line and inoperative until this evening ~ 10pm, since the rest of my mail was operating fine.
I hope you all enjoyed your present of "peaceful" day.
If you tried posting a message and it was rejected or you just did not see it, then just try again. I think all is back to the status quo.
Cheers... Nestor Your Friendly Neighborhood SysOp.
PS
Hmmm.. I wonder how many people noticed ? Naw.. I won't ask..
We have some spare equipment to dispose of to a good home, all in good working order.
Prices are negotiable, but you will have to collect.
1. De Vere 504 enlarger
2. Durst M35 colour enlarger
3. LKB knifemaker 7800 series (sensible offers on this one please)
4. Reichert OM U3 ultramicrotome
Hurry while stocks last!
Pete
Peter Bond Plymouth Electron Microscopy Unit University of Plymouth Drake Circus Plymouth Devon UK PL4 8AA Tel/Fax: 01752 233092 email: pbond-at-plymouth.ac.uk
A colleague of mine needs to replace an old keyboard for a Tracor Nothern TN 5500 EDS system. Does anyone have one to sell trade or give away. Please contact Charlie Cooney -at- CBC-at-post.queensu.ca Thank you Paul Nolan
I am looking for individuals interested in cryoplaning techniques (using a cryoultramicrotome to produce blocks with a very flat surface and examining this surface in a cryo-SEM). Please contact me back-channel if interested.
Best regards, Steven E. Slap, Vice-President ******************************************* Energy Beam Sciences, Inc. The Laboratory Microwave Company Adding Brilliance to Your Vision http://www.ebsciences.com {http://www.ebsciences.com} *******************************************
Wyeth-Ayerst Research, a major division of Fortune 100 American Home Products Corporation, has an opportunity at our pharmaceutical research facility in Chazy, New York for a Scientist in our Investigatory Pathology group.
JOB TITLE: Scientist II, JOB# C5780 DEPARTMENT: Investigatory Pathology LOCATION: Chazy, NY HR REPRESENTATIVE: B. Hebert HOURS: 8:00 a.m. - 4:30 p.m. POSITION REPORTS TO: Dr. Laura Patrone, Senior Research Scientist I
EDUCATION REQUIREMENTS: BS/BA degree in Biology, Biochemistry, or science related field or MS degree in science related field.
EXPERIENCE REQUIREMENTS: BS/BA candidates must have a minimum of 4 years experience developing immunohistological assays in an industrial, hospital or highly specialized academic setting; MS candidates must have a minimum of 2 years relevant pharmaceutical laboratory experience, or 2 years experience in a highly specialized relevant academic environment.
OTHER SKILLS REQUIRED: Additional histomorphological skills required; must have excellent oral and written communication skills as well as excellent interpersonal and organizational skills; some travel may be required.
PRINCIPAL DUTIES OF POSITION: Scientist II will perform immunohistochemistry and associated techniques, develop new panels of tests to characterize tissues and differentiate cell types; will provide technical support for method development in morphologic analytical techniques; perform data interpretation and assist in report writing; review literature for new technologies to expand and enhance research capabilities; will provide technical support for regulatory pathology area; will also perform departmental and facility support functions as needed (e.g., inventory, scheduling, purchasing).
Interested candidates must contact the Human Resources Representative: Barbara Hebert Office of Human Resources Wyeth-Ayerst Research 641 Ridge Road Chazy, NY 12921 Phone (518) 846-6237
Available - Balzers 360M Freeze Fracture System, 1970 vintage. Complete with E-Beam Pt-C gun, evaporative C electrodes, QSD-201D quartz crystal thickness monitor, commutator unit, and timer for C electrodes. Also included is extra EVM-052A E-Beam power supply, extra GA-1 control unit, extra ODP, extra main valve, extra baseplate, microtome, and feedthroughs, and many extra sample holders, feedthroughs, tools, etc. Must move immediately for best offer. Robert J. Munn rjmunn-at-ucdavis.edu
I am looking for a method to visualize the somata of live epithelial cells. I plan to inject these cells intracellularly, using a micropipette. What I need is a stain with the following characteristics: 1) it can be injected intracellularly via a micropipette, preferably using intophoresis. 2) it is NOT fluorescent 3) it is visible without the need to further process the tissue. This last point is important because I need to watch the injection under the LM and be able to tell when the cell is filled. --E. Peterson Ellengene H. Peterson Neurobiology Program & Department of Biological Sciences Ohio University Athens, OH 45701 740 + 593-2111 (tel) 740 + 593-0355 (fax) peterson-at-ohiou.edu
Select the online scheduler; username=guest, password=guest. You will be able to make reservations on the instrument "Dummy".
Unfortunately not all web browsers are created equal. You can use Navigator 4 (or higher) or IE 4 (or higher) on a PC, but only IE 4 (or higher) on a Mac. (It is a Java compatibility problem). You will also have to accept a cookie so that the server can recognize the platform you are running on.
cheers, Henk
At 06:15 PM 5/16/99 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
Hi everyone, First of all I would like to thank Dr. Simkin from Michigan University forthe information about Electron Channelling. I am going to get in touch with him. This time my question is about jet polishing solution of copper and copper and zinc alloys. Actually i found a solution that is sixty percent phosphoric acid and deionized water but the problem is that it is hard to get rid of that solution from the surface of specimen after you are done with jet polishing. If anybody has some idea about how to clean the surface after jet polishing i really appreciate it. Thanks
I'm looking for any input on the quality of Russian made LOMO (Leningrad Optical and Mechanical Organization) microscopes. Specifically the Biolam, Polam models and the stereomicroscope marketed here in the US.
I would be interested to hear from anyone who has experience using these scopes and commenting on quality of optics, mechanical, illumination,etc. Although you get alot for your money, I'm concerned about service, spare parts, and future support.
Much appreciated.
{italic} {color} {param} 8080,0000,0000 {/param} Raymond Nip, RRT, RCP
I would like to begin by telling you I work for and represent a manufacturer, Allied High Tech Products, Inc. We manufacture the MultiPrep=99 system.
The MultiPrep is a semi-automatic tool used for many applications including TEM sample preparation using the wedge technique. The MultiPrep's= =20 capabilities include parallel polishing, precise angle polishing (0.02 degree increments), site-specific polishing or any combination thereof. It provides reproducible results by eliminating inconsistencies between users. When using the MultiPrep only the specimen is in contact with the abrasive, eliminating the problem of excessive wear of the hand tool's feet, and unwanted faceting of your specimen. In fact, by using the MultiPrep, you can completely eliminate the need for hand-held polishing tools.
The MultiPrep allows you to set an angle and polish through the angle without it changing, and monitors the amount of material that has been removed from the sample (real time during the polishing operation) in 1-micron increments. =20
The MultiPrep system sells for $11,775.00 plus the accessories that are necessary for your individual applications. =20
The other applications of the tool are preparation of SEM cross sections, TEM sample Prep, Pre-FIB thinning, parallel polishing and de-layering of semiconductor devices or thin section preparation and backside sample preparation for semiconductor devices for emission spectroscopes.
If you have any questions please feel free to contact me, send mail to mailto:info-at-alliedhightech.com, or visit our web site at http://www.alliedhightech.com.=20
We also have an new 12 page brochure that can be emailed to you in a PDF format. It can be sent via snail mail too.
Thank you,
Ed Hirsch
} Hello } } We are to begin the process of using tripod polishing for preparing } TEM samples, and request vendors to contact me with regards to } types of polishers available, style, cost, ease of use. } } I also welcome users of this method to comment online (or email directly) } as to the ease of use and technique of sample preparation, since our lab } has not gone this route before. } } thanks in advance } } Fred } } } } } } ******************************************************** } Fred Pearson } Brockhouse Institute for Materials Research } McMaster University } 1280 Main St. West } Hamilton, Ontario=20 } Canada L8S 4M1 } } } email: eoptics-at-mcmaster.ca } phone: (905) 525-9140 ext. 24609 } fax: (905) 521-2773 } ******************************************************** } } } ************************************************* Edward A. Hirsch Product Application Specialist Allied High Tech Products 2376 East Pacifica Place Rancho Dominguez, CA 90220 ph: (919) 846-9628 vm:(800)675-1118 x245 fx: (310)762-6808 http://www.alliedhightech.com *************************************************
I have a JEOL JWS 7515 in England that need to be de-installed. This is a Critical Demension SEM. Does anyone have a lead on independent service companies that can do this type of work?
Regards,
Earl Weltmer Scanservice Corporation Third Party SEM Service USA
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Hi, Zia: The OL coil is OK for three reason: 1. you have told me that you found a short so that the OL coil was unconnected to the connector. It was meaning you found out the reason why the OL coil is open. 2. If the OL coil was broken or short inside, the OL resistance would be changed. But you know 5.2 ohms is OK. 3. The insulation of OL coil is relative with high voltage and the temperature of the OL coil only. If you keep it below 80 C degree, you can test it by DC power supply and flow 10 amps currents in 10 minutes . In fact, you have get right way and go on please . I do not know why you can not receive my e-mail and you can call me 65-8743253 ( office phone) at 9:00 pm Singapore time. Good luck! Regards Tiejun
to add a comment on the answers you've already got concerning your request on measurements of optical parameters for HRTEM simulation... Mike O'Keefe's answer is acceptable as a first try. If you want to measure some values more accurately (especially the Cs), just refer to the literature quoted by Jonathan Barnard. However, there is a little problem with the Cs measurement from HRTEM diffractograms obtained from an amorphous thin film ; the usual method assumes that the specimen behaves like a pure weak-phase object, which is generally not fulfilled : there is a phase shift due to the propagation of electrons throughout the object, which leads to a breakdown of the 'potential projection' hypothesis, which may lead to false values of defoci and Cs... To get more details on this problem raised by J.M. Gibson (Ultramicrosc., 1994), check the work below (where the complete reference from J.M.G. is also recalled) :
"Quantitative analysis of HRTEM images from amorphous materials. I : about the estimation of Cs and df from HRTEM diffractograms", H.S. BAIK, T. EPICIER, E. VAN CAPPELLEN, Eur. Phys. J. AP 4, 11-26, (1998)
As a consequence of the approach developed within this paper, you can also estimate rather precisely the other parameters (delta and alpha) if you know precisely the structure (and the thickness) of the amorphous film you're using for your measurement ; the treatment is however somewhat tedious, but it works (we have applied it successfully to such measurements for a JEOL2010F and a CM200FEG).
Good luck,
--------------------------------------------------------------------------- Dr. Thierry EPICIER, GEMPPM, umr CNRS 5510, INSA de LYON, Bat 502, 20, Av. Einstein, F69621 VILLEURBANNE CEDEX FRANCE
I need to characterise the microstructure and composition of soft electroceramic film(PZT) by using FEGTEM. When I put the specimen in to the beam path, I observed that there was a very strong interaction between the electron beam and specimen which can clearly be seen to be badly charged. I have been advised to coat carbon or gold on the surface of the specimen. I would like to know if there any other methods which could be used to avoid this charging problem? Do you have any further ideas or methods for this sort of material to prepare a specimen which is suitable for high resolution image and electron energy loss spectroscopy? Currently, I use a method of ion thinning by Gatan DuoMill and PIPS.
Dear members: I have placed an order for a dimpler and precision coring tool made by VCR company several months ago and am now waiting for their shipping. But I was told that VCR was bancrupt or will be sold by other company. I worry about the quality of the product and A/S. Could any of you send me the right story what is going on there? Any information will be appreciated. Best wishes, Jondo YunDepartment of Inorganic Materials EngineeringCenter for Instrumental AnalysisKyungnam University449 Weolyeong-dong, Masan, 631-701, Korea82-551-249-2697 (tel)82-551-248-5033 (fax)jdyun-at-hanma.kyungnam.ac.kr
A quick question: we have some blood samples which were received in "Trump's Fixative", described as phosphate-buffered gluteraldehyde and paraformaldehyde, pH 7.2. So far we've been unable to locate any reference to Trump's in any of our literature.
I expect it's perfectly fine to proceed with regular phosphate buffer washes, then postfixation, but thought I would check with collective microscopy mind out there first.
Earl Weltmer wrote: =============================================== I have a JEOL JWS 7515 in England that need to be de-installed. This is a Critical Dimension SEM. Does anyone have a lead on independent service companies that can do this type of work? ============================================== You can find listings for third party SEM service firms in the UK on our website URL http://www.2spi.com/hot-service5.html
This is part of our website directory of third party service providers on equipment in the microscopy and microanalysis world.
Either one of them could probably do what you want to have done.
Chuck
Disclaimer: We have no financial interest in either firm, both have good reputations, so far as we know.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
I have noticed that some non-conductors can be put into the microscope without the objective aperture and they can set up a steady state where they do not charge. Once you put the objective aperture in they continually charge. I recently noticed on a GaN on sapphire substrate imaged in a 120 kV machine where this behavior is reversed, i.e. charges without the aperture in and does not when it is in.
I have had some success with the small angle cleavage technique with glass substrates without coating. Most times the sample will not charge without coating, but some times it will. I try to minimize the amount of material potentially exposed to the beam by putting the silver epoxy as close to the tip area as possible.
However, I am using a 120kV machine for most of the time and I need to carbon coat to keep the glass from softening. I have less of a problem with this in a 200kV machine. I believe that there is more heating in the sample at 120kV than 200kV because more current is being deposited into the sample at the lower voltage. At higher voltages more electrons pass through the sample.
Incidentally, when I do coat with carbon, I can get by with about 200 A on each side. You can get away with about 400 A on one side but there are other reasons why I need to coat on both sides. This thickness is about the minimum for me to prevent softening of the glass. I think that you can get away with only about 200 A or less on just one side if your sample doesn't have this problem. There was a question some time ago that I asked when I first started working with glass about whether it matters if you coat just one side or two and if you coat one side does it matter if it is towards the beam. I think that you get a little better performance with the coating towards the beam, especially if the beam is exposed to any thick parts of the sample at lower magnifications.
These observations are all based on a limited experience of a little over a year on glass samples.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: P.Wang To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hi all
I need to characterise the microstructure and composition of soft electroceramic film(PZT) by using FEGTEM. When I put the specimen in to the beam path, I observed that there was a very strong interaction between the electron beam and specimen which can clearly be seen to be badly charged. I have been advised to coat carbon or gold on the surface of the specimen. I would like to know if there any other methods which could be used to avoid this charging problem? Do you have any further ideas or methods for this sort of material to prepare a specimen which is suitable for high resolution image and electron energy loss spectroscopy? Currently, I use a method of ion thinning by Gatan DuoMill and PIPS.
That is the way we do it all the time. You can find info on Trumps at this site by searching for such.
http://www.biotech.ufl.edu/~emcl/tips.html
At 08:23 AM 5/19/1999 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Don't you get Calcium Phosphate precipitates forming if you fix in a phosphate buffered aldehyde solution???
} A quick question: we have some blood samples which were received in "Trump's } Fixative", described as phosphate-buffered gluteraldehyde and } paraformaldehyde, pH 7.2. So far we've been unable to locate any reference } to Trump's in any of our literature. } } I expect it's perfectly fine to proceed with regular phosphate buffer } washes, then postfixation, but thought I would check with collective } microscopy mind out there first. Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
} From September 1999 there will be a 2 year post-doc position available in the electron microscopy group (Department of Physical Chemistry) at the University of Mainz, Germany. At present the group has 2 electron microscopes and a well-equipped specimen preparation laboratory. In September 1999 a new 300 kV Philips Tecnai 30 Electron Microscope with FEG and GIF will be installed. Applications from physicists and chemists with Ph.D experience in electron microscopy and a sound background in electron energy loss spectroscopy are welcome.
Please send applications to
Dr. I.G. Voigt-Martin Inst. f. Physikalische Chemie Jacob Welder Weg 11 55099 Mainz
Responding to the message of {4.1.19990518173013.030d9100-at-pop.er6.eng.ohio-state.edu} from "Hendrik O. Colijn" {colijn.1-at-osu.edu} :
} Feel free to check out our site. } } http://www.ceof.ohio-state.edu }
I tried, but it doesn't seem to like the single most popular browser/platform combination at our University - Netscape Navigator running on Mac OS.
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist Office:(612) 626-7594 CIE Characterization Facility, University of Minnesota Desk: (612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
Hi, I am looking for a low magnification oil immersion objective. I look at two scales in my work -- I use 100x oil for locating bacteria and 25x for the sediment grains they are attached to, and I need to be able to go back and forth. I do not need a high NA. This is for a Zeiss Universal, non-infinity corrected. From what I gather, older Leitz and perhaps olympus objectives are interchangeable. Thanks if you can help, Jill
------------------------------------------------------------------------------ Jill L. Schmidt Phone: (206) 685-1926 University of Washington FAX: (206) 543-0275 Oceanography Box 357940 e-mail: jills-at-ocean.washington.edu Seattle, WA 98195-7940 http://www.ocean.washington.edu/people/grads/jills/Schmidt.html ------------------------------------------------------------------------------
OK, I know asking this question of the group may seem strange, but this is the most densely populated area of microscopy wisdom in the universe and I need your global perspective.
I am the only person in a small EM lab that serves a primarily undergraduate campus. I am part of the technical staff, I am not a faculty member. We have doctorate programs and a few researchers using TEM, but no materials science types and no professional schools. I guess our TEM might be used 300 to 500 hrs/yr sometimes less, never more.
We have a JEOL 100B circa 1975. For most of our applications, it works fine. I have a whole other microscope to use for parts and a clever service provider who may be able to keep this thing going for a long time. In a darkened room, it is hard to tell it from a more modern instrument based on the images seen on the screen. When the lights go on, it looks 24 years old.
Recently a new faculty member asked when are we getting a new TEM, one with digital imaging. I choked, said I would check into it. So here are a few questions I need help with and I hope for some kindness and understanding in your replies. I do need your replies because as the only real EM person here I could use your help.
I have never gone out looking for $$ for a new microscope. Where does one look? Could we justify $$ based on our level of usage? What do you think? What are the criteria used by funding agencies when evaluating requests. Could you share you experiences?
Suppose we can't get the $$ for a whole new TEM. Would I be crazy to try to get $$ to equip our current TEM (remember 24 years old) with an add on digital camera? The ports are there and it could be done, but I'm not sure it is a good idea.
I hope you can give me some leads and encouragement. Most of the burden of doing this job will fall on me. Although often supportive in casual discussions, I don't expect much help from anyone else here.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I have looked in our archives and have found 2 solutions that Bernie Kest= el from Argonne National Lab has developed for copper which may be of use. =
This work was done with our Model 550 single vertical jet electropolisher=
so references to jet height would be unique to our system. The other parameters may need to be adjusted slightly depending on the type of jet polisher you are using. =
Recipe #2 150 ml HNO3 350 ml ethanol 40 ml butyl cellosolve
Temp: -20 degrees C Jet height: 3.9mm Pump setting: 2.5 Volts: 40 Current: 75mA NOTES: excellent polish. Lower temperature result in an uneven foil surface.
I hope this information helps. If I can be of any additional assistance,=
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Ozgul Keles } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Hi everyone, First of all I would like to thank Dr. Simkin from Michigan University forthe information about Electron Channelling. I am going to get in touch=
with him. This time my question is about jet polishing solution of copper and copper and zinc alloys. Actually i found a solution that is sixty percent=
phosphoric acid and deionized water but the problem is that it is hard t= o get rid of that solution from the surface of specimen after you are done with=
jet polishing. If anybody has some idea about how to clean the surface after jet polishing i really appreciate it. =
Does your current TEM meet the department's needs?
You said: "no materials science types and no professional schools." are you into biological TEM, engineering or physics?
A bit more information is needed to help you on the technicality.
As for age, I am not sure it is such a problem. We have a Philips EM400 (120 keV) bought back in 1984 (I think- I was 11 at the time!), which despite its age is fantastic to work with. The success of this machine is that we have always had a service contract with Philips on it. If your machine is fine at the moment why don't you get a good service contract, and maybe spend some money on retro fitted equipment, for example CCD camera for digital imaging.
If you need to extend the TEM capability you may want to invest in a second hand machine ($10,000's) that will be a bit more adaptable. If you want to do this, then I am sure someone will suggest a web page, so that you can have a look at the current price ranges. A safer bet is to contact a reputable TEM manufacturer (Philips, Jeol, Hitachi, .etc) and ask for a contact and ask if they will support it.
I am afraid you are going to have to do a bit of research about this, so it may take a couple of weeks to get an idea as to what your options are. If you do this you should find yourself with a TEM that is well supported (if things do go wrong), should be a delight to use and be at a good price. However if you don't you could end up with something that is a waste of cash, frustrating to use and out of date.
Good luck, Jon
P.S. I know at present Philips are taking a lot of orders for their new Tecnai's (fully integrated computer/TEM), so you may find that there are a lot of second hand TEMs being advertised over the next few years.
-- ***************************************** Dr Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
In a message dated 5/19/99 2:25:49 PM EST, jmkrupp-at-cats.ucsc.edu writes:
{ { I guess our TEM might be used 300 to 500 hrs/yr sometimes less, never more.
We have a JEOL 100B circa 1975. For most of our applications, it works fine. I have a whole other microscope to use for parts and a clever service provider who may be able to keep this thing going for a long time. In a darkened room, it is hard to tell it from a more modern instrument based on the images seen on the screen. When the lights go on, it looks 24 years old.
Recently a new faculty member asked when are we getting a new TEM, one with digital imaging. I choked, said I would check into it. So here are a few questions I need help with and I hope for some kindness and understanding in your replies. I do need your replies because as the only real EM person here I could use your help.
I have never gone out looking for $$ for a new microscope. Where does one look? Could we justify $$ based on our level of usage? What do you think? What are the criteria used by funding agencies when evaluating requests. Could you share you experiences?} }
The decision to buy another instrument would depend entirely on need for additiona; features. Sounds as though the current instrument works well for your current needs. However, you might consider upgrading to a more recent instrument with additional features - but look for a used EM. There always seems to be used EMs available at excellent prices.
{ {Suppose we can't get the $$ for a whole new TEM. Would I be crazy to try to get $$ to equip our current TEM (remember 24 years old) with an add on digital camera? The ports are there and it could be done, but I'm not sure it is a good idea.} }
If you have the ports then why not. So long as you have a good resolution image coming down the column then you can do what you want with it in terms of image storage.
The Department of Microscopy and Microanalysis at Abbott Laboratories i= s recruiting a senior level microscopist for its Materials Science group.=
Requirements include a Ph.D. in Materials Science, Chemistry, or relate= d field, and a working knowledge of most of the following instrumentation= : TEM, SEM, EDXS, XPS/SIMS, AFM, and light microscopy methods (polarized light= , fluorescence, and confocal microscopy). Familiarity with spectroscopic=
techniques such as FTIR and a strong background in physical and analyti= cal chemistry are desirable. Experience with biological systems would be = highly advantageous.
We are looking for a team player with excellent interpersonal skills an= d the ability to readily adjust to rapidly changing priorities. The successf= ul candidate will seek out and learn new technologies as needed to solve problems related to pharmaceutical and healthcare products. The abili= ty to communicate clearly, both verbally and in writing, is essential.
Essential job functions: Independently design and carry out experiments to evaluate materials specimens by microscopic and microanalytical techniques. Develop methods using multidisciplinary approaches. Interpret data and effectively communicate results. Review data and reports for scientific integrity and clarity. Mentor career development of junior scientists. Present data at scientific meetings and publish in peer-reviewed journa= ls. Provide technical and scientific leadership.
The Department of Microscopy and Microanalysis provides corporate-wide support in materials and biological microscopy to all Abbott Laboratori= es divisions. The facility houses two TEMs (a Philips CM12 STEM and a LEO= 910), two SEMs (a Philips XL30 and AMRAY 1830i), three EDXS systems, a BioRad= MRC 1024 UV confocal scanning laser microscope, a Digital Nanoscope III AFM= with Bioscope, a Physical Electronics 5600 XPS/SIMS, and the usual assortmen= t of light microscopes and sample preparatory equipment.
For further information, please contact:
Jane A. Fagerland, Ph.D. Abbott Laboratories D45M/AP31 200 Abbott Park Rd. Abbott Park IL 60064-6202
Hello dear all! I am looking for any information on the microscope, called Grumhauser and on the person who made it. The spelling I have might be incorrect. Best regards, Dima My email address is la_dima-at-hotmail.com
_______________________________________________________________ Get Free Email and Do More On The Web. Visit http://www.msn.com
You can test the reservation system by logging in as user=guest, password=guest. You can make reservations on the "dummy" machine. During the couse of writing the software, we discovered that not all web browsers are created equal. There seem to be some major differences in the level of Java support. Anyway, on a Mac you need IE 4.0 or higher, on a PC you need IE 4.0 or Navigator 4.0 or higher Mac Navigator doesn't work yet.)
Our system is running on a WinNT PC using Microsoft Access as a database backend. The front end was done with MS InterDev 6.0 and Borland JBuilder 2.0.
At 10:33 AM 5/15/99 -0500, you wrote: } } } We are also considering developing a web-based reservation system, and I } think many others would also be interested what other labs are doing in } this area. I would be interested in learning of any software which is } freely available or low cost and open source. } } We are in the very early stages of modifying an open source (Perl) } web-calender and scheduling system to suite our needs. The reference url } is: } http://curiosityshoppe.tierranet.com/framecal/index.shtml } } I found this and a few others by searching the web with I believe was } combinations of ("resource" and "scheduling" and "calenders") } } If we develop a system based on FrameCal or something else we will make } it freely available as licensing permits. } } Of course, if someone else has a suitable system available freely or at } low cost it would save us development time and costs. } } Jim Mabon } } } } } } I know there have been some discussions on this listserver in the past } } } about computer sign up systems for electron microscopes. Are there any } new } } programs out there? We are working on a web-based instrument sign up } } program to replace our existing sign up/microscope accounting system. } The } } old system is not Y2K friendly. Besides the Universty of Minnesota, } is } } there any other group which has a working web-based system? } } } } John C. Wheatley } } Lab Manager } } Arizona State University } } Center for Solid State Science } } PSA-213 } } BOX 871704 } } Tempe, AZ 85287-1704 } } } } Phone: (602) 965-3831 } } FAX: (602) 965-9004 } } John.Wheatley-at-ASU.Edu } } } Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility 116 W. 19th Ave. (614) 292-0674 "Nothing is as inevitable as a mistake whose time has come."
I have some crystals of a drug in aqueous suspension. They clump together. I would like to disperse the crystals. Sonication is about the only way I can think of to do it. Any ideas? If sonication, for how long should I do it (about 0.3 ml volume) and how long would it be likely to last before it all clumps again? I should add that I can't add anything to the solution and I don't know the chemistry of the drug!
Thanks, Diana
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
At 12:10 PM 5/19/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Consider other than a new scope. Digitize your existing scope for a fraction of the cost of a new scope. A good source is http://www.elmdas.com
They make a nice computer control and image analysis package at a decent price. POC is John Best.
One- or two-year POST-DOC position available at the Electron Microscopy for Materials Science (EMAT) group of the University of Antwerp, RUCA, Belgium. Do visit our website for more info on the working of the group (http://wcc.ruca.ua.ac.be/EMAT).
The position is part of a European TMR network (9/98 - 9/02) on Phase Transitions in Crystalline Solids and is open to inhabitants of member states of the European Community or associated countries. The network is a collaboration between experimental and theoretical groups all working on the understanding of microstructures resulting from phase transitions in solids (see also http://www.dmsa.unipd.it/tmr/). Subprojects on martensitic transformations in alloys, twinning in oxides, thin films structures etc. have already been chosen. The candidate should have a large experience in different TEM techniques for the characterisation of atomic and microstructures. The position is open as of October 1, 1999. Salary starts at 1.500 Euro (scholarship, no taxes deducted), depending on experience.
If you are interested or like more information, please contact Dr. D. Schryvers (tel.: 32-3-2180247, fax: 32-3-2180257, e-mail: schryver-at-ruca.ua.ac.be)
!!!!! NEW SERVER !!!!
DO CHECK MY E-MAIL ADDRESS: schryver-at-ruca.ua.ac.be (replies to old mails, e.g. from before March 15, will not work anymore!)
*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=* *=* *=* *=* Dr. D. Schryvers *=* *=* Electron Microscopy for Materials Research (EMAT) *=* *=* University of Antwerp, RUCA *=* *=* Groenenborgerlaan 171 *=* *=* B-2020 ANTWERP *=* *=* Belgium *=* *=* tel: 32-3-2180247 *=* *=* fax: 32-3-2180257 *=* *=* e-mail: schryver-at-ruca.ua.ac.be *=* *=* homepage: http://www.ruca.ua.ac.be/EMAT *=* *=* *=* *=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*
One- or two-year POST-DOC position available at the Electron = Microscopy for Materials Science (EMAT) group of the University of = Antwerp, RUCA, Belgium. Do visit our website for more info on the = working of the group (http://www.ruca.ua.ac.be/EMAT).
The position is part of a joint project between EMAT and the = photographic company Agfa-Gevaert (3/99 - 3/01) on new photographic = materials and is open to inhabitants of member states of the European = Community or associated states. The candidate should have a large = experience in different TEM techniques for the characterisation of = atomic and microstructures. The position is immediately available. = Salary starts at =B1 1.500 Euro (after tax deduction), depending on = experience.
If you are interested or like more information, please contact Dr. D. = Schryvers (tel.: 32-3-2180247, fax: 32-3-2180257, e-mail: = schryver-at-ruca.ua.ac.be)
!!!!! NEW SERVER !!!!
DO CHECK MY E-MAIL ADDRESS: schryver-at-ruca.ua.ac.be (replies to old = mails, e.g. from before March 15, will not work anymore!)
*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*= =3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D* *=3D* *=3D* *=3D* Dr. D. Schryvers *=3D* *=3D* Electron Microscopy for Materials Research (EMAT) *=3D* *=3D* University of Antwerp, RUCA *=3D* *=3D* Groenenborgerlaan 171 *=3D* *=3D* B-2020 ANTWERP *=3D* *=3D* Belgium *=3D* *=3D* tel: 32-3-2180247 *=3D* *=3D* fax: 32-3-2180257 *=3D* *=3D* e-mail: schryver-at-ruca.ua.ac.be *=3D* *=3D* homepage: http://www.ruca.ua.ac.be/EMAT *=3D* *=3D* *=3D* *=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*= =3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*
I have just started trying a different approach on our intranet. The intranet runs on Microsoft BackOffice 2.5 and includes Exchange Server 4.0. I have setup mailboxes for our HRTEM and Ion Mill and hope to make use of the group scheduling and calendaring (is that a word?) functions of Exchange to achieve the same end. Client side software will be Exchange client / Outlook 97. Users can book by sending appointment requests to the mailboxes which can be set to automatically accept / suggest free time slots. I know this has access restricted only to our intranet unlike web based programs, but then at present we do not have scientists outside the organization directly booking resources. At present I am trying to get over the problem of compatibility: exchange client users do not have access to the outlook calendar and outlook users have chosen not to use schedule 97 as default calendar. If there are others using this method, I would like to hear from them as well as about web based programs.
--- R Divakar PMS, IGCAR, Kalpakkam 603102, India ----
This is a repeat of an earlier message that might not have made it. ;)
I have three (3) Osram UV bulbs that were used in an older, large, stand-alone UV lamp (The box is labeled: OSRAM Quecksilber-Hochstdruck-Lampe HBO 200W) If you need more info, or have need of these contact me direct. Thanks, john
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu
We have a system that we have just implemented (Actually my colleague=20 Corinna Wauchope did it) that uses Filemaker Pro to book instrument=20 time, track our user information, keep the instrument logbooks and=20 bill our users. It is working quite well now. We plan to put it on=20 the Web when we have all the glitches ironed out and all the=20 instruments included on it and whenCorinna has some spare time to do=20 the necessary Web stuff ( :-) ).
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Please note new FAX number.
John Mansfield PhD CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
I run an EM lab at a small private college using a 1968 Hitachi Hu 125E and an old 1970 ETEC autoscan. !00% biological TEM & SEM, very few beam hours per year, no fees, no hassel. I've got spares in the closet, do 100% of my own service ~ with some help from my friends (THANKS to Allen Sampson).
Currently, I have said no to the question "buy a new scope", because what we have now has been working for 10 years, plus the money is not there. I feel that one is tied into a service contract with the purchase of a new instrument because of the complexity. If a new scope were purchased, no longer can you fix it with a relay, vacuum tube, or spare parts in the closet. Someday I will change my mind as the avaliability of parts dwindles and frustration levels go up.
I know our situation will not go on forever, and feel that planning for a new scope, service contract and in-house support should take place long before the old scopes become more trouble than they are worth. Your facutly members will have to be the ones to write grants to NSF programs and private foundations. Within the grant writing process, the questions that you ask will get answered as support either builds or dies for new instrumentation.
Robert
} } OK, I know asking this question of the group may seem strange, but this is } the most densely populated area of microscopy wisdom in the universe and I } need your global perspective. } } I am the only person in a small EM lab that serves a primarily } undergraduate campus. I am part of the technical staff, I am not a faculty } member. We have doctorate programs and a few researchers using TEM, but no } materials science types and no professional schools. I guess our TEM might } be used 300 to 500 hrs/yr sometimes less, never more. } } We have a JEOL 100B circa 1975. For most of our applications, it works } fine. I have a whole other microscope to use for parts and a clever service } provider who may be able to keep this thing going for a long time. In a } darkened room, it is hard to tell it from a more modern instrument based on } the images seen on the screen. When the lights go on, it looks 24 years } old. } } Recently a new faculty member asked when are we getting a new TEM, one with } digital imaging. I choked, said I would check into it. So here are a few } questions I need help with and I hope for some kindness and understanding } in your replies. I do need your replies because as the only real EM person } here I could use your help. } } I have never gone out looking for $$ for a new microscope. Where does one } look? Could we justify $$ based on our level of usage? What do you think? } What are the criteria used by funding agencies when evaluating requests. } Could you share you experiences? } } Suppose we can't get the $$ for a whole new TEM. Would I be crazy to try to } get $$ to equip our current TEM (remember 24 years old) with an add on } digital camera? The ports are there and it could be done, but I'm not sure } it is a good idea. } } I hope you can give me some leads and encouragement. Most of the burden of } doing this job will fall on me. Although often supportive in casual } discussions, I don't expect much help from anyone else here. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
Robert Fitton Luther College Department of Biology 700 College Drive Decorah, IA 52101
Voice 319-387-1559 FAX 319-387-1080
Enjoy a visit to our website: http://www.luther.edu/dept/bio.htm
There have been several questions recently concerning the status of VCR Group. I have heard many rumors and a lot of talk "around the water cooler". I would like to dispel any rumors and clarify the situation. South Bay Technology and VCR Group have come to an agreement that will ensure the continuation of the VCR Group product line. The VCR products are being incorporated into the South Bay Technology line of products an= d will be manufactured and marketed by South Bay Technology, Inc. Althoug= h VCR Group, Inc. will no longer exist, that does not mean that the product= s will die or that support for existing customers will not be available.
We are in the process of transferring VCR operations and manufacturing to=
our corporate offices in San Clemente, CA. We expect to be fully operational by June 1 and will be able to fulfill orders shortly thereafter. Please be assured that we are here and available to support your VCR products and will be well into the future. In addition, to ensure a smooth transition, South Bay Technology has hired Vince Carlino, President of VCR Group.
As communications with VCR have been spotty over the past few months, I would encourage anyone with any questions about the VCR Group or products to contact me immediately with the relevant details. I will ensure that your order or problem is dealt with promptly=
and that your order is shipped within a realistic timeframe.
I thank you for this opportunity to set the record straight and I welcome=
all of our new VCR customers to the South Bay Technology family - I will look forward to hearing from you!
Best regards-
David Henriks Vice President
South Bay Technology, Inc. TEL: 800-728-2233 (tollfr= ee in the USA) 1120 Via Callejon +1-949-492-260= 0 San Clemente, CA 92673 USA FAX: +1-949-492-1499
Randi We use Trump's fixative for routine TEM analysis of surgical pathology specimens or surgical biopsies. Basically it is 4% formaldehyde and 1% glutaraldehyde in phosphate buffer. One reference available is Arch. Pathol Lab Med 100:405, 1976, this is a comparison of several fixatives, If I can help further, feel free to contact me. Marge
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
Steven, I'm sure you are aware there is a room temperature equivalent to your message, but I want to make sure that others on the Listserver don't conclude that cryosectioning is the only way to do this. For many materials, e.g. softer metals and some polymers, one can section smoothly at RT with a normal microtome and diamond knife. For many other polymers (and most biological materials), a smooth surface is produced at RT, but smearing distorts the shape of structural features. For harder crystalline materials, e.g. steels or intermetallics, a surface usually is produced which is microscopically rough to a degree dependent on the shear/brittle fracture details of the system, no matter what the temperature. Even more complex is the case for embedded ceramic/mineral particulate, fibers and the like, where the critical dimension seems to be size. Such features a few microns or less in diameter tend to section smoothly at either cryo or RT, but much larger, and they start to section with a rough surface (which still might furnish much useful information, of course).
I am encountering a growing number of microtomy workshop students, both private and public sector, who are using sectioning as a prelude to scanned probe studies, as well as optical, SEM and TEM. Thus the SPM community would do well to consider what is meant by a 'smooth' or 'rough' specimen surface in terms of what types of information can be collected realistically from one extreme to the other.
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 613-992-8735 email: malis-at-nrcan.gc.ca
} ---------- } From: Slap, Steven[SMTP:SSlap-at-ebsciences.com] } Sent: May 18, 1999 10:34 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: cryoplaning } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear fellow microscopists, } } I am looking for individuals interested in cryoplaning techniques (using } a cryoultramicrotome to produce blocks with a very flat surface and } examining this surface in a cryo-SEM). Please contact me back-channel } if interested. } } Best regards, } Steven E. Slap, Vice-President } ******************************************* } Energy Beam Sciences, Inc. } The Laboratory Microwave Company } Adding Brilliance to Your Vision } http://www.ebsciences.com {http://www.ebsciences.com} } ******************************************* } } }
Maybe someone has already suggested this to you, but if digital imaging is the only thing you lack, the cheap (but effective) way to go might be to invest in a good quality scanner. You scan in the negatives you shoot on your present instrument and you have instant digital images, plus an archival, high quality film backup in your originals. This will cost a fraction of adding on digital capabilities to your scope, especially if you already have a decent computer to run it from.
I am a big advocate of capturing images on film, then manipulating those images digitally. Highest quality, lower cost, and platform/software independent archiving of images. My two cents.
OK, I know asking this question of the group may seem strange, but this is the most densely populated area of microscopy wisdom in the universe and I need your global perspective.
I am the only person in a small EM lab that serves a primarily undergraduate campus. I am part of the technical staff, I am not a faculty member. We have doctorate programs and a few researchers using TEM, but no materials science types and no professional schools. I guess our TEM might be used 300 to 500 hrs/yr sometimes less, never more.
We have a JEOL 100B circa 1975. For most of our applications, it works fine. I have a whole other microscope to use for parts and a clever service provider who may be able to keep this thing going for a long time. In a darkened room, it is hard to tell it from a more modern instrument based on the images seen on the screen. When the lights go on, it looks 24 years old.
Recently a new faculty member asked when are we getting a new TEM, one with digital imaging. I choked, said I would check into it. So here are a few questions I need help with and I hope for some kindness and understanding in your replies. I do need your replies because as the only real EM person here I could use your help.
I have never gone out looking for $$ for a new microscope. Where does one look? Could we justify $$ based on our level of usage? What do you think? What are the criteria used by funding agencies when evaluating requests. Could you share you experiences?
Suppose we can't get the $$ for a whole new TEM. Would I be crazy to try to get $$ to equip our current TEM (remember 24 years old) with an add on digital camera? The ports are there and it could be done, but I'm not sure it is a good idea.
I hope you can give me some leads and encouragement. Most of the burden of doing this job will fall on me. Although often supportive in casual discussions, I don't expect much help from anyone else here.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Biological Measurements and Forces in MAC Mode™ AFM Stuart Lindsay, Wenhai Han, and Yanzhang Liu; ASU
Biological force measurements and the AFM Protein unfolding may play a vital role in protein function (Soteriou, Clarke et al. 1993). Last year, Rief et al. (Rief, Gautel et al. 1997) demonstrated that the AFM could be used to follow the unfolding of a single protein molecule trapped between the AFM probe and a gold surface
I have a colleague who would like to label antibodies for fluorescence for use in leaves, but would like an alternative to fluorescein. I believe Molecular Probes has quite a selection... are there some in particular that users have been happy with?
thanks in advance for your help shea
Dr. S. Shea Miller Agriculture and Agri-Food Canada Eastern Cereal and Oilseed Research Centre 2068 K.W. Neatby Bldg Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 email: millers-at-em.agr.ca phone: 613-759-1760 fax: 613-759-1701 ! !
} -----Original Message----- } From: Howard Fielding } Sent: Monday, May 17, 1999 1:25 PM } To: 'microscopy-at-msa.microscopy.com' } Subject: Electron Microscope For Sale } } Our firm has a ISI Electron Microscope, model DS 130 C for sale. Any } interested parties may call Howard Fielding at 800 992 7844 ext 304 for } information or respond via e-mail to hfielding-at-mayflower-sac.com. We have } acquired this device from a warehouse customer who has defaulted on } storage charges and can make a very attractive price to prospective } purchasers.
Does anyone know if in situ's can be done on epoxy embedded sections? If yes, does the tissue have to be treated any differently during fixation and embedding?
Thanks for all the interest, but the bulbs have already found a good home. We are also looking to get rid of some vacuum tubes (assorted sizes and manufacturers). All free for the cost of shipping. Spring cleaning, y'know? john
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu
I am interested in exploring the feasibility of jet propane freezing our samples to minimize artifacts from ice formation, but am having a hard time finding a facility that has a rig. Any contact pointers would be very much appreciated, as would any clues on techniques used to minimize phase separation caused by the formation of ice. We are currently plunging our samples (colloidal dispersions) in liquid propane, but the size of the ice artifacts are on the order of our particle size.
I have been watching this discussion closely as I too am working with an antiquated scope, a 1975 Philips 201. I work only with clinical specimens, almost no research, and primarily muscle, nerve and renal biopsies. The 201 is functioning quite well also due in part to a master "Mr. Fixit". We have even more in common. I have a 201 parts scope sitting right next to my primary scope. I have sold the pathologists I do EM for on digital imaging, but due to tight purse strings am not going to be able to do what I would like to do. So, now to my question. I have been looking into the scanning possibilities and would love all the input you all could give me. I am looking to scan the standard Kodak 4489 negatives. What scanners are good for this? What price range am I looking at? Thanks in advance for all your input.
Douglas C. Rennie Electron microscopy lab coordinator University of Nebraska Medical Center Omaha Nebraska (402) 559-7729
We have an immediate opening in our facility for specialist in materials characterization. Areas of application include SEM/EDS, mProbe and ESCA/XPS. Background in inorganic chemistry, AA & ICAP are also desirable.
Breim Engineering is a laboratory environment that applies these techniques to metallurgical, materials characterization and failure analysis. Reply to: dlpark1-at-briemeng.com
********************************************************************** Donald L. Parker Briem Engineering, Inc 4134 Rider Trail North Earth City, MO 63045 [314] 298-3773, direct ext #27 FAX [314] 298-7097 **********************************************************************
We have a Hitachi HU12A to give away. It is a complete system with manuals. It is located at the Health Science Campus of USC in Los Angeles, CA. For additional information please email me, and I will get back to you asap. Thanks
At 09:36 AM 5/21/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Agfa Arcus II does 600x1200 dpi and runs about $850. The UMAX Powerlook III does 1200x2400 dpi and runs about $1200. Both do a good job. Higher resolution is good and the UMAX has a bit higher D range.
I need to find a light source, either a mirror or an electric lamp for several monocular light microscopes (Leitz). What would be the cost? Is there a generic brand that would work? Thanks for any suggestions!
Barbara Plowman Univ. of the Pacific School of Dentistry email: Bplowman-at-uop.edu
We are planing to immuno-gold label the melanocytes of equine melanoma. If you know of any commercial or non-commercial source of specific antibodies against melanocytes (such as Ab against melanosomes) please let us know. We would really appreciate that.
Regards
-Majid Ghoddusi
........................................................................... ........................................................................... ...... Majid Ghoddusi, PhD Centre for Microscopy & Microanalysis and Division of Veterinary Pathobiology & Anatomy The University of Queensland Qld 4072 Australia
To all who have replied to my inquiry, I want to thank you for being open and candidly sharing your experiences and expertise with an amateur like myself. It has been extremely helpful in clarifying my concerns.
I continue to welcome any additional input or comments.
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Without throwing the cat too heartily amongst the pigeons I would value your considered comment on the virtues of LVSEM against ESEM in a multi-user mainly biological EM facility.
Obviously I have investgated the subject as thoroughly as I can but there is nothing like a touch of personal experience to enrich this information for us,
Please send your responses directly to me by return or to bruton-at-emu.unp.ac.za. Our website (still under construction) address is provided beliow if you would like to know a little more about us.
I look forward to hearing your views.
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Tel +27 (0)331 260 5155 Fax +27 (0)331 260 5776 website:http:www.nu.ac.za (departments} units) Email:bruton-at-emu.unp.ac.za postal address; Private Bag X01, Scottsville, 3209 KwaZulu-Natal South Africa
Hi All: I am looking for a third party company that can offer me a service contract on a Philips 430 ST. Thank you so much for your ongoing assistance.
I recently bought a new (now discontinued) Wild M7S stereo Microscope, which included as part of the package a Wild model 10446174 trinocular phototube - this is the latest white version, 100% observation or 100% photo, built-in double iris diaphragm, 37mm photo port. This last part is driving me nuts, since all my photo equipment requires a 38mm photo port. Boring the phototube out to 38mm is not an option. What is an option is trading this to someone for an older black version with the 38mm photo port. The version that I have fits the M3 and M7, as well as all the current Leica stereo microscopes (MS5, MZ6, MZ8, etc.) - anything Wild or Leica except the M5 (this takes a different size, and they will not interchange). If you have a 38mm Wild phototube and are interested in becoming compatable with all of the new Leica digital and video adapters (which are 37mm, or so I understand), let me know and perhaps we can work out something.
Can anybody tell who are suppliers of gold-labelled Tumour Necrosis factor ? I don't seem to able to find anything in the catalogues I have at hand.
Thanks in advance
Cathy Gillespie
Cathy Gillespie Head Electron Microscopy/Histology/FACS Facility John Curtin School of Medical Research Australian National University Canberra Australia
Our Kevex 8000 system won't talk to an external computer and we would like to do peak to background analysis on spectra we have and will collect. It appears that Quantex will not give us straight P/B. Suggestions on how to proceed will be appreciated.
Dear Richard, I feel that it would be impossible to do in-situ hybridisation on conventional fixed and epoxy-embedded sections. There occur a few severe problems :
1.) Fixation: If you use conventional aldehyde-fixation, proteins as well= as nucleic acid will be crosslinked heavily and binding sites for your in-si= tu will be deleted
2.) Postfixation: Osmium will even increase the cross-linking and should = be avoided in any case
3.) Embedding: Using conventional hydrophobic epoxy resins you archive va= st three-dimensional cross linking of proteins, nucleic acids, amino-acids .= . Your in-situ probe will not have any access to binding sites.
If you have too much time/manpower or in situ hybridisation in absolutely necessary for your work you could try it with the following hints but I wouldn=B4t expect too much:
a) use a "soft" fixation (e.g. 2-4% Formaldehyd) b)don=B4t use osmium nor uranyl acetate c) don=B4t use epoxy resins, try more hydrophilic resins (e.g. LR-White o= r LR-Gold or the Lowicryls if you aren=B4t frightened of their toxic potential...) d) improve contrast of sections: strong postfixation and staining (why no= t including osmium)of sections after in-situ labelling
Another trial would be to try Ultracryotomy of cryo-protected deep frozen tissue (Tokaysu technique). But this will cost you a lot of equipment, ti= me and motivation ...
Good luck, with best regards Michael Reiner
Michael Reiner Department of Anatomy University of Cologne Germany e-mail: a2811111-at-smail.uni-koeln.de
Richard Gardiner schrieb:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------. } } Does anyone know if in situ's can be done on epoxy embedded sections? } If yes, does the tissue have to be treated any differently during } fixation and embedding? } } Richard Gardiner
Thanks very much to all who responded to my query about Trump's fixative. Got what I needed to finish processing the samples on hand.
Learned a couple of interesting things in the process: apparently there are two versions of Trump's---the standard one with paraformaldehyde and gluteraldehyde, and another one using formalin. The latter is probably a histology fixative, I guess. The other neat thing to know is that several people described Trump's as an excellent storage fixative. One person even said they'd had specimens in Trump's for nine years, with no noticeable deterioration!! (Don't try this at home folks?)
Anyway, thanks again. As usual, the list came through.
Dear Dr. Gauler, We got copy of your message and we wonder why you did not get in touch with our representative or with the field engineer in charge of your area. Eventually you can get in touch directly with us visiting our site www.leo.de We should like to help you to solve your problem, and to do that we need some information. Please, let us know: How long ago have been made the last service on the penning gauge. What is the vacuum reached in the column when the separation valve is closed. Where did you get the filaments you are using. How long ago did you get them. It could be helpful to know the serial number of your EM 10.
It may be that you have a vacuum problem or that something is wrong with your wolfram filaments. The all data we ask are useful for us to know the exact configuration of your microscope as well as to rebuild its all technical history. We are waiting for your answer in order to solve your problem. Thanks in advance for your cooperation.
LEO COE-TEM / Marco Arienti Carl-Zeiss Strasse, 56 D-73446 Oberkochen
Greetings to the Microscopy list. As a relatively new member of the list, this is my first request for help. I would like to know if is possible to obtain image of dendritic cells detected by elettron microscope. Furthermore if you know an internet site whit some informations related to these cells. Thanking you all in advance,
I would appreciate advice on the best way to obtain grain size statistics from a TEM image. Is there any software that will do this? Since we will be starting with a print from a TEM negative, are there any special scanner capabilities that are required? Any other suggestions on this process would also be appreciated.
Thank you for taking time to consider this request.
Sincerely,
Mick Thomas --------------------------------
Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
-----Original Message----- } From: Fagerland,Jane [mailto:jane.a.fagerland-at-abbott.com] Sent: Wednesday, May 19, 1999 5:59 PM To: Microscopy-at-sparc5.microscopy.com Cc: Ulrich,Roger
The Department of Microscopy and Microanalysis at Abbott Laboratories is recruiting a senior level microscopist for its Materials Science group.
Requirements include a Ph.D. in Materials Science, Chemistry, or related field, and a working knowledge of most of the following instrumentation: TEM, SEM, EDXS, XPS/SIMS, AFM, and light microscopy methods (polarized light, fluorescence, and confocal microscopy). Familiarity with spectroscopic techniques such as FTIR and a strong background in physical and analytical chemistry are desirable. Experience with biological systems would be highly advantageous.
We are looking for a team player with excellent interpersonal skills and the ability to readily adjust to rapidly changing priorities. The successful candidate will seek out and learn new technologies as needed to solve problems related to pharmaceutical and healthcare products. The ability to communicate clearly, both verbally and in writing, is essential.
Essential job functions: Independently design and carry out experiments to evaluate materials specimens by microscopic and microanalytical techniques. Develop methods using multidisciplinary approaches. Interpret data and effectively communicate results. Review data and reports for scientific integrity and clarity. Mentor career development of junior scientists. Present data at scientific meetings and publish in peer-reviewed journals. Provide technical and scientific leadership.
The Department of Microscopy and Microanalysis provides corporate-wide support in materials and biological microscopy to all Abbott Laboratories divisions. The facility houses two TEMs (a Philips CM12 STEM and a LEO 910), two SEMs (a Philips XL30 and AMRAY 1830i), three EDXS systems, a BioRad MRC 1024 UV confocal scanning laser microscope, a Digital Nanoscope III AFM with Bioscope, a Physical Electronics 5600 XPS/SIMS, and the usual assortment of light microscopes and sample preparatory equipment.
For further information, please contact:
Jane A. Fagerland, Ph.D. Abbott Laboratories D45M/AP31 200 Abbott Park Rd. Abbott Park IL 60064-6202
Be very, very careful concerning "software for grain size measurement from TEM images." Because grains seen in the TEM often contain dislocations, twins, stacking faults, high preferred orientation, etc; as well as having every shade of grey from B to W; ^ ANY ^ video-input/scanning based "automatic" grain size measuring machine will produce suspicious results. I didn't say they don't produce a "number," just that the "number" has little correlation with grain size. The human eye can easily see grains in a TEM image; machines can't.
What we do is measure the maximum diameter (chord) of about 200 grains in a series of TEM images using a digitizer. After entering the image's magnification into the PC connected to the digitizer, we 'click-on' each end of each grain's max chord. The PC calculates the chord length. When we think we have measured enough grains, we tell the PC to calculate the grain size. It calculates both the normal and lognormal grain sizes from the chord lengths (equations and correction factors from Shuckher's chapter in DeHoff & Rhines "Quantitative Microscopy"), tests which one better represents the measured distribution and tests whether we really have measured enough grains. Assuming it doesn't tell us to measure more grains, the computer plots a grain size histogram, overlays it with the calculated distribution for an eye-ball check, and reports all the normal and lognormal parameters (telling us which parameters are best). Besides using the human eye-brain for doing what it does best (judgement) and the computer for doing what it does best (crunching), the method as described beats automated systems time-wise (from walking in with an image to walking out with data), which we consider funny.
Having said all of this, I realise time marches on and things might have improved. If anyone out there has an automated machine that can measure grain size on my collection of images they are welcome try. They are: An 'easy' Mo grain size with no internal structure--just b/w grains; Cu grains with sharp b/w twins; ceramic grains with dislocation tangles and b/w gradation in the grains; An Al film with high preferred orientation such that neighbouring, similarly oriented, grains have nearly the same shade of grey; and a horrible duplex Cu distribution with a b/w gradiant across the image, twins, and dislocations.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Mick wrote:
Fellow microscopists,
I would appreciate advice on the best way to obtain grain size statistics from a TEM image. Is there any software that will do this? Since we will be starting with a print from a TEM negative, are there any special scanner capabilities that are required? Any other suggestions on this process would also be appreciated.
Thank you for taking time to consider this request.
Sincerely,
Mick Thomas --------------------------------
Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
If your 8000 has a printer, it is likely it will have (?) a 4 port serial card. The OSys "copy" command can be used to send a file out this port which can be received by a PC in the serial capture mode. Another option I have used is to equip a PC with a SCSI card and a drive like that used on the 8000 (I am now using Syquests, was using Bernoullii 44s previously). Using the program "RTCOPY" files saved in RT-11 format cam be copied into dos/windows on the pc.
EDS spectra can be saved as individual (ASCII - Lotus delineated) files using the command "SAV/EXT" and be exported using the above information. It will be up to Lotus / Excel or the like to do the P/B calculations. Alternately, you could save the raw spectrum, strip the background, then save the background subtracted version. the difference between them would yield the bkgn.
If memory serves: A simple alternate approach, if you only have 4 elements (8 windows max), is to paint "windows" (like selecting for external dot map) on the elements and adjacent background area. Save the window file, then recall/print windows. Intergals contained in each window should be printed.
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Our Kevex 8000 system won't talk to an external computer and we would like to do peak to background analysis on spectra we have and will collect. It appears that Quantex will not give us straight P/B. Suggestions on how to proceed will be appreciated.
by mail.mdacc.tmc.edu (8.8.5/8.8.5) with SMTP id LAA00748 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 25 May 1999 11:46:25 -0500 (CDT) Received: by utm-notes-m2.mdacc.tmc.edu(Lotus SMTP MTA v1.2 (600.1 3-26-1998)) id 8625677C.005D1064 ; Tue, 25 May 1999 11:56:30 -0500 X-Lotus-FromDomain: MDACC To: Microscopy-at-Sparc5.Microscopy.Com Message-ID: {8625677C.005C19C7.00-at-utm-notes-m2.mdacc.tmc.edu}
Douglas
I am currently getting good results wioth an older HP ScanJet 4c with a transparency adaptor attached to it. Unfortunatly, the transparency adaptor cost more than the Scanjet did in the first place.
Both of these pieces are over 3 years old and I am sure that there are newer, better and cheeper scanners with transparency adaptors on the market today.
Mannie steglich Tech Dir Path E M Lab M D Anderson Cancer Center
Does anyone out there know of an epoxy for TEM use that does not soften substantially at temperatures up to 500C? We want to do a high-temperature straining experiment but don't have too much material and would like to use a 3mm disk as the gauge with bits glued to the end for the 'grips'. Also, it would need to cure at less than 150C so we don't alter the microstructure before we start the test.
Thanks in advance.
Cheers, JSV *************************** John S. Vetrano Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
We've just started acquiring digital 16 bit x-ray radiographs.
1) Is it worth doing 16 bit radiographs or will 8 bit do? 2) The images are HUGE! Also, much of our processing software only works well with 8 bit. Would converting from 16 to 8 bit be acceptable? If you don't for x-ray, please give your opinion for normal microscopy work.
Dear Jacob, We have done this on our Kevex 8000 and we just marked Windows of equal size on the peak and background regions and then read off the Integral of the Window (lower right-hand of the screen) after the count and copied it down manually. Clumsy, but it worked. You wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
I agree with Ron, that grain measurements on TEM images are very hard to do and one should not trust fully automatic measurements just because they are done by a computer. The reasons are the contrast mechanisms in a TEM. Bend conours, orientation contrast, dislocations and other effects can lead to artifacts that can easily be interpreted as grain boundaries or confuse software with grains of different intensity levels.
However, we have developed software to analyze grains in SEMs, where we also have to deal with grains of different intensity. While I cannot promise that it will work on your TEM images in an automatic fashion, I would like to offer you a trial. Send me a couple of your images and I will try if the software can be used to measure the grain sizes. The software does intercept techniques, similar to what Ron describes, as well as fully planimetric measurements. It also offers a semiautomatic mode for grain boundary reconstruction, which might be useful in this case.
Again, send me those images and I'll give it a try.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: } "anderron-at-us.ibm.com"-at-sparc5.microscopy.com[SMTP:"anderron-at-us.ibm.com" } -at-sparc5.microscopy.com] } Sent: Tuesday, May 25, 1999 9:22 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Software for grain size statistics } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Mick, } } Be very, very careful concerning "software for grain size measurement } from TEM } images." Because grains seen in the TEM often contain dislocations, } twins, } stacking faults, high preferred orientation, etc; as well as having } every shade } of grey from B to W; ^ ANY ^ video-input/scanning based "automatic" } grain size } measuring machine will produce suspicious results. I didn't say they } don't } produce a "number," just that the "number" has little correlation with } grain } size. The human eye can easily see grains in a TEM image; machines } can't. } } What we do is measure the maximum diameter (chord) of about 200 grains } in a } series of TEM images using a digitizer. After entering the image's } magnification into the PC connected to the digitizer, we 'click-on' } each end of } each grain's max chord. The PC calculates the chord length. When we } think we } have measured enough grains, we tell the PC to calculate the grain } size. It } calculates both the normal and lognormal grain sizes from the chord } lengths } (equations and correction factors from Shuckher's chapter in DeHoff & } Rhines } "Quantitative Microscopy"), tests which one better represents the } measured } distribution and tests whether we really have measured enough grains. } Assuming } it doesn't tell us to measure more grains, the computer plots a } grain size } histogram, overlays it with the calculated distribution for an } eye-ball check, } and reports all the normal and lognormal parameters (telling us which } parameters } are best). Besides using the human eye-brain for doing what it does } best } (judgement) and the computer for doing what it does best (crunching), } the method } as described beats automated systems time-wise (from walking in with } an image to } walking out with data), which we consider funny. } } Having said all of this, I realise time marches on and things might } have } improved. If anyone out there has an automated machine that can } measure grain } size on my collection of images they are welcome try. They are: An } 'easy' Mo } grain size with no internal structure--just b/w grains; Cu grains with } sharp b/w } twins; ceramic grains with dislocation tangles and b/w gradation in } the grains; } An Al film with high preferred orientation such that neighbouring, } similarly } oriented, grains have nearly the same shade of grey; and a horrible } duplex Cu } distribution with a b/w gradiant across the image, twins, and } dislocations. } } } Ron Anderson, IBM, Hopewell Jct., New York, USA. } anderron-at-us.ibm.com } } IBM Analytical Services; http://www.chips.ibm.com/services/asg } } } Mick wrote: } } Fellow microscopists, } } I would appreciate advice on the best way to obtain grain size } statistics } from a TEM image. Is there any software that will do this? Since we } will } be starting with a print from a TEM negative, are there any special } scanner } capabilities that are required? Any other suggestions on this process } would also be appreciated. } } Thank you for taking time to consider this request. } } Sincerely, } } Mick Thomas } -------------------------------- } } Mick Thomas } UHV-STEM Laboratory } E-1 Clark Hall } Cornell University } Ithaca, NY 14853 } } Phone: 607-255-0650 } Fax: 607-255-7658 } e-mail: mgt3-at-msc.cornell.edu } } } } }
I believe I have the required software which can DO automatic GRAIN SIZING! (let us see the response to that... :)
However can it do exactly what you what have in mind ALL the time and automatically?
For that I would reserve judgment until I have seen an image what you are attempting to qualify.
Mr. Anderson has brought up a number of points that need to be kept in mind. I especially like the selective method of analysis - with the computer being used where it helped and not where it could/would not (be it time or method dependent).
} If anyone out there has an (should I add SEMI-? until I have seen the images) automated } machine that can measure grain size on my collection of images they are welcome try.
I accept your challenge and raise you =85.
Possibly the best way forward would be for both of you to send me "typical" images (of the best possible quality) and what you would like to measure. I can then see if the software can do what is required and send you (and who ever else may be interested) the results.
Barry T. Dudley DUDLEY-at-I-CUBEinc.com
PS - The software I have in mind is called Vision Gauge.
-- ************************************************************ B.T. DUDLEY I-CUBE http://www.i-cubeinc.com Ph 1-888-77-I-CUBE 301-858-0505 301-858-0615 (Fax) I-CUBE is a Systems Integrator and Value Added Reseller of image analysis and image processing products for scientific and industrial applications. We provide a single source for imaging products, using a consultative selling approach I-CUBE 2411 Crofton Lane; Suite 14A; Crofton; Maryland; 21114 ************************************************************
It is not an epoxy but we have used `Autostick' a high temperature adhesive (up to 1100 C) which will stick to keyless surfaces, eg. polished foils. I don't have the details to hand but if you need them let me know and I'll try to hunt them down.
Ron
} Does anyone out there know of an epoxy for TEM use that does not soften } substantially at temperatures up to 500C? We want to do a high-temperature } straining experiment but don't have too much material and would like to use a } 3mm disk as the gauge with bits glued to the end for the 'grips'. Also, it } would need to cure at less than 150C so we don't alter the microstructure before } we start the test.
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
} } To: Microscopy-at-Sparc5.Microscopy.Com } } Subject: TEM Availability? } } Content-Type: text/plain; charset=us-ascii } } Content-Transfer-Encoding: 7bit } } } } } I would appreciate information on availability of a used CM12/CM200 } } electron } } microscope. The ideal "candidate" would be equipped with a STEM unit, } } EDS, } } PEELS, CCD camera and located, preferably, in the U.S. Individual } } attachments listed would be considered as well. Please contact me off } } line } } if you can provide assistance in locating this equipment. } } } } Bob Roberts } } EM Lab Services, Inc. } } Tempe, Az 85282 } } } } }
I was visiting a colleague of mine and we discovered a cache of "obsolescent" computer goodies:
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Fellow Microscopists, We are presently measuring particle size and position by LM on small particles dispersed over a large area. This is accomplished by a frame by frame raster. We are in the process of upgrading our system to accomplish this. My question concerns the possibility of using a linear CCD array and scanning the sample under the array to cover our area of interest. This would involve specialized software to interact with the stage control and some dynamic particle identification scheme. Has anyone out there in microland attempted any similar procedure. Russ Gillmeister Xerox
Emitech make three types of cold stages two of these are also preparation stages. The stages suit most, if not all SEMs. You can read up on these in our online on page E10D. We are making additon to this page and more complete info will be up within a week. Please note that ProSciTech distribute these instruments in Australasia (south of Singapore) only. For agents elsewhere see Emitech's email contact on page E10. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
-----Original Message----- } From: Marisa Ahmad [SMTP:mahmad-at-semiconductor.com] Sent: Wednesday, 26 May 1999 01:30 To: 'MSA listserver'
Looking for parts for a Zeiss OpMi-1 stereomicroscope. Please contact me off-list. Thank you.
James Martin Director of Analytical Services & Research Williamstown Art Conservation Center http://members.tripod.com/~James_Martin/dasrhome.htm
Research Scientist in Chemistry Williams College (www.williams.edu) http://members.tripod.com/~James_Martin
I have been using FITC-dextran as a permeability marker in rabbit aorta. I have tried using molecular weights of 10,000 and 70,000 separately. The tissue was immersion fixed in glutaraldehyde and post-fixed in 1% potassium ferrocyanate mixed with osmium tetroxide. The result was disappointing. I have not been able to find any dextran-ferrocyanate deposits between any gap junctions nor glycogen. Would anyone have any suggestions what else I can try? I have used horse radish peroxidase and know that it worked but cost was a major issue. Thank you for any help offered.
Fanny Chu
Mark Elliott, PhD Research Associate UBC-Pulmonary Research Lab St. Paul's Hospital Vancouver, BC Canada V6Z 1Y6 604-631-5351 (FAX)
The Science Illustration program here needs help learning about the features available and criteria to use when selecting a set of stereo microscopes for their students. They are about to purchase 10 scopes to be used by graduate level science illustration students. They think they need drawing tubes/camera lucida systems and want to provide their students with the opportunity to have drawing experience using a 'real' microscope.
So, all you Rembrandts and Picasso's out there, here is your chance to help out some talented and devoted artists. Send me your advice and I will forward it to them. Help them make an informed decision that will keep them a fan of microscopy.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Remember the size of the files in bytes will be the x-size times the y-size times 1 for 8-bit images and 2 for 16-bit images. So by going to 8-bit images you could cut your file size in half. However, more gains might be made by reducing your x and y resolution. (You did not mention what resolution you are using.) How much do you really need? Images over 1024x1024 are not very useful unless you are digitally magnifying them to see the details.
16-bit imaging is helpful if you have a wide range of gray scale. You can concentrate on bright areas of the image and adjust the brightness and contrast locally to see the details you want without having the gray level steps showing up. At 16-bit (256 gray levels) you will almost certainly see the steps after much less contrast enhancement.
You might want to keep your original image "as is" and pull out smaller regions that can be enhanced and save those enhanced files as 1024x1024 8-bit images. I venture to guess that the aggregate size of those "region of interest" images would be much less than your original.
At 01:39 PM 5/25/1999 -0500, you wrote: } We've just started acquiring digital 16 bit x-ray radiographs. } } 1) Is it worth doing 16 bit radiographs or will 8 bit do? } 2) The images are HUGE! Also, much of our processing software only works } well with 8 bit. Would converting from 16 to 8 bit be acceptable? If you } don't for x-ray, please give your opinion for normal microscopy work.
I am looking for eyepiece reticles/graticles for a Nikon SMZ-10 dissecting scope. In my catalogs I can find the most common diameters (19 and 21 mm), which are too small for my eyepiece, which measures about 24 mm inner diameter.
thanks in advance
Steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Chairman, Educational Outreach subcommittee promoting access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
Dear Microscopists, We would like to get a UPS for our TEM to bridge the gap between the instant there is a loss of power and the time when the backup generator takes over (a minute or so??). We have some information about APC and Tripp Lite UPS units, but we are concerned that their experience is limited to computer applications. We would like information on the types of UPS employed by EM users. We would also appreciate suggestions on how to chose the appropriate size. The power consumption of our TEM is about 3 kilowatts (instuction manual). Many Thanks in advance. Vachik Hacopian
I am pretty sure what you have in mind can be done, but what advantages do you think this would provide? For example, we have software that interfaces with stage controllers and allows you to do exactly what you want without having to develop new software. We do so by controlling the stage, set up a raster for image acquisition, then automatically move the stage, acquire an image, montage the images and do the analysis on them. Alternatively you can evaluate the image separately. For accurate statistics you need to set up the acquisition withput overlap in the latter case.
Unless there is an advantage for using a linescan CCD array (is there?) I would consider using off-the-shelf parts. Of course, I have a vested interest in this, as we are selling them, but I am sure there are other vendors out there as well.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
} ---------- } From: Gillmeister, Russ[SMTP:RGillmeister-at-sdms.usa.xerox.com] } Sent: Wednesday, May 26, 1999 7:37 AM } To: 'MSA' } Subject: Particle Analysis } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Fellow Microscopists, } We are presently measuring particle size and position by LM on small } particles dispersed over a large area. This is accomplished by a frame } by } frame raster. We are in the process of upgrading our system to } accomplish } this. My question concerns the possibility of using a linear CCD } array and } scanning the sample under the array to cover our area of interest. } This } would involve specialized software to interact with the stage control } and } some dynamic particle identification scheme. Has anyone out there in } microland attempted any similar procedure. } Russ Gillmeister } Xerox }
The graticule usually sits losely on a ledge within the ocular. So one mm less than the inner tube diameter is acceptable, three appears a bit much. Graticules are frequently made to order and the non standard sizes just cost a little more. Our info is online on page S3. I am happy to acknowledge that other suppliers likewise would be able to procure other sizes. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
-----Original Message----- } From: Steve Barlow [SMTP:sbarlow-at-sunstroke.sdsu.edu] Sent: Thursday, 27 May 1999 06:19 To: microscopy-at-sparc5.microscopy.com
Dear members,
I am working in KRISS(korea research institute of standards and science) and studying micoelectromechanical systems characterization by home-built photothermal microscopy and calibrated atomic force microscope. I am seeking to find infrared microscope( not FTIR) which views temperature differences of micromechanical systems specimen at roomtemperature with a resolution of micrometer. Also I am seeking infrared detector array which can be used to make infrared microscopy.
=09As there have been several enquiries for details of=20 high temperature adhesive I am posting details to the list=20 rather than direct replies.
I originally used Autostick some 25 years ago and now=20 discover that I am no longer able to obtain it. The last=20 time I needed any I used the `Sensor and heating cement'=20 from Oxford Instruments (Fax +44 1865 393333 - part no TGZ=20 0005 costs about =A36 for 50 gm pot). This seemed to be very=20 similar and was successful in my use of sitcking heating=20 elements onto a hot stage. It does dry in air but needs to be outgassed before use in=20 the microscope to prevent contamination.
=09I believe that the original manufacturer of=20 Autostick was Cotronics Corporation of Brooklyn, NY 11235.=20 They have advertised high temp resins (up to 700F) and=20 cements (up to 3000F) as late as 1992 but in pint or quart=20 quantities. They did not include Autostick in the catalogue=20 but had a larger range of cements for even higher=20 temperatures.
Regards, Ron
I have no financial interest in either company.
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
E. Fjeld Co., Inc. has been manufacturing custom stages for Amray, JEOL, = and Hitachi SEM's for over 20 years. I have purchased several of their custom stages or stage accessories over the last 15 years and have been delighte= d with the performance. They are located at 3 Executive Park Drive, North Billerica, MA and their telephone number is (978)667-1416. Mark Reynolds,= VP of manufacturing, is a good listener and very helpful guide through the design and manufacture of whatever stage or SEM add-on you may be interes= ted in.
Regards,
/richard
-----Original Message----- } From: Marisa Ahmad [mailto:mahmad-at-semiconductor.com] Sent: Tuesday, May 25, 1999 11:30 AM To: 'MSA listserver'
Who out there makes custom stages for SEM's (specifically JEOL)?=A0 Pleas= e contact me if you do, or know of anyone who does.=A0 Thank-you!
Robert, In situ cleaning is probably not a viable procedure. I would think if someone would build a jig for supporting the cantilever then Co2 cleaning may be a viable, off line solution. The greater concern may be how the tips may survive the thermal shock. Russ Gilmeister Xerox
-----Original Message----- } From: Co2clean-at-aol.com [mailto:Co2clean-at-aol.com] Sent: Thursday, May 27, 1999 8:22 AM To: spm-at-di.com
Hi,
You can try G-1 epoxy which is heat curing epoxy designed for high temperature application. Gatan produces this epoxy. The price in the UK is around =A370. Studies at GATAN indicate, a technical information of G-1 said, that G-1 bound interfaces remain intact in a TE= M hot stage at temperatures in excess of 1000 C.
Good Luck.
P Wang Department of Engineering Materials University of Sheffield Sir Robert Hadfield Building Mappin Street Sheffield S1 3JD UK Tel: 0114 222 5973(O) 0114 222 5519(L) Fax: 0114 222 5943 E-mail: p.wang-at-sheffield.ac.uk
we want to embedd a small sample of polysulphone, but metacrylate changes the sample. Epoxide resins are also not possible, because we want to dissolve away the resin after cutting. Are any resins or embedding materials known, which are practicable for our effort?
Kind regards
Rainer Ziel
------------------------------------------------------------- Dipl.-Phys. Rainer Ziel Acordis Research GmbH Obernburg ACR-O/RMG-EM 63784 Obernburg Germany
At 03:28 PM 5/27/99 +0100, you wrote: } } Hi, } } You can try G-1 epoxy which is heat curing epoxy designed for=20 } high temperature application. Gatan produces this epoxy.
Gatan *SELLS* the epoxy. It bears a strong resemblance to EpoTek 353ND. =20
The price in=20 } the UK is around =A370. Studies at GATAN indicate, a technical=20 } information of G-1 said, that G-1 bound interfaces remain intact in a TEM= =20 } hot stage at temperatures in=20 } excess of 1000 C.=20 } } Good Luck. } } P Wang } Department of Engineering Materials } University of Sheffield } Sir Robert Hadfield Building } Mappin Street } Sheffield S1 3JD } UK } Tel: 0114 222 5973(O) } 0114 222 5519(L) } Fax: 0114 222 5943 } E-mail: p.wang-at-sheffield.ac.uk
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University=09 (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
I'm looking for a small wafer of Ge. Something in the order of 1/2 to 1" in diameter would do. I want to use it as a target and sputter it in my BalTec ion mill to make a Ge film. I don't really care how pure it is or if it is polycrystalline. Does anybody have a small piece that they could spare?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
I assume that you tried Gatan's G1 (same as Epoxy Technologies EPO-TEK 353ND) If I remember right, Gatan rates this up to 500C, but I think that people have pushed it higher for heating experiments in the TEM.
Check out Fiore and Herring's paper in the first MRS TEM sample Prep series book (MRS vol 115, p125). They use Ceramabond 569 from Aremco.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Vetrano, John S To: 'Microscopy Listserver' -----------------------------------------------------------------------.
Hi all;
Does anyone out there know of an epoxy for TEM use that does not soften substantially at temperatures up to 500C? We want to do a high-temperature straining experiment but don't have too much material and would like to use a 3mm disk as the gauge with bits glued to the end for the 'grips'. Also, it would need to cure at less than 150C so we don't alter the microstructure before we start the test.
Thanks in advance.
Cheers, JSV *************************** John S. Vetrano Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
Does anyone know how I can contact Virtual Laboratories (distributor of Desktop Microscopist). Their web-site is gone, and their phone has been disconnected. I am looking for a Mac-based version of the Electron Diffraction Database to link with DM. Thanks in advance.
Jim Cole
Jim Cole Argonne National Laboratory-West PO Box 2528 Idaho Falls, ID 83403-2528 (208) 533-7165 Fax (208) 533-7863
Thank you for the proofreading, Woody. You are correct that I meant that at the lower 8-bit, 256-gray-level resolution that steps in brightness would begin to appear after relatively little contrast enhancement.
At 07:35 AM 5/27/1999 -0500, "White, Woody N" {Woody.N.White-at-mcdermott.com} wrote: } Hello Warren, } } Good answer, but could you have meant 16 bit is 65.5K and 8 bit is 256 } levels? } ..Or did I miss something? Woody } } SNIP } steps showing up. At 16-bit (256 gray levels) you will almost certainly see } the steps after much less contrast enhancement. } SNIP
Hi All: I am trying to digitize and capture images from my 845 JEOL SEM. Has anyone done this themselves? What issues do I need to worry about? What capture cards are best? I am looking to do this with off-the-shelf components. Thanks and regards,
} ---... } } Good answer, but could you have meant 16 bit is 65.5K and 8 bit is 256 } levels? ..Or did I miss something? Woody } } SNIP } steps showing up. At 16-bit (256 gray levels) you will almost } certainly see } the steps after much less contrast enhancement. } SNIP }
Computer displays are not capable of displaying 65.5 gray levels. You are manipilating 65.5k gray levels but the display hardware is still showing you only 8bit grays. This fact doesn't take anything away from the fact you manipulate 16bit grays with better control, resulting in less gray level posterization.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
A major reason for using 16-bit grayscale images (or 12-bit ones from today's 36-bit color flatbed scanners for that matter) is for image processing before converting the images to 8 bits (256 grays) for printing. Applying a series of image processing operations such as background subtraction, filtering, and brightness/contrast adjustments to an 8-bit image causes posterization (loss of gray levels) seen as stairstepped contrast in the image. This problem arises from doing integer math starting with only 256 numbers. Processing images at 16 bits avoids these problems.
Several common image processing programs have 16-bit capabilities. Photoshop 5 is one. It allows tonal adjustments at 16 bits and can be extended with third-party plug-ins to allow filtering (e.g., unsharp masking) at 16 bits. After processing, you can easily convert to 8 bits. Another feature of Photoshop worth a microscopist's understanding is using adjustment layers to process 8-bit images with minimal data losses.
As to the suggestion to decrease file sizes by cutting the image resolution to 1024 x 1024 pixels, this is OK for final processed images but will eliminate the possibility of later magnifying the images to observe fine details. Saving files with a compressed format such as JPEG will greatly reduce file sizes without affecting the resolution or greatly affecting the grayscale information content. There may also be file compression methods applicable for 16-bit images, but I'm not familiar with any.
My suggestion would be to acquire and process for background leveling, sharpening and tonal correction at 16 bits, then convert to 8 bits and archive at full pixel resolution, possibly in a compressed format such as JPEG to save space.
---------- From: Warren E Straszheim Sent: Wednesday, May 26, 1999 12:12 PM To: Microscopy-at-Sparc5.Microscopy.Com Subject: Re: 8 bit versus 16 bit images
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Remember the size of the files in bytes will be the x-size times the y-size times 1 for 8-bit images and 2 for 16-bit images. So by going to 8-bit images you could cut your file size in half. However, more gains might be made by reducing your x and y resolution. (You did not mention what resolution you are using.) How much do you really need? Images over 1024x1024 are not very useful unless you are digitally magnifying them to see the details.
16-bit imaging is helpful if you have a wide range of gray scale. You can concentrate on bright areas of the image and adjust the brightness and contrast locally to see the details you want without having the gray level steps showing up. At 16-bit (256 gray levels) you will almost certainly see the steps after much less contrast enhancement.
You might want to keep your original image "as is" and pull out smaller regions that can be enhanced and save those enhanced files as 1024x1024 8-bit images. I venture to guess that the aggregate size of those "region of interest" images would be much less than your original.
At 01:39 PM 5/25/1999 -0500, you wrote: } We've just started acquiring digital 16 bit x-ray radiographs. } } 1) Is it worth doing 16 bit radiographs or will 8 bit do? } 2) The images are HUGE! Also, much of our processing software only works } well with 8 bit. Would converting from 16 to 8 bit be acceptable? If you } don't for x-ray, please give your opinion for normal microscopy work.
I need to colorize some SEM photos (digital). I have Adobe Photo Deluxe Home Version but I hear that CorrelDraw or Adobe Photo Shop might be better. What does everyone else use? If anyone has any special techniques to share I'd love to hear them.
I have a microscope lens for our Agema (recently bought out by FLIR) Thermovision 900 Infrared Camera with spatial resolution of 25 microns but a MFOV (Measurement Field of View) of 250 microns. In other words, although I can see 25 micron objects, the temperature measurement is from a much larger area. I'm not sure if there are any IR microscopes out there that are much better.
Also, Barnes used to make infrared microscopes. I believe they're now QFI - try http://www.quantumfocus.com/
I am working in KRISS(korea research institute of standards and science) and studying micoelectromechanical systems characterization by home-built photothermal microscopy and calibrated atomic force microscope. I am seeking to find infrared microscope( not FTIR) which views temperature differences of micromechanical systems specimen at roomtemperature with a resolution of micrometer. Also I am seeking infrared detector array which can be used to make infrared microscopy.
by mailhub.iastate.edu (8.9.3/8.9.3) with SMTP id SAA00465 for {Microscopy-at-msa.microscopy.com} ; Thu, 27 May 1999 18:02:39 -0500 (CDT) Message-Id: {4.1.19990527175423.00986e20-at-pop-2.iastate.edu} X-Sender: wesaia-at-pop-2.iastate.edu X-Mailer: QUALCOMM Windows Eudora Pro Version 4.1
Our JEOL 840A has been digitized for years. I think there are already connections inside to take active control of the beam. We ended up with another panel to sort out the control signals since we had a LeMont Scientific image analyzer, a Kevex Delta EDS, and a JEOL add-on imaging system all with the ability to take control of the beam.
Currently, the LeMont is gone and the Kevex has been replace by an IXRF Systems EDS box which we now use to digitize images. It works well.
We also have a Quartz PCI passive system down the hall on our Hitachi 2460N. It works real well but is a bit pricy. It should work just as well on the JEOL.
While you might be able to do the work yourself using off-the-shelf parts (or get a grad student to do it (or maybe you are a grad student)), you probably don't want to. I have done a fair amount of hardware and software work in my time, but I am losing my stomach for it as I get older. The system I might create probably would lack much compared to the commercial units available.
So much for my 2 cents worth. Hope it helped.
Warren
At 01:38 PM 5/27/1999 -0500, you wrote: } } Hi All: } I am trying to digitize and capture images from my 845 JEOL SEM. Has } anyone done this themselves? What issues do I need to worry about? } What capture cards are best? I am looking to do this with off-the-shelf } components. } Thanks and regards, } } Mike Coviello } UT Arlington }
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Diane Ciaburri wrote: ================================================= I need to colorize some SEM photos (digital). I have Adobe Photo Deluxe Home Version but I hear that CorrelDraw or Adobe Photo Shop might be better. What does everyone else use? If anyone has any special techniques to share I'd love to hear them. ================================================== There is a product called "Spectrum Color" and you can find full details on it on the SPI Supplies website at URL http://www.2spi.com/catalog/software/spectrum-2.html
It is very low cost and very effective.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Assuming that the internal hooks exist, as Warren notes, or can be obtained from the manufacturer, as in some Hitachi units, then a home grown device requires 2 channels of D/A and one of A/D. The D/As should be 16 bit devices though 14 is probably adequate. The A/D should be at least 8. Note that A/D speed is not a crucial variable since you can sit as long as you need to on a single point. (Though other dwell considerations obviously apply.)
Sorry if this seems to state the obvious. The hardware isn't the problem, it is the time to get it working satisfactorily.
Rich } } } Our JEOL 840A has been digitized for years. I think there are already } connections inside to take active control of the beam. We ended up with } another panel to sort out the control signals since we had a LeMont
} Dear all, } } we want to embedd a small sample of polysulphone, but metacrylate changes } the sample. Epoxide resins } are also not possible, because we want to dissolve away the resin after } cutting. Are any resins or embedding materials known, which are } practicable for our effort? } } Kind regards } } Rainer Ziel } } ------------------------------------------------------------- } Dipl.-Phys. Rainer Ziel } Acordis Research GmbH Obernburg } ACR-O/RMG-EM } 63784 Obernburg } Germany } } Tel: +49 (0) 6022 / 81-2645 } Fax: +49 (0) 6022 / 81-2896 } E-mail: Rainer.Ziel-at-AkzoNobel.com } } } } }
} Does anyone know how I can contact Virtual Laboratories (distributor of } Desktop } Microscopist). Their web-site is gone, and their phone has been } disconnected. I am looking } for a Mac-based version of the Electron Diffraction Database to link with } DM. } Thanks in advance. } } Jim Cole
Jim and all,
Maybe you tried the old web address at Rt66.com. They moved ISP and are now at http://www.easystreet.com/~lacuna/FrontPage.new/FrontPage.html It took me a bit of web searching to find this out for ourselves a few months ago.
By the way, maybe those who maintain lists of www addresses for microscopy companies (e.g. EMYP at Lausanne and Nestor's list at ANL) should update this link.
Anyway, I hope this helps.
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren Department of Experimental Physics Chalmers University of Technology S-412 96 G=F6teborg Sweden Tel: +46 31 772 36 33 FAX: +46 31 772 32 24 email: maclaren-at-fy.chalmers.se or: ianmaclaren-at-hotmail.com Research Group Homepage: http://fy.chalmers.se/microscopy/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
} } My suggestion would be to acquire and process for background leveling, } sharpening and tonal correction at 16 bits, then convert to 8 bits and archive } at full pixel resolution, possibly in a compressed format such as JPEG to save } space. } From: Warren E Straszheim
Once you have decimated the data either by lowering the resolution or the number of bits you can never get it back. CD ROM's are cheap so I save as much data as I can. It has proved very worth while if I have to go back and fix something later. For the same reason I always save the data in a raw format just as it comes from the source.
Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
} From: RCHIOVETTI-at-aol.com } Received: from RCHIOVETTI-at-aol.com (3950) } by imo23.mx.aol.com (IMOv20) id 8GTGa11104; } Wed, 26 May 1999 20:50:16 -0400 (EDT) } Message-ID: {85f34a05.247df0c7-at-aol.com} } Date: Wed, 26 May 1999 20:50:15 EDT } Subject: Re: eyepiece reticles/graticles } To: sbarlow-at-sunstroke.sdsu.edu, microscopy-at-sparc5.microscopy.com } MIME-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } Content-Transfer-Encoding: 7bit } X-Mailer: AOL 3.0 16-bit for Windows sub 38
} } Hi Steve, } } My favorite company for rulings, reticles, stage micrometers, etc. is as } follows: } } Klarmann Rulings, Inc. } P.O. Box 4795 } Manchester, NH 03108 } Tel. (800) 252-2401 } Fax (603) 424-0970 } http://www.reticles.com } } They have every kind of ruling and reticle imaginable, all sizes, custom } ones } as well, and if you need it, they can provide scales which are traceable to } NIST. } } Cheers, } } Bob } ****************************** } Robert (Bob) Chiovetti, Ph.D. } Microimaging Technologies, Inc. } Tucson, Arizona USA } Tel./Fax (520) 546-4986 } rchiovetti-at-aol.com } Manufacturers' Representatives } Systems Integrators } Analog & Digital Imaging Systems } Clinical & Research Microscopy } Cytology/Histology/Pathology/EM } *******************************
Hi. I would like to get 0.5% ruthenium tetroxide in ETHANOL for my freeze substition experiment. Does anybody know where I can get it? I appreciate if somebody can provide the information about where I can get ruthenium tetroxide solution (in ethanol) or ruthenium tetroxide crystal. Thanks.
At 03:29 PM 5/27/99 -0700, you wrote: } } Several common image processing programs have 16-bit capabilities. } Photoshop 5 } is one. It allows tonal adjustments at 16 bits and can be extended with } third-party plug-ins to allow filtering (e.g., unsharp masking) at 16 bits.
I certainly agree as to the necessity of keeping as much image depth as possible during the various processing steps. I would like to know what plug-ins will allow 16-bit manipulations? I've got John Russ's IP toolkit, but those only seem to work on 8-bit images.
} possibility of later magnifying the images to observe fine details. Saving } files with a compressed format such as JPEG will greatly reduce file sizes } without affecting the resolution or greatly affecting the grayscale information } content. There may also be file compression methods applicable for 16-bit
I would hesitate to use JPEG for pre-presentation images. Although the eye will often not see the degradation in JPEG images, it is definitely there. The various IP algorithms will sometimes enhance the JPEG artifacts. e.g. Try running an edge filter in a JPEG. You will see the squares that JPEG creates during the compression process ( I can send you a powerpoint slide illustrating this!)
That said, I often use JPEG compression after all my processing is done and I have my image set to its final size and resolution. Unless you enlarge the image, the eye won't see most of the JPEG artifacts.
Cheers, Henk
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu "The wise are pleased when they discover truth, fools when they discover falsehood."
I have inherited an OLD International Equipment Co. IEC CTF microtome cryostat Model# AB4757, minus documentation and chucks. It is apparently in working order, but its previous users are no longer around. If anyone knows where I could obtain parts and a manual, please let me know. Thanks in advance. Regards,
Andrew Ochalski, Microscopy Technician, University of Ottawa Dept. of Biology, Room 108, Gendron Bldg. 613-562-5800 x6343
Here's an interesting note from kodak. Basically they are saying that for digital images one of the biggest advantages (now they are selling a film scanner here) of this scanner is that you can store (or archive) your images on FILM and then scan them only when you need them. So the big boys in digital imaging, are actually agreeing with some thing we as microscopists have been saying for awhile: The best archiving of images is actually on FILM (which we know has a life expectancy of over 165 years).
Taken from: http://www.kodak.com/cluster/global/en/service/faqs/faq1035.shtml
1.What is a film drive? The KODAK ADVANTIX Film Drive FD 300 is a desktop scanner designed to scan film in Advanced Photo System (APS) film cassettes.
The name "film drive" indicates a new technological concept that uses the film cassette as an image storage device. Rather than take valuable space storing image data on the computer's hard drive, Zip, Jazz drives, or floppy drives, the `film drive' scans APS film,DIGITIZES THE IMAGE FOR CURRENT WORK, AND THE IMAGE DATA REMAINS ON THE FILM IN THE CASSETTE.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
Interestingly, we asked that question on our Cell Biology survey this past December. The results were a staggering 46 % of the total population (495 microscopists) said that they use Adobe Photoshop. When the data is analyzed in terms of the number of responses to that particular question (it was part of a special section on image analysis, so not everyone answered), the number jumps to a whopping 86%! Guess that answers your question.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 04:52 PM 5/27/99 -0500, diane.a.ciaburri-at-gdds.com"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Time is drawing near for finalizing the next issue of MSAs newsletter, "The MSA Bulletin" (the listserver would not let me use the title in the subject line, since it thinks it is apparently full of Bull). If you have anything that you would like to see included in this version of the MSA Newsletter, (and thereby prove the computer wrong) I need to have it in my possession by June 4th.
You can submit material to me electronically (preferred) or by hardcopy at the address shown below.
Thank you for your attention,
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist Office:(612) 626-7594 CIE Characterization Facility, University of Minnesota Desk: (612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by "Ian MacLaren" } Jim and all,
Maybe you tried the old web address at Rt66.com. They moved ISP and are now at http://www.easystreet.com/~lacuna/FrontPage.new/FrontPage.html It took me a bit of web searching to find this out for ourselves a few months ago.
By the way, maybe those who maintain lists of www addresses for microscopy companies (e.g. EMYP at Lausanne and Nestor's list at ANL) should update this link.
Anyway, I hope this helps.
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren Department of Experimental Physics {
} ...Saving files with a compressed format such as JPEG...
JPEG is a very lossy algorithm. Even at a "high" quality setting, a good deal of information will be lost. Resolution will be noticeably reduced, and grayscale values will be changed enough that the image will be unsuitable for some modes of processing and many types of digital analysis. LZW compression, while not providing as small a file size as JPEG, is lossless, as are the algorithms applied to images in Photoshop's native format.
Thanks to all those who responded to my question on scanning EM negatives. UMAX seems to be most commonly used.
I would like to ask a few more questions.
1. A person here who has done some negative scanning found the biggest problem not to be resolution, but grey scale replication. Is reproduction likely to be similar for most scanners, or are some better than others?
2. The Agfa DuoScan has been suggested to us here, though no one mentioned it on the server. I wondered if anyone had experience with it.
3. Ideally, we would like something that can also do a good job on 35mm slides, is reasonably easy to use and is not too slow.--(not asking for much!). I know there are some nice 35mm scanners, but are flat bed scanners able to do both sizes well?
Hi All, We are looking for a sample holder capable of rotational and tilt movements, thats fits on a Zeiss 10 TEM. Would be very gratefull to hear from anyone who has one or has leads to a used or new one. Thank you.
The plug-ins I was referring to are the Deep Bit Filters from ImageExpress (http://www.scanprep.com/). You can get a trial copy of the filters from their webside.
Must admit I probably went too far recommending JPEG compression for archival images. I wouldn't want to mislead anyone about the losses involved. In Robin's case of 16-bit x-ray images, I could easily imagine 50MB or larger files that would "quickly" fill a CD.
Larry
---------- From: Hendrik O. Colijn Sent: Friday, May 28, 1999 6:30 AM To: Microscopy-at-Sparc5.Microscopy.Com; Thomas, Larry Subject: RE: 8 bit versus 16 bit images
Hi Larry,
At 03:29 PM 5/27/99 -0700, you wrote: } } Several common image processing programs have 16-bit capabilities. } Photoshop 5 } is one. It allows tonal adjustments at 16 bits and can be extended with } third-party plug-ins to allow filtering (e.g., unsharp masking) at 16 bits.
I certainly agree as to the necessity of keeping as much image depth as possible during the various processing steps. I would like to know what plug-ins will allow 16-bit manipulations? I've got John Russ's IP toolkit, but those only seem to work on 8-bit images.
} possibility of later magnifying the images to observe fine details. Saving } files with a compressed format such as JPEG will greatly reduce file sizes } without affecting the resolution or greatly affecting the grayscale information } content. There may also be file compression methods applicable for 16-bit
I would hesitate to use JPEG for pre-presentation images. Although the eye will often not see the degradation in JPEG images, it is definitely there. The various IP algorithms will sometimes enhance the JPEG artifacts. e.g. Try running an edge filter in a JPEG. You will see the squares that JPEG creates during the compression process ( I can send you a powerpoint slide illustrating this!)
That said, I often use JPEG compression after all my processing is done and I have my image set to its final size and resolution. Unless you enlarge the image, the eye won't see most of the JPEG artifacts.
Cheers, Henk
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu "The wise are pleased when they discover truth, fools when they discover falsehood."
Shanling Shi wrote: ================================================= Hi. I would like to get 0.5% ruthenium tetroxide in ETHANOL for my freeze substitution experiment. Does anybody know where I can get it? I appreciate if somebody can provide the information about where I can get ruthenium tetroxide solution (in ethanol) or ruthenium tetroxide crystal. Thanks. ================================================== RuO4 is quite explosive. Indeed a relatively small amount when it goes off can be quite devastating. One precious metals supplier (in the USA) about twenty five years ago, had some go off while in their storage facility and it caused quite a mess and a huge amount of damage.
Polysciences at one time offered it as a solution in chloroform, however I don't know if they are still doing that or not. It was quite unstable in this form as well, however.
Because of the extreme explosive hazard, there are major restrictions on shipping the material in solid crystals form. I have been out of touch with the shipping aspect of the problem so I can't not speak with authority on today's regulations. Even if it could be shipped,we ourselves would have to give it a careful look in terms of the potential liability exposure should some kind of accident occur.
We have had reports of users of our ruthenium tetroxide kits generating the tetroxide, and then collecting some of the vapors on a cold finger suspended over the aqueous solution. Once a given amount was collected, it was then immersed in carbon tetrachloride, warmed slightly, and the result was a carbon tet solution. I don't know about the solubility of RuO4 in ethanol, but if there was some solubility, then I presume an alcohol solution could be prepared in a similar way.
More information about ruthenium tetroxide and its dangers can be found on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
} Must admit I probably went too far recommending JPEG compression for archival } images. I wouldn't want to mislead anyone about the losses involved. In } Robin's case of 16-bit x-ray images, I could easily imagine 50MB or larger } files that would "quickly" fill a CD. } } Larry
We don't know how long silver images will last at least 150 years or more. You can view them with out any hardware and you can always scan them in again when computer get more powerful. Nearly everybody is set up to handle them.
They do take up more space then digital but you can always get to them and they will always be compatible. A 4X5 negitive stores a lot more information than you can scan in.
IMHO Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
I recently had to re-order Ruthenium powder and my less than exhaustive search and interview with an msds and the manufacturer found no mention or link to the explosive nature that I'd heard about here. The received the order by Fed X the next morning. It may be that this particular compound is not as unstable. Please help me by sending me a reference to this 'explosive risk'. The powder is very hydroscopic and must be kept desiccated, I believe. I'll look up the source and post next week. Jeff Day/'JD'
We have just started to use PhotoShop LE after previously using Adobe Photo Deluxe Home Version. It appears to be fairly easy to use.
J. Roy Nelson Material Testing Lab. Pennington, NJ
Hi all! I need to colorize some SEM photos (digital). I have Adobe Photo Deluxe Home Version but I hear that CorrelDraw or Adobe Photo Shop might be better. What does everyone else use? If anyone has any special techniques to share I'd love to hear them.
Thanks to all who answered my plead for MSmouse. That problem is resolved. The same computer (can not operate old video boards with PII or PIII processors!), is now really sick. It is a tower with magitronic BIOS and generic 486/66 mother board and Intel processor. The RAM sockets are defective and there may be a crack on the mother board. I need at least 3 ISA 16 bits slots and 2 8 bits. The slots should NOT extend over the microprocessor because the boards need sitting low and extend the entire length of the mother board. If anyone has information on how to obtain similar or equivalent mother board please let me know. Additionally, someone may have an old 486 like it collecting dust and I would like to have it. Call me if you do. Thanks.
*Disclaimer: Whatever... is not Tulane opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web:[ http://www.tmc.tulane.edu/ferminlab/] or [http://www.tmc.tulane.edu/imaging/] Internet: [cfermin-at-mailhost.tcs.tulane.edu] 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
Who built the first TEM in North America? Dr. Anderson of Washington State university built one in 1935, but I have heard that a university in Torono may have had one before WSU. Toronto is usually credited with the first microscope in 1939, but the microscope on display in the museum at WSU is dated 1935. Thanks, Will Garrett ssrs396-at-wsu.edu
Postdoctoral Position Available: For the further development of a low temperature AFM. The cryo-AFM is near completion, but additional modifications are necessary. The structure and interaction of molecules will be studied with cryo-AFM. Candidates are preferred to have experience in instrument design and scanning probe microscopy. Good candidates with experience in other relevant areas will also be considered. The completion of the doctoral degree is necessary. The starting date is 1 August, 1999. Please send CV with a list of publications and description of research experience to: Prof. G. Dietler Institut de Physique de la Matiere Condensee (IPMC), BSP Universite de Lausanne CH-1015 Lausanne, Switzerland Tel +41 21 692 3663 Fax +41 21 692 3635 Email Giovanni.Dietler-at-ipmc.unil.ch http://www.unil.ch/ipmc/docs/gd/index.html
Giovanni Dietler Institut de Physique de la Matiere Condensee Universite de Lausanne CH-1015 Lausanne Switzerland Phone: (+41) 21 692 3663 Fax : (+41) 21 692 3635 Email: Giovanni.Dietler-at-ipmc.unil.ch
Will, The first North American TEM was designed by Albert Prebus and James Hillier in the Dept. of Physics at the University of Toronto over the 1937-38 Christmas holidays and was built during the first four months of 1938. This info is from a great commemorative article by U.M. Franklin, G.C. Weatherly, and G.T. Simon in ELECTRON MICROSCOPY 1978, VOL. III, STATE OF THE ART SYMPOSIA, Papers presented in Symposia at the Ninth International Congress on Electron Microscopy held in Toronto August 1-9, 1978, edited by J.M. Sturgess. Other good sources for TEM development are EARLY HISTORY OF THE ELECTRON MICROSCOPE by L. Marton, San Francisto Press, CA, 1968 and Reviews of Modern Physics, Vol. 59(3), July 1987. Enjoy! Bob Santoianni Electron Microscopy Laboratory Emory University Hospital Atlanta, GA
} } } William R Garrett {ssrs396-at-mail.wsu.edu} 05/30/99 03:04am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Who built the first TEM in North America? Dr. Anderson of Washington State university built one in 1935, but I have heard that a university in Torono may have had one before WSU. Toronto is usually credited with the first microscope in 1939, but the microscope on display in the museum at WSU is dated 1935. Thanks, Will Garrett ssrs396-at-wsu.edu
May I ask why you need 5 ISA slots? That sounds like you have a lot of "legacy" cards, which you mey be able to replace. 5 ISA slots are not easy to come by. Most Pentium and higher computers have a few PCI slots and 2 or 3 ISA slots. An "industrial" PC may provide a solution, but as posted below, that won't be cheap. Can you perhaps replace some of the ISA cards with PCI cards?
If you are looking for Motherboards, I would suggest you look at some = of the vendors and manufacturers websites. They usually have dimensions = and pictures of the boards posted. That way you can also make sure, that = you can plug in full size cards. Check out the ATX boards. They are smaller and tend not to place parts behind the slots.=20
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition=20 and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
-----Original Message----- } From: Exchange Administrator=20 Sent: Tuesday, June 01, 1999 6:12 AM To: Michael Bode
Mortherboard with 5 ISA slots ist not usual in commercial PCs . You = have to looked for in the market of industrial PC. Here you have the web of a supplier www.indcompsrc.com Tlf. (619) 677 0877 Fax: 619 677 0897 . Anyway, prices of industrial computers are much higher than commercial ones and 486 motherboards could be not easy to find. May be it=B4s worthy for you to get a new motherboard with Pentium/AMD 200/333 Mhz and that supports DIMM 100 Mhz RAM.
JL Bello FNMT Email: jlbello-at-fnmt.es c/ Jorge Juan 106 28009 Madrid Spain ----- Original Message ----- } From: Cesar D. Fermin Ph.D. {cfermin-at-mailhost.tcs.tulane.edu} To: Users Distirubute to list {Microscopy-at-Sparc5.Microscopy.Com} Sent: Sunday, May 30, 1999 1:57 AM
I am preparing a list of Mike Stobbs's publications and conference abstracts for a memorial issue of Ultramicroscopy but a few are proving very elusive. If you can supply any extra details of the following items, I shall be most grateful. Please reply to hawkes-at-cemes.fr
1. Stobbs, W.M. TEM approaches to the characterisation of interfaces and multilayers Les Houches NATO & French IoP, March 1986 [All details of this meeting wanted, were there proceedings?]
2. Stobbs, W.M. TEM techniques for the atomic level characterisation of nanometre scale multilayers, ICSCM, Montpellier, June 1987 [What was "ICSCM"? Were there proceedings? Is this Stobbs paper in them?]
3. Stobbs, W.M. The use of TEM for the characterisation of inhomogeneities NATO Summer School on Inhomogeneities, Cambridge 1987. [Any details welcome]
4. Stobbs, W.M. New TEM methods for the characterisation of interfaces SERC LDS Discussion Meeting, May 1987 [Is there any record of this meeting?]
5. Stobbs, W.M. The use of TEM for the assessment of interphase boundaries Interfaces and Ledges Symposium, TMS, Indianapolis 1989 [Any details welcome]
6. Richards, C.G. and Stobbs, W.M. Moir=E9 method for the measurement of misfits "Lattice distortions", J=FClich, April 1974. [Any extra details welcome]
JL writes .... } } } Motherboard with 5 ISA slots is not usual in commercial PCs } ...
I since replaced a older Pentium mobo with a new PC100 mobo, and it was quite difficult to find one with 3 ISA slots. When it come to replacing mobos which have cards needed for intrumentation, I can well imagine your needs ... but 5 ISA slots?? Are you sure you can't buy new cards which are not dedicated to instrumentation which would go into AGP and PCI slots??? (e.g. video, modem, ethernet ...)
(... BTW, the mobo I found was an ASUS P2B which can still be found on the market ... but it has been discontinued with the P2B-F (2 ISA slots) ...)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
The members of the listserver have been wonderfully helpful in the past, and I'm hoping someone can come to my aid again.
We have a Philips 430 being installed in our lab, and it turns out we need to get into the high-tension tank - which means I need more SF6, (needless to say, on a rather limited budget). I have three questions. First, is there anyone out there who has any SF6 they don't want (perhaps left over from a machine they don't have any more) that we could buy off them? Second, where is the best/most economical commercial source to buy SF6 from? Third, what grade of SF6 is it that is used in these applications?
I would really appreciate your input on this. Many thanks in advance.
Gill
Dr Gillian M. Bond Department of Materials & Metallurgical Engineering New Mexico Tech Socorro, NM 87801
Ah yes... the famous Haefely tank SF6 dump. Every time you pull the cable from the HT tank, you dump the load of SF6. Fortunately, the new HT tanks manufactured by Philips remain sealed upon removing the cable.
Philips specifies 99.9% SF6 for the tanks. This purity is unavailable as a standard grade in the U.S. You can purchase commercial grade (99.8%) or instrument grade (99.99%) SF6. The commercial grade SF6 runs about half the price of the instrument grade. I have heard of people using the commercial grade successfully. However, Philips MAY not honor a warranty or service contract if you use the lower grade of SF6. So far, they haven't responded to my queries on the subject.
I just purchased a K-size (115 lb, I think) cylinder and was quoted the following prices for instrument grade:
Praxair $4750 (a "preferred" supplier price for the university!) AGA $2750
cheers, Henk
At 11:25 AM 6/1/99 -0600, you wrote: } } Hi everyone, } } The members of the listserver have been wonderfully helpful in the past, } and I'm hoping someone can come to my aid again. } } We have a Philips 430 being installed in our lab, and it turns out we need } to get into the high-tension tank - which means I need more SF6, (needless } to say, on a rather limited budget). I have three questions. First, is } there anyone out there who has any SF6 they don't want (perhaps left over } from a machine they don't have any more) that we could buy off them? } Second, where is the best/most economical commercial source to buy SF6 } from? Third, what grade of SF6 is it that is used in these applications? } } I would really appreciate your input on this. Many thanks in advance. } } Gill } } Dr Gillian M. Bond } Department of Materials & Metallurgical Engineering } New Mexico Tech } Socorro, NM 87801 }
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
Dear Gillian, We use SF6 in both our IVEM and HVEM. The manufacturers reccommend the highest (instrument?) grade, but we find that use of a lower grade is OK. Two caviats are that our IVEM is a JEOL, so the gaps etc. may be different and lower grade SF6 may not work with the Phillips set-up, and there is a gas handling plant on the HVEM with a loop for circulating the SF6 through filters which remove particulates, water, and other impur- ities. The gas handling plant can also liquify and store much more than the 12 large cy- linders of SF6 it takes to fill our tanks. I definitely reccommend such a unit given the very large price increases in SF6. Good luck. Yours, Bill Tivol
I had the 'pleasure' of upgrading an OEM computer with lots of required legacy cards. Not only did the hardware need to remain supported, but the antiquated OS as well (OS/2 v1.3 from Microsoft). The cards that could be switched to PCI were (network, video). I still needed a 4 ISA slot motherboard. These can be found. The industrial path is certainly viable, and a good choice for easy conversion. There are also ISA bus extension boxes which work fine (add 4-8 full length slots to any motherboard with 1 ISA slot).
Keep in mind also that if you decide to upgrade your motherboard to a Pentium level processor that even if you can replace some of the legacy cards with PCI cards to reduce the ISA slot requirement, the BIOS may not support your ISA cards. Several important factors to consider and features to look for in the BIOS and motherboard are:
Support for a Memory Hole between 15-16M if memory mapping is needed (ISA cards don't map above this, and some BIOSs don't support this)
Make sure that in addition to the long cards not hitting the CPU and cooler, a deep skirt on the card does not hit other board components. This is harder to judge without trying it out. (If your case can accomodate the height, short ISA extension cards could raise them enough.
Control of the ISA clock in the BIOS is another thing to look for. That way you can upgrade to a decent processor and still run the legacy cards at 8 MHz.
Don't rely on the (ISA) card manufacturer to know anything about the card. If it is more than a generation old it has likely been forgotten about and is unsupported. If they do tell you anything, doubt it. Same for resellers of the motherboard knowing anything about the ISA slots - they likely won't. Go somewhere to look at the motherboard and read the manual to see if it will do the job.
The BIOS from AWARD was found to be particularly complete with regard to the above concerns. Computers from HP are particularly lacking in this area.
Good luck.
Jeffrey R. Kingsley, Ph.D. Analytical Fellow, Evans Sunnyvale Charles Evans & Associates 240 Santa Ana Ct. Sunnyvale, CA 94086
I had the 'pleasure' of upgrading an OEM computer with lots of required legacy cards. Not only did the hardware need to remain supported, but the antiquated OS as well (OS/2 v1.3 from Microsoft!). The cards that could be switched to PCI were (network, video). I still needed a 4 ISA slot motherboard. These can be found. The industrial path is certainly viable, and a good choice for easy conversion. There are also ISA bus extension boxes which work fine (add 4-8 full length slots to any motherboard with 1 ISA slot).
Keep in mind also that if you decide to upgrade your motherboard to a Pentium level processor that even if you can replace some of the legacy cards with PCI cards to reduce the ISA slot requirement, the BIOS may not support your ISA cards. Several important factors to consider and features to look for in the BIOS and motherboard are:
Support for a Memory Hole between 15-16M if memory mapping is needed (ISA cards don't map above this, and some BIOSs don't support this)
Make sure that in addition to the long cards not hitting the CPU and cooler, a deep skirt on the card does not hit other board components. This is harder to judge without trying it out. (If your case can accomodate the height, short ISA extension cards could raise them enough.
Control of the ISA clock in the BIOS is another thing to look for. That way you can upgrade to a decent processor and still run the legacy cards at 8 MHz.
Don't rely on the (ISA) card manufacturer to know anything about the card. If it is more than a generation old it has likely been forgotten about and is unsupported. If they do tell you anything, doubt it. Same for resellers of the motherboard knowing anything about the ISA slots - they likely won't. Go somewhere to look at the motherboard and read the manual to see if it will do the job.
The BIOS from AWARD was found to be particularly complete with regard to the above concerns. Computers from HP are particularly lacking in this area.
Good luck.
Jeffrey R. Kingsley, Ph.D. Analytical Fellow, Evans Sunnyvale Charles Evans & Associates 240 Santa Ana Ct. Sunnyvale, CA 94086
Responses came from a good cross section of the microscopy community. Individuals included professional microscopists, dealers, service and technical people, hobbyists. Some selected comments follow:
If you are looking for very inexpensive, solid construction, a little bit old fashioned microscope, the LOMO's microscope may be the choice.
Optically, and mechanically, the best of what comes out of the factory is not bad. But with used zeiss, leitz, wild, and even american optical 'scopes out there
in the market, why would one want to buy LOMO?
They do indeed seem to be a lot of microscope for the buck - but only if you are willing to put up with the problems you cited - I'd buy one for my personal use.
The optics seemed to be OK if a little dated - they were not up to a new Nikon 800 standards. But then they cost only about 1/10th as much. If price is your biggest hurdle you probably can't do better but be prepared to have a few hassels getting everything right.
I have a LOMO Biolam Researcher. I paid $950 for it and no scope I've
seen comes close to it for less than $4000. The old design Zena condenser
is fabulous.
As a dealer of LOMO Microscopes I can assure you the availability of
service and parts since the exclusive importer (LOMO America, Chicago,
IL) stocks parts and services the instruments.
The quality (mechanical) is good with some "rough edges" = could be
refined in some parts. The optical quality is very good, however some
designs are from the 50ties. The company is refining constantly.
They are basically good scopes at bargain prices!
Mechanically it is horrible, but I am the only user, and know its quirks. Optically it is great, as long as you don't mind post-war optical technology (e.g., no eye-popping binocular widefield).
{/smaller} {smaller} So, the bottom line is good micrsocopes , bad service.
******************************
} From the various input, my own {bold} {italic} personal {/italic} {/bold} conclusions are that:
1. Quality of scopes are variable. Quality control at the factory may be a problem. Optics are dated but good. Mechanical systems may be variable.
2. Be cautious. Check out the dealer. Get written warranties. Best to actually see the units in person at a dealer location and check everyting out before buying. Take it home yourself. Avoid shipping hassles and damage unless it is fully insured.
3. Depending on what you want to do, it may be better to pay a premium and have more dependable service.
4. Sounds like a good scope to have if money is the major hurdle. Get alot for your money. Good for the hobbyist looking for value for the dollar. On the other hand, there are many used scopes on the market made by well known manufacturers and reputable dealers that are readily available for service.
5. You can draw your own conclusions based on the comments above. I have not made up my mind about buying one yet. The jury is still out on this one.
Thanks again for all who have contributed to this dialogue.
{bold} {italic} This summary is neither an endorsement nor a complete review of LOMO scopes. List participants are encouraged to research this on their own and draw their own conclusions before committing $$$$. Thanks.
I just had someone inquire about short courses for light microscopy, especially including polarized light methods and principles. I know I saw some courses listed earlier but as I had no plans to attend, I didn't keep notes.
The interested person is Dr. Elena Vittadini: email to: evittadini-at-smtp1.cultorfs.com !!!! Please reply to her directly since she is not on this list.
Thanks in advance for the help!
Dale Callaham
+++++++++++++++++ Dale A. Callaham Senior Microscopist, Stuck Jar Lid Opener, Odd Job Bear Central Microscopy Facility The University of Massachusetts Amherst, MA 01007 ------------------------- Phone 413-545-3751 FAX 413-545-3243 email dac-at-bio.umass.edu
I have seen some traffic here about various SEM digital capture. I don't really know where my "contribution" fits in, but rather than sit on something that may be useful, I will give a brief description and let interested individuals get back to me.
I have built an "adapter" for PASSIVE capture of SEM images on a JEOL JSM-5400 scanning microscope. It is pretty simple but does a pretty nice job (1920x1440 pixel images, or 640x480 pixel images). Since I figure that there are lots of this series of microscopes out there in low-end installations that, like our facility, would not otherwise have good digital imaging, I hope to share this if people are interested. My cost was { $250 ($200 for an off-the-shelf DMA card) plus a 100MHz Pentium computer: I built a small interface board. It would probably cost others a bit more, depending on how much they can do themselves. The concept might even be useful on some other types of machines - I think it is a very versatile system.
I have one full resolution TIFF image (the file format used) - 2.8Mb!!!! and some reduced size/resolution/compressed images also at our website:
http://www.bio.umass.edu/microscopy
(look for the "Image Gallery" link). Note: some browsers are not set up for viewing TIFF images so you may need to point yours to some viewing application. Otherwise choose "Save to File" and then view it outside of the browser.
Rather than tie up internet bandwidth with pages of info, I will be glad to reply to anyone interested, and try to help as best I can with my limited time. I know of one vendor who may be interested in setting these up for those who can't do it themselves, although the setup is very simple.
Cheers!
Dale Callaham
+++++++++++++++++ Dale A. Callaham Senior Microscopist, Stuck Jar Lid Opener, Odd Job Bear Central Microscopy Facility The University of Massachusetts Amherst, MA 01007 ------------------------- Phone 413-545-3751 FAX 413-545-3243 email dac-at-bio.umass.edu
Thank you all that answered my inquire on BSE with no BSE detector under = a LEO 440. The suggestion to set the collector bias to zero or negative wor= ked fine. I have just tried it, so only today I could test your suggestions.
S=E9rvio T=FAlio Pires Amarante
serviopa-at-usp.br
Museu de Zoologia da Universidade de S=E3o Paulo Caixa Postal 42694-970 04299-970 S=E3o Paulo BRASIL
} Will, } The first North American TEM was designed by Albert Prebus and James } Hillier in the Dept. of Physics at the University of Toronto over the } 1937-38 Christmas holidays and was built during the first four months of } 1938. This info is from a great commemorative article by U.M. Franklin, } G.C. Weatherly, and G.T. Simon in ELECTRON MICROSCOPY 1978, VOL. III, } STATE OF THE ART SYMPOSIA, Papers presented in Symposia at the Ninth } International Congress on Electron Microscopy held in Toronto August } 1-9, 1978, edited by J.M. Sturgess. } I have a snapshot of that U of Toronto 'scope which now resides in the Ontario Science Centre. If anyone is interested, I can provide a .tiff or something of it. (Warning: The esthetic appeal of the picture is somewhat marred by yours truly standing beside the scope:-))
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia B2Y 4A2
McCrone Research Institute offers many courses on light microscopy incl= uding polarized light microscopy. More information is available on their web= page http://www.mcri.org/.
Regards,
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com =
We have the Ferrups unit from Best Power on several instruments. There are two things that I would keep in mind:
1. make sure that you have the capability to handle the power surges that come from motors switching on and off (ie, pumps and compressors). For Best Power, this is the DVR option.
2. make sure that your physical plant people RTFM when installing the unit. For example, Best requires grounding cable that is *at least* the same gauge as the hot and common wiring. This requirement is more stringent than the typical electrical code.
Hope this helps.
} Dear Microscopists, } We would like to get a UPS for our TEM to bridge the gap between the } instant there is a loss of power and the time when the backup generator } takes over (a minute or so??). We have some information about APC and Tripp } Lite UPS units, but we are concerned that their experience is limited to } computer applications. We would like information on the types of UPS } employed by EM users. We would also appreciate suggestions on how to chose } the appropriate size. The power consumption of our TEM is about 3 kilowatts } (instuction manual). } Many Thanks in advance. } Vachik Hacopian
-------------------------- John Bonevich NIST, Metallurgy, Stop 8554 100 Bureau Drive Gaithersburg, MD 20899 USA TEL: (301) 975-5428 FAX: (301) 975-4553
Dear Colleagues! I have the following problem: I don't know how to treat diffraction patterns from Quasi-Crystals. For example, is there analogue Bragg reflection formula in this case? I would be very thankful for references on any fruitful works about this problem.
Please send your message to my e-mail address: DOMAN-at-orm.irtm.kiae.ru
Thank you very much for attention to my question!
Alexandr Domantovsky Moscow, RRC "Kurchatov institute", Russia
Hi Gillian: We are also in the process of installing a Philips 430. I called around, and found a grade that meet Philips specs for $1417/110 lb cylinder (insulator grade 3) from BOC Gases -at- 800-262-4273 X1455 ask for Bob Korniche. I hope this helps. Regards,
Michael Coviello EM Lab Manager UT Arlington, Arlington, TX
Gillian Bond wrote:------------------------------------------------------------------------
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi everyone, } } The members of the listserver have been wonderfully helpful in the past, } and I'm hoping someone can come to my aid again. } } We have a Philips 430 being installed in our lab, and it turns out we need } to get into the high-tension tank - which means I need more SF6, (needless } to say, on a rather limited budget). I have three questions. First, is } there anyone out there who has any SF6 they don't want (perhaps left over } from a machine they don't have any more) that we could buy off them? } Second, where is the best/most economical commercial source to buy SF6 } from? Third, what grade of SF6 is it that is used in these applications? } } I would really appreciate your input on this. Many thanks in advance. } } Gill } } Dr Gillian M. Bond } Department of Materials & Metallurgical Engineering } New Mexico Tech } Socorro, NM 87801
} I have the following problem: I don't know how to treat diffraction } patterns from Quasi-Crystals. For example, is there analogue Bragg } reflection formula in this case? } I would be very thankful for references on any fruitful } works about this problem.
Dear Alexandr, I reccommend the article "High-resolution electron microscopy of quasicrystals" by Kenji Hiraga published in 1997 in Advances in Imaging and Electron Physics, Vol. 101, pp 37-98. There is a good introduction to quasiperiodic lattices and a section on electron diffraction which is clear and informative.
Greetings Friends, This was probably covered before, but I am having trouble getting freshly isolated myocytes to stick to Aclar. I'm willing to coat with laminin or Fibronectin etc. Does anyone have a tried and true protocol? or an arrow that points in the right direction? Hope to try this after a much needed vacation in the Northwoods of Wisconsin, so I won't be signing off during this time in order to receive replys. Thanks, Linda Fox lfox1-at-wpo.luc.edu
------------------------------------------------------------------------------- FUNDAMENTALS AND APPLICATIONS OF LIGHT MICROSCOPY -- FOURTH ANNUAL SHORT COURSE ------------------------------------------------------------------------------- June 20-25,1999, Wellesley College Science Center, Wellesley Massachusetts
A 5-day hands-on course for achieving the maximum information from light microscopy. The course will cover the principals of light microscopy, adjustments of the microscope for optimumcontrast and resolution, image capture and interpretation of images in terms of light-matter interactions. Contrast techniques, including polarized light, differential interference contrast, phase contrast, Hoffman Modulation coutrast and fluorescence microscopy will be included. A full range of reflected and transmitted light microscopes, as well as contrast equipment, will beprovided for use by the students. Students are encouraged to bring their own samples for exploration.
Organized by McCann Imaging For further information: see http://www.microscopyed.com For course brochure, contact Mary McCann, McCann Imaging 161 Claflin Street Belmont MA 02478 e-mail: mccanns-at-tiac.net Phone (617)-484-7865 Fax: (617) 484-2490
As an alternative to going to McCrone, MME will develop a customized, on-site workshop in this area. Our webpage info is listed below.
Best regards,
Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 07:36 AM 6/2/99 -0500, Neilly,Joseph wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Dear Colleagues! } I have the following problem: I don't know how to treat diffraction } patterns from Quasi-Crystals. For example, is there analogue Bragg } reflection formula in this case? } I would be very thankful for references on any fruitful } works about this problem.
Hello Dr. Domantovsky, There are numerous papers in the literature on indexing quasicrystal diffraction patterns. Bear in mind there are several different types of quasicrystals (icosahedral phase, four different decagonal phases, octahedral phase, dodecagonal phase, etc...), each with their own indexing system. In fact for the icosahedral phase there are several different (yet equivalent) indexing schemes.
For the icosahedral phase, all of the diffraction peaks and zone axes can be indexed with three orthogonal vectors pointing to three perpendicular two-fold edge centers of an icosahedron (J. W. Cahn, D. Shechtman, and D. Gratias, J. Mat. Res. 11, 12 1986. and D. S. Rokhsar, N. D. Mermin, and D. Wright, Phys. Rev. 135, 5487 1987). If the icosahedron has edge length 2, then the coordinates of the five-fold vertices are given by the cyclic permutations of ( +-1 , +-tau , 0), where the irrational number tau is defined from the equation tau^2=tau+1. Alternatively, the icosahedral phase can be indexed with six independent vectors with integer indices. The particular choice for the six vectors is a matter of convention. Elser (Phys. Rev. B32, 4892 1985) chose to align the icosahedron with a five-fold axis along the z-axis, and chose the basis as the six vectors pointing toward the upper vertices of the icosahedron.
I hope that you find this information of use.
_____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Job Opening at Intel: Sr.TEM Engineer/Materials Scientist Job Description and Responsibility: The successful candidate for this position will perform advanced imaging and microanalysis of Si-based integrated circuits using transmission electron microscopy and provide in-depth materials analysis and interpretation of the results for various processing related problems found in process development. A Ph.D. degree in Materials Science or Applied Physics is required for this position. The successful candidate should have a solid understanding of electron-solid interaction and basic knowledge of electron optics. The candidate should be experienced in utilizing TEM for various materials analysis and problem solving and should also have extensive knowledge and experience in x-ray energy-dispersive spectrometry and/or electron energy loss spectrometry. Some knowledge/experience in the area of semiconductor processing or electronic materials is highly preferred. Must possess good written and verbal communication skills and problem solving techniques. Able to work in a team environment. Work Site: Oregon Contact: Hiring Manager: Ma,Zhiyong Phone: (503)613-6480 Fax: (503)613-5907 Mailing Address: Intel Corporation Components Technology Development M/S: RA1-329 2501 NW 229th St. Hillsboro, OR 97124
If you find any technique to colorize the SEM images can you be so kind to share it with the list?
Gary.
On Thu, 27 May 1999 diane.a.ciaburri-at-gdds.com-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi all! } } I need to colorize some SEM photos (digital). I have Adobe Photo Deluxe Home } Version but I hear that CorrelDraw or Adobe Photo Shop might be better. What } does everyone else use? If anyone has any special techniques to share I'd love } to hear them. } } Thanks, } } Diane Ciaburri } }
Does anyone have a reliable "recipe" for preparing holey carbon grids. I'm using a glycerol-formvar mixture, however my holes are not very homogeneous in size and are very sparse. Maybe there's something I'm missing - any advice? Does anyone know if these solutions store well?
Your advice would be greatly appreciated. Thanks. Minh ^_^
Minh, I find that the best recipe for holey grids is to order them already made from someone like EMS. They're not that expensive, especially if you consider what your time is worth. Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
At 06:09 PM 6/3/99 +0930, Trinh, Minh-Uyen - TRIMY002 wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Recently, graduates in physics or biophysics are encouraged to apply for a post doctoral position in the-state-of-the-art time-resolved imaging laboratory at Microcosm. The successful candidate will be part of a team developing photonics technologies for solid state and biomedical applications. Hands-on experience with femetosecond lasers, time-resolved and imaging instrumentation is essential. This position is only for US citizens.
Please fax or e-mail your resume to Dr. H. Malak. ________________________________________________________ Dr. Henryk Malak Director of Research Microcosm, Inc. | 9140 Guilford Road, Suite O |Columbia, MD 21046 Phone: (301) 725-2775, Fax: (301) 725-2941 Our web site is located at: http://www.microcosm.com {http://www.microcosm.com} ________________________________________________________
{html} We would like to post the following position on your list server. If you need any additional information, please let me know. Thank you. {br} {br} UNIVERSITY OF CENTRAL FLORIDA {br} Advanced Materials Processing and Analysis Center {br} Faculty Position in Advanced Electron Microscopy {br} {br} As part of the partnership between the University of Central Florida and Lucent Technologies, a position in advanced electron microscopy of materials is expected to be available to begin Fall of 1999. The position will be within the Advanced Materials Processing and Analysis Center (AMPAC) at the level of a tenure-track Assistant Professor in Physics. Applicants should have a Ph.D. degree in Physics, Materials Science, or closely related field, and should have demonstrated expertise in atomic resolution scanning transmission electron microscopy, annular dark-field imaging, Z-contrast imaging, energy loss microscopy, analytical microscopy, and similar techniques. Send a copy of a current vita, including a research and teaching plan, along with a minimum of three letters of reference, to: {br} {br} {div align="center"} Dr. Vimal Desai, Director {br} AMPAC {br} University of Central Florida {br} 12424 Research Parkway {br} Suite 408 {br} Orlando, FL 32826 {br} {br} {/div} The University of Central Florida is an equal opportunity, affirmative action employer. Screening of applications will begin May 25 and remain open until position is filled. {br} {BR} {/html}
TECHNICAL DIRECTOR Center for Microscopy and Microanalysis Department of Biological Sciences Bowling Green State University
Responsible for overseeing the day-to-day operations of the Center; maintaining equipment in operating order, scheduling and monitoring users of the facilities, participating in educational and research activities consistent with the University's mission, assisting faculty and staff in instrument use, performing contracted work for others, and organizing/conducting tours of the Center for student fieldtrips and site visits of grant funding agencies. Master of Science degree and 2 yrs. experience in the operation of maintenance of electron microscopes required. Full-time administrative staff position. Administrative Grade Level 14, minimum salary $32,348. Salary is commensurate with education and experience. Full benefit package available. Submit letter of application, resume, and names/addresses/telephone numbers of three professional references postmarked by June 18, 1999 to: Ofc. of Human Resources (Search S-036), 100 College Park Ofc. Bldg., Bowling Green State University, Bowling Green, OH 43403. BGSU is an EEO/AA employer/educator.
I have written a very mini-tutorial on using Photoshop for colorizing SEM images. It will appear in the June issue of Microscopy Today. I'm not giving away all my secrets, but it should give you a start! Why not use what you learn and challenge me in the Just For Fun contest that Microscopy Today is having at MSA in Portland in August? Do you think I can win First and Second prizes again?! You can subscribe to Microscopy Today at their web site http://www.microscopy-today.com and it's free.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} Does anyone have a reliable "recipe" for preparing holey carbon grids. I'm } using a glycerol-formvar mixture, however my holes are not very homogeneous in } size and are very sparse. Maybe there's something I'm missing - any advice? } Does anyone know if these solutions store well? } } Your advice would be greatly appreciated. Thanks.
Dear Minh, We use 45 ml of actetone plus 45 ml of chloroform to which 0.2 g of formvar and between 0.82 and 1.5 ml 90% glycerine are added. The formvar is added to the CHCl3 and shaken until the formvar dissolves, then the acetone and glycerol are added and skaken vigorously for ~10 min. The mixture is sonicated for 5 min with a probe sonicator and must be used within a few hours or re-sonicated. If the mix is used immediately, I get a very large number of small holes (~2 um), and if the mix stands for ~1 hr, I get a large number of larger holes (~5 um). I almost always get "wall-to-wall" holes with very thin bars of formvar remaining. Good luck.
I always seem to have trouble floating the films off the slides; does anyone have any tips? Also, I'd like to get 10-20 um holes; any tips there?
Dear Bill, Thanks for your advice - much appreciated ^_^ Regarding the floating of films - I found it is important for the slides to have a clean/hydrophilic surface. If the slides are new, it is usually ok to just rinse the slide with a bit of detergent and wiping it dry with lint free tissue. However, if you don't have clean slides I find the following method for cleaning surfaces 100% successful - the film easily floats off (and if you're skilled enough you can float both sides off at the same time).
*Place slides in a beaker containing conc. HNO3 (~60%) and boil for 1 hr, rinse with copious amounts of Milli-Q water several times. *Rinse with a bit of detergent and wipe dry before dipping into mixture.
To get larger holes I presume you would need to add more glycerol to the mixture.
Regards, Minh T ^_^
-----Original Message----- From: William Tivol [SMTP:tivol-at-wadsworth.org] Sent: Friday, June 04, 1999 6:52 AM To: Trinh, Minh-Uyen - TRIMY002; microscopy-at-sparc5.microscopy.com Subject: Re: TEM need help with preparing holey carbon grids
Trinh, Minh-Uyen - TRIMY002 wrote:
} Does anyone have a reliable "recipe" for preparing holey carbon grids. I'm } using a glycerol-formvar mixture, however my holes are not very homogeneous in } size and are very sparse. Maybe there's something I'm missing - any advice? } Does anyone know if these solutions store well? } } Your advice would be greatly appreciated. Thanks.
Dear Minh, We use 45 ml of actetone plus 45 ml of chloroform to which 0.2 g of formvar and between 0.82 and 1.5 ml 90% glycerine are added. The formvar is added to the CHCl3 and shaken until the formvar dissolves, then the acetone and glycerol are added and skaken vigorously for ~10 min. The mixture is sonicated for 5 min with a probe sonicator and must be used within a few hours or re-sonicated. If the mix is used immediately, I get a very large number of small holes (~2 um), and if the mix stands for ~1 hr, I get a large number of larger holes (~5 um). I almost always get "wall-to-wall" holes with very thin bars of formvar remaining. Good luck.
I always seem to have trouble floating the films off the slides; does anyone have any tips? Also, I'd like to get 10-20 um holes; any tips there?
Minh, For making holey carbon films, I use polystyrene films casted from glass slides the same way as formvar. Polystyrene films are easy to float them off after simple Kleenex cleaning of glass slides or mica sheets. ~0.25% w/v solution of polystyrene in amyl actate results in ~ silver film. Breathing once on a drying film will give you a whole range of diameters from a few nm to microns. Adding a few drops of water to polystyrene solution and sonication before casting will give you very reproducible and uniform holes. You can evaporate carbon onto filmed grids or onto films on slides followed by floating off carbon coated polystyrene films.
You can easily remove polystyrene by keeping the grids on a wire net or on a filter paper over the amyl acetate to have clean holey carbon films only.
Let me know if you need more details. Marek.
} Date: Fri, 4 Jun 1999 10:25:02 +0930 } From: "Trinh, Minh-Uyen - TRIMY002" {TRIMY002-at-students.unisa.edu.au} } Subject: Re: TEM need help with preparing holey carbon grids } To: "'William Tivol'" {tivol-at-wadsworth.org} , } "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-Sparc5.Microscopy.Com} } MIME-Version: 1.0 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all, Is there anybody out there using an Agfa Duoscan for TEM negatives?
Our lab has just bought one (the T2500) but the negative holder (60 by 90) does not quite fit our TEM negatives (64 by 88 mm).
What have others done about this? Manually adjusting the holder? Having a special holder made?
We would appreciate any ideas that you can share.
Yours sincerely
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren Department of Experimental Physics Chalmers University of Technology S-412 96 G=F6teborg Sweden Tel: +46 31 772 36 33 FAX: +46 31 772 32 24 email: maclaren-at-fy.chalmers.se or: ianmaclaren-at-hotmail.com Research Group Homepage: http://fy.chalmers.se/microscopy/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Hello, we've been using the Duoscan for about 3 years now, and believe it or not, we use scotch tape to hold TEM as well as various other negs to the glass tray which comes with the scanner. The scotch tape holds them flat quite well, and not very much is needed. We've also had some succes placing the negatives on the glass tray and keeping them flat with an 8x10 sheet of glass on top. If this double glass is used you have to be very careful when scanning, as the Duoscan is great at picking up Newton's Rings.
Good Luck
---------------------------------------------------------------------- Adam Baker Carleton University Biology Department Email address: adbaker-at-ccs.carleton.ca ----------------------------------------------------------------------
I have a question regarding SEM/EPMA analysis. I cannot find information about EPMA analysis on the web. Can anyone tell me what it is? Is it an acronym for Electron Probe Microanalysis? Does anyone have any information on an EMAX3700 mounted on a Hitachi SEM. I have a customer who is asking about this, and would like to find out some information. ANY information would be greatly appreciated.
Thanks in advance,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
There ought to be something about EPMA on the web. There is a mailing list at microprobe-at-www.microanalysis.org, but I think you have to be subscribed to post to it. I can get you details.
You are right about the acronym. Often, it refers to wavelength-dispersive spectrometers with crytals instead of the energy-dispersive Si and Ge solid state detectors. But some can get sloppy with the nomenclature. (I still prefer to refer to EDS than to EDAX. Nothing against EDAX, the company, but I prefer not to give them free advertising.)
I am not familiar with EXAX3700. What does you client want to know or do? Do they want low detection limits, high resolution scans, etc.?
At 09:47 AM 6/4/1999 -0400, you wrote: } } Hi all, } } I have a question regarding SEM/EPMA analysis. I cannot find information } about EPMA analysis on the web. Can anyone tell me what it is? Is it an } acronym for Electron Probe Microanalysis? Does anyone have any information } on an EMAX3700 mounted on a Hitachi SEM. I have a customer who is asking } about this, and would like to find out some information. ANY information } would be greatly appreciated. } } Thanks in advance, } } David Bell } Scientist } Electron Microscopy Lab } Millipore Corporation
Here are some posts that may be helpful on the subject from my archives:
I know of two methods of preparing holey films. The first is to shake up a mixture of liquid soap and water and Collodion until it froths, then drop a drop or two on the surface of distilled water. Lay the grids on top of the Collodion layer, pick up with a filter paper, dry and carbon coat. Put the grids in a Jaffe washer with cloroform for 48 hours to remove the Collodion. The other method, if you have Nucleopore type filters, is to carbon coat a strip of Nucleopore filter film, place a square of the coated filter on the grid sitting on a nickel mesh on the surface of the Jaffe washer full of cloroform, wait 48 hours to dissolve the Nucleopore, dry grids. The holes in the Nucleopore will be now be holes in the carbon coat. I must admit I have not tried the first method myself, but someone in my lab did, years ago.
I use a method by Kuga and Brown from Philips Electron Optics Bulletin v126, p.19 (1989) to make lacy formvar grids. First a note of caution. I use dichloroethane as my solvent for the formvar resin. After having much difficulty in preparing my films, I was told that when exposed to light, dichloroethane slowly forms HCl in the solution. The HCl destroys the integrity of the formvar polymer. Solution: store the formvar solution in a brown bottle in a dark cabinet.
Method:
- I use 0.25% formvar solution in dichloroethane and freshly cleaved mica sheets as a substrate. The formvar lifts off the mica much easier than from glass microscope slides.
- Dip the mica into the formvar solution, wick off excess on a paper towel, then breathe heavily on the mica for about 5 seconds while it is still wet. The moisture in your breath condenses in the solution. Caution: Don't inhale!
- When the mica has dried, score the edges and float off on water.
- Place 200 mesh TEM grids on the floating film. The area with the best holes will appear milky.
- I then take saran wrap, stretch it tightly across the mouth of a small (~100ml beaker), and press it down at a slight angle onto the floating formvar film. The film will stick to saran wrap and you can easily pick it up off the surface of the water.
- After the film has mostly dried, pick up the grids from the saran wrap "drumhead" and place them on filter paper.
- The film will have many "pseudoholes" that have a thin residual film across them.
- Following the method of Kuga and Brown, by heating the film to about the T(g) temperature you can break these holes open. The time and temperature are critical. I place the bare filter paper in a small lab oven (not in a petri dish; it has too much thermal mass) at 110C for 12 minutes.
- I then coat both sides with carbon to stabilize the formvar lace.
I can easily make 50-100 grids in an hour (exclusive of the carbon coating). These grids make wonderful supports for looking at fine particulate dispersions.
Cheers, Henk Henk Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility
The question of how to make holey carbon films came up on the list server two years ago. At that time John Gabrovsek (gabrovj-at-ccsmtp.ccf.org) gave the following reference: Baumeister & Seredynsky, Micron 1976, Vol 7, p. 49, and Jane Fagerland (fagerland.jane-at-igate.abbott.com)gave this one: Elsner, Proceedings, 29th EMSA Meeting, p. 460. Both methods were said to work satisfactorily. You might try contacting these people for more details.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu;
Sara,
Here is a method I have used in the past. It's a hassle, believe me, but it usually works. It also makes you realize why purchasing holey films commercially costs so much---they're not easy to make.
1) If you can find it, purchase a product called "Victawet" (I believe Electron Microscopy Sciences carries it). This comes in very small vials and it lasts forever. It serves as a lubricant to release plastic films from glass slides in the following steps.
2) Get some high quality glass microscope slides. We have found that Esco brand slides work more reliably than any other kind. Don't know why. The ones with one frosted end are useful for keeping orientation.
3)Polish these slides until they gleam and show NO contamination upon close examination. Do this even if they are "precleaned".
4)Take a piece of Victawet about the size of a cooked grain of rice and put it in a tungsten basket in a vacuum evaporator. Arrange as many CLEAN glass slides around it facing the basket. A distance of several to many centimeters is fine----i.e., distance is not too critical. It may be possible to make a custom rack for this purpose out of plastic or metal---the material used should not outgas too much.
5) Pump down the evaporator and when high vacuum has been reached, gently increase current to the basket. At a certain point the victawet will start to melt and you will see a fog developing on the slides. (If you use too much current, the piece of victawet may jump out of the basket and you have to start over, so be patient.) This "fog" is what you want. No need to overdo it, a little bit will work fine.
6) Repeat Step 5 until you have as many slides as you need. Use a new piece of victawet for each batch. Store these slides in a container for later use, since they keep for a few weeks.
7) Get some formvar (some people use butvar) in 1,2 dichloroethane (ethylene dichloride). 0.5% or 0.25% usually works. You can order 1% and dilute it, too. Keep this solution in a dessicator. Water in the solution causes problems, so only open it when necessary and for as brief a time as possible. Pour this in a tube wide enough to hold a slide, to a depth just short of the frosted end of the slide. (You can conserve formvar by using a special tube with a constricted lower end. One product is called "Dip Miser" and I think it's sold by Ted Pella. We had a special dipping tube made by a glassblower from a regular glass cylinder fused with a very wide constricted bottom.)
8) Cover the top of the tube with the formvar with something so that the atmosphere inside becomes saturated with the evaporating dichloroethane.
9) Get a container big enough to hold water to a depth of several inches. It should be 8-10 inches in diameter for comfortable working. Fill it to the top with, preferably, double-distilled water.
10) Now you're ready. Take the victawet coated slides and polish them again. They should feel slippery. Clean them until they gleam. Clip the frosted end of the slide with something to hold it, like a paper clamp attached to a piece of wire, and put it in the formvar solution in the tube. Keep it there for about 5-10 seconds, then pull it up and let it drain INSIDE THE TUBE. Only leave the top of the tube uncovered while putting the slide in and taking it out, in order to keep the atmosphere saturated. While the slide is draining, keep something over the tube opening, allowing only enough opening for the wire. The length of time you let the slide drain determines the final thickness of the formvar film. Start at about 10 seconds. If you desire a thinner film, increase the draining time up to about 15-20 seconds. (This is why the atmosphere must remain saturated with dichloroethane inside the tube. If it is not, the formvar just dries with draining properly.)
11) To make the holes, expose the slide to moisture IMMEDIATELY upon removing the wet slide from the dipping tube. We would breathe on it, but passing it quickly over a bath of steaming water might do the same thing. The microdroplets of water in the steam or in your breath make the holes. To repeat, this must be done IMMEDIATELY before the slide has time to begin drying. This is also a good time for prayer, invocations to the ancestors, luck rabbit's feet, and anything else you might find effective in appeasing the gods of holey films.
12) Lean the coated slide up against something in a dust-free area to dry for at least two minutes.
13) Back to the basin of water---take a clean laboratory tissue paper and drag the surface of the water to rid it of oil and dust. You can also put a drop of collodion in amyl acetate on the water, let it form a film, then pick this up and discard it to clean the surface.
14) Take the slide and cut around the outside perimeter of the film with a clean razor blade or scalpel. Then take the slide and, holding the frosted end, SLOWLY dip it into the water. If all goes well, the victawet was good, and you have been virtuous recently, the film will separate from the slide and float on the surface of the water. (HINT: a friend of mine---thanks, Steve---discovered that heating the water really helps in releasing the film. Don't boil it, just warm it up.)
15) You can now take your clean TEM grids and place them carefully on the floating film. When you have enough, take another CLEAN glass slide (no need for victawet this time) and scoop the film up. This is tricky and hard to describe: the idea is to catch the end of the slide on the end of the floating film so that when the slide is pushed down into the water the film will be pulled down with it and against it. When you finish, the grids should be between the formvar film and the glass slide. Put it somewhere dust free to dry. When dry, you place the slides in a vacuum evaporator and coat them with 100-300 angstroms of carbon. Then, you can GENTLY AND CAREFULLY push on the edge of the grid to pop it loose from the film. At this point, you will hopefully have a carbon-coated holey film on a grid, ready for use.
Alternatively, you can use a "domino rack", a piece of metal with small holes in it and two bent ends that serve as legs (kind of like a little bench with holes a few millimeters larger than TEM grids). These can be purchased commercially, or made yourself if you can find the right kind of metal with holes. When the film is floating on the water, before putting the grids on it, slowly lower the CLEAN domino rack onto the film, which will adhere to it by surface tension. Lift it out and put it in a dust free place to dry. The result is formvar film covering a bunch of little holes. The grids are later set upon these films by placing a drop of double-distilled water on the film, placing the grid on the drop, and letting it dry down. The film with grids can then be carbon-coated, or you can coat the individual grids later.
I warned you. This is a good starting procedure and variations can be done at several points, depending upon your circumstances. Things can go wrong at any stage, affected by humidity especially. It's a frustrating procedure and I would welcome any suggestions for making it easier, but it does work if you're patient and willing to experiment a little. If you do get a batch to work, I suggest making a whole bunch at once while luck is with you. You may not be so fortunate next week.
Hope this helps. Let me know if you have any questions, and I'll try to help more.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
There has been some recent discussion about the desire to purchase ruthenium tetroxide and also to put it into an alcoholic solution. There was also a possible suggestion that it was being shipped by normal shipping channels used for hazmat materials. Or at least that was the way some had interpreted it to mean.
I contacted one of my friends who is a precious metals chemist and expert in this area, his response follows: ======================================= Regarding the hazards, ruthenium tetroxide consists of golden-yellow monoclinic prisms. It is very volatile, sublimes at room temperature. It should be handled in a hood only. Vapors will irritate the eyes and respiratory tract. mp 25.4 degrees. bp 40 degrees. It reacts explosively with alcohol and filter paper. Uses: Oxidizing agent similar to osmium tetroxide, but more difficult to handle. The solvents normally employed in OsO4 oxidations (either, benzene, pyridine) cannot be used because of their violent reaction with RuO4. Only CCl4 is recommended.
We are unaware of the legal aspects and transportation methods for shipping RuO4. Because of the high volatility shipment may not be possible. ===================================================== So the point is that this is a very dangerous material, I am unaware of anyone who is brave enough to sell the tetroxide form of ruthenium as crystals (of course, the dioxide, ruthenium red, the metal, etc., there is no problem) and in any case, one can not put it into alcohol safely, and CCl4 (carbon tetrachloride) seems to be the only solvent that can handle it safely.
After the first posting, I suggested a way, privately, to a few who asked, that one could collect small amounts of ruthenium and dissolve it in alcohol . However, in view of the above information, I want to retract even the suggestion that could be done.
I stand behind my original statement that this material can not be shipped safely, and that is why it is not available in crystal form. To my knowledge, those using ruthenium tetroxide in electron microscopy, are generating it from a kit such as is described on our URL http://www.2spi.com/catalog/chem/chem1c.html
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi3spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
There is still one hole remaining for sponsership at the annual golf tournament during the M & M meeting in Portland. Hole sponsership is $65.00. Each golfer will recieve a gift bag containing gifts from various sponsers including:
JEOL FEI EVEX DENTON HITACHI PELLA
Please contact John Arnott at Ladd (see below) as soon as possible to participate in this annual event schedueled for Aug. 1, 1999 in Portland. There are still a few slots open for players as well.
Thanks,
John Arnott -- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
Thanks for your comments. Actually, the client already has data and a report, and wanted to know more about the technique. From what I'm learning, though, the data looks more like elemental mapping than what I would expect for EPMA. It seems that whomever supplied the report is doing just as you suggested: being sloppy with the nomenclature. I think what they really want is for us to be able and willing to provide similar analysis in house, but before I jumped to any conclusions or agreed to anything, I thought I'd learn as much as possible about the "alleged" technique. If anyone is familiar with the specifics of the EMAX3700 system, I would appreciate getting some details.
Thanks again.
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Warren E Straszheim {wesaia-at-iastate.edu} on 06/04/99 10:14:25 AM
To: Microscopy-at-Sparc5.Microscopy.Com cc: (bcc: David Bell/NA/Millipore)
There ought to be something about EPMA on the web. There is a mailing list at microprobe-at-www.microanalysis.org, but I think you have to be subscribed to post to it. I can get you details.
You are right about the acronym. Often, it refers to wavelength-dispersive spectrometers with crytals instead of the energy-dispersive Si and Ge solid state detectors. But some can get sloppy with the nomenclature. (I still prefer to refer to EDS than to EDAX. Nothing against EDAX, the company, but I prefer not to give them free advertising.)
I am not familiar with EXAX3700. What does you client want to know or do? Do they want low detection limits, high resolution scans, etc.?
At 09:47 AM 6/4/1999 -0400, you wrote: } } Hi all, } } I have a question regarding SEM/EPMA analysis. I cannot find information } about EPMA analysis on the web. Can anyone tell me what it is? Is it an } acronym for Electron Probe Microanalysis? Does anyone have any information } on an EMAX3700 mounted on a Hitachi SEM. I have a customer who is asking } about this, and would like to find out some information. ANY information } would be greatly appreciated. } } Thanks in advance, } } David Bell } Scientist } Electron Microscopy Lab } Millipore Corporation
Dear All: Thank you for your assistance w/ the companies that offer independent service contracts for TEM. For anyone who is interested, I can forward the info to them. This forum is a great resource for all of us. Thanks
Regards, Mike Coviello EM Lab Manager Materials Science & Engineering UT Arlington Arlington, TX
I'd like to get input from any users who have evaluated the differences in the ease of use/quality of output of a passive vs. an active digital acquisition system add-on that is available from a multitude of vendors for upgrading analog SEM's. I would also invite recommendations of either type of system that you have purchased and are happy with. We would like to put the system on our JEOL 845 SEM. Any input would be greatly appreciated.
Regards, Mike Coviello EM Lab Manager Materials Science & Engineering UT Arlington Arlington, TX
This is an advertisement for an imaging position. Nina Stromgren Allen
TECHNICAL DIRECTOR Center for Cellular and Molecular Imaging (CMIF) Department of Botany North Carolina State University, Raleigh, North Carolina
Responsibilites include overseeing the day-to-day operation of a light microscopy facility, instructing and collaborating with investigators, participating in education activities including organizing and conducting tours of the facility and participating in microscopy course teaching in conjunction with a second technician in the facility. This requires detailed knowledge of laser scanning confocal microscopy, video microsopy and image analysis with computers, digital imaging methods and the use of Photoshop etc., computer networking, as well as high competence in microinjection methods. Some knowledge of computer programming is very desirable. Part of the job will involve participation in grant writing and research in the area of plant cell biology. Ph.D and a minimum of 2 years experience is required. Minimum salary is $35,000. Full benefit package is available. Applicants should submit their curriculum vitae and reprints with a cover letter to Dr. Nina S. Allen, Department of Botany, Box 7612, North Carolina State University, Raleigh, NC 27695-7612 by June 18, 1999. Also please request that three professionals send reference letters to Dr. Nina S. Allen.
North Carolina State University is an Equal Opportunity Employer and operates under Affirmative Action Policy. The University strongly encourages all qualified applicants.
Nina Stromgren Allen Professor, Department of Botany; Director, Cellular and Molecular Imaging Facility Box 7612 North Carolina State University Raleigh, NC 27695-7612 Phone: 919-515-8382 (Office), 515-3525 (Lab), 515-2727 (Department Secretary) Fax: 919-515-3436
Dear colleagues, A few months ago I put up the question of how to quantify the instrument peak broadening of a x-ray diffractometer. I feel obliged to report the result.
I was put onto the NIST Standard Reference Material (SRM) #660 described in the very useful NIST's SRM web page at: http://ts.nist.gov/ts/htdocs/230/232/232.htm The specific web page for x-ray diffraction calibration standards is: http://ois.nist.gov/srmcatalog/tables/209-1.htm
The calibration material for line broadening is LaB6 in powder form (?).
Some suggested a single crystal wafer of Si and pointed out some problems to watch out for. According to my colleague, whom I was originally asking this question for, the peak broadening for a single crystal is not what he is after. He has polycrystalline specimens and he wants to determine the base or instrumental broadening without contributions due to the broadening from the thickness, grain size or internal stresses of the specimen. He was looking for a powder of a specific crystallite size, spread to certain thickness to satisfy the above criteria.
Thank you, for all of your contributions.
-- Steven Celotto Netherlands Institute of Metals Research (NIMR) Department of Applied Physics, University of Groningen Nijenborgh 4, 9747 AG Groningen, The Netherlands Ph: +31 50 363 4344 Fax: +31 50 363 4881 email: s.celotto-at-phys.rug.nl http://www.phys.rug.nl/mk/people/celotto/celotto.html http://www.nimr.nl/
Dave, The answer to your question is yes. I have usually seen reference to EMPA, electron microprobe analysis. A microprobe is not much more than an SEM with one or more wavelength spectrometers attached plus lots of expensive software. I'm not familiar with a EMAX3700. You may find some of the SEM vendors may be able to help you. Russ, Xerox
-----Original Message----- } From: "David_Bell-at-Millipore.com"-at-sparc5.microscopy.com [mailto:"David_Bell-at-Millipore.com"-at-sparc5.microscopy.com] Sent: Friday, June 04, 1999 9:47 AM To: microscopy-at-sparc5.microscopy.com
Hi all,
I have a question regarding SEM/EPMA analysis. I cannot find information about EPMA analysis on the web. Can anyone tell me what it is? Is it an acronym for Electron Probe Microanalysis? Does anyone have any information on an EMAX3700 mounted on a Hitachi SEM. I have a customer who is asking about this, and would like to find out some information. ANY information would be greatly appreciated.
Thanks in advance,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
A few months ago I asked about how to deal with magnetic specimens in the TEM. Recently another microscopist put up the same question. The series of books called Transmission Electron Microscopy by Williams and Carter give some good advice on pages 534 to 535 [Williams, D.B. and Carter, C.B, Transmission Electron Microscopy, No. 3, Imaging, Plenum, Press, 1996]. One good piece of advice they gave was to switch to aluminium specimens.
I have collated the advice I received and seen listed on the listserver, from what I have read and what experience I have gathered so far. What follows is a very brief summary and I can send the full information for those who are interested. I am interested in any further ideas or suggestions, so please inform me.
Suggestions for dealing with magnetic Fe specimens in the TEM:
1. Specimen preparation - minimise the amount of magnetic specimen in the microscope - Small flake specimens. - Window polishing technique - Tripod polishing - Self-supporting disks less than 50=B5m thick, 30=B5m is best. - Grinding down to 100-200=B5m - Chemical polish down to 30-50=B5m - Electropolish\ion-beam thin to perforation - 1mm disk of the steel is punched out, this is inserted into a 3mm disk of another material with a 1mm hole punched out.
2. In the microscope. - Do not use c-shaped spring clips, screw-down holder mechanisms are best. - Turn off the objective lens when: - Inserting the specimen - Moving to another cradle (if the holder has more than one) - Going through zero tilt - Removing the specimen probe. - Tilting in general for extreme problems when cradle will not tilt back. - Reduce the acceleration voltage -} reduces objective lens excitation -} reduces magnetic forces on specimen. - Increase the acceleration voltage -} increases momentum of electrons -} reduces deflection of electron beam. - Finding the eucentric height =96 - If by rotating back and forth the microscope goniometer, stick to one side of the zero tilt. - Use a non-magnetic sample to find the objective lens current for focus at eucentric height, place the magnetic sample in the microscope focus with the height control. Most modern microscopes now tell you what the current objective lens is for the eucentric height. - Aligning the microscope beforehand with a nonmagnetic sample and using a non-magnetic sample to get a firm idea where the transmitted beam should be on the screen (e.g. relative to the focus spot on the phosphor screen). - Use the dark-field mode - there is supposed to be more range in the beam deflection (and the bright-field settings will not set too far off for nonmagnetic specimen users). - Re-align the beam focus deflection compensators, current centre, voltage centre and objective stigmator whenever you have tilted or translated the specimen. Also, your co-workers will appreciate it if you restore normal settings when you finish. - The sample can be ripped out of the holder or will reorient the holder tilt mechanism so the holder cannot be withdrawn without scratching the OL polepieces. If this happens, don't try to remove the holder. Just turn off the OL, open the OL chamber, and carefully free the stuck parts.
For some of this I have cut & pasted from several emails. Thanks to those who replied to my question.
-- Steven Celotto Netherlands Institute of Metals Research (NIMR) Department of Applied Physics, University of Groningen Nijenborgh 4, 9747 AG Groningen, The Netherlands Ph: +31 50 363 4344 Fax: +31 50 363 4881 email: s.celotto-at-phys.rug.nl http://www.phys.rug.nl/mk/people/celotto/celotto.html http://www.nimr.nl/
On Friday, June 04, 1999 9:47 AM, "David_Bell-at-Millipore.com"-at-sparc5.microscopy.com [SMTP:"David_Bell-at-Millipore.com"-at-sparc5.microscopy.com] wrote: } ------------------------------------------------------------------------ } } } Hi all, } } I have a question regarding SEM/EPMA analysis. I cannot find information } about EPMA analysis on the web. Can anyone tell me what it is? Is it an } acronym for Electron Probe Microanalysis? Does anyone have any information } on an EMAX3700 mounted on a Hitachi SEM. I have a customer who is asking } about this, and would like to find out some information. ANY information } would be greatly appreciated. } } Thanks in advance, } } David Bell } Scientist } Electron Microscopy Lab } Millipore Corporation } 80 Ashby Road } Bedford, MA 01730 } (781) 533-2108 }
David,
Yes, EPMA stands for electron probe microanalysis. I describe it a bit on our web page:
I am not familiar with the Hitachi system you mention. You might want to contact Hitachi in Gaithersburg, MD for info. If you want to know more about EPMA, there are books by K. Heinrich, S.J. Reed, and numerous others. There is also a separate society, The Microbeam Analysis Society, that has bunches of EPMA specialists.
Hope this helps,
Jim McGee
{} {} {} {} {} {} {} {} {} {} {} {} {} James J. McGee (jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
Anyone interested in receiving a bid request form for acquiring our CAMECA Microprobe with:
3 ea. WDS spectrometers Regular & Biological Stage TN5000 converted to PC with software from Scripps Institute of Oceanography All Electronics Full set of manuals and documentation etc.
Please contact me based on information shown below:
Since I received several requests for the list of independents and because some direct e-mail was returned, I am sending them to the entire listserver community. If you are uninterested in this, please disregard this.
I received recommendations of independent EM service contractors who would offer a service contract on our Philips 430. I believe they said they would all service other brands.
1) Alex Green at Scientific Instrument Services out of Austin, TX. His number is 512-282-5507.
2) Vitaly Feingold out of Atlanta, vitalylazar-at-worldnet.att.net -at- 770-232-7785 or 770-605-6105.
3) Pesto Inc., P.O. Box 648, Gwynedd Valley, PA 19437-0648 215-699-6160 FAX 215-699-5275.
I hope this helps.
Regards and good luck, Mike Coviello EM Lab Manager Materials Science & Engineering UT Arlington Arlington, TX
In my electron microscopy laboratory I do not work with biological materials and hence have no biological specimens.
Many students and people interested in electron microscopy visit me from time to time and I would love to have some tissue sections to show them.
Would anyone out there be willing to send me a few grids that I could use? I would particularly like some sections of liver (my original area of study), kidney and skin.
If you contact me off-list I can let you have my address and FedEx number etc.
My early inquire on BSE with no BSE detector resulted in some suggestions that included sputter coating. Well, were we use to coat our specimens, when it is allowed, simply with gold; but I have heard that other methods could render better results. I guess that the gold coating could be regarded fine for temporary preparations, so it not lasts long and can be removed from the sample. However, gold coated specimens become useless if examined for long periods of time or in multiple SEM sessions. I heard that coating samples first with carbon and then with gold would result in long lasting preparations, which fits well in our purposes to preserve specimens in a entomological collection. My question is on what method is really the more appropriate and results in better images, both under BSE and SE scanning? And, what about gold/palladium coating, there are detectable differences between it and gold?
Thanks
Servio
S=E9rvio T=FAlio Pires Amarante serviopa-at-usp.br Museu de Zoologia - USP Caixa Postal 42 694 04299-970 Sao Paulo, SP BRASIL
I am an educator in a community college Medical Assistant program and have a an AO One Sixty microscope that is in need of condenser lens repair. Can anyone advise me of a company that might be able to help? I have been unable to locate any information from the web on the American Optical company that was at one time part of Warner-Lambert Technologies.
If you have any information please reply to: bphilpo-at-kirkwood.cc.ia.us
I have a University Safety Officer who has enquired about safety issues with regard to electron microscopy.
Can anybody recommend suitable books that cover both issues regarding the instrumentation itself and those relating to specimen preparation and handling?
Best regards,
-- Larry Stoter JEOL (UK) Ltd Silver Court, Watchmead, Welwyn Garden City, AL7 1LT, United Kingdom tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
Bill Hatcher Lab Optical Service 20222 Avondale Road Abingdon VA 24210-6942 phone: 540-676-7280 (ask for Bill Hatcher)
I believe that this phone number is current, but if you have problems getting a hold of Bill, let me know. The area code may have changed.
I have used Bill for many years to refurbish my A/O Microscopes, and he is excellent.
Ford Royer Analytical Instruments (Refurbished Laboratory Equipment) 9921 13th Ave. N. Minneapolis MN 55441 (800) 565-1895 (612) 929-1865 fax
Ian Philpott wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am an educator in a community college Medical Assistant program and } have a an AO One Sixty microscope that is in need of condenser lens } repair. Can anyone advise me of a company that might be able to help? } I have been unable to locate any information from the web on the } American Optical company that was at one time part of Warner-Lambert } Technologies. } } If you have any information please reply to: } bphilpo-at-kirkwood.cc.ia.us } } Bev Philpott
In regards to Chuck Garber's series of messages on Ruthenium Tetroxide, please note that he specifically refers to the crystalline form. As he has indicated, sources variously indicate ruthenium tetroxide reacts "violently", "explosively", or "it may explode". Other vendors do sell ruthenium tetroxide in aqueous form if you don't want mess with Garber's kit.
My thanks to Chuck, though, for publicly noting the hazards of ruthenium tetroxide.
I was wondering if you might be able to give me some help on finding references for specific properties of gold that might be relevant for in-situ TEM work on amorphous carbon grids. Basically, I'd like to know about the following things:
Solubility of carbon in gold, esp. at high T Vapor pressure of both liquid and solid gold near the m.p. (1064 C) Wetting properties of liquid gold on both solid gold and amorphous carbon Phase diagram of gold
Any help you could give me would be greatly appreciated. Please send responses to me (phantom-at-owlnet.rice.edu), and I'll summarize to the list if there's interest in me doing so.
Thanks for your time.
Steven Robertson The Colvin Group phantom-at-owlnet.rice.edu nanonet-at-ruf.rice.edu http://www.owlnet.rice.edu/~phantom http://nanonet.rice.edu
I am not a user of digital aquisition systems for an SEM, but we produce them. So perhaps I can shed a little light on this issue. I am pretty sure that other manufacturers of these systems would agree with what I am saying below.
Passive systems:
Advantages Easy setup (there is usually no modification of the microscope necessary) Easy image acquisition. Since the digital system "listens" to the SEM signal, all you need to do usually is to tell it to record the image or freeze it. No interference with EDS systems
Disadvantages Cannot extend functionality of microscope, only provides digitization of SEM microscopes No other image size than available on SEM No acquisition of X-ray maps possible, because the system has no control over beam. No control over e-beam
Active systems
Advantages More versatile, since beam can be controlled by PC. Control of beam position and dwell time Possible to acquire digital X-ray maps (requires EDS system) Any image size possible, up to 4k x 4k (or higher with diminishing returns) Other scans possible (backward, sideways, whatever), providing more freedom for scan and measurements In combination with EDS system allows for combined morphological and chemical analysis (aquire image, find particles, move beam, acquire EDS data).
Disadvantages Installation more complicated may interfere with existing EDS beam control system
I am not sure, these are all the differences. There may be more. I would take a good look at what you want to accomplish with the digitizer. If all you want to do is to get the images into a PC, a passive system might do. If you think, you ever need beam control, I would look for an active system. Of course, our systems are upgradeable, and for a relatively small amount you can go from a passive system to an passive+active one, giving you all the advantages of either. I am sure, other vendors do the same. And once the acquisition channels are set up and defined, you can switch from passive to active, from fast scan to high-resolution scan to EDS acquistion by just pushing a button.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
} ---------- } From: Michael Coviello[SMTP:COVIELLO-at-MAE.UTA.EDU] } Sent: Friday, June 04, 1999 1:01:50 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM-Passive vs. Active digital acquisition systems } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi All:
I'd like to get input from any users who have evaluated the differences in the ease of use/quality of output of a passive vs. an active digital acquisition system add-on that is available from a multitude of vendors for upgrading analog SEM's. I would also invite recommendations of either type of system that you have purchased and are happy with. We would like to put the system on our JEOL 845 SEM. Any input would be greatly appreciated.
Regards, Mike Coviello EM Lab Manager Materials Science & Engineering UT Arlington Arlington, TX
I've built and used both kinds, and while I agree with some of what Michael Bode says there are a couple of other points worth making
1. It certainly is possible to collect X-ray maps in passive mode. Depending on your multichannel analyzer you may be able to get 1, or several elements at once. 2. Many SEM scans are not perfectly linear. The manufacturers have presumably already tweaked their electronics so that the displayed images are correct, even if this means that the voltage driving the scan coils isn't linear. If you don't know what the nonlinearities are, it can be very difficult to set up an active system to get undistorted images. On the other hand, if the passive system is sitting on the display signals it doesn't care about this and the images come out right. 3. A really good passive system will even let you scan at high rates and average the values at each pixel, filling in the image as it can grab values (i.e., it reads X, Y and brightness and adds the values into memory, and after some time (perhaps 10 seconds is typical) the entire image is filled in with several readings everywhere and the software can divide everything down and give you a low noise image. This fast scanning capability can be very useful when you have charging problems. It is not easy to make an active system do that.
Servio is asking several questions basics to SEM specimen coating.
For conventional SEM sputter coating of Au is by far the most used. Au/Pd was shown many years ago to be slightly finer, but in practice its difficult to note any difference. Au/Pd targets tend to be more expensive because of greater assaying cost and the huge variety of target sizes has killed any economies of scale.
Carbon/graphite is an inferior conductor, but scatters well during evaporation, so specimens with complex structures are better covered. Carbon coatings are the choice material for microanalysis, but quite inferior for imaging since resolution greatly increases with the atomic number of the coating and also the A.N. of the specimen below the coating. A double coating with C/Au if fine, but a lot of trouble for little gain. These days using a lower accelerating voltage or sputtering the specimen twice from two sites are the better ways to prevent specimen charging.
Durability of the coating largely depends on: 1 Dry storage - get a sizeable desiccating cabinet. 2 Poor microscope vacuum conditions 3 Beam damage from an intense beam, this is only marginally related to coating quality. After WDS analysis, which requires high counts, a spot will be damaged regardless of coating material used.
If a coating is unsatisfactory another can be applied on top, if the specimen is used for medium or low power observation. "Thick" coatings obscure finest structures; its much like a dusting of snow on the ground leaves pebbles and other small features visible, whereas a metre of snow reveals only major topography. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Saturday, June 05, 1999 1:33 PM, Servio Tulio Pires Amarante [SMTP:serviopa-at-usp.br] wrote:
} Dear all } } My early inquire on BSE with no BSE detector resulted in some suggestions } that included sputter coating. Well, were we use to coat our specimens, when } it is allowed, simply with gold; but I have heard that other methods could } render better results. I guess that the gold coating could be regarded fine } for temporary preparations, so it not lasts long and can be removed from the } sample. However, gold coated specimens become useless if examined for long } periods of time or in multiple SEM sessions. I heard that coating samples } first with carbon and then with gold would result in long lasting } preparations, which fits well in our purposes to preserve specimens in a } entomological collection. My question is on what method is really the more } appropriate and results in better images, both under BSE and SE scanning? } And, what about gold/palladium coating, there are detectable differences } between it and gold? } } Thanks } } Servio } } Servio Tulio Pires Amarante } serviopa-at-usp.br } Museu de Zoologia - USP } Caixa Postal 42 694 } 04299-970 } Sao Paulo, SP } BRASIL } }
Chao-ying Ni wrote: ================================================ I've heard that Au/Pd coating is more uniform and the particles in the film are smaller, which are beneficial especially to high resolution SEM.
-cy ================================================ This is almost certainly true for vacuum evaporated 60%Au/40% Pd vs. 100% Au (the alloy composition normally implied when mention is made of "Au/Pd"). The smaller grain size was first reported in the mid-1960's. But these were the days before sputter coaters. Or at least the days before sputter coating approaches were being used in the SEM laboratory (which did not really start until about 1972 or so).
Has anyone ever shown, however, that sputter coated Au/Pd results in a grain size that is smaller than sputter coated pure gold ? We ourselves tried some years ago to see if there was a difference but could not find any difference.
If this is true, then would not the use of the alloy result in longer sputtering times but without any real benefit? The metal value of the alloy is somewhat less than for a gold cathode of identical dimensions, but the fabrication charges to make the alloy are highter, so the final selling prices are not all that much different. However, if the sputter time for fragile and heat sensitive samples is increased, then this would be a drawback for use of the alloy.
If by "high resolution" you are referring to FESEM instruments, then the use of either gold or gold palladium, either in a conventional or ion beam sputter coater will, in terms of grain size, still fall short of several other alternatives, such as chromium coaters or osmium coaters.
Disclaimer: SPI Supplies is a manufacturer and distributor of conventional, chromium, as well as osmium coaters for SEM lab applications.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
On all of our microscopes with scanning we have fitted with ImageSlaves for digitisation. Polaroid and roll film recording are now historical in our lab.
These are passive acquisition boards which detect and digitise the video record signal.
They work very simply. On pressing the Photo record button the image is digitised. There is an auto save feature which makes the entire process extremely simple.
Pixel resolution is 1024 x 1024 maximum. I think they will record images at 12 bits.
U.S.A. Contact Jim Hilton Advanced Database Systems 7931 S. Broadway #322 Littleton CO 80122 U.S.A. Tel: + 1 303 761-5635 Fax: + 1 303 761-592 } } ***************************************************** Mel Dickson, Deputy Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
We have both in our lab. I find a few things to consider.
1) It is my impression is that the passive system yields a better image in the same amount of time. It seems the active system has a fair amount of overhead involved in positioning the beam. Thus only a fraction of the total time is spent collecting image data.
2) The active system affords a point-and-click x-ray analysis option. Since the computer system knows where the image came from, it can reposition the beam to locations in the image for x-ray collection. That is not easy or perhaps even possible when the computer is riding along. I have found this to be a very big aid to productivity.
3) The software implementation will have as much to do with satisfaction as does the technology. Both systems can be great or terrible depending on how well the software is written. You should try for a hands-on demo to see how the software "feels" before committing.
So we have and use both systems. If we only need an image (no x-rays), I use the passive system for better quality in less time. If I need to do x-ray analyses, I use the active system.
Good hunting.
At 02:01 PM 6/4/1999 -0500, you wrote: } Hi All: } } I'd like to get input from any users who have evaluated the differences } in the ease of use/quality of output of a passive vs. an active digital } acquisition system add-on that is available from a multitude of vendors } for upgrading analog SEM's. I would also invite recommendations of } either type of system that you have purchased and are happy with. We } would like to put the system on our JEOL 845 SEM. Any input would be } greatly appreciated. } } Regards, } Mike Coviello } EM Lab Manager } Materials Science & Engineering } UT Arlington } Arlington, TX } } }
Would You please review the below copy to post the group.
Hi Larry,
A good reference on EM safety is the book Electron Microscopy Safety = Handbook (2nd edition) by V. C. Barber and J. A. Mascorro. Published byb = San Francisco Press, Inc. =20 ISBN 0-911302-72-7
Some of EM supply houses carry this edition, or can be ordered from = publisher.
Best of Luck,
Ed } } } Larry Stoter {LPS-at-teknesis.demon.co.uk} 06/05 2:49 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html=20=
I have a University Safety Officer who has enquired about safety issues with regard to electron microscopy.
Can anybody recommend suitable books that cover both issues regarding the instrumentation itself and those relating to specimen preparation and handling?
Best regards,
-- Larry Stoter JEOL (UK) Ltd Silver Court, Watchmead, Welwyn Garden City, AL7 1LT, United Kingdom tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.c= om=20
Edward P. Calomeni Dept Pathology - EM Lab 3000 Arlington Ave. Toledo, OH 43614
Regarding the distortions: Normally the scan generator provides nothing but a linear saw-tooth voltage for the scanning in x and y direction. This is then fed into the scan amplifier, which takes care of getting the voltages right for the appropriate magnifications. Most systems that I know, and certainly ours, replace the scan generator, but not the scan amplifier. The manufacturers have to tweak the scan amplifier for the non-linearities, as they may be different for different mags. So by feeding the signal in BETWEEN generator and amplifier, we take advantage of any kind of tweaking the manufacturers have done. This means, that an active system does not really have to worry about the non-linearities.
Averaging: Of course you can average a number of images on a snapshot, say 128 frames. This can be done of course with active and passive systems. In active systems you have the additional option of increasing the dwell time to reduce noise, which is equivalent to changing the scan time on the microscope.
X-ray mapping: I agree, that it is possible to acquire X-ray maps with a passive system. If you have no active component in your system (neither digitizer nor EDS beam control) and the digitizer just looks at the video signals, you would have to scan at a very low rate and perhaps run multiple images for averaging and noise reduction. You would have to deal with image shifts between the exposures, etc. With an active system, you simply set the dwell time per pixel and have the system scan the area, acquiring SE or BSE images at the same time X-rays are counted (with a pulse counter in our case). So, if there is a shift during exposure, it will lead to slight distortions of both the SE image and the X-ray maps, but the distortions will be the same, so that the data can be correlated without problems.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
} ---------- } From: "DrJohnRuss-at-aol.com"-at-sparc5.microscopy.com[SMTP:"DRJOHNRUSS-at-AOL.COM"-at-SPA RC5.MICROSCOPY.COM] } Sent: Saturday, June 05, 1999 5:41:20 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: RE: SEM-Passive vs. Active digital acquisition systems } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I've built and used both kinds, and while I agree with some of what Michael Bode says there are a couple of other points worth making
1. It certainly is possible to collect X-ray maps in passive mode. Depending on your multichannel analyzer you may be able to get 1, or several elements at once. 2. Many SEM scans are not perfectly linear. The manufacturers have presumably already tweaked their electronics so that the displayed images are correct, even if this means that the voltage driving the scan coils isn't linear. If you don't know what the nonlinearities are, it can be very difficult to set up an active system to get undistorted images. On the other hand, if the
passive system is sitting on the display signals it doesn't care about this and the images come out right. 3. A really good passive system will even let you scan at high rates and
average the values at each pixel, filling in the image as it can grab values (i.e., it reads X, Y and brightness and adds the values into memory, and
after some time (perhaps 10 seconds is typical) the entire image is filled in with several readings everywhere and the software can divide everything down and give you a low noise image. This fast scanning capability can be very useful when you have charging problems. It is not easy to make an active
This person should be encouraged! Please help him out. Does anyone have the email or website for ISI, which I believe is now TOPCON?
Please respond DIRECTLY to Eric-at-fotherby.freeserve.co.uk Re, Scanning electron microscope. I have at home in my living area a scanning electron microscope which I have restored.
MODEL ISI SUPER 3A. ( sms 3a ) which is very similar to ISI MODEL 40. Manufactured FEB 1977. Serial number 510028. I purchased the SEM a couple of years ago from a army surplus depot --------however one part missing, the oil filled electron gun high voltage power supply / filament supply. I have converted a electron beam welder power supply, this works but I really need a circuit diagram of the original gun power supply, so I can find the values of bias resistance to use. I have obtained full service manuals of the SEM from two sources but in both cases the high voltage electron gun power supply details are missing------circuit diagram number N83HA08. The internet ( which I am new to ) points me in your direction, can you please advise ? any information is better than none! My address, 117, PEASEHILL, RIPLEY, DERBYSHIRE, DE5-3JN, My EMAIL ADDRESS, Eric-at-fotherby.freeserve.co.uk Thankyou. Yours faithfully, Eric Fotherby.
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Chairman, Educational Outreach subcommittee promoting access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
This is the list I have compiled. I hope the people who requested a copy, were not expecting dozens of companies. Since I received numerous requests for the list of independents and because some direct e-mail was returned, I am sending them to the entire listserver community. If you are uninterested in this, please disregard this.
I received recommendations of independent EM service contractors who would offer a service contract on our Philips 430ST. I believe they said they would all service other brands.
1) Alex Green at Scientific Instrument Services out of Austin, TX. His number is 512-282-5507.
2) Vitaly Feingold out of Atlanta, vitalylazar-at-worldnet.att.net -at- 770-232-7785 or 770-605-6105.
3) Pesto Inc., P.O. Box 648, Gwynedd Valley, PA 19437-0648 215-699-6160 FAX 215-699-5275.
I hope this helps.
Regards and good luck, Mike Coviello EM Lab Manager Materials Science & Engineering UT Arlington Arlington, TX
I have heard to grow Mo crystals on a TEM grid, I should burn a piece of Mo and pass a holey carbon grid through the smoke. Is this toxic? Can I use MoO3 powder in a crucible instead of a piece of foil?
} We have both in our lab. I find a few things to consider. } } 1) It is my impression is that the passive system yields a better image in } the same amount of time. It seems the active system has a fair amount of } overhead involved in positioning the beam. Thus only a fraction of the } total time is spent collecting image data. }
(etc.)
While any particular system may produce better or worse images than another, I don't believe there's any inherent magic in passive scanning that produces better images. I was involved in the design of an active system a few years ago, and we spent a great deal of time worrying about the issue of beam settling times and efficiency of sampling. Passive systems still have to be concerned with flyback settling times, for example, or the first few samples in the line will be trash. You have no idea where the beam is for some period of time at the beginning of each scan line; how long you have to wait depends on design choices by the EM manufacturer.
Probably the main determining factors of image quality in either type of system are the speed, linearity and noise performance of the digitizing ADC, and the equivalent parameters for the position DACs (active system) or raster timing circuitry (passive system).
You get much nicer MoO3 crystals if you put ammonium molybdate in a small crucible, leave the lid slightly ajar and gently warm the crucible with a bunsen burner. The MoO3 crystals will condense under the lid of the crucible and can be washed off with methanol. You can then put a drop of the suspension on a holey formvar grid.
This recipe is from J.W. Edington "Practical Electron Microscopy in Materials Science" and produces much better crystals than the old Moly smoke method.
Cheers, Henk
At 03:05 PM 6/7/99 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
You can also take a Mo boat used for deposition in an evaporator system and just heat it in the evaporator. I only recently saw Tom Nuhfer at Carnegie Mellon University heat a Mo wire in a propane torch and when it was hot enough, take it away from the flame and wave the wire under the grid. No special precautions were taken and both Tom and I walked away healthy (although I'm still overweight.)
I wouldn't intentionally breath the smoke -its easy to avoid. The MoO3 smoke pretty much goes straight up and you can do all of this at arm's length. If you are worried about it you can do this last procedure in a hood. You could also just use a particulate mask which would probably also give you a sense of security.
I would also suggest that when you have a question like this, you should also consult an MSDS for the compound and then use your best judgment on how to proceed.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: jennifer taylor To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
I have heard to grow Mo crystals on a TEM grid, I should burn a piece of Mo and pass a holey carbon grid through the smoke. Is this toxic? Can I use MoO3 powder in a crucible instead of a piece of foil?
Dear list members, Let us please put an end to this as a subject heading, as it is not a heading and does not relate to subject of ones text. The purpose of the subject heading is to get your message or questions to people who are interested in that topic. So, let's be open about what and why we are posting to the group. Nestor, I would like to suggest that this phrase be a cue for rejection of posting.
Regards,
Images & Info at http://www.molbio.princeton.edu/confocal {http://www.molbio.princeton.edu/confocal}
Joe Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University jgoodhouse-at-molbio.princeton.edu {mailto:Jgoodhouse-at-molbio.princeton.edu}
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
These crystals can be purchased on carbon films. An advanced version has the crystals on a grating replica so that magnification and rotation calibration can be done on the same image-a neat time saver. The first item is part No. 625, while the grating replica version is No. 625-B.
by mozart.cems.umn.edu (8.9.3/8.9.3) with SMTP id RAA05790 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 7 Jun 1999 17:50:17 -0500 (CDT)
Responding to the message of {70A79B42AEE7D21181B300805FA9180F15D80E-at-molbio.Princeton.EDU} from "Goodhouse, Joseph" {jgoodhouse-at-molbio.princeton.edu} : [...] } Dear list members, } Let us please put an end to this as a subject heading, as it is not } a heading and does not relate to subject of ones text. The purpose of the } subject heading is to get your message or questions to people who are } interested in that topic. So, let's be open about what and why we are } posting to the group. Nestor, I would like to suggest that this phrase be a } cue for rejection of posting. } Unfortunately, as one who has had mail rejected by the listserver for inappropriate subject contents (MSA Bulletin) I have to say that the phrase "I am not spam" is actually sugested by the listserver as an alternative to the rejected subject. Having discovered that, I had more sympathy for those using this as a subject line. However, it is clearly not very helpful in identifying the message contents, and I tend to automatically delete such messages.
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist Office:(612) 626-7594 CIE Characterization Facility, University of Minnesota Desk: (612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
You forget. Nestor put this in for people who get rejected by his SPAM filter. They are to reply as per his instructions. This happened to me when he put the filter in and it got fixed. It turned out to be a problem with information that my company sends out with all Email messages in the message header. If I remember correctly, it took him a couple of tries to fix why I (and a few others) were getting rejected. There are Listserver messages under those subject lines, "I am not spam", or there should be.
Patience. Nestor is doing a great job. Let's do it his way.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Goodhouse, Joseph To: 'Microscopy-reguest' -----------------------------------------------------------------------.
Dear list members, Let us please put an end to this as a subject heading, as it is not a heading and does not relate to subject of ones text. The purpose of the subject heading is to get your message or questions to people who are interested in that topic. So, let's be open about what and why we are posting to the group. Nestor, I would like to suggest that this phrase be a cue for rejection of posting.
Regards,
Images & Info at http://www.molbio.princeton.edu/confocal {http://www.molbio.princeton.edu/confocal}
Joe Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University jgoodhouse-at-molbio.princeton.edu {mailto:Jgoodhouse-at-molbio.princeton.edu}
I would also like to point out that the rotation calibration, magnification calibration over an extended range, and camera constant can all be done using the MAG-I-CAL sample. There are a number of sources for this sample. I bought mine from South Bay Technology (1-800-728-2233). I believe that electron Microscopy Sciences and SPI both carry it.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Bernard Kestel To: Microscopy Listserver -----------------------------------------------------------------------.
These crystals can be purchased on carbon films. An advanced version has the crystals on a grating replica so that magnification and rotation calibration can be done on the same image-a neat time saver. The first item is part No. 625, while the grating replica version is No. 625-B.
At 15:05 7/06/1999 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi, You just put a (small) piece of moly foil from a discarded aperture or heater strip in the bottom of a small test tube.
Heat the foil strongly in a bunsen flame.
You will see a mist of crystals evolve from the foil.
Hold filmed grids in the smoke. Make lots and you have a lifetime supply.
If you want, you can also let the crystals settle on the wall of the tube.
Wash them off with distilled water. Put a drop of the suspension on the filmed grids.
That way the crystals will lie flat on the film.
No information about toxicity, but do it in the fume hood to be cautious.
***************************************************** Mel Dickson, Deputy Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Mike, There are several other independents that were factory trained to work on the EM430: Sam Amtower in New England (amtec-at-neca.com) and myself, Ron Veil (veilcs-at-earthlink.net - URL: http://home.earthlink.net/~veilcs/)
I have a list of 17 independent field service techs should anyone be interested. It is in Win-95/Excel format, so I can send it as an attachment.
Ron Veil V.E.I.L. (Veil Electron Instrument Lab) Customer Services 173 Santa Inez Ave. San Bruno, CA. 94066 (650) 952-3099 FAX (650) 869-4978 Pgr. (650) 205-0302
On Mon, 07 Jun 1999 11:39:28 -0500 Michael Coviello {coviello-at-mae.uta.edu} writes: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America
___________________________________________________________________ Get the Internet just the way you want it. Free software, free e-mail, and free Internet access for a month! Try Juno Web: http://dl.www.juno.com/dynoget/tagj.
The Hitachi H-7000 TEM is fitted with an RS232 interface for external I/O communication.
Does anyone out there know if it can be used to issue commands to control functions of the microscope?
We would like to adjust focus & image shift & stigmation on out H-7000. All these are digitally controlled within the microscope.
The next model produced, the H-7100 DOES have this facility and there is a book of control codes for the H-7100. Officially, the H-7000 does not have the facility and the RS232 interface is variously described as being for use by service engineer, or for inserting alphanumerics on the film. Is there any unofficial experience out there we can tap into?
***************************************************** Mel Dickson, Deputy Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
The subject of film scanners have come up a number of times. I've mentioned in the past that I'm pretty happy with the Polaroid SprintScan 45.
I just came across a catalog from Publishing Perfection (Vol 17) that has a Nikon LS-4500 for $6500 and a Minolta Dimage Scan Multi for $2500. The Minolta write-up specifically mentions TEM negatives. Their number is 1-800-852-2348 or 1-800-782-5974 and their web site is www.publishingperfection.com
I have no financial interests in any of these companies.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Hi, This heading is REQUIRED by Nestor for people posting from domains that otherwise would get filtered out from the list because of the few bad apples that may be spamming from said domain. I personally feel it is a little humiliating to put that in the subject line of my posts, since my domain is filtered. I will do very little posting as a result. Cheers, Dave Harrison
I wish people would read directions, but perhaps that is too much to ask some times.
When a message is received that is SUSPECT as SPAM, then the sender gets an AUTOMATIC message saying there is a problem. The sender is then instructed to reply to POSTMASTER-at-MSA.MICROSCOPY.COM with the subject line I AM NOT SPAM, and then I personnally review the message and fix the problem. They are instructed not to send the mail back to the Listserver.
Unfortunately, this last set of problems arose simply because people did not follow the directions!!!!!!!!!!
I have just implemented yet another code modification on the filter to make this clear, and with this change to the filter I do not think any "I AM NOT SPAM" messages will get through now. But if people can't follow simple directions there is not much I can do.
Right... I'm going to get a beer and a late dinner...
Looking for any information/experience on interfacing CCD camera model # TE3N/A. Camera was manufactured in 1994 by ASTROMED (UK?). The CCD sensor is KODAK KAF 0400-C1, Peltier cooled. Any info such as interface type, control cards vendors, and software vendors will be very helpful.
Please reply to this e-mail address: vitalylazar-at-worldnet.att.net
Vitaly Feingold Scientific Instruments and Applications Duluth, GA (770)605-6105 (770)232-7785
I deeply agrre with others, My message was rejected as a spam, despite that my server ans.com.au was never involved it any spam,.. I stop posting messages to microscopy, this is first one after 5 months.. Even this 3 lines was rejected a a spam and I am sending it again
Hi all! Does anybody have experience in cutting (nondecalcified) teeth for transmission electron microscopy. Is it necessary to use diamond knives or is a sapphire knife good enough? How can cutting be done without getting a lot of fractures in the enamel and how do the knives tolerate the cutting? Is there any difference in cutting human vs.rodent teeth? Are there any special methods or tricks in embedding the teeth? I would appreciate any information, hints, or advice regarding this matter. Thank you very much. marketta Hormia
___________________________________________
Marketta Hormia University of Turku Institute of Dentistry Lemmink=E4isenkatu 2 FIN-20520 Turku Tel +358-2-333 8330 Fax +358-2-333 8356 E-mail: marketta.hormia-at-utu.fi
Dear all, In relation to the MoO3 rotation calibration question, I have just done a set of these on our microscope but I have one question.
To determine whether there is a 180=B0 inversion, Williams and Carter say to defocus the diffraction pattern slightly. If the diffraction focus is adjusted, however, there is a 180=B0 inversion of the image in the transmitted spot between underfocus and overfocus (as you would expect with lenses). Which side of focus should I use for finding out whether or not there is a 180=B0 inversion between image and diffraction?
Hope someone understands my question and can help.
Thanks
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren Department of Experimental Physics Chalmers University of Technology S-412 96 G=F6teborg Sweden Tel: +46 31 772 36 33 FAX: +46 31 772 32 24 email: maclaren-at-fy.chalmers.se or: ianmaclaren-at-hotmail.com Research Group Homepage: http://fy.chalmers.se/microscopy/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Thank you VERY much for the great responses from everyone! The report my client had received was from Japan and was translated/written rather poorly, but with your help we were finally able to figure out what was done. I can't emphasize enough what a great resource this listserve is, despite some of its foibles. Thank you Nestor for doing a terrific job. (hope the beer and late dinner was relaxing!)
Thanks again,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781)533-2108
I am looking for a flatbed scanner that can scan an A3 X-ray film in one pass (size A3, approx. 11.7x15.7 inches) in transparent mode. It should be able to handle densities up to 3.0 at least. A few months ago an Umax Mirage II A3 flatbed scanner was bought. The density range of this scanner should go down to 3.3D according to its specifications. However according to our measurements (tested with Kodak Wratten filters, checked with a density meter !) it only reaches about 2.5D.
After a preview with maximum gray range (0-255) settings one can adjust the lower (0) and upper (255) input range limits. If the upper limit is lowered below 100 or even below 50 (input range for scanning 0 to 50) very peculiar effects take place. Is there anyone out there who owns such a scanner and has the same experience ?
Any remarks on this subject or info on other brands of A3 tranparency flatbed scanners are much appreciated.
Thank you very much for your time.
Please mail directly to : vanderwulp-at-pml.tno.nl
Kees van der Wulp TNO Prins Maurits Laboratory P.O.Box 45 2280 AA Rijswijk Netherlands
You need to weaken the diffraction (1st intermediate lens) to get the non-inverted image within the diffraction pattern.
Henk
At 02:14 PM 6/8/99 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University=09 (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
Please Unsuscribe for a while, I=B4m going to be out my lab. for a month.
Thanks a lot and best regards. M.C. Ma. Guadalupe Nieto Lopez Laboratorio de Microscopia Electronica ECOSUR Tapachula Carr. Ant. Aeropuerto Km. 2.5 30700 Tapachula, Chis. Tel. (962) 81077 y 81103 Fax. (962) 81015
For years, we sputter coated in air but recently switched to argon when a tank was given to us. The tank is getting low and I'm getting poor results. Research with the supplier leads me to believe it is grade 2.2 (99.95%). they also have grade 4.8(99.99 %) in the small tank size. However the price goes from $18 to $69 with the jump in grades.
What grade argon is normally used in sputter coating? If I stick with the 2.2 am I going to cause problems somewhere else?
Go to a convergent beam and go to diffraction mode, i.e. form a CBED pattern. Decrease the Condenser lens strength (Brightness, CCW) and = you will form a shadow image of the sample in the BF disk. The diffraction pattern and this image are not rotated with respect to each other. You = can then relate that to your image mode and diffraction pattern. -Scott
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---------- } From: Ian MacLaren To: Microscopy = -----------------------------------------------------------------------.=
Dear all, In relation to the MoO3 rotation calibration question, I have just done a set of these on our microscope but I have one question.
To determine whether there is a 180=B0 inversion, Williams and Carter say to defocus the diffraction pattern slightly. If the diffraction focus is adjusted, however, there is a 180=B0 inversion of the image in the transmitted spot between underfocus and overfocus (as you would expect with lenses). Which side of focus should I use for finding out whether or not there is a 180=B0 inversion between image and diffraction?
Hope someone understands my question and can help.
Thanks
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren Department of Experimental Physics Chalmers University of Technology S-412 96 G=F6teborg Sweden Tel: +46 31 772 36 33 FAX: +46 31 772 32 24 email: maclaren-at-fy.chalmers.se or: ianmaclaren-at-hotmail.com Research Group Homepage: http://fy.chalmers.se/microscopy/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
A diamond knife is probably required for cutting undecalcified dental enamel. Try the sapphire and see what happens. Since enamel is 97% mineralized it is impossible to get much epoxy into it. Try using Spurrs resin and infiltrating for a week or so and maybe even using vacuum to help the infiltration. Do this embedding with a 100 micrometer or thinner slice of tooth. It helps to pre-trim the sample so that you will cut a very small cross-section, less than 0.1mm. When sectioning you may have to take very small steps, less than 100nm, to "polish" the sample and than take one section at about 200nm. Pick the section up right away before the surface tension forces of the knife boat water break the section up on a grid with a support film. In the very best case after many attempts you may get some useable pieces to image in the TEM. Then, interpretation of the images will require some knowledge of cyrstals and crystallography. One more trick to try and keep the fractures together (you are fracturing not really cutting) is--after "polishing" the sample surface as above, put a drop of Formvar or collodion or your favorite support material on the face of the sample and let it dry then cut one section and pick it up. Rat teeth are not much different from human teeth is this respect. What are you trying to study? If you decalcify the enamel slightly embedding and sectioning become more likely. Another approach is to use ion milling. Find a good materials prep lab and try it out. I had some luck with a crude ion mill and using a 1000kV microscope tho 100 kv gave some information. Caveat: I did this work more than 20 years ago so may have forgotten some details.
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
I am posting this message for a local hospital that is closing their TEM lab and surplusing some pieces of equipment. Their lab manager has indicated that they would be free to anyone who would come to get them, or pay for shipping them. For additional information on the equipment, please contact them directly. Contact person, address and telephone number is:
Karen Michalski Cytogenetics Laboratory Franciscan Shared Laboratories St. Joseph's Hospital 5000 West Chambers Street Milwaukee, Wisconsin 53210 USA 414-447-2883
Item #1 Hitachi HU12A Transmission Electron Microscope - Purchased in "late 70's or early 80's". I have seen the microscope, it has been run using city water to chill it. It had a preventative maintenence performed on it within the past calendar year, and the last time I talked with the electron microscopist (before his retirement), the microscope was working.
Item #2 PelAire Station - I turned it on, and it appears to work O.K.
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
Dear Mel, having used an Hitachi HF2000 in the past I can say that it is most likely that you can use an external computer to change your lenses.=20 The way we did this in the past was to use a small BASIC program on a PC, which talked to the TEM through the RS232 port. The program prompted you for two input numbers (both are 4 digit hexadecimal) which were then sent to the TEM.
The first number is the `address=B4 number which tells the TEM computer whic= h lens it is, the second is the value you wish to put in ranging from 0000 to FFFF. The program we used also had the ability to read the settings, so we could check the lenses to see if manually changing them altered the values.= =20
Because of the way this works you are going to have to calibrate the settings, which is no mean feat.=20
I could try to get hold of the program, but it will take a week or so to find out if my old TEM lab can a/ find it and b/ let me take a copy (I do not see why not).
Since I do not have access to this machine any more I cannot remember too much about it except that it was a pain to use. The reason I wanted to use it was for performing a tilt series.=20
My other line of suggestion is to find someone who knows how to interface a computer to the RS232 port. You should be able to find the addresses in the Technical manual of the H7000.=20
Good Luck, Jonathan ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The =C5ngstr=F6m Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
We are running an HF-2000 remotely and we have done it for several years. However, we go through Gatan's software DigitalMicrograph. Otherwise, the control through the RS232 interface should be as expected (i.e., is standard for an RS232). It is worthwhile to note that the magnification can be set to the same values as if turning the magnification nob directly. It is not necessary to set each lens directly, thus avoiding calibration problems. Hitachi has given us the manuals without a problem.
Hope this helps...
Edgar
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Does anyone know of any other software other than EL/P that can open an EL/P file? DTSA perhaps? Or is it easy/preferable to export a spectra from EL/P as two-column ascii or otherwise?
-a colleague has sent me an EL/P file to show me a spectra. I have EmiSpec's ES-vision software for handling my PEELS as well as other graphing software and have not used in EL/P in several years.
Thanks, Brendan Foran Materials Analysis / Internal Technical Support SEMATECH Austin, Texas
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We have a Hitachi H 9000 with a Gatan 622 video camera. Some things = that we look at such as crack propagation in the straining stage and the = melting of precipitates during implantation seem to happen on a time scale = that is too rapid for the standard 30 frames per second video camera. = Have there been any applications of high speed video cameras on TEMs? By = high speed I would mean up to 250 frames per second or more.
Anthony W. McCormick Materials Science Division Argonne National Lab. 9700 S. Cass Ave. Argonne, IL 60439
Someone wants to take color pictures using my Nikon UFX 35mm camera system and a stereo scope. It has been so long since I used the system, that I have forgotten the tricks. Our first set of pictures came out expposed correctly, but with a distinctly yellow cast, such that whites were yellow and yellow was orange-red. Is this the film type (we used Kodak 100 daylight)? Should we have used tungsten film? The light source was a fiber optic illuminator. Thanks-Dave
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Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax)
DTSA won't open an EL/P file. Your colleague could save the spectrum in EL/P as an ASCII file, even as an ASCII file in the MSA/MAS format (i.e., with a header). ---------------------------------- } } } To those doing electron energy loss spectroscopy } } Does anyone know of any other software other than EL/P that can open an EL/P } file? DTSA perhaps? } Or is it easy/preferable to export a spectra from EL/P as two-column ascii or } otherwise? } } -a colleague has sent me an EL/P file to show me a spectra. I have EmiSpec's } ES-vision software for handling my PEELS as well as other graphing } software and } have not used in EL/P in several years. } } Thanks, } Brendan Foran } Materials Analysis / Internal Technical Support } SEMATECH } Austin, Texas
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798 cook-at-aaem.amc.anl.gov
Hello Ron and Others, The argon we have been using cost $15.00 for the small tank. I have never specified any grade; but according to your findings and if price has anything to do with it, it must be the lower grade. It has done well by us.
Hope this helps, Sandra
At 10:40 AM 6/8/99 -0400, you wrote: } } Dear listmembers, } } For years, we sputter coated in air but recently switched to argon when a } tank was given to us. The tank is getting low and I'm getting poor results. } Research with the supplier leads me to believe it is grade 2.2 (99.95%). } they also have grade 4.8(99.99 %) in the small tank size. However the price } goes from $18 to $69 with the jump in grades. } } What grade argon is normally used in sputter coating? If I stick with the } 2.2 am I going to cause problems somewhere else? } } Thanks. } } Ron } lherault-at-bu.edu } } } Sandra F. Zane, EM Tech. sfzane-at-email.uncc.edu Dept. of Biology, UNC-Charlotte Ph.(704)547-4051 9201 University City Blvd. Fax (704)547-3128 Charlotte, NC 28223
Folks, I need some advice on the practicality of providing backup power supply for a SEM. The power here drops out about six times a year and can last from a few minutes to hours and of course can occur days, nights, weekends... I recall this being discussed in past year, but I wasn't sure if there was feeling as to the success of attempting to provide power for an orderly shutdown or just trying to hang on until power comes back on. The only time I operated a scope with a propane generator backup, in the five years I was there, the power never went out so I don't know if that would have handled a real test.
Any help would be appreciated.
Dave Audette OSRAM Sylvania Beverly, MA david.audette-at-sylvania.com
Use the purest argon available. If you are going to try refractory metals you will need the best grade AND make sure you have metal to metalconnections from the bottle to the coater. If you are only going to do noble metals and their alloys maybe you can get away with cooking style argon, but to continue the metaphore, we always use argon equivalent to a First Growth Claret.
Patrick Echlin CambridgeOn Tue, 8 Jun 1999, lherault wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear listmembers, } } For years, we sputter coated in air but recently switched to argon when a } tank was given to us. The tank is getting low and I'm getting poor results. } Research with the supplier leads me to believe it is grade 2.2 (99.95%). } they also have grade 4.8(99.99 %) in the small tank size. However the price } goes from $18 to $69 with the jump in grades. } } What grade argon is normally used in sputter coating? If I stick with the } 2.2 am I going to cause problems somewhere else? } } Thanks. } } Ron } lherault-at-bu.edu } } } }
Microcosm, Inc. is looking for Zeiss LSM-410 microscopes. We will offer a fair price.
If you have LSM-410 for sale or you are planning to sale one in near future please contact me at (301) 725-2775.
Dr. Henryk Malak ________________________________________________________ Dr. Henryk Malak Director of Research Microcosm, Inc. | 9140 Guilford Road, Suite O |Columbia, MD 21046 Phone: (301) 725-2775, Fax: (301) 725-2941 Our web site is located at: http://www.microcosm.com {http://www.microcosm.com} ________________________________________________________
Rememeber, you can use a terminal emulator or a dumb terminal to experiment with such ports in the early stages. The first challenge will be to get the communication parameters correct (baud rate, stop bits, etc.) and to make sure that the cable is correct. A null modem or other specialized cable may be necessary. A lot of instruments with RS-232 interfaces don't totally follow the standard, or maybe I should say that the standard allows for a variety of flow-control options that one cannot tell ahead of time what the settings will be. A good manual from the manufacturer will go a long way to help. You may also want to borrow a computer geek to get the communication set up.
The next issue is getting the right command syntax. I would probably try the 7100 codes and see if they work. You should be able to send commands manually from a terminal to see if they work at all. Then you could see about packaging them up in a program such as QBASIC as was already suggested.
At 10:10 AM 6/8/1999 +1000, you wrote: } } Fellow listers.... } } The Hitachi H-7000 TEM is fitted with an RS232 interface for external I/O } communication. } } Does anyone out there know if it can be used to issue commands to control } functions of the microscope? } } We would like to adjust focus & image shift & stigmation on out H-7000. } All these are digitally controlled within the microscope. } } The next model produced, the H-7100 DOES have this facility and there is a } book of control codes for the H-7100. Officially, the H-7000 does not have } the facility and the RS232 interface is variously described as being for } use by service engineer, or for inserting alphanumerics on the film. Is } there any unofficial experience out there we can tap into? }
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
I pretty much agree with Rick Mott's comments. There are a host of factors involved and clever engineers can do much to minimize or even eliminate the differences. Conceptually, I would think an active system need not wait for the beam to completely settle before sampling the signal. The sample would simply be a mix of the signal from the area under the beam as it settled in. That could do a lot to reduce the overhead of active beam control. And frankly, I don't know whether our system overlaps beam position and sampling or not.
The disadvantage of our active systems is a limited number of choices for dwell times. We have choices of 10, 100, 200, 400, 800 us, etc. We would really like to have more choices between 10 and 100 us in order to match the collection time of our passive system. We can get a good passive image in 40 seconds. While 10 us yields an active image quickly, it is too noisy. A 100 us dwell produces a good image, but it takes more than 80 sec. It shouldn't be a big issue, but we collect thousands of images and the time adds up. I guess there could be similar problems with passive systems. You are constrained by the available scan rates on the microscope. Hopefully, you have a good set of choices.
So I guess I will summarize by saying the devil is in the details of the implementation - as always.
At 04:12 PM 6/7/1999 -0400, you wrote: } Warren E Straszheim wrote: } } } We have both in our lab. I find a few things to consider. } } } } 1) It is my impression is that the passive system yields a better image in } } the same amount of time. It seems the active system has a fair amount of } } overhead involved in positioning the beam. Thus only a fraction of the } } total time is spent collecting image data. } } } } (etc.) } } While any particular system may produce better or worse images than } another, I don't believe there's any inherent magic in passive scanning } that produces better images. I was involved in the design of an active } system a few years ago, and we spent a great deal of time worrying } about the issue of beam settling times and efficiency of sampling. } Passive systems still have to be concerned with flyback settling times, } for example, or the first few samples in the line will be trash. You have } no idea where the beam is for some period of time at the beginning of } each scan line; how long you have to wait depends on design choices } by the EM manufacturer. } } Probably the main determining factors of image quality in either type of } system are the speed, linearity and noise performance of the digitizing } ADC, and the equivalent parameters for the position DACs (active } system) or raster timing circuitry (passive system). } } Rick Mott }
You bet, the daylight film is less sensitive to blue than tungsten balanced film, so if you use it with tungsten lighting that has less blue than daylight, the images will have the yellow shift. Even when you use the tungsten balanced film, you may need the test the lamp voltage. The color of the illumination will change dependent on the voltage. Higher voltage the more blue, lower voltage more yellow. Some lamps will still are not in the range of the tungsten film and require color compensating filters. Also, if you only have daylight film, you can use a 80A or 80B color conversion filter to balance the light to daylight.
Bob Derm Imaging Center
On Wed, 9 Jun 1999, David Knecht wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Someone wants to take color pictures using my Nikon UFX 35mm camera system } and a stereo scope. It has been so long since I used the system, that I } have forgotten the tricks. Our first set of pictures came out expposed } correctly, but with a distinctly yellow cast, such that whites were yellow } and yellow was orange-red. Is this the film type (we used Kodak 100 } daylight)? Should we have used tungsten film? The light source was a } fiber optic illuminator. Thanks-Dave } } Home of the 1999 NCAA Basketball National Champion HUSKIES !!! } ************************************************************ } } Dr. David Knecht } Department of Molecular and Cell Biology } University of Connecticut } 75 N. Eagleville Rd. U-125 } Storrs, CT 06269 } Knecht-at-uconnvm.uconn.edu } 860-486-2200 860-486-4331 (fax) } } ************************************************************ } } } }
I'm a Newcastle Brown Ale kinda guy myself but I have decided to try the higher grade of Argon, since it is used so slowly and should give the best results. I have not had any problems I can associate with using tygon tubing to deliver the gas so I probably will not get into modification of the delivery system at this time.
Thanks to all who have contributed information.
Ron L -----Original Message----- } From: Dr P. Echlin {pe13-at-cus.cam.ac.uk} To: lherault {lherault-at-bu.edu} Cc: Microscopy-at-Sparc5.Microscopy.Com {Microscopy-at-Sparc5.Microscopy.Com}
Hi Ya'll: We are looking for user experiences on the Vitalscan imaging system when used specifically on the JEOL 840 series SEM. We are looking for ease of use/archiving/x-ray acquisition/software stability/image quality, etc. Any experiences/recommendations would be greatly appreciated. Tahnk you in advance.
When using most fiber optic systems (that use a projector type bulb with reflector), you should turn the bulb up to nearly full brightness to get the proper color temperature. Even then, you may need to use a blue filter in the system (80A, I believe). You could use tungsten film such as Fuji 64T but you will need to still adjust the brightness of the bulb so that it is at the proper color temperature.
I would recommend the following: use a tungsten film and take an exposure series varying the bulb brightness from the half-way position up to max. Then evaluate the images, picking the optimal one. Now, ALWAYS set the bulb to this same setting. If the lamp is too bright, then use a neutral density filter rather than turning the bulb down since this will change the color temperature.
The ideal solution would be to use a color temperature meter to set the bulb to the proper temperature each time you shoot the images but they are expensive.
John
} Someone wants to take color pictures using my Nikon UFX 35mm camera system } and a stereo scope. It has been so long since I used the system, that I } have forgotten the tricks. Our first set of pictures came out expposed } correctly, but with a distinctly yellow cast, such that whites were yellow } and yellow was orange-red. Is this the film type (we used Kodak 100 } daylight)? Should we have used tungsten film? The light source was a } fiber optic illuminator. Thanks-Dave
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
This yellow color is usually derived from the illumination you are using. Try an 82B filter (there is often a blue filter included with your microscope). The problem is related to what is called "color temperature". There are a number of good, basic discussions. For example, see: 1) Delly, John. Photography through the microscope (Kodak publication, available at most good camera stores) 2) Photomicrography (Polaroid) 3) Optimizing Light Microscopy (available through our web site - see below).
Basic principle for solving color correction ("CC"): make a circle; divide into 6 segments. Label alternating segments with primary colors (red, green, blue); label the other three with secondary colors (between red and blue = magenta; between blue and green= cyan; between red and gree = yellow).
To suppress a color (ex: the yellow you saw or the green tinge which often accompanies instant prints), try a filter of the opposite sector.
Kodak used to have a neat set of gelatin "CC" filters available in a little booklet, again through a good camera supply store. When you get the print, just hold a filter up in front of it under similar lighting to see which one will work best then insert that filter in the light path.
Good luck!
Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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At 09:43 AM 6/9/99 -0400, David Knecht wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes, you have to use Tungsten film (unless you're shooting color negatives & printing, then the color balancing is done during printing). The other trick is to make sure the lamp is turned up bright. The scope should have some guide associated with this to indicate when the lamp is bright enough (in the photography range, much brighter than the eyeball range). This is necessary, because if the lamp is run too low, it will be too red--the black-body temperature (aka color-temperature) will be too low. If this is too bright for what you're doing, *don't* lower the brightness of the lamp, instead insert one or more neutral-density filters.
The other trick to balance the color if you only have daylight film is to use an "80" series 35mm camera filter. Put it over the window for the field diaphragm, below the condensor. Usually 80A, but B or C might be needed (the filters are denser, going from A to C). There are also Color Compensating (CC) filters that can be used, but these are designed for say color photography under fluorescent lights. The 80 series works fine usually. Using Tungsten film works better.
(Note, Tungsten film has to be used for halogen lights, not just tungsten bulbs. The difference between tungsten & daylight isn't wire filament or the sun, but rather color temperature, and halogen lights are closer to tungsten bulbs in color.)
Phil
} Someone wants to take color pictures using my Nikon UFX 35mm camera system } and a stereo scope. It has been so long since I used the system, that I } have forgotten the tricks. Our first set of pictures came out expposed } correctly, but with a distinctly yellow cast, such that whites were yellow } and yellow was orange-red. Is this the film type (we used Kodak 100 } daylight)? Should we have used tungsten film? The light source was a } fiber optic illuminator. Thanks-Dave } } Home of the 1999 NCAA Basketball National Champion HUSKIES !!! } ************************************************************ } } Dr. David Knecht } Department of Molecular and Cell Biology } University of Connecticut } 75 N. Eagleville Rd. U-125 } Storrs, CT 06269 } Knecht-at-uconnvm.uconn.edu } 860-486-2200 860-486-4331 (fax) } } ************************************************************
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The H 9000 that we have does not have a scanning mode so the electron = beam is fixed at one point on the sample. The scan originates in the = Gatan video camera.
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We also just bought an Agfa Duoscan 2500.
If you turn the 3 1/4 x 4 TEM negs sideways they just fit in the 4 x 5 hol.der. You can also use the glass holder, although I realize that glas= s is undesireable. I bought an extra 4 x 5 holder to modify it, but found that since the neg fit in the 4 x 5 holder sideways that works also. If you make a modified holder to hold 4 TEM negs, I would appreciate the design. That would be extremely handy.
Thanks Judy Murphy
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
Ian MacLaren wrote:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------= =2E } } Dear all, } Is there anybody out there using an Agfa Duoscan for TEM negatives? } } Our lab has just bought one (the T2500) but the negative holder (60 by } 90) does not quite fit our TEM negatives (64 by 88 mm). } } What have others done about this? } Manually adjusting the holder? Having a special holder made? } } We would appreciate any ideas that you can share. } } Yours sincerely } } +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Ian MacLaren } Department of Experimental Physics } Chalmers University of Technology } S-412 96 G=F6teborg } Sweden } Tel: +46 31 772 36 33 } FAX: +46 31 772 32 24 } email: maclaren-at-fy.chalmers.se } or: ianmaclaren-at-hotmail.com } Research Group Homepage: http://fy.chalmers.se/microscopy/ } +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
I had problems with both Spectra and ratio images from the GIF with respect to taking my data home where I do not have EL/P or Digital Micrograph; in fact I don't own a Mac. Here's what works best for me.
EELS SPECTRA For spectra, save it in both the MSA format (I think that it is still called EMSA format in the program) and two column ASCII. Both are ASCII. The MSA format will have all of the header information about the spectrum that you need, however, the data comes out in groups of rows of 5 numbers and these groups are separated by a blank row. There may be groups of 10 rows, but I'm not exactly sure here. I use the MSA format header data for information and the ASCII data graphs very quickly in Excel or any graphing program. I just have not gotten around to writing a small Basic program to decipher the MSA format data into x,y pairs. There is information in the header of the MSA that tells how many columns of data there are, so this should be fairly straightforward to do it. I've been playing with learning Visual Basic specifically to do this and will have it done before too long.
RATIO IMAGES With respect to ratio images you can run into problems with how you store the TIF images. Digital Micrograph has the option of saving images in an 8 bit mode (I can't remember what they call it) or Raw mode which is 16 bit. If you take a regular image and save it in the raw TIF mode, there is no problem in opening it in Photoshop or other software such as Thumbs Plus. However, if any pixel value is negative when the difference of the two images is taken for the ratio image, you will not be able to open it in Photoshop. It gives an error about non-standard format or some such message. I could not find any software that would open the file other than Digital Micrograph. Even NIH image would not do it. There are no problems if you save the ratio images in the 8 bit format except you had better have the contrast and brightness of the image set before saving the image. PC USERS, DON'T FORGET TO ADD THAT EXTENSION TO THE FILENAME!
I'm sure that this is more than what you needed, but I just recently had to deal with these problems and it is not too much of a problem for me now that I know what's going on.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: "Brendan.Foran-at-sematech.org"-at-Sparc5.Microscopy.Com To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
To those doing electron energy loss spectroscopy
Does anyone know of any other software other than EL/P that can open an EL/P file? DTSA perhaps? Or is it easy/preferable to export a spectra from EL/P as two-column ascii or otherwise?
-a colleague has sent me an EL/P file to show me a spectra. I have EmiSpec's ES-vision software for handling my PEELS as well as other graphing software and have not used in EL/P in several years.
Thanks, Brendan Foran Materials Analysis / Internal Technical Support SEMATECH Austin, Texas
At 11:31 AM 6/9/99 , you wrote: } At 09:43 AM 6/9/99 -0400, David Knecht wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You need a color temperature correction filter that changes the 2950K color temp of the halogen lamp to 5500K of daylight film. Tungsten film will not match your source if it is quartz halogen. In this case, use a Wratten 80A (blue) filter or what Olympus calls an LBD-2....light balancing daylight.
The color temperature of the halogen lamp does change with applied voltage so without a color temperature meter, you might have to do a small amount of experimentation to be sure. you will either need an 80A or 80B filter. Use chrome film for adjusting perfect color balance. C-41 has quite a wide latitude viz color balance.
Molecular Imaging, a leader in high resolution biological AFM research and manufacturing, will be visiting UCSF campus Friday, June 11th. An informal demo session will be open to the public 9am to 12 pm, in Science 420. During the demo, a protein sample (ferritin) will be imaged in solution. We will be happy to discuss any questions/comments related to the application of AFM in your research projects.
Posters and Images at: http://www.molec.com/biology/index.html
I am trying to label one or more reptor(s) using polyclonal antibodies (that we know works at the light level) on tissue that needs to also be treated for the DAB reaction product. We are attacking this by pre-embedding and post embedding. One of our concerns is the effect of the hydrogen peroxide and DAB on the antigenic sites, and the other is the fixative. During pre-embedding, should we do the DAB reactions first then the immunogold or vice versa? For the post embedding ,using LRW, can you perform the DAB reaction on the grids? Creating the next question, will nickel grids work with the peroxide?
We used a mixture of 4% paraformaldehyde, 0.5% glut, plus 10%v/v picric acid (PA), all in Sorensons buffer. What effect could the PA have on the DAB reaction? Several papers recomend PA for IEM, but I am wondering now about the PA because I had a previous DAB experiment not turn out the way we thought (neg. to poor reaction). I notice that most papers use Tris buffers at 0.05M for the DAB but a few do mention phosphate buffer? Is there a problem not using Tris?
If someone out there has had experience with combining DAB and immunogold labeling I would like to here from them.
-----Original Message----- } From: Dr. Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Thursday, June 10, 1999 9:28 AM To: yanga-at-em.agr.ca Cc: MSA listserver
At 06:25 PM 6/9/99 , you wrote: } I am going for holidays on May10 and will be back on June 7. } } Ann Fook
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes, the distinct yellow cast that you found on your first roll of film is indeed due to using daylight balanced film with a tungsten halogen light source. Most fiber optic illuminators use tungsten halogen projector lamps. Tungsten Halogen lamps usually run at a color temerature of up to 3200K or 3400K, depending on their wattage. The words "up to" are important, because the total light output AND the color temperature are dependent on the voltage of the lamp. Raising the voltage will increase the output and increase the color temperature of the lamp, causing the output to be richer in blue light.
You have two choices to correct the problem, you can use daylight balanced film with filtration, or tungsten balanced film with little or no filtration.
If you must use daylight film, you will need the equivalent of a Wratten 80A filter (for 3200K) or a Wratten 80B filter (for the higher wattage 3400K light sources). The glass filters usually supplied with your compound microscopes are closest to the Wratten 80A filter.
Alternatively, you can use daylight balanced film with little or no filtration. The higher wattage tungsten halogen lamps operating at 3400K at full voltage would need a small correction with a Wratten 81A filter. However, if you do a voltage series, you may find that reducing the voltage of the lamp slightly gives you the right color temp for no filtration with tungsten balanced film.
Most fiber optic illuminators that I have seen do not have slots for filtering the light. I usually ended up cutting small pieces of a Wratten filter and securing it to the end of the fiber optic nearest the specimen. Alternatively, I have secured a clean, flat filter over the objectives of the stereo microscope and focussed through the glass without any noticeable degradation of the image.
The wratten color correction (CC) filters are not approprate filters for light balancing for color temperature. They absorb a smaller portion of the spectrum than do the color conversion Wratten 80 and 85 series, or the light balancing Wratten 82 and 81 series. The absorption curves of these filters are available in the very helpful Kodak publication B-3, "Kodak Filters for Scientific and Technical Uses"
I have a question for the microscopists who may have experimented with color temperature vs. lamp voltage for tungsten and tungsten halogen lamps at the compound microscope. I found that reducing the voltage of a tungsten lamp (which might operate at up to 2800K) led to a very large difference in both output and color temperature of the lamp. When I made the same changes with a tungsten halogen lamp, I found that the variation of output and color temperature were far less pronounced. Does anyone know why this may be the case?
Mary McCann McCann Imaging Ph: 617-484-7865 Fax:617-484-2490 email: mccanns-at-tiac.net http://www.microscopyed.com
} } Someone wants to take color pictures using my Nikon UFX 35mm camera system } and a stereo scope. It has been so long since I used the system, that I } have forgotten the tricks. Our first set of pictures came out expposed } correctly, but with a distinctly yellow cast, such that whites were yellow } and yellow was orange-red. Is this the film type (we used Kodak 100 } daylight)? Should we have used tungsten film? The light source was a } fiber optic illuminator. Thanks-Dave } } Home of the 1999 NCAA Basketball National Champion HUSKIES !!! } ************************************************************ } } Dr. David Knecht } Department of Molecular and Cell Biology } University of Connecticut } 75 N. Eagleville Rd. U-125 } Storrs, CT 06269 } Knecht-at-uconnvm.uconn.edu } 860-486-2200 860-486-4331 (fax) } } ************************************************************
Apparently Ann Fook has setup an auto-reply message during her absence from office. I have seen many others do that and wish they would not, at least if they remain subscribed to mailing lists. It means that when you and I send a post to the list, their auto-reply kicks in and we as the poster gets to hear about their vacation, again. Realizing that, I just shake my head and hit the delete button.
I suggest that if someone wants to use such an auto-reply function, they should set the nomail option on their lists (if available) or else temporarily unsubscribe. Otherwise, if they want to remain on the lists, they should avoid using the auto-reply. Or finally, they might learn how to use filters to only send the auto-reply to personal communications.
Warren
At 02:50 PM 6/10/1999 +0800, you wrote: } } I hope that I do not have to see this again!! } } Catherine } } -----Original Message----- } } At 06:25 PM 6/9/99 , you wrote: } } I am going for holidays on May10 and will be back on June 7. } } } } Ann Fook
Re, SEM 3A REQUEST FOR INFO REGUARDING HT OIL TANK.
I feel that I should say a big thank you to everyone who answered my Email concerning the above subject. Hopefully I will receive the info I need in the next few days. THANK YOU EVERY ONE. Yours faithfully, Eric Fotherby.
Trying to collect more data... If you have specification/sales brochures for Hitachi, Jeol, Philips, or Amray SEMs covering models, from about 1990 up to (but not including) the current offerings, I would dearly love to obtain a copy.
Thanks, Woody ---------------------- Woody White McDermott Technology, Inc Lynchburg Research Center P.O. Box 11165 Lynchburg, VA 24506
My colleague resent me the data saved from EL/P as "xy-ascii" which was of course easily graphed. Something apparently was wrong with the EL/P file originally sent to me.
To avoid discrediting EmiSpec, and as Gerd had surmised, EmiSpec's software is set up to import EL/P ".tad" files; I hadn't realized that at first, and then still it didn't work for the file originally sent to me (could be that we forgot to specify that the file was text format in moving it from the Mac to the PC- and we are going to re-try it though going the route of ascii is easy enough.
Thanks again for the advice,
Brendan
Brendan Foran, Ph.D. Materials Analysis Group SEMATECH Austin, TX
-----Original Message----- } From: Walck. Scott D. [mailto:walck-at-ppg.com] Sent: Wednesday, June 09, 1999 5:35 PM To: 'Brendan.Foran-at-sematech.org'; Micro
I had problems with both Spectra and ratio images from the GIF with respect to taking my data home where I do not have EL/P or Digital Micrograph; in fact I don't own a Mac. Here's what works best for me.
EELS SPECTRA For spectra, save it in both the MSA format (I think that it is still called EMSA format in the program) and two column ASCII. Both are ASCII. The MSA format will have all of the header information about the spectrum that you need, however, the data comes out in groups of rows of 5 numbers and these groups are separated by a blank row. There may be groups of 10 rows, but I'm not exactly sure here. I use the MSA format header data for information and the ASCII data graphs very quickly in Excel or any graphing program. I just have not gotten around to writing a small Basic program to decipher the MSA format data into x,y pairs. There is information in the header of the MSA that tells how many columns of data there are, so this should be fairly straightforward to do it. I've been playing with learning Visual Basic specifically to do this and will have it done before too long.
RATIO IMAGES With respect to ratio images you can run into problems with how you store the TIF images. Digital Micrograph has the option of saving images in an 8 bit mode (I can't remember what they call it) or Raw mode which is 16 bit. If you take a regular image and save it in the raw TIF mode, there is no problem in opening it in Photoshop or other software such as Thumbs Plus. However, if any pixel value is negative when the difference of the two images is taken for the ratio image, you will not be able to open it in Photoshop. It gives an error about non-standard format or some such message. I could not find any software that would open the file other than Digital Micrograph. Even NIH image would not do it. There are no problems if you save the ratio images in the 8 bit format except you had better have the contrast and brightness of the image set before saving the image. PC USERS, DON'T FORGET TO ADD THAT EXTENSION TO THE FILENAME!
I'm sure that this is more than what you needed, but I just recently had to deal with these problems and it is not too much of a problem for me now that I know what's going on.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: "Brendan.Foran-at-sematech.org"-at-Sparc5.Microscopy.Com To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
To those doing electron energy loss spectroscopy
Does anyone know of any other software other than EL/P that can open an EL/P file? DTSA perhaps? Or is it easy/preferable to export a spectra from EL/P as two-column ascii or otherwise?
-a colleague has sent me an EL/P file to show me a spectra. I have EmiSpec's ES-vision software for handling my PEELS as well as other graphing software and have not used in EL/P in several years.
Thanks, Brendan Foran Materials Analysis / Internal Technical Support SEMATECH Austin, Texas
The best source for refractive index oils of all sorts is Cargille Labs in Cedar Grove, NJ: 973-239-6633
They also have a very good reference booklet by Dr. Robert Allen on Refractometry. We use it for all our Am. Chem. Soc. courses.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 10:16 PM 6/9/99 -0600, wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I received several questions regarding color LM microphotography. Here is a short compilation of the individual posts for everyone's general use as may be beneficial:
If you are using daylight film, use the LBD-2 filter. Depending on which lamp house and bulb it uses, there still may be a slight color shift. If the results are warm (on the red, yellow side), increase the color temperature of the source by increasing the lamp voltage. Usually there is a setting for taking photos. However, on some scopes, this setting is too warm and there will be a color shift.
The LBD-IF filter can be used for tungsten film but your best results and controls are achieved with daylight film. And there are many more choices as well. There are two types of tungsten film: A and B. A is rated at 3,200K while B is rated at 3,400K. Some tungsten films do not directly indicate which kind they are; or the maker makes it really a challenge to find out which kind it is. It really does make a difference. If you are shooting slides, use Fuji Provia (RDP-II) at ISO 100 or Provia 400 (RHP) at ISO 400. The grain increases of course with faster film.
If you use C-41 negative film, I would recommend Fuji NPS shot at ISO 100. It is a 4-layer emulsion film and is good for a little extra light correction.
Remember that it is much easier to adjust the final color balance when making prints from negative (C-41) films. This is done during printing and if the neg is decently exposed (highlights not blown out) then good results can be had. Printing from E-6 slides is a different process (Ilfochrome/Cibachrome) and is much more expensive and difficult to control/vary.
If you are interested in color temperature of various bulb sources, check out this following URL:
http://photoweb.net/pw_tech/floures1.html
Here you will find the color temp of different light sources and the filters needed to correct them to daylight film. Without correction, even tungsten A or B film will still have a cast. Especially if you use Type B tungsten film. The mired shift is enough between 2900K of the halogen source and the rated 3200K of the film to show up as a subtle warming cast. Daylight C-41 film will also require severe color correction without an LB filter. There are actually two types of filters required for color compensation. The LB filter is the light balancing filter. The CC filter is the color correction filter. In reality, the LB is the main filter which adjusts the red/blue ratio. The CC filter handles the green. Continuous sources, like tungsten and halogen bulbs, really only need LB compensation. The CC filter comes into play when using non-continuous spectra sources like flourescent or halide lamps.
The best way, short of having a color meter, to balance color temperatures of source and film is to shoot chromes and adjust source voltage and/or filtering to achieve a neutral, real color result. With a typical halogen source, this means using for example an Olympus LBD or LBD-2N filter or an 80A blue filter.
The other option aside from printing is to digitally scan both or either negs or chromes (slides). This opens up many useful features. For example, if you are shooting C-41 negs, you can calibrate the scanner to the color cast of the backing and have the software automatically remove that after a preview scan. This will auto correct the color balance of the film frame to as close as daylight as can be done. Furthermore, you can play with the color settings yourself to remove yellow/red by adding blue and/or cyan. These scanners will also sharpen your images, auto adjust highlights or shadows and with separate software plugins to Photoshop (like Extensis Intellihance) automatically perform up to 20 separate steps to optimize each scanned image.
Finally, there are basically two types of color meters. Type 2 and Type 3. Really all this means is whether the meter has 2 sensors (red + blue) or 3 sensors (red + blue + green). There are numerous older models of Type 2 meters that have two sensors, a pair of filters and a simple D'arsonval meter movement to indicate the color temperature. The newer digital meters are much better but also more expensive.
Our 20 year old freeze drier is dying and I am looking to buy a new one. It will be used for standard SEM prep stuff of biological material. I am looking for anyone's recent experience with new models. Vendors in the UK are welcome to reply. It might be preferable to reply directly to me rather than the list. As usual I will be happy to post a digest if there is interest. Many thanks
Chris
Chris Gilpin Experimental Officer Biological Sciences EM Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 0161 275 5170 Fax +44 0161 275 5171 Chris Gilpin http://www.empgu.man.ac.uk
I am s-o-r-r-y! The auto-reply has been taken out.
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } Warren E Straszheim {wesaia-at-iastate.edu} 06/10 8:27 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Apparently Ann Fook has setup an auto-reply message during her absence from office. I have seen many others do that and wish they would not, at least if they remain subscribed to mailing lists. It means that when you and I send a post to the list, their auto-reply kicks in and we as the poster gets to hear about their vacation, again. Realizing that, I just shake my head and hit the delete button.
I suggest that if someone wants to use such an auto-reply function, they should set the nomail option on their lists (if available) or else temporarily unsubscribe. Otherwise, if they want to remain on the lists, they should avoid using the auto-reply. Or finally, they might learn how to use filters to only send the auto-reply to personal communications.
Warren
At 02:50 PM 6/10/1999 +0800, you wrote: } } I hope that I do not have to see this again!! } } Catherine } } -----Original Message----- } } At 06:25 PM 6/9/99 , you wrote: } } I am going for holidays on May10 and will be back on June 7. } } } } Ann Fook
} I received several questions regarding color LM microphotography. Here is } a short compilation of the individual posts for everyone's general use as may } be beneficial:
} { stuff deleted in the interest of brevity}
Dr. Gaugler makes some excellent points, only one of which I disagree with. If you shoot print film, the automatic printing machine, not being familiar with images shoot through the microscope, will rarely give accurate color (in my experience) so you may have to pay for custom printing. Expensive. If you shoot slide film you can select the best image and have it printed, either from an internegative, by Ilfochrome or by scanning. Now you also have a slide for presentation at seminars or meetings. Also you can experiment and fine tune the light source you are using by using varing light intensities, color correction filters and making accurate notes. Since the color on the slides is not adjusted in printing you can find the right setting/filter for your equipment and subject matter. Just my $.02 worth.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
The Imaging and Analysis Center at Princeton Materials Institute, Princeton University has an open position for a Specialist. The successful candidate will instruct students in the operation of instruments for imaging (electron, optical, and scanning probe microscopies), analysis, and diffraction; maintain and repair these instruments; oversee upkeep of the center; support faculty and students; analyze samples; supervise students; maintain a safe environment; and other assigned duties. Bachelor degree in physical science and/or 4+ years-related work experience required. Candidates should be familiar with the instruments in the facility, mechanical and electronic equipment, vacuum systems, PC and/o SUN and EDS/WDS systems. The candidate should be competent with sample preparation techniques, including ion milling, ultra-thin sectioning, staining, and coating of samples. Good communication and interpersonal skills are essential.
Interested candidates should send application to: Susan Calvetto, Business Manager, Princeton Materials Institute, Princeton University, 70 Prospect Ave. Princeton, NJ 08540, calvetto-at-princeton.edu. Applications should include a resume, letter of application and three letters of recommendation. Princeton University offers excellent benefits
Is there software available that will display graphical output from the various EDS file formats? I am using a Link Oxford system. I would like to access spectra at my desk. I can export to EMSA/MAS format.
I am looking for a chemical etch that would etch my substrate (MgAl2O4) but not my film (CoFe2O4). I would very much appreciate any suggestions. Thanks for taking time to consider this request.
Sincerely,
Mick Thomas ---------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
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sure...mike we have one for you... -----Original Message----- } From: Ingram, Mike {MIngram-at-rodel.com} To: 'Microscopy-at-MSA.Microscopy.com' {Microscopy-at-sparc5.microscopy.com}
Does anyone know any company or individual who services Reichert Ultramicrotomes. I live in Tampa, Florida and would prefer finding one who is relatively near. Thank you very much. Margie Bryant
JEOL JSM - 35U Scanning Electron Microscope for sale. It was on JEOL Service Contract from purchase date (1976) until March 31, 1999. It still takes excellent pictures easily at mags below 10,000x's and nice pictures with manipulation at mags up to 20,000x's. I feel as if I am selling my best friend. Best offer.
Phoebe J. Doss, Manager/Adjunct Instructor Electron Microscope Lab Oklahoma State University (405)744-6765
I recently have come across literature from Instrumedics, Inc. on their device for cutting and mounting cryo sections without passing sections through aqueous solutions and without melting the sections onto the slide.
The system attaches sections to the slides using a UV-cured adhesive. It sounds great, but at a $6,600 price tag (you supply the cryostat) I need to know whether this really works. Are there people out there who have used this? Please forward your comments to me.
Thanks.
Don
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
Geoff brings up an interesting point. Automatic printing machines will certainly try to adjust color balance to levels that they "think" are best, regardless of what the researcher wants. Depending on the lab you use, you may or may not be able to specify color corrections and hand prints back to be redone. Some labs just won't or can't do it.
Similarly, these machines will give you brightness/darkness levels they "feel" are appropriate. They don't know that the area of tissue you really wanted to see was that really dark patch in the center, instead of the perfectly exposed surrounding matrix, for example. They try to average brightness values across the whole negative to come up with a reasonable compromise, which works fine for most family picnic pictures.
Slides, as Geoff says, overcome these problems since there is no machine interpretation of the image. The final image you see is the original piece of film exposed in the camera---just the way you exposed it in terms of color and brightness. But you can take it and specify improvements when having prints made, or you can scan it in and do anything you want digitally.
Slide films come in various color balances, just like print films, and you can use the same filters. They are very useful for these types of applications.
I have been asked by a faculty member to fix rat brains, cut and measure the dendrites of a neuron. He has given me three articles but I they are dated (1972). I was hoping for some direction of the current protocols used.
Any assistance would be greatly appreciated. Debbie Lietz
Debbie Lietz Electron Microscopy Suite Department of Biology Trent University Peterborough, Ontario K9J 7B8 Telephone: (705)748-1486 Fax: (705) 748-1205 email: dlietz-at-trentu.ca
South Bay Technology, Inc. will be offering another in its ongoing series=
of Tripod Polisher Workshops on September 24-25, 1999 in San Clemente, CA= . =
New at this workshop will be additional sections on low energy ion milling, plasma cleaning and ion beam sputtering/etching. For a course description and on-line registration information, please go to our websit= e at www.southbaytech.com and select the "Workshops" icon.
We need to prepare some TEM samples of a powder metallurgy sample of roughly
Pb 8% Sn 1.5% Cu (?) Al balance
Any suggestions on preparation? We were thinking of trying Nitric/Methanol (the usual Al electropolish).
Thanks, Henk Colijn
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
I have a JEOL IC845 SEM. When recording images on film, I get evenly spaced light and dark areas (similar to fringes) on my micrographs. It does not appear on screen when viewing an image in normal viewing mode, but in watching the photo scan line it does appear to be present, but difficult to see. Does anyone have an idea as to what is going on?
Which way are the lines oriented and what is their spacing on the film? If the lines run vertical on Polaroids, it could be due to gunk on the rollers or the smoothness of pulling the film from the camera.
At 04:49 PM 6/11/1999 -0400, you wrote: } } I have a JEOL IC845 SEM. When recording images on film, I get evenly spaced } light and dark areas (similar to fringes) on my micrographs. It does not } appear on screen when viewing an image in normal viewing mode, but in } watching the photo scan line it does appear to be present, but difficult to } see. Does anyone have an idea as to what is going on?
I don't know how many of you may have heard of the death of Arthur Cohen this past week. He was a pioneer in the preparation techniques used in scanning electron microscopy and critical point drying. He was Professor Emeritus of Botany at Washington State University, and carried out his research and electron microscopy interests long after his retirement.
I was fortunate enough to spend some time in his company over the last few years, and learn a few things which have enhanced my career.
Franklin Bailey Electron Microscopy Center University of Idaho Moscow, ID 83844-2204
UNIVERSITY OF CALIFORNIA, SAN DIEGO Scripps Institution of Oceanography, June 1999
Scripps Institution of Oceanography=92s Analytical Facility announces the= ir AFM/STM services lab. Unlike SEM, AFM has the unique capability to provid= e high resolution imaging of biological samples in solution to maintain the= ir structural integrity. We can also vary the solution (pH, buffers, etc.) t= o examine the behavior of samples in vitro.
The total image size can be as high as 30um x 30um(lateral) and 7um(heigh= t). High resolution images can produce feature sizes as low as ~5nm(laterally= ) and sub-angstrom height resolution. Our capabilities for biological sampl= es in solution and samples under environmental control (electrochemistry, humidity, and temperature) make us a unique facility compared to other contract AFM labs.
For more information, please contact Kevin Walda, PhD, Manager SIO Analytical Facility. Tel: (619) 534-3558. Email: kwalda-at-ucsd.edu.
Review article on SPM in Biology! First draft, posted for comments/feedback only, final version will be published by John Wiley in the book "Scanning Tunneling Microscopy and related techniques" ed. D. Bonnell.
Author: Prof. Stuart Lindsay, ASU
View the Article here. http://green.la.asu.edu/index.html
At 01:49 PM 6/11/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would guess that there is a bad or dirty relay contact in your mag circuit. This of course assumes that the scope uses relays to control lens current.
Scopes usually have separate circuits for display and record CRTs. If the display is OK but the record is bad, look for the control relays that are related to the record CRT. If the display is bad and the record is OK, look at the display relays. If both displays are bad....well, move back to the scan generator as a start.
someone initially wrote ... } } } } I have a JEOL IC845 SEM. When recording images on film, I get evenly spaced } } light and dark areas (similar to fringes) on my micrographs. It does not } } appear on screen when viewing an image in normal viewing mode, but in } } watching the photo scan line it does appear to be present, but difficult to } } see. Does anyone have an idea as to what is going on? } } ...
Maybe we should first eliminate the obvious. Tell us more about the spacing and how many lines you see ... after all, these could be the photo CRT's own scan lines which are only visible on the film(???)
Ingram, Mike wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have a JEOL IC845 SEM. When recording images on film, I get evenly spaced } light and dark areas (similar to fringes) on my micrographs. It does not } appear on screen when viewing an image in normal viewing mode, but in } watching the photo scan line it does appear to be present, but difficult to } see. Does anyone have an idea as to what is going on?
Could be dirty (poor contact) on the thumbwheel scan speed switch for photo mode. Try a different scan speed by changing the photo speed setting or replacing the switches. Other possibilities include charging sample, defective scintillator, dying filament. Try taking a blank raster filament off) and see if the lines are still present. If they are still present the problem is in the recording system.
Dear sir or madam: I need to get an idea of how much tissue is processed in the USA per year for electron microscopy. Could you please point me towards a source, person, group etc. that might be able to help me get such an estimate? Any help at all would be greatly appreciated. Thanking you in advance, Regards, Tom Kuwahara -- ******************************* Thomas J. Kuwahara Senior Immunohistochemist Advanced Pathology Systems 3801 Sacramento St. suite 621 San Francisco, CA 94118 415 750 6800 x23067 tel 415 750 2332 fax tom-at-adpath.com
Re Franklin Bailey E mail Dr Arthur Cohen It with regret that I have heard of the death of Arthur Cohen . Having met him on a visit from England a lot of years ago at an EMSA meeting , his work in my humble judgement was excellent and we still use his reference papers in our work and for users of CPD . We shall continue to do so , and his name lives on .
Respectfully David Robinson Emitech England In message {991D50EBD-at-oak.csrv.uidaho.edu} , J.F.Bailey {JFB-at-novell.uidaho.edu} writes } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I used to operate a JEOL 840A and have seen what sounds like this problem, where you see about 15 equally spaced vertical lines on the tv screen and the photos. The service group recommended I shut down the console, pull the boards and, using a pencil eraser, clean the contacts (fingers) on the three boards in the image bin on the upper console rack. I assumed one board was the culprit, but did the bunch rather than experiment. This was the left most bin on the 840A and has the boards that control the screen, alpha numerics, brightness, and such. Apparently it was an every several years maintaince recommendation. If this sounds like your problem, it might be worth a try.
Dave Audette OSRAM Sylvania Beverly, MA david.audette-at-sylvania.com
} -----Original Message----- } From: Ingram, Mike [SMTP:MIngram-at-rodel.com] } Sent: Friday, June 11, 1999 4:49 PM } To: 'Microscopy-at-MSA.Microscopy.com' } Subject: Problems with Photo Scan on JEOL JSM 845 SEM } Importance: High } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have a JEOL IC845 SEM. When recording images on film, I get evenly } spaced } light and dark areas (similar to fringes) on my micrographs. It does not } appear on screen when viewing an image in normal viewing mode, but in } watching the photo scan line it does appear to be present, but difficult } to } see. Does anyone have an idea as to what is going on? }
I will be needing a set of scan selector switches for a Hitachi S450. These are the buttons we push to select scan speeds {the old gal is stuck on slow scan, I hope this is the only problem!}. Individual buttons would be fine but if there is a complete board out there it would save me some soldering time.
One feature of assessing performance of objectives that I have never really understood is the non-financial cost of using a phase objective for either bright-field, fluorescence or DIC. How much does the phase ring & coating they use to correct for differences in intensity really affect the performance of these objectives when they are used in non-phase types of optics. Thanks for any comments. Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I have been asked to serial section areas of retina 5 mm and more in length, for 3-D reconstruction, and then be able to produce TEM of selected sections. Is there a practical way to do this and hopefully a reference? 10um paraffin sections and re-embedment in epon or 2um epon sections seem the most likely way.
Also, is there a good way to mark the tissue so that the source of a section can be located on a gross photo of the tissue?
I'm using NIH image right now, it's freeware on the net, but I always get garbage scans. Does anyone know of any digitizing software where I can scan a TEM negative and analyze it? I need to take a density count of copper oxide islands (heteropataxial growth) and doing it with a light table and a magnifying glass is not fun. thanks
DEADLINE FOR APPLICATIONS EXTENDED TO FRIDAY JUNE 19, 1999.
Summer School on Computing in Electron Microscopy slotted for August 9-13, 1999 , Berkeley, California
(Berkeley, CA) The seventh annual Summer School on Computer-Interactive HRTEM Image Acquisition, Processing and Simulation will be held at the National Center for Electron Microscopy (NCEM), Lawrence Berkeley National Laboratory, University of California, Berkeley from August 9 through August 13, 1999.
The curriculum will focus on training participants in techniques of computer-assisted acquisition and interpretation of high-resolution electron microscope images, including remote-control microscopy. Participants will learn general principles and apply them to specific cases. Instruction on use of computer assistance to obtain images on NCEM microscopes will be followed by training in the use of specific application programs for image interpretation by image processing and simulation.
Participants who wish to apply newly acquired techniques to their own projects are encouraged to extend their visit at NCEM into the next week. Please note: this type of arrangement requires advance submission of a proposal. Projects may involve prepared specimens for microscopy, images and diffraction patterns for processing, or crystal and defect data for simulations. The fee of $375 will cover all materials, instruction, continental breakfast daily, two lunches and an evening reception. Deadline for applications is June 19, 1999. For more information and downloadable application materials contact:
We are looking for a recent Ph.D. with experience in electron microscopy and/or modern light microscopy to work on a new project examining lineage specific transcription events inside the intestine of the developing Caenorhabditis elegans embryo. This will be a joint project between the
laboratories of Drs. David Bazett-Jones and Jim McGhee in the Department
of Biochemistry and Molecular Biology at the University of Calgary. The
developing C. elegans gut provides a unique opportunity to combine structural and ultrastructural analyses of a fundamental problem in biology. For background details of the experimental system, please visit the web site at "www.ucalgary.ca/~jmcghee". For further details, please contact either:
Dr. David P. Bazett-Jones (bazett-at-ucalgary.ca)
or
Dr. James D. McGhee (jmcghee-at-ucalgary.ca)
Department of Biochemistry and Molecular Biology, University of Calgary, Faculty of Medicine, 3330 Hospital Drive, Calgary, Alberta T2N 4N1 CANADA
} Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } Date: Mon, 14 Jun 1999 17:34:39 -0400 } To: Microscopy-at-sparc5.microscopy.com } Subject: JEOL 200CX TEM free
} } A JEOL 200CX TEM is available to anybody who wants it, at no cost for } purchase, with the taker responsible for all moving costs and any repairs. } This is a high-resolution 200kV transmission electron microscope. The } microscope is located at MCNC in the Research Triangle Park, North } Carolina. It is currently in operation. The microscope has high-resolution } pole pieces, a top-entry stage, and a LaB6 filament. It is presently } working only with untilted samples because the tilt mechanism is broken. } All other major components of the microscope are presently working. Anyone } interested in the microscope should contact me by e-mail or phone. } } Michael Lamvik } {mlamvik-at-mcnc.org} } Phone 919-248-1909 } Fax 919-248-1455 } }
There seems to be a lot if interest in the use of focussed ion beam milling for TEM sample prep. I have put up a short tutorial covering the basic steps on our web site. You can access the tutorial via http://www.cmp-cientifica.com/FIB/temprep.htm
During a recent one week "Intensive SEM" coarse, that took place in Johannesburg South Africa, the students came up with a superb cleaning technique that they wished to report upon. The results are, or should be= , interesting to everyone who has to clean a cathode assembly!
=46rom my side the course contains a number of carefully structured practicals that are designed to present the operating variables to the students in the clearest possible fashion. From time to time one finds oneself developing practicals "on the run" to suit the situation. It was=
such a case that is reported below; the filament failed!
A Rapid Cleaning Technique For EM Cathode Assemblies
} From Errol Kelly - University of Fort Hare Belinda White - University of Natal, Pietermaritzburg Allan Hall - University of Pretoria Akos Szabo - Rand Afrikaans University Neville Baker - Anaspec
Introduction The most time consuming operation during the routine use of a SEM or TEM = is often the cleaning of a cathode assembly. The procedure outlined require= s little operator intervention, is free from possible cathode contamination=
by the cleaning media and takes comparatively very little time.
History There is a vast array of cleaning media used by laboratories to clean the=
cathode assemblies of electron microscopes. With many of these the bigge= st failing is the difficulty in removing completely the cleaning media leadi= ng to excessive contamination within the system. This problem is further complicated by the human hazards associated with some of the solvents bei= ng used. In some countries acetone and ether are not permitted in the laboratory!
Steve Chapman has been using and teaching an ultrasonic cleaning techniqu= e he developed in 1964. The procedure took advantage of tungsten being soluble in an ammonia solution (NH4OH) and combined this media with any metal polish that was also soluble in ammonia. The technique used an ammonia solution that had been diluted from a stock solution down to 10 t= o 15 parts water to 1 part ammonia. A range of metal polishing media had been used dictated by their availability in various countries of the worl= d. This cleaning procedure relied more upon the abrasive effect of the meta= l polishing media rather than the chemical attack from the ammonia. =
Subsequent to the metal polish ultrasonic cleaning period of about 30 minutes, the polishing media was removed by way of two further 5 minute ultrasonic cleaning periods in the dilute ammonia alone, to ensure comple= te removal of the metal polishing media. The components were then washed in=
alcohol and dried with a hot air blower. With severely contaminated cathodes, as would be typical of a SEM used at high emission currents, a degree of manual cleaning was often required in the "burnt on" tungsten areas around the cathode aperture. That could be prior to or after the initial ultrasonic cleaning procedure. =
The New Procedure The cathode assembly was placed, aperture face upwards, in a beaker of stock ammonia solution diluted 3 parts ammonia to one part water. The stock solution was thought to be about 40% ammonia. After 15 minutes in the ultrasonic cleaner the beaker was placed under running water and thoroughly flushed through. Care was taken to ensure that none of the clamping or alignment screws had fallen out of the cathode assembly and could be flushed away! The cathode was then washed with alcohol before being dried with a hair drier. A new filament was fitted and centered. =
The assembly was checked for cleanliness by observing with a 20X lens pri= or to re installation in the microscope. Total time for this procedure less=
than 25 minutes.
Safety Great care was taken not to allow the ammonia solution to make contact wi= th the skin or eyes of the operator. When flushing the solution through wit= h water its flow was set so as not to splash the solution over the operator=
prior to placing the beaker under the flow.
Observations The procedure was used on a severely contaminated SEM cathode and a catho= de assembly from an electron probe. Contamination rate had been noted as an=
earlier part of the course so we are able to state that observations in t= he SEM before and after cleaning indicated little or no increase in contamination levels.
--------- Back to me-
We were all amazed at the results, a totally wet cleaning method, a perfectly clean cathode assembly with apparently no instrument problems? = I do not know how ammonia reacts with tantalum (the Philips cathode apertur= e) but I would guess this technique would be applicable to any SEM, TEM, or probe?
What do your members think?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Steve, Errol, Belinda, Allen, Akos, Neville, Thanks for the cleaning procedure. It sound great. I'm looking forward to trying it. One item I would like to mention in favor of some polishing. Over time small irregularities develop on the surfaces through handling or arcing, etc. We even polish new apertures to confirm roundness and smootheness. Possibly some water based abrasive could be used in conjunction with your ammonia technique. Thanks, Russ, Xerox
-----Original Message----- } From: Steve Chapman [mailto:PROTRAIN-at-CompuServe.COM] Sent: Tuesday, June 15, 1999 3:35 AM To: American; MSSAfrica
Hi to Electron Microscopists,
During a recent one week "Intensive SEM" coarse, that took place in Johannesburg South Africa, the students came up with a superb cleaning technique that they wished to report upon. The results are, or should be, interesting to everyone who has to clean a cathode assembly!
} From my side the course contains a number of carefully structured practicals that are designed to present the operating variables to the students in the clearest possible fashion. From time to time one finds oneself developing practicals "on the run" to suit the situation. It was such a case that is reported below; the filament failed!
A Rapid Cleaning Technique For EM Cathode Assemblies
} From Errol Kelly - University of Fort Hare Belinda White - University of Natal, Pietermaritzburg Allan Hall - University of Pretoria Akos Szabo - Rand Afrikaans University Neville Baker - Anaspec
Introduction The most time consuming operation during the routine use of a SEM or TEM is often the cleaning of a cathode assembly. The procedure outlined requires little operator intervention, is free from possible cathode contamination by the cleaning media and takes comparatively very little time.
History There is a vast array of cleaning media used by laboratories to clean the cathode assemblies of electron microscopes. With many of these the biggest failing is the difficulty in removing completely the cleaning media leading to excessive contamination within the system. This problem is further complicated by the human hazards associated with some of the solvents being used. In some countries acetone and ether are not permitted in the laboratory!
Steve Chapman has been using and teaching an ultrasonic cleaning technique he developed in 1964. The procedure took advantage of tungsten being soluble in an ammonia solution (NH4OH) and combined this media with any metal polish that was also soluble in ammonia. The technique used an ammonia solution that had been diluted from a stock solution down to 10 to 15 parts water to 1 part ammonia. A range of metal polishing media had been used dictated by their availability in various countries of the world. This cleaning procedure relied more upon the abrasive effect of the metal polishing media rather than the chemical attack from the ammonia. Subsequent to the metal polish ultrasonic cleaning period of about 30 minutes, the polishing media was removed by way of two further 5 minute ultrasonic cleaning periods in the dilute ammonia alone, to ensure complete removal of the metal polishing media. The components were then washed in alcohol and dried with a hot air blower. With severely contaminated cathodes, as would be typical of a SEM used at high emission currents, a degree of manual cleaning was often required in the "burnt on" tungsten areas around the cathode aperture. That could be prior to or after the initial ultrasonic cleaning procedure.
The New Procedure The cathode assembly was placed, aperture face upwards, in a beaker of stock ammonia solution diluted 3 parts ammonia to one part water. The stock solution was thought to be about 40% ammonia. After 15 minutes in the ultrasonic cleaner the beaker was placed under running water and thoroughly flushed through. Care was taken to ensure that none of the clamping or alignment screws had fallen out of the cathode assembly and could be flushed away! The cathode was then washed with alcohol before being dried with a hair drier. A new filament was fitted and centered. The assembly was checked for cleanliness by observing with a 20X lens prior to re installation in the microscope. Total time for this procedure less than 25 minutes.
Safety Great care was taken not to allow the ammonia solution to make contact with the skin or eyes of the operator. When flushing the solution through with water its flow was set so as not to splash the solution over the operator prior to placing the beaker under the flow.
Observations The procedure was used on a severely contaminated SEM cathode and a cathode assembly from an electron probe. Contamination rate had been noted as an earlier part of the course so we are able to state that observations in the SEM before and after cleaning indicated little or no increase in contamination levels.
--------- Back to me-
We were all amazed at the results, a totally wet cleaning method, a perfectly clean cathode assembly with apparently no instrument problems? I do not know how ammonia reacts with tantalum (the Philips cathode aperture) but I would guess this technique would be applicable to any SEM, TEM, or probe?
What do your members think?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
For stainless steel (304) cathodes, I have had customers used "ammonium hydeoxide hydrogen peroxide" 30%. I am not a chemist but leaving the wehnelt caps overnight leaves a bright, clean finish. It is my understanding that the solution attacks most everything except the stainless. A further advantage is that there is no abrasive used, so no erosion of the wehnelt assembly.
Earl Weltmer
Steve Chapman wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi to Electron Microscopists, } } During a recent one week "Intensive SEM" coarse, that took place in } Johannesburg South Africa, the students came up with a superb cleaning } technique that they wished to report upon. The results are, or should be, } interesting to everyone who has to clean a cathode assembly! } } } From my side the course contains a number of carefully structured } practicals that are designed to present the operating variables to the } students in the clearest possible fashion. From time to time one finds } oneself developing practicals "on the run" to suit the situation. It was } such a case that is reported below; the filament failed! } } A Rapid Cleaning Technique For EM Cathode Assemblies } } } From } Errol Kelly - University of Fort Hare } Belinda White - University of Natal, Pietermaritzburg } Allan Hall - University of Pretoria } Akos Szabo - Rand Afrikaans University } Neville Baker - Anaspec } } Introduction } The most time consuming operation during the routine use of a SEM or TEM is } often the cleaning of a cathode assembly. The procedure outlined requires } little operator intervention, is free from possible cathode contamination } by the cleaning media and takes comparatively very little time. } } History } There is a vast array of cleaning media used by laboratories to clean the } cathode assemblies of electron microscopes. With many of these the biggest } failing is the difficulty in removing completely the cleaning media leading } to excessive contamination within the system. This problem is further } complicated by the human hazards associated with some of the solvents being } used. In some countries acetone and ether are not permitted in the } laboratory! } } Steve Chapman has been using and teaching an ultrasonic cleaning technique } he developed in 1964. The procedure took advantage of tungsten being } soluble in an ammonia solution (NH4OH) and combined this media with any } metal polish that was also soluble in ammonia. The technique used an } ammonia solution that had been diluted from a stock solution down to 10 to } 15 parts water to 1 part ammonia. A range of metal polishing media had } been used dictated by their availability in various countries of the world. } This cleaning procedure relied more upon the abrasive effect of the metal } polishing media rather than the chemical attack from the ammonia. } Subsequent to the metal polish ultrasonic cleaning period of about 30 } minutes, the polishing media was removed by way of two further 5 minute } ultrasonic cleaning periods in the dilute ammonia alone, to ensure complete } removal of the metal polishing media. The components were then washed in } alcohol and dried with a hot air blower. With severely contaminated } cathodes, as would be typical of a SEM used at high emission currents, a } degree of manual cleaning was often required in the "burnt on" tungsten } areas around the cathode aperture. That could be prior to or after the } initial ultrasonic cleaning procedure. } } The New Procedure } The cathode assembly was placed, aperture face upwards, in a beaker of } stock ammonia solution diluted 3 parts ammonia to one part water. The } stock solution was thought to be about 40% ammonia. After 15 minutes in } the ultrasonic cleaner the beaker was placed under running water and } thoroughly flushed through. Care was taken to ensure that none of the } clamping or alignment screws had fallen out of the cathode assembly and } could be flushed away! The cathode was then washed with alcohol before } being dried with a hair drier. A new filament was fitted and centered. } The assembly was checked for cleanliness by observing with a 20X lens prior } to re installation in the microscope. Total time for this procedure less } than 25 minutes. } } Safety } Great care was taken not to allow the ammonia solution to make contact with } the skin or eyes of the operator. When flushing the solution through with } water its flow was set so as not to splash the solution over the operator } prior to placing the beaker under the flow. } } Observations } The procedure was used on a severely contaminated SEM cathode and a cathode } assembly from an electron probe. Contamination rate had been noted as an } earlier part of the course so we are able to state that observations in the } SEM before and after cleaning indicated little or no increase in } contamination levels. } } --------- } Back to me- } } We were all amazed at the results, a totally wet cleaning method, a } perfectly clean cathode assembly with apparently no instrument problems? I } do not know how ammonia reacts with tantalum (the Philips cathode aperture) } but I would guess this technique would be applicable to any SEM, TEM, or } probe? } } What do your members think? } } Steve Chapman } } Senior Consultant E.M. } Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. } Tel & Fax 44 (0)1844 353161 } Web Site - http://ourworld.compuserve.com/homepages/protrain } For Consultancy and Courses in Electron Microscopy World Wide
Our EM facility will be moving to a new building currently under construction. We have just learned that a major feed line consisting of six 3 inch conduits carrying three 400 amp parallel lines (1200 A total) has been installed 11 feet from where our highest resolution TEM is going. We have been asked to supply a minimum acceptable distance that these lines could be from the microscopes, since it will be quite expensive to reroute them (they are in concrete). My specific questions are:
What is the appropriate formula to compute the falloff of the field with distance?
Does this theoretical formula adequately predict the field in a real situation?
How does the orientation of the lines affect the calculation?
Thanks for your help. Any references would also be appreciated.
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-4861936
Sometime in the last two years I remember a thread on preparing CD's for examination in the SEM. Can anyone remember what solvent was used to remove the polymer coating from CD's so imaging of the data is possible. Please respond to me directly at the address below. Thanks.
Owen
============================= Owen P. Mills Electron Optics Facility Engineer Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
As my first post to this wonderful newsgroup, I would like your professional opinions as to which term applies to which techniques. Spectroscopy and Spectrometry, though these terms are used interchangeably in many texts and journals, they must have precise definitions. While Spectroscopy is generally used for techniques utilizing electromagnetic spectra components, what about Spectrometry? As I am currently writing a manuscript on Spectroscopies, Spectrometries, and Microscopies, I wanted to give the reader a good definition of each. Any information you can give me will be much appreciated. I have enjoyed this newsgroup, and gained several good tidbits of info from it. Thanks, Robert Dr. Robert K. Pope CORE/NRL Postdoctoral Fellow Naval Research Laboratory Code 7303, Building 1105 Stennis Space Center, MS 39529-5004 Tel: 228-688-5105 Fax: 228-688-5379 e-mail "rpope-at-nrlssc.navy.mil"
Hi All, A few weeks ago i posted a note here asking for help in regard to filaments burning too fast on our Zeiss TEM. After asking around we have finally found the solution to this problem. It turned out to be that 2 capacitors needed to be changed on the filament control board. If anyone is interested in more details we'll be happy to provide.
There has been a lot of traffic lately regarding IR microscopy. Does anyone know whether similar techniques have been tried with stereomicroscopy? One of my contacts has a project that could benefit from such a technique.
TIA for any advice or suggestions.
Cheers,
Bob
Bob Chiovetti GTI Microsystems rchiovetti-at-aol.com Tucson, Arizona USA
I have a Cambridge S360 SEM that I would like to modernize. I currently take pictures with Polaroid film and I want to go digital. I have a new Dell Workstation adjacent to the SEM. I need advise on the best way to interface the new Dell (with all of its peculiarities) to the old SEM. All I want is a passive system that grabs what I see on my imaging screen. LEO (who now owns Leica, formerly Cambridge) does not have the hardware/software for this interface. I need your expert advise and soon.
} We were all amazed at the results, a totally wet cleaning method, a } perfectly clean cathode assembly with apparently no instrument problems? I } do not know how ammonia reacts with tantalum (the Philips cathode aperture) } but I would guess this technique would be applicable to any SEM, TEM, or } probe? } } What do your members think?
Dear Steve, The Handbook of Chemistry and Physics says that alkalis attack tantalum only slowly, below 150 deg C. Even ~10 M NH4OH should be safe.
Ok, I am sitting in my lab at the terminal, scratching my head on the difference between spectrometry and spectroscopy. I have a couple thoughts.
First, I think spectrometry is the use and practice of an instrument to measure waves, and I think spectroscopy is the study of material systems and their response to harmonic vibrations. Spectrometry emphasizes the instrumentation, its operation, calibration, and the radiation properties=
under measurement. Spectroscopy emphasizes the study of the sample, such=
as the exhaust gases in an automobile emission analyzer or the colored dy= e reagent in an over-the-counter blood glucose monitor.
The topic is not limited to electromagnetic waves, as there are studies i= n other areas like acoustics and ultrasonics which use terms like ultrasoni= c spectroscopy (and even ultrasonic microscopy) are used. I imagine studying the fundamental vibrations of a building also use some kind of spectrometer. Waves, spectrum, spectrometer, spectroscopy, spectrum analyzer, etc.
When I see the word spectrometry in the title of a seminar or paper, I anticipate detailed discussion of the instrumentation and method, with careful quantitation and attention to the intensity, wavelength, polarization, and spatial dependence of the propagating field. I also throw into spectrometry the study of larger samples, with little attentio= n to detailed, atomic level characterization. One example that comes to mi= nd is the infrared picture of a hot iron, or IR imaging altogether. Little interest in the atomic structure of the metal, but lots of interest in radiant intensities. =
Another spectrometry example is the cosmic microwave background black bod= y radiation curve measured by the COBE satellite. This emission from 1 to = 20 cm-1 in the far-infrared region is measured with extrordinary attention t= o intensity and polarization over the entire sky. The emphasis is upon the=
measurement. The radiation itself is associated with the formation of atoms and decoupling of matter and radiation in the Big Bang cosmology model. The variations in this radiation are associated with the "lumpiness" in the early universe, and are not related to specific energy=
levels of atomic systems.
Wow. Now I do need more coffee.
As a suggestion, you might contact the National Institute on Standards an= d Testing (?), or NIST in Colorado. I bet they have someone who has a much=
shorter answer. Another resource might be someone at the Society of Photo-Optical Instrumentation Engineers, or SPIE, at www.spie.org.
} Our EM facility will be moving to a new building currently under } construction. We have just learned that a major feed line consisting of } six 3 inch conduits carrying three 400 amp parallel lines (1200 A total) } has been installed 11 feet from where our highest resolution TEM is going. } We have been asked to supply a minimum acceptable distance that these lines } could be from the microscopes, since it will be quite expensive to reroute } them (they are in concrete). My specific questions are: } } What is the appropriate formula to compute the falloff of the field with } distance?
The formula for the magnetic field from a straight wire of infinite length is H = i/2a(pi), where i is the current, and a is the distance to the wire. This is for steady current, but will also apply for AC. The field is normal to the plane containing the wire and the point where the field is measured. B = mu H is another formula which is significant, since this gives the factor for shielding by substances of high magnetic permeability (such as mu metal), and should be con- sidered if there is any ferromagnetic material between the wires and the scope; e.g., if the wires are in a steel conduit. In the latter case, you have to calculate the field at the outside of the con- duit, then calculate an equivalent current. Since most substances have a magnetic permeability about the same as that of space, you need to worry about this only if there is iron in between the wires and the scope.
} Does this theoretical formula adequately predict the field in a real situation?
Probably. However, you could make measurements at a few distances from the wires to confirm the actual fields.
} } } How does the orientation of the lines affect the calculation? }
For straight wires, the orientation only affects the direction of the field, but for curved wires, you have to calculate the field generated by each (infinitesimal) segment of wire and integrate. Since you possably don't know either the exact path of the wires or the material surrounding them, you'll have to try several models and see which ones match best. If the phase of the AC on each of the wires is the same, you can probably treat the three wires as the equivalent of a single wire (unless they are spaced far enough apart that the distances from each to the scope are significantly different), but if the phases are not the same, you have to calculate an equivalent current. As Dave Barnard pointed out to me, the lines may be designed with three-phase cancellation to reduce the resulting fields.
} Thanks for your help. Any references would also be appreciated. }
I used my old physics text by Frank, Introduction to Electricity and Optics. Yours, Bill Tivol
} } As my first post to this wonderful newsgroup, I would like your } professional opinions as to which term applies to which techniques. } Spectroscopy and Spectrometry, though these terms are used interchangeably } in many texts and journals, they must have precise definitions. While } Spectroscopy is generally used for techniques utilizing electromagnetic } spectra components, what about Spectrometry? As I am currently writing a } manuscript on Spectroscopies, Spectrometries, and Microscopies, I wanted to } give the reader a good definition of each. Any information you can give me } will be much appreciated. I have enjoyed this newsgroup, and gained } several good tidbits of info from it. }
My dictionary defines a spectroscope as a device to separate spectra into various wavelengths for measuring or recording and a spectrometer as a spectro- scope with scales to determine the positions of the peaks. I am not sure how this compares with technical usage, but the terms are nearly interchangable. Yours, Bill Tivol
Bill Tivol's equation was essentially correct, BUT you have to be careful of the units. In SI units (International System of Units), the Biot-Savart Law which Bill quoted becomes B = (2x10^-7)I/D, where B is in tesla, I in amperes, and D in meters. ===================================== } Our EM facility will be moving to a new building currently under } construction. We have just learned that a major feed line consisting of } six 3 inch conduits carrying three 400 amp parallel lines (1200 A total) } has been installed 11 feet from where our highest resolution TEM is going. } We have been asked to supply a minimum acceptable distance that these lines } could be from the microscopes, since it will be quite expensive to reroute } them (they are in concrete). My specific questions are: } } What is the appropriate formula to compute the falloff of the field with } distance? } } Does this theoretical formula adequately predict the field in a real } situation? } } How does the orientation of the lines affect the calculation? } } Thanks for your help. Any references would also be appreciated. } } Dr. Marie E. Cantino } Dept. of Physiology and Neurobiology } University of Connecticut } Storrs, CT 06269-2131 } Phone: 860-486-3588 } Fax: 860-4861936
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798 cook-at-aaem.amc.anl.gov
Spectroscopy: electromagnetic radiation, i.e. photons Spectrometry: Mass to charge ration of particles.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Dr. Robert K. Pope To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hi all.
As my first post to this wonderful newsgroup, I would like your professional opinions as to which term applies to which techniques. Spectroscopy and Spectrometry, though these terms are used interchangeably in many texts and journals, they must have precise definitions. While Spectroscopy is generally used for techniques utilizing electromagnetic spectra components, what about Spectrometry? As I am currently writing a manuscript on Spectroscopies, Spectrometries, and Microscopies, I wanted to give the reader a good definition of each. Any information you can give me will be much appreciated. I have enjoyed this newsgroup, and gained several good tidbits of info from it. Thanks, Robert Dr. Robert K. Pope CORE/NRL Postdoctoral Fellow Naval Research Laboratory Code 7303, Building 1105 Stennis Space Center, MS 39529-5004 Tel: 228-688-5105 Fax: 228-688-5379 e-mail "rpope-at-nrlssc.navy.mil"
I think LEO's statement needs to be qualified. They may not offer or support passive systems for the 360, but I think a third-party system ought to work just fine. There should be no modifications needed to attach a passive system. Installation simply involves tapping off the X, Y, and Z signals in the right place. We had a JEOL JSM-U3 hooked up for active control almost 20 years ago. That was much trickier than hooking up a passive system today.
We have the Quartz PCI passive system, but there are many fine systems out there, and someone recently posted about building their own. Good luck in picking one.
At 01:34 PM 6/15/1999 -0400, you wrote: } To all SEM experts: } } I have a Cambridge S360 SEM that I would like to modernize. I currently } take pictures with Polaroid film and I want to go digital. I have a new } Dell Workstation adjacent to the SEM. I need advise on the best way to } interface the new Dell (with all of its peculiarities) to the old SEM. All } I want is a passive system that grabs what I see on my imaging screen. LEO } (who now owns Leica, formerly Cambridge) does not have the hardware/software } for this interface. I need your expert advise and soon. } } Thanking you in advance, } } Sheila R-W
I've been cleaning the Wehnelt caps and cathodes of two SEM's and one TEM for at least 10 years using 30-40% ammonium hydroxide solution seemingly without any problems. After sonicating the parts in "ammonia" for several minutes, we rinse them well in tap water, and then sonicate in absolute ethanol for several more minutes. Lastly, the cap and/or cathode are air-dried. This eliminates all the problems associated with removing metal polish from the filament assembly and only requires the appropriate safety precautions taken with the use of strong ammonium hydroxide solutions.
Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124-4305
} ---------- } From: Steve Chapman[SMTP:PROTRAIN-at-CompuServe.COM] } Sent: Tuesday, June 15, 1999 2:34 AM } To: American; MSSAfrica } Subject: An EM Dream? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi to Electron Microscopists, } } During a recent one week "Intensive SEM" coarse, that took place in } Johannesburg South Africa, the students came up with a superb cleaning } technique that they wished to report upon. The results are, or should be, } interesting to everyone who has to clean a cathode assembly! } } From my side the course contains a number of carefully structured } practicals that are designed to present the operating variables to the } students in the clearest possible fashion. From time to time one finds } oneself developing practicals "on the run" to suit the situation. It was } such a case that is reported below; the filament failed! } } A Rapid Cleaning Technique For EM Cathode Assemblies } } } From } Errol Kelly - University of Fort Hare } Belinda White - University of Natal, Pietermaritzburg } Allan Hall - University of Pretoria } Akos Szabo - Rand Afrikaans University } Neville Baker - Anaspec } } Introduction } The most time consuming operation during the routine use of a SEM or TEM } is } often the cleaning of a cathode assembly. The procedure outlined requires } little operator intervention, is free from possible cathode contamination } by the cleaning media and takes comparatively very little time. } } History } There is a vast array of cleaning media used by laboratories to clean the } cathode assemblies of electron microscopes. With many of these the } biggest } failing is the difficulty in removing completely the cleaning media } leading } to excessive contamination within the system. This problem is further } complicated by the human hazards associated with some of the solvents } being } used. In some countries acetone and ether are not permitted in the } laboratory! } } Steve Chapman has been using and teaching an ultrasonic cleaning technique } he developed in 1964. The procedure took advantage of tungsten being } soluble in an ammonia solution (NH4OH) and combined this media with any } metal polish that was also soluble in ammonia. The technique used an } ammonia solution that had been diluted from a stock solution down to 10 to } 15 parts water to 1 part ammonia. A range of metal polishing media had } been used dictated by their availability in various countries of the } world. } This cleaning procedure relied more upon the abrasive effect of the metal } polishing media rather than the chemical attack from the ammonia. } Subsequent to the metal polish ultrasonic cleaning period of about 30 } minutes, the polishing media was removed by way of two further 5 minute } ultrasonic cleaning periods in the dilute ammonia alone, to ensure } complete } removal of the metal polishing media. The components were then washed in } alcohol and dried with a hot air blower. With severely contaminated } cathodes, as would be typical of a SEM used at high emission currents, a } degree of manual cleaning was often required in the "burnt on" tungsten } areas around the cathode aperture. That could be prior to or after the } initial ultrasonic cleaning procedure. } } The New Procedure } The cathode assembly was placed, aperture face upwards, in a beaker of } stock ammonia solution diluted 3 parts ammonia to one part water. The } stock solution was thought to be about 40% ammonia. After 15 minutes in } the ultrasonic cleaner the beaker was placed under running water and } thoroughly flushed through. Care was taken to ensure that none of the } clamping or alignment screws had fallen out of the cathode assembly and } could be flushed away! The cathode was then washed with alcohol before } being dried with a hair drier. A new filament was fitted and centered. } The assembly was checked for cleanliness by observing with a 20X lens } prior } to re installation in the microscope. Total time for this procedure less } than 25 minutes. } } Safety } Great care was taken not to allow the ammonia solution to make contact } with } the skin or eyes of the operator. When flushing the solution through with } water its flow was set so as not to splash the solution over the operator } prior to placing the beaker under the flow. } } Observations } The procedure was used on a severely contaminated SEM cathode and a } cathode } assembly from an electron probe. Contamination rate had been noted as an } earlier part of the course so we are able to state that observations in } the } SEM before and after cleaning indicated little or no increase in } contamination levels. } } } } --------- } Back to me- } } We were all amazed at the results, a totally wet cleaning method, a } perfectly clean cathode assembly with apparently no instrument problems? } I } do not know how ammonia reacts with tantalum (the Philips cathode } aperture) } but I would guess this technique would be applicable to any SEM, TEM, or } probe? } } What do your members think? } } Steve Chapman } } Senior Consultant E.M. } Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. } Tel & Fax 44 (0)1844 353161 } Web Site - http://ourworld.compuserve.com/homepages/protrain } For Consultancy and Courses in Electron Microscopy World Wide }
} I hope you will all please excuse me, but I'm looking for a home for our } JEOL 100S TEM. It is available to anyone for the cost of removal and } shipping. It was on a JEOL service contrct up until this year and is in } excellent condition. It would be great for teaching and/or simple } biological work. It is located in the Dept. of Ophthalmology at the } University of Washington in Seattle, WA. } } Please contact me via email or telephone: } } Daniel Possin {danpossn-at-u.washington.edu} } 206-685-7241 } 206-543-3883 } } Thank you for your patient attention, } } Dan % Daniel Possin Voice: 206/ 221-3845 % % Dept. of Ophthalmology FAX: 206/ 543-4414 % % Box 35 6485 - C209 HSB Email: danpossn-at-u.washington.edu % % University of Washington % % Seattle WA 98195 USA % % % % "Orchids may bloom where the sun never shines and rain never falls". %
} ... } } Spectroscopy and Spectrometry, though these terms are used } interchangeably in many texts and journals, they must have } precise definitions. ...
I have my own connotations, and I may look some of these up and reply later with respect to being precise ... but I believe some of the terminology will break down in terms of "qualitative" and "quantitative" ... and for the sake of including all terminology, you should need to consider "spectrophotometry" as well ... you just know some student will ask :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
} } Date: Tue, 15 Jun 1999 11:03:22 -0500 } To: {microscopy-at-Sparc5.Microscopy.Com} } From: "Dr. Robert K. Pope" {rpope-at-nrlssc.navy.mil} } Subject: Spectrometry vs Spectroscopy } } } Hi all. } } As my first post to this wonderful newsgroup, I would like your } professional opinions as to which term applies to which techniques. } Spectroscopy and Spectrometry, though these terms are used interchangeabl } in many texts and journals, they must have precise definitions. While } Spectroscopy is generally used for techniques utilizing electromagnetic } spectra components, what about Spectrometry? As I am currently writing a } manuscript on Spectroscopies, Spectrometries, and Microscopies, I wanted t } give the reader a good definition of each. Any information you can give m } will be much appreciated. I have enjoyed this newsgroup, and gained } several good tidbits of info from it. } Thanks, } Robert } Dr. Robert K. Pope } CORE/NRL Postdoctoral Fellow } Naval Research Laboratory } Code 7303, Building 1105 } Stennis Space Center, MS 39529-5004 } Tel: 228-688-5105 } Fax: 228-688-5379 } e-mail "rpope-at-nrlssc.navy.mil" } Robert, My primary experience is in mass spectroscopy though I have worked quite a bit with microscopy and visible light spectrometers.
In mass spectroscopy, one originally (and perhaps still in a few labs) had the choice of instruments where the detected signal was detected electronically (i.e., metered) at a single mass position in a mass spectromoeter; or the ions were spread out in space and a wide mass range was collected on a photo plate (and now on some form of CCD) and the instrument was called a spectrograph. The same situation, I believe, prevailed in emission spectroscopy. So spectrometry and spectrography are sub titles under the overall title of spectroscopy, depending on whether the beam is collected at a single point/value or over a wide range.
Spectrometers have exit slits and spectrographs do not.
I hope you will summarize the results of this inquiry for all our benefit.
Thanks.
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology "A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
I wouldn't recommend paraffin. I've done re-embedding of retina from paraffin to Epon and the results were consistently horrible. Start with Epon. I've cut serial 1um Epon sections of retina by the hundred. Fortunately I didn't have to collect them all. It's easy, but BORING. I haven't taken them to EM; that would be the difficult part.
I presume as a marker you want something that will enable you to translate from the cross section of resin embedded tissue to the original unembedded retina photographed as a flat mount? If so, your best bet may be (depending on the species) to use the retinal blood vessels as markers, preferably stained. They're easily seen in both cross section and flat mount. Do you have to be exact in positioning? You can measure the diameter of the retina and keep track of how many sections have been cut - thus how much of the retina has been cut away. Obviously the optic nerve provides a nice centre. Just make sure one edge is notched for initial orientation. If you can't flat mount the retina, but have to keep it it whole, like a cut ball, it's more difficult, but the latter method should still work.
Diana
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
Sheila R-W wrote: ============================================== I have a Cambridge S360 SEM that I would like to modernize. I currently take pictures with Polaroid film and I want to go digital. I have a new Dell Workstation adjacent to the SEM. I need advise on the best way to interface the new Dell (with all of its peculiarities) to the old SEM. All I want is a passive system that grabs what I see on my imaging screen. LEO (who now owns Leica, formerly Cambridge) does not have the hardware/software for this interface. I need your expert advise and soon. ================================================== You might want to take a look at the product called Spectrum Mono™ which is described on the SPI website at URL http://www.2spi.com/catalog/software/spectrum.html
This is a "passive" system and has been interfaced to many different aging analog SEMs, including old Cambridge instruments.
Disclaimer: SPI distributes this product worldwide so we are not exactly a disinterested third party.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi3spi-at-2spi.com
Look for us! ############################ WWW: www.spi.cc ############################ ==================================================
My experience in installing electron microscopes goes back over 33 years and covers many countries where the problem you outline was and is very common. I have not tried to calculate the size of the problem, always using a meter to judge the best place for a microscope. Even in these cases when an empty laboratory was fine, time and time again a laboratory=
in action proved to be a far bigger problem.
If I was in your position I would not consider the location you describe for ANY electron optical instrument. Screening etc etc does not often do=
the job! You should not compromise as you may well find you have a wonderful new laboratory which will be providing a very poor service spoi= lt by magnetic fields, they do not go away just get worse!
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
For SEM you can strip the data layer using adhesive tape. Score the supercoat film first round the area you want to examine. Choose an adhesive tape with a powerrful bond strength, and strip it from the disc with sudden force at right angles to the surface.
Writeable and rewritable discs strip more easily than hot-pressed CDs
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Sometime in the last two years I remember a thread on preparing CD's for } examination in the SEM. Can anyone remember what solvent was used to } remove the polymer coating from CD's so imaging of the data is possible. } Please respond to me directly at the address below. Thanks. } } Owen } } } } ============================= } Owen P. Mills } Electron Optics Facility Engineer } Michigan Technological University } Metallurgical & Materials Engineering } Rm 512 MME Building } Houghton, MI 49931 } 906-487-2002 } 906-487-2934 FAX } opmills-at-mtu.edu } }
===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
Wiliams and Carter's book on TEM (1996 edition) states (p 276) "The RDF = can be obtained directly from DPs ... software ... listed in section = 1.5"
How is this actually done? I need to understand the principle. I would = appreciate further information on this as explanation / references / = software sources.
Best wishes, --- R Divakar PMS, IGCAR, Kalpakkam 603102, India ----
When I joined STL Harlow in 1976, my boss showed me how to clean Wehnelts with conc. ammonia solution. He used a 15 min soak in 35% (or 880, as we called it, SG of .880, I think), followed by rinse in DI water and then IPA. Stubborn stains could be removed using "Wenol" type polish on a cotton tip bud or using silver wadding polish like "Duraglit". Care was taken to remove any fibres then ultrasonic rinse in IPA. I still use this method and have passed it on to my engineers.
Barry
} -----Original Message----- } From: Gillmeister, Russ [SMTP:RGillmeister-at-sdms.usa.xerox.com] } Sent: Tuesday, June 15, 1999 1:57 PM } To: 'Steve Chapman' } Cc: 'MSA' } Subject: RE: An EM Dream? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Steve, Errol, Belinda, Allen, Akos, Neville, Thanks for the cleaning } procedure. It sound great. I'm looking forward to trying it. One item I } would like to mention in favor of some polishing. Over time small } irregularities develop on the surfaces through handling or arcing, etc. We } even polish new apertures to confirm roundness and smootheness. Possibly } some water based abrasive could be used in conjunction with your ammonia } technique. Thanks, Russ, Xerox } } } -----Original Message----- } } From: Steve Chapman [mailto:PROTRAIN-at-CompuServe.COM] } Sent: Tuesday, June 15, 1999 3:35 AM } To: American; MSSAfrica } Subject: An EM Dream? } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi to Electron Microscopists, } } During a recent one week "Intensive SEM" coarse, that took place in } Johannesburg South Africa, the students came up with a superb cleaning } technique that they wished to report upon. The results are, or should be, } interesting to everyone who has to clean a cathode assembly! } } } From my side the course contains a number of carefully structured } practicals that are designed to present the operating variables to the } students in the clearest possible fashion. From time to time one finds } oneself developing practicals "on the run" to suit the situation. It was } such a case that is reported below; the filament failed! } } A Rapid Cleaning Technique For EM Cathode Assemblies } } } From } Errol Kelly - University of Fort Hare } Belinda White - University of Natal, Pietermaritzburg } Allan Hall - University of Pretoria } Akos Szabo - Rand Afrikaans University } Neville Baker - Anaspec } } Introduction } The most time consuming operation during the routine use of a SEM or TEM } is } often the cleaning of a cathode assembly. The procedure outlined requires } little operator intervention, is free from possible cathode contamination } by the cleaning media and takes comparatively very little time. } } History } There is a vast array of cleaning media used by laboratories to clean the } cathode assemblies of electron microscopes. With many of these the } biggest } failing is the difficulty in removing completely the cleaning media } leading } to excessive contamination within the system. This problem is further } complicated by the human hazards associated with some of the solvents } being } used. In some countries acetone and ether are not permitted in the } laboratory! } } Steve Chapman has been using and teaching an ultrasonic cleaning technique } he developed in 1964. The procedure took advantage of tungsten being } soluble in an ammonia solution (NH4OH) and combined this media with any } metal polish that was also soluble in ammonia. The technique used an } ammonia solution that had been diluted from a stock solution down to 10 to } 15 parts water to 1 part ammonia. A range of metal polishing media had } been used dictated by their availability in various countries of the } world. } This cleaning procedure relied more upon the abrasive effect of the metal } polishing media rather than the chemical attack from the ammonia. } Subsequent to the metal polish ultrasonic cleaning period of about 30 } minutes, the polishing media was removed by way of two further 5 minute } ultrasonic cleaning periods in the dilute ammonia alone, to ensure } complete } removal of the metal polishing media. The components were then washed in } alcohol and dried with a hot air blower. With severely contaminated } cathodes, as would be typical of a SEM used at high emission currents, a } degree of manual cleaning was often required in the "burnt on" tungsten } areas around the cathode aperture. That could be prior to or after the } initial ultrasonic cleaning procedure. } } The New Procedure } The cathode assembly was placed, aperture face upwards, in a beaker of } stock ammonia solution diluted 3 parts ammonia to one part water. The } stock solution was thought to be about 40% ammonia. After 15 minutes in } the ultrasonic cleaner the beaker was placed under running water and } thoroughly flushed through. Care was taken to ensure that none of the } clamping or alignment screws had fallen out of the cathode assembly and } could be flushed away! The cathode was then washed with alcohol before } being dried with a hair drier. A new filament was fitted and centered. } The assembly was checked for cleanliness by observing with a 20X lens } prior } to re installation in the microscope. Total time for this procedure less } than 25 minutes. } } Safety } Great care was taken not to allow the ammonia solution to make contact } with } the skin or eyes of the operator. When flushing the solution through with } water its flow was set so as not to splash the solution over the operator } prior to placing the beaker under the flow. } } Observations } The procedure was used on a severely contaminated SEM cathode and a } cathode } assembly from an electron probe. Contamination rate had been noted as an } earlier part of the course so we are able to state that observations in } the } SEM before and after cleaning indicated little or no increase in } contamination levels. } } } } --------- } Back to me- } } We were all amazed at the results, a totally wet cleaning method, a } perfectly clean cathode assembly with apparently no instrument problems? } I } do not know how ammonia reacts with tantalum (the Philips cathode } aperture) } but I would guess this technique would be applicable to any SEM, TEM, or } probe? } } What do your members think? } } Steve Chapman } } Senior Consultant E.M. } Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. } Tel & Fax 44 (0)1844 353161 } Web Site - http://ourworld.compuserve.com/homepages/protrain } For Consultancy and Courses in Electron Microscopy World Wide }
} -----Original Message----- } From: Rosenfield, Sheila A. [SMTP:SARosenf-at-rmc.com] } Sent: Tuesday, June 15, 1999 6:35 PM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: SEM Image Capture } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To all SEM experts: } } I have a Cambridge S360 SEM that I would like to modernize. I currently } take pictures with Polaroid film and I want to go digital. I have a new } Dell Workstation adjacent to the SEM. I need advise on the best way to } interface the new Dell (with all of its peculiarities) to the old SEM. } All } I want is a passive system that grabs what I see on my imaging screen. LEO } (who now owns Leica, formerly Cambridge) does not have the } hardware/software } for this interface. I need your expert advise and soon. } } Thanking you in advance, } } Sheila R-W }
Two quick questions for my British Colleagues on digital imaging terminology. I am looking to see if there are accepted differences between "British English" and "American English" which I am not aware of (I am reviewing a paper for a British Journal and wish to be accurate). Specifically which of the following terms is correct in British English:
(1) "Grey images" or "Grey scale images"?
(2) "256 Greyness levels" or "256 Grey levels"?
Thanks you and Tally ho, eh?
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
We used an oxygen plasma technique to remove the top surface lacquer with the label printed on. The exposed Al layer was removed in conc HCl with a few drops of hydrogen peroxide added, leaving the plastic surface with the data pits imprinted. The sample was gold coated before SEM examination.
Hope it's of some use
Barry } -----Original Message----- } From: Owen P. Mills [SMTP:opmills-at-mtu.edu] } Sent: Tuesday, June 15, 1999 4:40 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: imaging CD sectors } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Sometime in the last two years I remember a thread on preparing CD's for } examination in the SEM. Can anyone remember what solvent was used to } remove the polymer coating from CD's so imaging of the data is possible. } Please respond to me directly at the address below. Thanks. } } Owen } } } } ============================= } Owen P. Mills } Electron Optics Facility Engineer } Michigan Technological University } Metallurgical & Materials Engineering } Rm 512 MME Building } Houghton, MI 49931 } 906-487-2002 } 906-487-2934 FAX } opmills-at-mtu.edu } }
Hi Chuck, You're right! Should have said I'd only use the ammonium hydroxide solution on solid stainless steel parts. I wouldn't use it on the filament base holders for example (not that I clean them anyway) or anything with brass in it.
Bruce
Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124-4305
} ---------- } From: Garber, Charles A.[SMTP:cgarber-at-2spi.com] } Sent: Wednesday, June 16, 1999 12:14 AM } To: Ingber, Bruce F. } Subject: 2 good to B true: An EM Dream? } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Bruce, Would you mind if I asked you a question about this? Some years } ago, } more than I will admit, when I was myself a graduate student at Case in } Cleveland, they had a JEOL JEM 6 or 7 and a Hitachi HU11-A. I used both } of } them, and I can not remember which it was, but one of them had a wehnelt } cap } , for example, that was plated (it looked like chrome) on some other } metal, } perhaps brass. } } The caps had some amount of use, e. g. they were not brand new and the } technician at the time in charge of running the laboratory tried some } ammonia technique. And what happened was this: There was a reaction with } the brass through the porosity in the chromium layer and that cap was } pretty } much destroyed. } } So I just wondered if the materials of construction as solid stainless } steel } , in which case there would be not problem or could any of them be plated, } with a substrate that had a copper containing alloy. If the latter, then I } would think that, based on my own experience in Cleveland, this technique } might have more lurking dangers that is appreciated. } } Chuck } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi3spi-at-2spi.com } ================================================== }
Purely speaking, spectroscopy is the general study or observation of spectral responses while spectrometry is the actual measurement of absorbance, transmittance, or reflectance at specific locations in the spectrum. As a past microspectrometry specialist at Zeiss and Cambridge (now Leica), it has been my experience that most microscopists are interested in the latter. We routinely do microfluorimetry (point measurements of fluorescence intensity) as well as microspectrometry in the UV-visible, Raman, and FT-IR ranges. We also have spectrometers attached to electron microscopes. For more information that area, research the terms "energy dispersive spectrometry (EDS)", "wavelength dispersive spectrometry (WDS)", and "X-ray fluorescence". In these last disciplines, spectrometry and spectroscopy may be used more interchangeabley because they produce chemical maps as well as doing specific measurement. Princeton Gamma-Tech, EDAX, Oxford Instruments, and Noran all have a good supply of application notes which can orient you to these areas.
Thanks for trying to bring some order to this area. I'd be delighted to see a manuscript when it is near completion.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 09:05 PM 6/15/99 -0400, donald j marshall wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Probably the best person to answer this question is John Reffner: John Reffner {jareffner-at-compuserve.com} He is currently working with Sensor Technologies in Danbury: 203-207-9700. They have a neat new IR microscope.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 12:32 PM 6/15/99 EDT, RCHIOVETTI-at-aol.com"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Having been trained initially by the RMS, I am all too familiar with your dilemma.
The second of terms in each question are the typical terms used in this country.
You might want to check with Dr. Savile Bradbury (retired, Oxford) as to their correct British counterparts. (Please send him my regards)
Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 06:54 AM 6/16/99 -0500, edelmare-at-casmail.muohio.edu"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Wiliams and Carter's book on TEM (1996 edition) states (p 276) "The RDF } can be obtained directly from DPs ... software ... listed in section 1.5" } } How is this actually done? I need to understand the principle. I would } appreciate further information on this as explanation / references / } software sources.
A detailed treatment of the diffraction theory may be found in Chap. 10 of _X-Ray Diffraction_ by B. E. Warren, Dover Inc, New York 1990.
For a quick and dirty calculation, Eq. 18.10 in Williams and Carter tells you most of what you need to know. That equation could be executed by most any software that will do image Fourier transforms. I would use Digital Micrograph to rotationally average the diffraction pattern then do the FT, but NIH Image can probably do the same thing for much less money.
Of course, people can and do work much harder to get a low-noise, high accuracy RDF from diffraction data. A recent and extremely thorough example of calculating the RDF for amorphous silicon from x-ray diffraction can be found in "High Resolution Radial Distribution Function of Pure Amorphous Silicon" Khalid Laaziri, S. Kycia, et. al. Phys. Rev. Letts. Vol. 82, p. 3460 (1999). On the web (AIP web site, available only with subscription):
An older example is "Structural investigation of hydrogenated amorphous silicon by X-ray diffraction" W. Schulke, Phil. Mag. B Vol. 43, p. 451, (1981).
Obtaining the need diffraction data in a TEM is possible, but may be difficult if you need high accuracy. Quantitative data at high scattering vector (up to 55 1/A according to Laaziri et al) is necessary which means you need a very high dynamic range detector. Imaging plates will produce the best results. Stitching together several CCD images at different exposures might also work, although you have to be carefully about saturated pixels at low k "blooming" - spreading intensity over a large number of adjoining pixels.
Good luck, Paul
Paul Voyles (609) 951-2627 voyles-at-research.nj.nec.com NEC Research Institute, 4 Independence Way, Princeton, NJ 08540
Whoa!!! I wouldn't allow this to happen!! We have had considerable experience with stray fields affecting our FESEMs and causing picture wobble. I always use a search coil (a TV degaussing coil, calibrated against a "Gauss Maus" field strength meter) to determine the intensity and direction of the field. I found fields } 99milligauss at five yards from a 600A cable buried one yard below the ground (I think 5 milligauss is the max. tolerable). You can't always rely on calculations, it's much better to measure the fields yourself. As for shielding, it's almost impossible due to the gaps in the shielding. The best method, if you have no choice, is an active cancellation system. I have seen a case when one phase went down (it was pulling little power, anyway) and the resulting imbalance had a dramatic effect on all the high res monitors and, of course, the SEM. For the cables you describe, I would say at least a distance of eight yards is required, if not ten!!
Good luck!
Barry
} -----Original Message----- } From: Marie E. Cantino [SMTP:cantino-at-uconnvm.uconn.edu] } Sent: Tuesday, June 15, 1999 4:48 PM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: calculating magnetic fields } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Our EM facility will be moving to a new building currently under } construction. We have just learned that a major feed line consisting of } six 3 inch conduits carrying three 400 amp parallel lines (1200 A total) } has been installed 11 feet from where our highest resolution TEM is going. } We have been asked to supply a minimum acceptable distance that these } lines } could be from the microscopes, since it will be quite expensive to reroute } them (they are in concrete). My specific questions are: } } What is the appropriate formula to compute the falloff of the field with } distance? } } Does this theoretical formula adequately predict the field in a real } situation? } } How does the orientation of the lines affect the calculation? } } Thanks for your help. Any references would also be appreciated. } } Dr. Marie E. Cantino } Dept. of Physiology and Neurobiology } University of Connecticut } Storrs, CT 06269-2131 } Phone: 860-486-3588 } Fax: 860-4861936 } } }
Thanks to everyone who has responded to my question. Unfortunately I did not make clear in my first post that direct field measurements, the best way of determining the extent of the problem, are not an option because the building is still under construction; there will be no power in those lines until it is too late to move them (which is going to require jackhammering the concrete floor). So I need to tell the contractor what a "safe" distance is.
} From various comments and other information we have gathered, it seems that predicting the field at a particular distance is not simple because the fields in lines traveling in opposite directions will cancel out, and the drop off with r will be faster than 1/r. At this point we are looking for an analogous current-conduit configuration to try to get an empirical measurement, but if anyone has any other ideas, let me know. Thanks.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-4861936
In all of the enthusiasm for low labor cleaning methods, please do not extend this method to copper alloy parts. Any copper or brass stands a good chance of reacting with ammonia. There are two results: there is a tarnish film that may form rather rapidly; its removal will require just the elbow grease that you were trying to avoid. More important, brass will run the risk of an interesting phenomenon called stress corrosion cracking, which may destroy the component.
I still prefer to use Pol or Wenol and elbow grease.
Disclaimer I: my employer, Structure Probe, Inc., sells Pol (tm) and Wenol (tm) through its SPI Supplies (tm) Division. We do not sell ammonia.
Disclaimer II: I get to call myself "Dr." because of a study of stress corrosion cracking of copper alloys done in the 60s. We still get failure analysis assignments because the phenomenon is not widely known.
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
Dear Owen, I was successful in cutting out a one cm. chunk of a hot-pressed CD and leaving it in chlorothene overnight. Unfortunately, we can't use that anymore. Any good plastic solvent should work, even acetone. You are left with a wisp of very thin aluminum foil with the dints in it visible at about 10,000X. The CD-Rs are a very different technology. At 11:39 AM 6/15/99 -0400, you wrote: } } Sometime in the last two years I remember a thread on preparing CD's for } examination in the SEM. Can anyone remember what solvent was used to } remove the polymer coating from CD's so imaging of the data is possible. } Please respond to me directly at the address below. Thanks. } } Owen Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
At 11:56 PM 6/16/1999 +0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
The following are excerpts or paraphrases from Heinz-Helmut Perkampus, Encyclopedia of Spectroscopy, VCH publishers. Although a translation, the parts I have read are literate and helpful.
Spectrometer, an instrument for making relative measurements in the optical spectral region, especially for analytical applications. The concept has become established outside optical spectroscopy . . . AA, AE, UV-VIS, IR, F, Raman, microwave, ESR, NMR, photoelectron, mass, Moessbauer, photoacoustic, reflectance. . . . [block diagram showing similarities and differences]
Spectrophotometer, a photometer coupled with a spectrometer. . .
Spectroscopy, all the methods in the field of electromagnetic radiation which are important for research and applications. The word is derived from Latin, spectrum = ghost, and Greek, skopos = watcher. In this sense, a spectroscopist is a ghost watcher. . . . [Newton coinage] . . . the region of the electromagnetic spectrum accessible . . . covers a range of 12 to 14 powers of ten in energy units . . . .
Leonard Corwin Research Chemist Fort Dodge Animal Health Princeton, NJ 08543-0400
We use 1 part reagent strength HCL to 4 parts water by volume.
We got this recipe from the listserver some time back.
Cheers, Henk
At 11:56 PM 6/16/99 +0800, Keith Moulding wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
A retired EM person several years ago swore by BonAmi which is a water = soluble abrasive for polishing pots and pans. It was removed readily = with warm water. I used for several years without any adverse effects. = Once I ran out of my supply I stopped using it. I have asked several = service engineers about it and none of them knew of any adverse effects. = Though I never saw any evidence of scratching I was worried about the = particle size of the abrasive.
Hank Adams Integrated Microscopy Core Cell Biology Baylor College of Medicine=20 Houston, Tx
-----Original Message----- } From: Blackwood, Andrew [SMTP:ablackwood-at-2spi.com] Sent: Wednesday, June 16, 1999 12:42 PM To: microscopy-at-sparc5.microscopy.com
-- [ From: Blackwood, Andrew * EMC.Ver #3.1 ] --
16 June 1999
Greetings:
In all of the enthusiasm for low labor cleaning methods, please do not extend this method to copper alloy parts. Any copper or brass stands a = good chance of reacting with ammonia. There are two results: there is a = tarnish film that may form rather rapidly; its removal will require just the = elbow grease that you were trying to avoid. More important, brass will run the risk of an interesting phenomenon called stress corrosion cracking, = which may destroy the component.
I still prefer to use Pol or Wenol and elbow grease.
Disclaimer I: my employer, Structure Probe, Inc., sells Pol (tm) and = Wenol=20 (tm) through its SPI Supplies (tm) Division. We do not sell ammonia.
Disclaimer II: I get to call myself "Dr." because of a study of stress corrosion cracking of copper alloys done in the 60s. We still get = failure analysis assignments because the phenomenon is not widely known.
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
Dear microscopists, I realize that this subject may be out of the realm of microscopy but I thought maybe one of you could help. Our lab generates alot of poster titles for various meetings. The posters are then mounted in the departmental hallways for a few months so they need to survive for some time. I use a double sided adhesive tape made by Letraset to put the title(printed on card stock) on posterboard. I have just found out that the company is no longer making the tape and I can't find a decent substitute. The double sided scotch tape isn't strong enough; the spray photo mount is very difficult to work with in a lab and doesn't hold well either. It leaves a sticky residue on anything nearby. Does anyone know of a very high tack tape similar to the Letraset tape? Thanks in advance,
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
We are looking to clear out electronics that we have had in storage, ie. various read outs (scalers, pulse-height analyzers, etc), power supplies, controllers, etc. These are mostly probe parts. Come and take them, as-is, where-is, otherwise they go to the scrap yard.
We also have an older ARL-EMX/SM microprobe for sale. It was working when taken out of service several years ago. We have spare parts, crystals, sample holders, etc. It also has an EDS detector and electronics, but we are now using the computer analyzer with our SEM. Make us a modest offer for this nostalgic instrument.
Alan Stone ASTON Metallurgical Services 4201 North Ravenswood Ave Chicago, IL 60613
Hi! Instead of double sticky tape, I use Perma/mount 2. They're sheets of double sided sticky stuff you can cut to any size. I think I got them from some photo supplier, but I also saw them in the EMS catalog, as well as another kind of tape substitute . They're a little pricey, but very easy to use, and hold real well. EMS phone is 1-800-523-5874. I'm sure other catalogs have them as well. Nancy
I'm not familiar with the Letraset tape but would double-sticky carpet tape work? This comes in quite wide lengths and is very tenaceous (perhaps too much so). You might check at local hobby shops as well for these sorts of tape.
Finally, check with a framing shop (one where you would have artwork mounted and framed). I just had some chinese scrolls mounted on large posterboards and they did an excellent job. I believe they used the equivalent of wheat paste (a.k.a wallpaper paste) and did an absolutely perfect job.
Let us know what you find out.
John
} I realize that this subject may be out of the realm of microscopy but I } thought maybe one of you could help. Our lab generates alot of poster } titles for various meetings. The posters are then mounted in the } departmental hallways for a few months so they need to survive for some } time. I use a double sided adhesive tape made by Letraset to put the } title(printed on card stock) on posterboard. I have just found out that the } company is no longer making the tape and I can't find a decent substitute. } The double sided scotch tape isn't strong enough; the spray photo mount is } very difficult to work with in a lab and doesn't hold well either. It } leaves a sticky residue on anything nearby. } Does anyone know of a very high tack tape similar to the Letraset tape?
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
} Our lab generates alot of poster } titles for various meetings. The posters are then mounted in the } departmental hallways for a few months so they need to survive for some } time. I use a double sided adhesive tape made by Letraset to put the } title(printed on card stock) on posterboard. I have just found out that the } company is no longer making the tape and I can't find a decent substitute. } The double sided scotch tape isn't strong enough; the spray photo mount is } very difficult to work with in a lab and doesn't hold well either. It } leaves a sticky residue on anything nearby. } Does anyone know of a very high tack tape similar to the Letraset tape? } Thanks in advance,
Panel-beaters (maybe you call them something different ----- the people who repair smashed-up cars) use very strong double-sided tape for fastening trim to exterior panels.
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Responding to the message of {3.0.5.32.19990616153149.00802450-at-pop.uky.edu} from Mary Gail Engle {mgengle-at-pop.uky.edu} : } } Dear microscopists, } I realize that this subject may be out of the realm of microscopy but I } thought maybe one of you could help. Our lab generates alot of poster } titles for various meetings. The posters are then mounted in the } departmental hallways for a few months so they need to survive for some } time. I use a double sided adhesive tape made by Letraset to put the } title(printed on card stock) on posterboard. I have just found out that the } company is no longer making the tape and I can't find a decent substitute. } The double sided scotch tape isn't strong enough; the spray photo mount is } very difficult to work with in a lab and doesn't hold well either. It } leaves a sticky residue on anything nearby. } Does anyone know of a very high tack tape similar to the Letraset tape? } Thanks in advance, } } Mary Gail Engle } Electron Microscopy & Imaging Facility } University of Kentucky
I've used a product called Dry Bond, by Chartpak (#DBS25), which is a dry adhesive that comes on sheets (11x14") bound in a booklet. You press your title cards, photos, etc onto the sheet and peel it off and the dry adhesive comes with it. You then press the work onto your mounting board and thats it. The work is removeable, that is, repositionable.
It will last long enough for your 3 months, but after a year or so, it does tend to loosen up, bubble, up, etc. The adhesive probably dries out and the bond is weakened.
Just my 2.5 cents worth, good luck.
Gib
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
There is a much more insidious problem associated with power cables than you might expect based simply on their current capacity, and that is residual unbalanced current.
What happens is something like this: Someone has a remote piece of equipment - lets say a furnace - which is, of course, grounded to the supply ground (as electrical regulations require). However, the user has heeded the institution's requirements to plumb in the water cooling with metal pipe. Now the supply ground is connected to the water ground. Nobody worries about this because, far from being hazardous, it actually improves the personal safety of the electrical system.
Well, all is fine provided there are no connections between the live circuits and the ground, right?
WRONG!! There are always capacitive connections between conductors and nearby metal parts. In any one piece of equipment they may be small, but they add up. I have known one system with a beautifully engineered isolation transformer, with a solid copper electrostatic shield between the primary and secondary, with a leakage current of close to 1mA, which had to be capacitance because the DC resistance between the primary and the frame was unmeasurably high. This current then flows to ground through your water pipes and, hey presto - you now have an unbalanced current in the supply - and you only need a few mA unbalance to get a disastrous effect.
What is the bottom line? Calculations of fields from theoretical first principles don't work. Do not allow you main power supply to be anywhere near your electron microscopes. They *WILL* interfere.
One person's actual experience - I have a single 200A three-phase feeder about 20 feet from the closest point of a STEM and can see the interference. (A STEM is no more prone to EM interference than a CTEM - you can just see the interference more easily)
Good luck!
Tony Garratt-Reed.
At 11:48 AM 6/15/99 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Folks; We have an S360 SEM and have used several image capture systems over the years. There are problems associated with passive capture on the S360. First the S360 is a PAL (European video 740x576 pixels) so many frame grabber boards will not give the correct aspect ratio for images. The voltage on the image board of the 360 in only + or - 1 volt so many systems will not sync to the signal from the image transfer board. We have had great success with the Orion system from E.L.I. sprl in Belgium. Their web page is www.OrionMicroscopy.com . As a passive system we feel they are the best solution for the S360. On our other machines we use the Quartz PCI which is a great system for SEM's particularly, Hitachi, JOEL. The PCI woks well with the Cambridge S250 as well. When we first started doing image capture, the system we built took advantage of the S360's noise reduction and frame integration. We simple took the frozen image signal from the mono monitor output on the back of the machine, fed the signal into a broad bandwidth video distribution amplifier and then fed the output of the amp to a Matrox P8 frame grabber using the old Snapshot software. SEM signals were also distributed to our Compix Image Analyser and Codonics video printer all at the same time. The image size was only 740x576 put the images were noise free and quite useful as we distributed them over our video network and workers at remote sites could view samples being examined in relative real time. This method is not passive capture but real frame grabbing. The down side to this method is the image grey scale is an average of all the pixels in the image including the black header and thus the numerics in the header will not be pure white as they are with the Orion passive capture system. Hope this is of some use. Jon McGovern J. P. McGovern and Associates www.microscopy.net jon-at-microscopy.net
The best stuff that I have found for mounting pictures and posters and it really holds up for a very long time is 3M 568 positionable mounting adhesive. This stuff is used by graphic art shops and it is really easy to work with. I've been using it for about 15 years. It holds better than the spray mount and if you are nominally careful, there is no mess.
I believe the 568 number is for the roll format that I have. It is 11 inches wide by many feet long. I know that there is an 18 inch wide version with another number because I have used it for 16 x 20 prints.
You put your artwork on the material pressing slightly. Then cut it out with an Exacto knife along the edges of your artwork. Using the scraper supplied with the PMA, you press the adhesive onto the back of the artwork. Peel the backing off of the artwork and the adhesive is transferred to the back. The artwork can be laid gently down and repositioned until it is aligned correctly. (Try that with spray adhesive.) Then using the scraper again with a protective sheet over the artwork, press over the entire artwork. This will stay in place for years. I have only found one type of sublimation dye printer paper that I had trouble with on the first stage. Much harder pressure than I normally use fixed that.
You can get it from photographic shops, but they may have to order it. If you try this once, you'll never go back to the spray.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Mary Gail Engle To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Dear microscopists, I realize that this subject may be out of the realm of microscopy but I thought maybe one of you could help. Our lab generates alot of poster titles for various meetings. The posters are then mounted in the departmental hallways for a few months so they need to survive for some time. I use a double sided adhesive tape made by Letraset to put the title(printed on card stock) on posterboard. I have just found out that the company is no longer making the tape and I can't find a decent substitute. The double sided scotch tape isn't strong enough; the spray photo mount is very difficult to work with in a lab and doesn't hold well either. It leaves a sticky residue on anything nearby. Does anyone know of a very high tack tape similar to the Letraset tape? Thanks in advance,
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
I put a cleaning procedure on the web some time ago it came from the team=
in the EM unit in Canberra.
LANTHANOM HEXABORIDE Technique developed by E.M. Unit Canberra
Clean the cathode with 25% hydrochloric acid in water by immersing for 60=
seconds and then cleaning with a weak alkaline (ammonia or sodium hydroxide). Wash with water and then alcohol before drying.
LaB6 sources should last a long time (1000 hours plus) but they do need a= n intermediate cleaning session about every 250 to 350 hours. Some people amaze us by getting away with 1100 hours without cleaning but this is the=
exception not the rule.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Some of the replies to our posting seem to have run a little off track.
The ammonia clean is intended to be used on the stainless steel gun components only, no mention of any other areas! A few points are set out=
below in relation to those who have replied to our request for comment.
1. Three people, two from the USA, one from South Africa already use=
this technique to whom due respect is given, they have not seen any problems.
2. There are those who use manual techniques that they claim are far=
quicker. The point of this technique is to remove the manual component from cleaning as the methods and media used are often the most common reasons for increased contamination within a column. Hand cleaning methods, particularly electro mechanical methods, are also far more damaging to the cathode than a wet technique. Mis shaped cathode apertur= es have been found to be the cause of terminal astigmatism! A liquid clean = is a preferred route as it removes the metal polish and cotton, or tissue, that are used to apply the polish. All three media are difficult to remove and may cause contamination and discharge in an electron gun if th= ey are not completely removed. A higher level of technical and observationa= l skills are also required with manual techniques.
3 There are some who have commented that the occasional hand polish= , or better still in the ultrasonic cleaner with a metal polish, is useful = as it removes any irregularities caused by discharge. I would agree with them, perhaps one in three cleans should follow this route in a TEM. The= re should be less of a problem in a SEM.
4 It has been mentioned that plated cathodes may be damaged if the plating is incomplete. I do not know of any plated cathodes as far as I have seen they are all stainless steel with some using tantalum apertures= . =
Take care if you clean a SEM anode as some are made of aluminium alloy an= d they will go black when placed in ammonia solution!
5 Tantalum so I am told, is only mildly attacked by alkalis, needin= g to heated above 150 deg C to encourage unacceptable attack. Thanks to Bi= ll Tivol for that information.
6 The suggestion of using plasma etching as the ideal cleaning devi= ce for all components is not challenged, however the ammonia method describe= d is cheap, simple and almost always available in the EM laboratory world wide.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
The fields will drop off with the square of the distance. i.e.....
1 Ft 1 Gauss 2 1/4 G 0.25 4 1/16 G 0.625 8 1/32 G 0.03125
The fields you're describing could be significant. Be careful, even in calculating the potential problem. If it develops into an image disturbance problem you may be able to cancel the fields. For more information on site selection pitfalls see http://www.vibeng.com or call.
Craig Franklin Vibration Engineering Consultants 303-689-2224 } } } Our EM facility will be moving to a new building currently under } construction. We have just learned that a major feed line consisting of } six 3 inch conduits carrying three 400 amp parallel lines (1200 A total) } has been installed 11 feet from where our highest resolution TEM is going. } We have been asked to supply a minimum acceptable distance that these lines } could be from the microscopes, since it will be quite expensive to reroute } them (they are in concrete). My specific questions are: } } What is the appropriate formula to compute the falloff of the field with } distance? } } Does this theoretical formula adequately predict the field in a real situation? } } How does the orientation of the lines affect the calculation? } } Thanks for your help. Any references would also be appreciated. } } Dr. Marie E. Cantino } Dept. of Physiology and Neurobiology } University of Connecticut } Storrs, CT 06269-2131 } Phone: 860-486-3588 } Fax: 860-4861936 } } } }
try using a sticky tab on a stub, placed around a small pre-scored section on the label side of the CD. just push it on hard and pull it off and you should get a little piece of the aluminum film. works quick and easy... } } Dear Owen, } I was successful in cutting out a one cm. chunk of a hot-pressed CD and } leaving it in chlorothene overnight. Unfortunately, we can't use that } anymore. Any good plastic solvent should work, even acetone. You are left } with a wisp of very thin aluminum foil with the dints in it visible at about } 10,000X. The CD-Rs are a very different technology. } At 11:39 AM 6/15/99 -0400, you wrote: } } } } Sometime in the last two years I remember a thread on preparing CD's for } } examination in the SEM. Can anyone remember what solvent was used to } } remove the polymer coating from CD's so imaging of the data is possible. } } Please respond to me directly at the address below. Thanks. } } } } Owen } Regards, } Mary } } Mary Mager } Electron Microscopist } Metals and Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } e-mail: mager-at-interchg.ubc.ca } } } } -=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=- Brian McIntyre University of Rochester Electron Microscopy Lab 716-275-3058 716-244-4936 (fax)
Does anyone know of a supplier for an anti chicken-25nm gold conjugate that is ideally raised in a goat, for the EM. I have found a rabbit anti chicken-25nm conjugate from Airon, but rabbit seems to routinely cause us problems Thanks, Susie Nilsson
Seeking advice on the use of electrochemical solutions for use in the Tenupol Apparatus for preparing thin TEM samples. Is anyone using a laboratory prepared mixture of conc nitric acid (30%) and methanol (70%) in the apparatus? If so, what are the documentation / policy requirements for the training, use, storage, handling disposal of this mixture in your facility? What alternative commercially available chemical solutions are currently being used in the Tenupol apparatus for low alloy steels? Barry EM UNIT UNSW
The best way to affix posterboard or paper to another piece of posterboard or paper is to use drymount tissue. This essentially is tissue paper impregnated with a heat-sensitive adhesive. The piece of tissue is trimmed to exactly the same size as the piece to be attached. The two pieces then are fused by pressing them in a drymount press (a huge press with a heated platen, the same device use to iron on t-shirt transfers) for about 45 seconds. The bond is permanent (in fact you cannot separate the two pieces without destroying one).
The tissue is available at photo supply and art supply stores/catalogues. A widely-available brand is by ColorMount by Seal, though I know there are other manufacuturers. The down side of this technique is that one needs the press. The media centers and art departments at most academic institutions will have one. (If you are going to mount resin coated photos, be careful to get the drymount tissue with the proper melting point, otherwise you will ruin your photos).
Nothing holds as well as drymount tissue; this is what all the professionals use. All other methods that I have tried will bleed through paper, form bubbles/ripples between the two surfaces, and/or will eventually peel away, particularly with changes in relative humidity.
DL ______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
Disclaimer: E.L.I. Belgium distributes a passive grabbing system worldwid= e so we are not exactly a disinterested third party.
Choosing an active or passive system is a difficult task and obviously th= e choice is a function of the =93external=94 operating conditions like imag= e size needs (must be 512x512 for image analysis for example) or EDX mapping tas= ks (be able to scan a specific area), but is also a function of the SEM itself.
The SEM itself is important in choosing an active or passive system if th= e SEM scanning generator provides enough lines, then there is probably no need for an active drive; a passive system is always easier and simpler to install an= d configure than an active one. It is also easier to use because its is considered a pure slave of the SEM scanning generator. And a grabbing sys= tem must deliver images with the lowest possible amount of noise and the mini= mal distortion: the pixels must be square.=20
Our Cy (E.L.I. sprl in Belgium) designed a very powerful passive system (Orion) with four ideas in mind:
- the system must be easy to use:
Getting the image from the SEM is the last step and must be done efficien= tly, so the number of clicks and actions must be reduced to the minimum. For example, in the Orion system, you need only one mouse click to get the im= age from the SEM, and one to get a printout! So we directly help the user red= ucing his overhead time just by optimizing the user interface.
- it must provide an accurate copy of the image produced on the SEM scree= n:
During the system installation, it is important to properly configure and calibrate the grabbing system in order to avoid image distortions. The oversampling factor can be optimized for any scanning mode. The result is= a clean, jitter-free image immediately ready for processing, storage or printing.
- it must include functions that are in direct relation with the job done= on the SEM:
For example, a very useful function inside the Orion system is the abilit= y of reconstructing a micron bar when you extract an area from a grabbed image= ; this prevent the loss of the distance calibration information. We have the policy of constantly adding new functions inside the software, which are directly helping the system user producing images. Other functions include: very efficient distance measurement, line/rectangle drawing, contrast / brightness adjustment, etc.=20
- Some functions are available as options because some user=92s don't nee= d them and won't have to pay for them. We currently have four options:
- the 3D, anaglyph construction, which includes a tool for automatic plan= e alignment. This compensates for any stage drift that can occur during til= t; - The EDX, which allows you to mix EDX mappings with grey scale images;=20 - The DSM option, which optimizes the Orion software for the LEO DSM 94x/= 95x series - The Macro command processor, for linking buttons with repetitive tasks stored in text files.
Furthermore, the Orion system has a feature which is unique on the market= : the analog part (which is connected to the SEM electronics) is completely flo= ating (isolated) from the digital part (connected to the PC processor).=20
This very particular design, although very technical, provides the Orion system with two unique advantages:
- A security advantage: your SEM is fully protected against electrical disturbances that could occur in your computer or network and is fully =93floating=94. If a fault occurs in the computer electrical system, the = SEM is protected. - A bigger noise insensitivity: by breaking the ground loop between the S= EM and the computer, we make the Orion system virtually =93jitter=94 free and noise-free.=20
By the way, I take this opportunity to let you know that we are currently looking for highly qualified representatives in different countries. If you are interested, please contact me.
At 14:01 04/06/99 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Paul Vanderlinden. Sales Manager.
*********************************************************************=20 See our web site: http://www.orionmicroscopy.com To contact us:
Good Morning, I'm wondering if someone could give me information about who to contact to post a position we have available here at UCSD in the Nuerosciences dept? It's a research position, specifically looking for someone with microsopy background. Please let me know. Thank you, Susi Goodman, Staffing Dept, UCSD
I'm sorry I didn't get your name! Here is an initial description. But I also want to put together a formal job announcement as a MS Word Attachment, that includes all the information re: how to apply for position. I'll send that this afternoon. Thank you, Susi Goodman
116509-S STAFF RESEARCH ASSOCIATE I Scientific & Laboratory Research Neurosciences Open Until Filled $2289-$2742 STAFF RESEARCH ASSOC I 100% Career Perform immunolocalization studies on protein kinases and related proteins in the nervous system and other tissues. Technician will collaborate with researchers from several laboratories as part of a program project. Duties will include specimen preparation for light and electron microscopic immunocytochemistry. Other duties will include perfusion fixation of laboratory rats and other small animals, sectioning tissue for light microscopic examination using both the cryostat and Vibratome, examination and analysis of patterns of immunostaining at both the LM and EM levels, flat embedding Vibratome sections for electron microscopy, production of ultrathin sections, and semi-thin sections for conventional and high voltage EM, examination and analysis of sections under both conventional and intermediate high voltage electronic microscopes, and production of publication quality light and EM micrographs. REQUIREMENTS: Familiarity and skill with transmitted light microscopy, laser scanning confocal microscopy, conventional and high voltage electron microscopy. Ability to produce high quality thin and semi-thin sections for electron microscopy. Experience with high-resolution immunolocalization techniques for electronic microscopy, including colloidal gold detection. Understanding of immunocytochemical theory and practice. Ability to conduct a research study with minimal supervision. Ability to understand and evaluate scientific literature. Must be able to juggle several projects at once and work collegially with collaborators. Theoretical background in molecular and cell biology or related field.
Good Morning, I'm wondering if someone could give me information about who to contact to post a position we have available here at UCSD in the Nuerosciences dept? It's a research position, specifically looking for someone with microsopy background. Please let me know. Thank you, Susi Goodman, Staffing Dept, UCSD
Just a quick reply about the double sided tape questions. I don't know if this is the same as the tape you were using, but scotch also makes what they call "Permanent-Linerless Double coated Tape". It's 3/4 inches wide and extremely sticky. We've had great success with this tape, to the point that Posters that have been made a year and a half ago are still hanging strong.
thanks
Adam
---------------------------------------------------------------------- Adam Baker Carleton University Biology Department Email address: adbaker-at-ccs.carleton.ca ----------------------------------------------------------------------
{bold} {fontfamily} {param} Times {/param} {bigger} {bigger} Title {/bigger} {/big= ger} {/fontfamily} {/bold} {fontfamily} {param} Times {/param} {bigger} {bigger} : Nitrogen-carbon bonding in new materials - Experimental and theoretical investigations
{bold} Job description {/bold} :=20
Within the framework of a 4-year TMR research network,=20
=AB synthesis, structure and properties of new carbon-based hard materials =BB ( {/bigger} {/bigger} {/fontfamily} http://www.physto.se/tmr {fontfamily} {para= m} Times {/param} {bigger} {bigger} / ) initiated in 1998, the CEMES,=20
situated in Toulouse (France), offers a post-doctoral position.=20
The Toulouse group (which is included in a partnership encompassing two other groups:=20
Solid-State Physics Lab in Orsay and ONERA-Chatillon) is specialized in the implementation
and development of analytical techniques in electron microscopy to explore the structure and
the local chemical and electronic properties of new materials.
The research to be carried out includes EELS (Electron Energy Loss Spectroscopy) and EFD (Elastic Filtered Electron Diffraction) experimental investigations, as well as theoretical calculations of the optical and electronic properties using CASTEP (pseudopotential plane-wave code).
The position is limited to European Community citizens (excluding French).
Duration: 1 year.
{bold} Fields {/bold} : Solid state physics, prior experience in EELS and/or
carbon-based materials would be an advantage.
{bold} Application deadline {/bold} : September 1, 1999
{bold} Place of work {/bold} : Centre d'Elaboration des Mat=E9riaux et Etudes Structurales
(CEMES) du CNRS, Toulouse France, Electron Microscopy and Analysis
group.
{bold} Contact person {/bold} : Virginie SERIN
E-mail : serin-at-cemes.fr
Phone : +33 (0)5 62 25 78 67
Fax: +33 (0) 5 62 25 79 99
Address: CEMES/CNRS
29 rue J. Marvig, BP4347, 31055 Toulouse Cedex, France
I just want to put in my 2 cents worth regarding digital image acquisition from the LEO 360s:
As Jon describes below, there are different methods for acquiring images:
Via a simple frame grabber Passive acquisition Active acquisition
There was a thread recently that discussed the differences between passive and active acquisition, so I won't say anything about that.
acquisition via frame grabber: this is possible if the system has a TV output. This is not necessarily a TV-rate scanning, but must be a true TV signal. The signals are usually either NTSC or PAL, NTSC being the US and Japanese standard, PAL the European. Unfortunately, they are not compatible. Both use the same bandwidth, but PAL sacrifices repetition rate for resolution (NTSC: 30 frames per second, 640x480, PAL: 25 fps, 768x576). Most frame grabbers can either be bought for one of the signals or can digitize both. However, acquiring TV images from an SEM is probably not the best solution. The images are low in resolution, and they are only 8 bit. The display is fast, though, and through averaging a good S/N ratio can be reached.
Regarding the active/passive difference, please check out the previous thread. For active acquisition from the 360 series it is necessary to have a beam control interface installed. That option is available from LEO.
Of course, if you select an acquisition system that is based on a frame grabber, like ours, you can have all three options available. A TV rate acquisition for finding and focussing, and then a passive or active acquisition for high resolution images.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
} ---------- } From: Jonathan P. McGovern[SMTP:SEMRUS-at-TELUSPLANET.NET] } Sent: Wednesday, June 16, 1999 6:38:22 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: S360 Image capture } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Folks; We have an S360 SEM and have used several image capture systems over the years. There are problems associated with passive capture on the S360. First the S360 is a PAL (European video 740x576 pixels) so many frame grabber boards will not give the correct aspect ratio for images. The voltage on the image board of the 360 in only + or - 1 volt so many systems will not sync to the signal from the image transfer board. We have had great success with the Orion system from E.L.I. sprl in Belgium. Their web page is www.OrionMicroscopy.com . As a passive system we feel they are the best solution for the S360. On our other machines we use the Quartz PCI which is a great system for SEM's particularly, Hitachi, JOEL. The PCI woks well with the Cambridge S250 as well. When we first started doing image capture, the system we built took advantage of the S360's noise reduction and frame integration. We simple took the frozen image signal from the mono monitor output on the back of the machine, fed the signal into a broad bandwidth video distribution amplifier and then fed the output of the amp to a Matrox P8 frame grabber using the old Snapshot software. SEM signals were also distributed to our Compix Image Analyser and Codonics video printer all at the same time. The image size was only 740x576 put the images were noise free and quite useful as we distributed them over our video network and workers at remote sites could view samples being examined in relative real time. This method is not passive capture but real frame grabbing. The down side to this method is the image grey scale is an average of all the pixels in the image including the black header and thus the numerics in the header will not be pure white as they are with the Orion passive capture system. Hope this is of some use. Jon McGovern J. P. McGovern and Associates www.microscopy.net jon-at-microscopy.net
Has anyone looked or researched the tribology and or tribochemistry (e.g. chemical mechanical polishing) involved in either or both dimpling and tripod polishing?
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:L-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
L.D. Marks wrote: -----------------------------------------------------------------------.
Has anyone looked or researched the tribology and or tribochemistry (e.g. chemical mechanical polishing) involved in either or both dimpling and tripod polishing?
++++++++++++++++++++++++++++++++++++++++++++++++ If you mean has anyone done a serious research project (as in a graduate degree thesis project) of these prep method's tribology/chemistry? I would say no, not that I'm aware of (and I do follow these subjects closely).
However, one of the reasons why tripod polishing (and the related and evolved tools) have been so successful is that they came along at the same time some remarkable polishing compounds appeared on the scene. All through the 1980s the semiconductor industry was really gearing up for the manufacture of large-wafer integrated circuits, mostly on Si but to some extent on III-V and II-VI compounds as well. These wafer surfaces had to be nearly atomically smooth and free of polishing damage. A HUGE effort went into the creation of colloidal polishing compounds (colloidal silica as the best example) among others to meet the semiconductor industry's polishing needs. There must have been a large number of tribological and or tribochemistry studies performed on the effect of colloidal products polishing silicon at the time. I use the phrase "must have been" in recognition that many projects like these in semiconductor manufacturing facilities were deemed proprietary by the companies involved and few papers were written. But, good grief, enough time has passed and they should be in the literature by now!
Tripod polishing was the beneficiary of these efforts as Klepeis and Benedict here in IBM had access to these media in our facility when the method was being invented. Certainly the clout generated by our needing a high tech polishing media for making TEM specimens would not have accounted for beans.
So, are there studies of colloidal media polishing silicon, etc available? Certainly. Are these studies applicable to tripod polishing TEM specimens? Sure. Are they back-applicable to dimpling when the dimplers use these media? Sure. Is there room for a new study specifically directed to the tribology and or tribochemistry of making TEM specimens? ABSOLUTELY! Should such a study occur you can count on our help.
Ron
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Having been in tribology and having done work with these materials in the TEM, I think that I can be reasonably certain that there hasn't been any investigations in these areas of TEM preparation. One reason might be that there simply aren't any economic reasons to do it.
However, the ball cratering device that has been used for dimpling TEM samples is a common device that is used for examining and testing hard coating on tool steels and other substrates.
There have been examinations of wear-tested films of solid film lubricants (MoS2) as I am pretty sure that you are aware. I presented some work at MSA in 1993. There was a paper by the French group in the MRS Journal (1992 or 1993, First author: Mosely?) I have made samples of MoS2 film samples using both techniques as well as the small angle cleavage technique. I've also prepared WS2 by these techniques as well as FIB. When MoS2 is wear-tested, one of the tribochemical products is MoO3. I've never seen any evidence of this phase in the as-deposited films examined in the TEM. Pulsed Laser deposited films of MoS2 go down amorphous and when wear-tested, they generate both the MoS2 and MoO3 phases. You can wave a carbon coated grid in the wear track and you will find both phases. In the as-deposited samples prepared by these various techniques, again, I have not detected these phases. I should point out however that the dimpled samples and tripod polished samples have all been finished with ion milling. Those prepared by cleavage and FIB have not been touched by abrasive.
Of course, the testing that we did would have been done with a pin-on-disk and without lubricant. Dimpling and tripod polishing both have the complication that you would have to consider the abrasive/lubricant and sample wear couple.
Hard coatings may be a different matter however. There may be some studies on wear-tested tool steels that may be analogous to the sample preparation techniques. But, again, I think any tribochemical changes will get wiped out by ion milling. About 2 or 3 years ago, Ainissa Ramirez from the Stanford group presented some of her Ph.D. work on tribo-mechanical changes in DLC films. I'm not sure where they published, but try the ICMCTF meeting papers published in Thin Solid Films or Surface and Coatings Technology.
I once examined a tripod polished single crystal SiC sample without the cleanup ion mill and saw an interesting defect structure from the polishing on both sides. Although you could see the scratches and dislocations in the sample, you could still see the single crystal structure of the material. Short ion milling totally cleaned the sample. I'm sure there are a number of such examples for silicon for both dimpling and tripod polishing.
I think that an interesting avenue for research on tribochemical changes could be performed on already prepared TEM samples in which the samples are "wear-tested" with a contact mode AFM in an electron transparent area. You could then have an area that is wear-tested adjacent to an area that is not.
I probably didn't answer your question, but there may be some overlaps in my ramblings.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: L. D. Marks To: Microscopy List -----------------------------------------------------------------------.
Has anyone looked or researched the tribology and or tribochemistry (e.g. chemical mechanical polishing) involved in either or both dimpling and tripod polishing?
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:L-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Dear all The Web site for our Electron Microscope Unit at the University of the Witwatersrand is a as follow
http://www.wits.ac.za/emu
Mr. S H Coetzee Electron Microscope Unit Private bag X3 Wits Johannesburg 2050 Tell: +27 11 716 2419 Fax : +27 11 339 3407 E-mail stephan-at-gecko.biol.wits.ac.za
May I remind the list about the one day Advanced School to be held on Tuesday 24 August 1999 in Sheffield, UK?
The subject is High Resolution Electron Microscopy and, unlike a recent Sheffield movie, the school aims to provide the full coverage. Field emission sources, computer-aided alignment, image reconstruction from focal or tilt series, energy filtering and more.
The four speakers are well known in the field: John Hutchison, Thierry Epicier, Chris Boothroyd and Angus Kirkland. There will also be demonstrations on a FEGTEM and a 300kV HREM, and participants will take away a complete set of notes.
N.B. Fast approaching is 25 June 1999 - the deadline for EMAG bursary applications for students and young scholars who are members of the Institute of Physics.
For details, go to: http://www.iop.org/IOP/Confs/EMAG or write to: c.j.hetherington-at-sheffield.ac.uk
I recently acquired a Wild M21 polarized light microscope. Does anyone have a manual that they could lend to me (I would copy it and then return it) or forward a copy. I will pay any associated fees.
Thank you.
Alan Stone ASTON Metallurgical Services Co., Inc. 4201 North Ravenswood Ave Chicago, IL 60613 773/528-9830
For more information on this position, and guidance on how to apply, please visit our website at www-hr.ucsd.edu. Look under "Job Bulletin", "employment opportunities", and "scietific & laboratory research." Thank you.
116509-S STAFF RESEARCH ASSOCIATE I Scientific & Laboratory Research Neurosciences Open Until Filled $2289-$2742 STAFF RESEARCH ASSOC I 100% Career Perform immunolocalization studies on protein kinases and related proteins in the nervous system and other tissues. Technician will collaborate with researchers from several laboratories as part of a program project. Duties will include specimen preparation for light and electron microscopic immunocytochemistry. Other duties will include perfusion fixation of laboratory rats and other small animals, sectioning tissue for light microscopic examination using both the cryostat and Vibratome, examination and analysis of patterns of immunostaining at both the LM and EM levels, flat embedding Vibratome sections for electron microscopy, production of ultrathin sections, and semi-thin sections for conventional and high voltage EM, examination and analysis of sections under both conventional and intermediate high voltage electronic microscopes, and production of publication quality light and EM micrographs. REQUIREMENTS: Familiarity and skill with transmitted light microscopy, laser scanning confocal microscopy, conventional and high voltage electron microscopy. Ability to produce high quality thin and semi-thin sections for electron microscopy. Experience with high-resolution immunolocalization techniques for electronic microscopy, including colloidal gold detection. Understanding of immunocytochemical theory and practice. Ability to conduct a research study with minimal supervision. Ability to understand and evaluate scientific literature. Must be able to juggle several projects at once and work collegially with collaborators. Theoretical background in molecular and cell biology or related field.
Good Morning, I'm wondering if someone could give me information about who to contact to post a position we have available here at UCSD in the Nuerosciences dept? It's a research position, specifically looking for someone with microsopy background. Please let me know. Thank you, Susi Goodman, Staffing Dept, UCSD
Midwest Microscopy and Microanalysis Society (MMMS) Members:
We are soliciting nominations for people to serve as officers of MMMS f= or the 2000 society year. Nominees should be active members in good standing = of MMMS. Elected positions include President-elect, Treasurer, Recording Secretary, Life Sciences Director, Materials Sciences Director, and Pro= gram Co-ordinator. Duties for these officers are described in the MMMS Constitution and Bylaws, which can be accessed at http://www.msa.microscopy.com/MSALAS/MMMS/MMMSConstitution.html.
All suggestions for nominations should be sent to the Recording Secreta= ry, Linda LaRonge-Snow (linda.snow.nmi-at-abbott.com) and should be received b= y Friday, June 25. Please include a brief description of you or your nominee's qualifications for the position. Final decisions on nominati= ons will be made by the MMMS Executive Council. This is your chance to bec= ome involved and have a voice in your society, so please let us know if you= are willing to serve!
Jerry Gagne (gerard.d.gagne-at-abbott.com) President, MMMS =
I am considering purchasing a scanner for scanning TEM negatives into electronic form. I have reviewed some of the brands and came up with a couple of candidates, and I wonder if any of the microscopy listserv members have used these or other scanners for TEM negatives, and could give me some advice.
I am considering Microtek ScanMaker5, and the UMAX Powerlook III. Both scanners work in transparency and reflection mode, and come with a SCSI interface and lots of software, and can handle various sizes. The Scanmaker5 is listed at 1000 x 2000 dpi optical resolution, and 3.6 maximum optical density (Dmax). It costs about $2400 The UMAX Powerlook III is listed as 1200 x 2400 dpi optical resolution and 3.4 maximum optical density. The price is about $1200.
The difference between the two appears to be that the Scanmaker has no glass in the optical path, thus avoiding Newton rings and possibly other distortions (as well as dust, etc). The salesman at UMAX claimed that their geometry, although it has glass in the optical path, avoids Newton rings because a film holder causes the negatives not to sit on the glass.
I would clearly prefer to pay $1200 than $2400, but since this is a large purchase, I would like to get it right, and will find the extra money if necessary.
Have any of you direct experience with these scanners for TEM negatives that you could advise me as to which would be better? I would appreciate any feedback (including suggestions for other scanners), and if there is enough interest, I propose to summarize the advice I get in a later posting to this group.
Thank you,
Arthur Motta *************************************************************************** Arthur T.Motta Department of Nuclear Engineering 814-865-0036 The Pennsylvania State University fax: 814-865-8499 231 Sackett Bldg, University Park, PA 16802-1408 http://www.nuce.psu.edu
The University of Souther California, School of Engineering is seeking an experienced electron microscopist to fill the position of Laboratory Manager in the Center for Electron Microscopy and Microanalysis (CEMMA), who will oversee the operation and maintenance of the Center.
The Laboratory Manager is expected to:
Provide technical laboratory expertise to faculty, research staff and graduate or undergraduate students in the design and execution of experiments, participate in educational and research activities consistent with the University's mission. This individual should be self motivated and resourceful, possess good organizational and strong computer skills. The ability to instruct students and researchers in the operation of microscopes and ancillary equipment is important. He needs to schedule users and maintain equipment in proper operating condition.
Qualifications:
B.S. in one of the Physical Sciences or equivalent. Experience in TEM/SEM/EDS required. Familiar with mechanical and electronic equipment, and vacuum systems.
Contact:
Florian Mansfeld University of Southern California Chair/Department of Materials Science Los Angeles, CA 90089-0241 (213)740-3016 mansfeld-at-mizar.usc.edu
Salary Range:
Commensurate with education and experience.
Jack Worrall Director of Operations Univ. of Southern Cal. CEMMA Los Angeles, CA 90089-0101
Accidentally delete files folders? =46disk or =46ormat the wrong drives? Lose data because of a virus? Are you or have you seen errors like, invalid drive specification, invalid media type or error reading drive c:? Hard drives are getting cheaper and cheaper, but the data on those drives can be invaluable. However you have options. You can send the drive to a Data Recovery Professional and pay the price (thousands). You can buy an over the counter retail product which may due more harm than good, or you can use the software that the professionals use for this type of recovery. We are for a limited time, making the software we sell to the professionals available to the general public to handle the out cry for data recovery software we have experienced due to virus infection, and the simple fact that almost everyone now has a computer. Virus scanners are great, but the fact is they can not update virus signatures as fast as people are producing viruses. More and more people are learning how to program. Unfortunately that means there are more programmers capable of producing viruses. Major companies spending millions of dollars on security and virus protection have lost data due to virus infection. It will get worse before it gets better. You don=92t have t= o spend thousands to recover your data if you have the right software. We are making this software available to you now, at a very reasonable cost! Act now and receive unlimited free technical support ! This won=92t last long!
=46or more information please reply to: mailto:merch2-at-bigfoot.com?subject=3Dmore-info
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Has it always been difficult obtaining schematics for Zeiss SEM's? I've been referred by a LEO serviceman...to a LEO manager...to a german engineer, who has been unresponsive to my requests for documentation on behalf of a customer.
If anyone can help obtain copies of the DSM-960 schematics, particularly the scan generators and video, I'd certainly be in your debt.
A few years back I remember a CD ROM that I saw at the Microscopy and Analysis meeting which was I think called "The Virtual Microscope". It covered basic light microscopy, optics and physics. It also had a virtual microscope upon which users could practice their aligning skills. I am in need of locating this software for a possible class offering at the graduate level. Does anybody know of where this program can now be purchased?
ZEISS did not include the schemnatics in the manual so that you have to = get them for service. It is their SECRET. =20 It would be easier to get it if you had made it as a condition before = purchasing. We have learned that this condition must be included in all = equipment purchase.
Ann Fook
} } } jbest {jbest-at-elmdas.com} 06/19 7:28 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html=20=
Has it always been difficult obtaining schematics for Zeiss SEM's? I've = been referred by a LEO serviceman...to a LEO manager...to a german engineer, = who has been unresponsive to my requests for documentation on behalf of a = customer.
If anyone can help obtain copies of the DSM-960 schematics, particularly = the scan generators and video, I'd certainly be in your debt.
An investigator is trying to look at some cross bridges in the germaria of = a mutant Drosophila. We have fixed the tissue in 2% Glut in 0.1M cacodylate= =20 buffer, post fixed in 1% OSO4 in 0.lM cacodyalte, treated with 2% Tannic = Acid in the same buffer, and en bloc staining with 2% aqueous uranyl acetate,=20=
followed by routine dehydration. The tissue is embedded in Spurrs's = media. Sections were cut in the 60 nm range. Stained sections on formvar coated grids with 7% UA in absolute methanol, followed by Lead Citrate. Also stained sections on plain grids with = routine UA and LC. We liked the plain grids better but the intensity of the stain = was not as great as we would like. The structures of interest are ER-like = cisternae, and are very hard to see.
Does anyone have any ideas about how to improve the intensity of the stain or suggestions on how to improve our techniques in order to see our tiny structures in the germaria?
George Lawton Chief Electron Microscopist Microscopy and Imaging Service Center UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, Tx 75235-9039 Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
I have a Carl Zeiss Microscope ll which is equiped for fluorscence microscopy. The bulb that I have is very old. Does anyone know where I can get a Philips 126184 C.S.I.250 watt H7 mercury bulb?
Thanks Debbie
Debbie Lietz Electron Microscopy Suite Department of Biology Trent University Peterborough, Ontario K9J 7B8 Telephone: (705)748-1486 Fax: (705) 748-1205 email: dlietz-at-trentu.ca
I recall some discussions on Snow Cleaning a while back. Does anyone have any information about the technique and how it works ? Any information would be greatly appreciated.
Thanks,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
Back when I was a graduate student some 50 years ago the term 'Spectroscopy' was used to refer to measurements made with an instrument in which the separated spectral components were measured or recorded either visually or photographically (thus the 'scope' component), while the term 'Spectrometry' generally was used to refer to measurements made with a device that measured the separated spectral components with a phototube and displayed the results on a meter or via a strip chart recorder (i.e. a metering device).
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
A technical position is available in the newly established Microscopy Center at Mount Sinai School of Medicine in New York. We are seeking an experienced Electron Microscopy technician with a BS/BA or MS in Biology/Life Sciences. Applicants should have excellent communication and organizational skills, an understanding of basic laboratory procedures, and the ability to manage a large and varied workload. The successful candidate will participate in ultrastructural studies of various biological systems. Qualifications include at least 2 years of experience in routine transmission electron microscopy procedures, ultramicrotomy, immunogold labelling, specimen preparation, photographic darkroom work, and routine maintenance of equipment. Experience with immunofluorescence and confocal microscopy is an asset.
We offer a salary commensurate with experience and excellent benefits. For consideration, please mail/email your resume to:
Scott Henderson, Ph.D., Director, Microscopy Center, Box 1007, Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, NY 10029-6574
We are an equal opportunity employer fostering diversity in the workplace.
As noted on p. 72 of my book Vacuum Methods in Electron Microscopy (http://www.2spi.com/catalog/books/book48.html), Peter B. Sewell, of LAB-6 Inc., recommends cleaning lanthanum hexaboride deposits from Wehnelt cylinders and apertures by soaking them for about a minute in a solution consisting of 1 part by volume of concentrated hydrochloric acid and 4 parts water, then rinsing sequentially with water, dilute ammonia, deionized water, and isopropyl alcohol, and then drying with a blast of clean warm air or with a gas blaster.
A guy who works in the company that makes LaB6 products should know, so I suggest you try his method.
Good luck!
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
On April 7, 1999, I responded to a need that had become apparent on this list, i.e., a source for instruction manuals for older instruments. I suggested that "It would be a real service to our community if a master compilation of these manuals, with a suitable index and regular updates, could be put together."
I could not volunteer personally for this task but fortunately several people have responded to this discussion and I thought this would be an appropriate time to summarize the comments. I have not figured out all the company/institutional affiliations for the respondents but the emails are given. I believe that all of these respondents have sent copies of their replies to the entire list so they should be receptive to direct inquires.
Yvan Lindekens suggested large manufacturers might not support idea and there might be a problem with copyrights. I suspect that the copyrights will not be too much of a problem, especially since we do not propose to make any money on the deal. Speaking for my own situation, a request for a manual for an older instrument causes us a lot of trouble in searching out information and never pays for itself directly.
Dmitri Sokolov (sokolov-at-ryouko.rciqe.hokudai.ac.jp) suggested that the idea can be considered a part of his about global knowledge base as self-growing unified searching source of information, as described in his home page.
Uri Watson (uri-at-watson.ibm.com) planning to scan a few manuals and put them on Yvan LIndekens web site.
During the last 2 1/2 months or so there have been numerous other postings askinig for specific manuals and I suspect that many of these may have been responded to privately.
So here is a general framing of the problem.
How to do it? Scanning or copying or a combination.
When to do it? Shgould it be doneas needed/requested or should we try to collect as much as possible at the outset. In either case, there will need to be a master list posted somewhere to show what is available and how to get it, cost, etc.
Where would the effort be centered? We need a site which will be as close to permanent as possible so that the work will not be jeopardized by job changes, etc. Barbara Foster.MME, has volunteered for part of this, though clearly this is subject to change when we see how big a task it turns into.
Reimbursement. Some form of reimbursement would be needed to at least cover direct costs - though reimbursement for time spent would be prohibitive.
Copyrights. Theoretically a problem but maybe not a real problem in practice. A good place to start would be to see what kind of copyright notice, if any, is included in prospective manuals and then we could contact manufacturers as needed. If any manufacturer's representative reading this discussion would like to volunteer their position, it would be helpful.
What would we call the service? Here's a chance for all the punsters/acronymers to jump aboard. How about "MOD squad" for Manuals On Demand?
Volunteers Ed Sharpe, COURYHOUSE-at-aol.com, had server drive space available and is going to scan in the ones that he has.
Barbara Foster, mme-at-map.com, has resources for archiving, copying, shipping. Also specific manuals - see later section.
Yvan Lindekens, yvan.lindekens-at-skynet.be volunteer to help.
Manuals for specific items
Coolwell Model SE chiller, Valdemar Furdanowicz, rwafu-at-bsco.com
Reichert, Barbara Foster
Zeiss, Barbara Foster
Reichert Zetopan, Yvan Lindekens web site
SEM, TEM, and EDX manuals, over 100 instruction sheets, going back to the late 80's, Steve Chapman, Protrain, PROTRAIN-at-compuserve.com
Comments???
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology "A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
} A few years back I remember a CD ROM that I saw at the Microscopy and } Analysis meeting which was I think called "The Virtual Microscope". It } covered basic light microscopy, optics and physics. It also had a virtual } microscope upon which users could practice their aligning skills. I am in } need of locating this software for a possible class offering at the } graduate level. Does anybody know of where this program can now be } purchased?
} Larry -
Perhaps you saw it at a RMS meeting. There's a "forthcoming" English CD with that title, but I've had no response from them to several Emails. There is an alternative, done at the University of Washington; it's good, and I believe that it has been improved since I wrote this review for the Project MICRO bibliography:
Pagliaro, l., Murray, C., Curran, G., Orkand, A., and Astion, M. 1997 Microscopy-Tutor. ISBN 0-7817-1217-3 $195.00 plus shipping; distributed by Lippincott-Raven Publishers, 12105 Insurance Way, Hagerstown, MD 21740, 800-638-3030. For Macintosh and Windows 3.1 or 95/NT; easy installation. Developed by the Department of Laboratory Medecine and the Center for Bioengineering at the University of Washington, Seattle, this is a college-level introduction to the use of a research-quality compound microscope. Its extensive use of QuickTime animation to illustrate alignment steps and optical principles make it much more than a "book on a CD"; moving ripples on a pond really do make it easier to understand wave theory! Kohler illumination is emphasized and explained. Most, but not all, terminology is defined; a glossary would help beginners. Proper care of lenses is covered well, but there's no instruction on how to use immersion oil properly. There is a brief concluding self-test that is more of a review of important information than a quiz. Although it's a good CD, it's definitely not appropriate for precollege microscopists. Adult.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
For increased staining intensity with Spurr's resin, I routinely use an approximation of the double lead stain technique as published by Daddow in J. Submicrosc. Cytol. 18:221-224 (1986).
The method is as follows: I. Stain sections for 30 secs - 2 mins in lead citrate (either Reynolds or Venable and Coggeshall recipe). Rinse in distilled water and let the sections air dry. 2. Stain with whatever recipe uranyl acetate you normally use for 1-3 mins.; the article describes saturated aqueous, but I use the method found in J.E.M.Technique16:81-82 (1990) . Rinse in distilled water. 3. Immediately transfer the rinsed, but not dry, grids to a fresh drop of the lead citrate and stain for another 30 secs to 2 mins. Rinse in distilled water and air dry.
That's it! It's fast and effective.
M. Nesson--
_______________________________________________________________________ Michael Nesson, Ph.D. Department of Biochemistry & Biophysics 2011 Ag&LS, Oregon State University, Corvallis, OR 97331-7305 (541)737-5245 FAX:(541)737-0481 nessonm-at-ucs.orst.edu
Hello : We want to determine the distribution of PVA in a composite containing Nylon, glass fibers , PET and paper fibers either by OM or SEM. We were hoping to use image analysis, but are not able to apply segmentation without any further contrast enhancement through staining. We can selectively stain the nylon , and the glass fibers are no problem, but we have not been able to selectively stain the PVA. Any suggestions would be appreciated.
Does anyone have a manual that could forward it to me (I would be a copy). Please, tell me how I can pay any associated fees. It was old (maybe 20 years)and has a label with "Carl Zeiss Jena, made in DDR". It has 4 lens under the table that rotate (360 grades). Thanks,
Rejane Pimentel Universidade Federal Rural de Pernambuco-Brazil Functional Plant Anatomy Av. Boa Viagem, 6592/602 CEP 51130-000, Recife, Pernambuco, Brasil
Virtual SEM available from Brendan Griffin (bjg-at-cyllene.uwa.edu.au) at UWA Perth he has been giving this away for duplication costs.
He is also co-author of Virtual EDS with Clive Nockolds (clive-at-emu.usyd.edu.au) , that one is for sale at some nominal amount to cover their expenses. I don't know the exact amount but it is relatively cheap ~ $100-200.
I've used both and they are well done.
You should contact Brendan or Clive directly for more information.
Dear Listers, Does anyone have a Hitachi S-450, working or not, for sale? Or the main parts of one? Please reply to me privately.
Thanks, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } An investigator is trying to look at some cross bridges in the germaria of a } mutant Drosophila. We have fixed the tissue in 2% Glut in 0.1M cacodylate } buffer, post fixed in 1% OSO4 in 0.lM cacodyalte, treated with 2% Tannic Acid } in the same buffer, and en bloc staining with 2% aqueous uranyl acetate, } followed by routine dehydration. The tissue is embedded in Spurrs's media. } Sections were cut in the 60 nm range. } Stained sections on formvar coated grids with 7% UA in absolute methanol, } followed by Lead Citrate. Also stained sections on plain grids with routine } UA and LC. We liked the plain grids better but the intensity of the stain was } not as great as we would like. The structures of interest are ER-like cisternae, } and are very hard to see. } } Does anyone have any ideas about how to improve the intensity of the stain } or suggestions on how to improve our techniques in order to see our tiny } structures in the germaria? } } } George Lawton } Chief Electron Microscopist } Microscopy and Imaging Service Center } UT Southwestern Medical Center at Dallas } 5323 Harry Hines Blvd. } Dallas, Tx 75235-9039 } Phone: 214-648-7291 } eMail: George.Lawton-at-email.swmed.edu } } Hi,
Your sections are too thin! And then they are on films! No wonder you cannot see anything. If you cannot use thicker sections and do away with films on grids, try the following (all tested successfully in our lab) Keep the tissue in osmium overnight in the refrigerator. Keep the tissue in the enbloc UA overnight in the refrigerator. Do not embed in Spurr's. Aside from its dangerous toxicity, it has the lowest contrast of the common epoxies. Use Luft (1961), perhaps medium hard (any textbook) and you will see an immediate increase in contrast. Spurr's is so highly crosslinked that poststains penetrate poorly. "Overexpose" your negatives in the TEM! That is, increase the negative density. This often works like charm. Then print on Grade 1 or 2 paper. If you need 3 paper, you need more negative density. Bye, Hildy
In the May 1999 Microscopy Today (#99-4), Gary W. Gill of Diagnostic Cytology Laboratories, Inc., has a short article entitled "Cover Glass Perspectives." Although the article has a lot of interesting information, at one point the author states that one should not use No. 1 1/2 coverglasses even tho they have a nominal thickness of 0.16 to 0.19. He says to use No. 1 coverglass to make up for the "substantial and variable thickness of the mounting medium." This is counter to everything I was ever taught. I always use the minimum mounting media possible and press the slide down firmly on to the coverslip to ensure this is kept as thin as possible.
Isn't standard to use #1 1/2 coverslips? Is there really a school of thought that one should use #1's?
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
RESEARCH RESOURCES CENTER, UNIVERSITY OF ILLINOIS AT CHICAGO
Position Title: MICROSCOPY SPECIALIST
The Electron Microscopy Facility of the Research Resources Center (RRC) at the University of Illinois at Chicago has an open position for a Microscopy Specialist. The facility provides electron and laser/ light microscopy services for the university research community and external organisations from two sites on the campus. The open position is in the recently completed RRC-East facility which specialises in physical and materials science fields. The east side facility includes JEOL JEM-3010 and JEOL JEM-100CX TEMs, JEOL JEM-2010F and VG HB501 STEMs, a JEOL JXA733 Microprobe and a Renishaw Raman Spectrometer.
The person we are looking for should have a bachelors's degree minimum and preferably a master's degree or higher in physical sciences, engineering or a related field, with at least four years experience in laser and electron microscopy. They will supervise the operation of the expanding laser microscopy area (Raman Spectroscopy and Laser Scanning Confocal Microscopy), including record keeping and maintenance and will assist the staff in the day to day running of the Electron Microscopes and preparation area. Interpersonal/ Communications skills are important as this individual will work with users, provide technical advice and demonstrate how microscopy can advance their research.
Further information may be obtained by consulting the following website: http://www.rrc.uic.edu/
For consideration, interested parties should send an application letter, complete curriculum vitae, and the names and addresses of three references to:
Gordon L. Humphrey, Ph.D. (gordo-at-uic.edu) Research Resources Center (m/c 937) University of Illinois at Chicago 835 S. Wolcott Ave. Chicago, Illinois 60612-7341
Voice: (312) 996-7600 Fax: (312) 996-0539
Alan W Nicholls, PhD Manager - Electron Microscopy Service Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Dear Friends' I am looking information about a scanner for 3.25X4inch TEM sheet film negatives. I shall very much appreciate any information from users of this size or multiple size negative scanner. Thanking you C.Singla.
I am interested in analyzing air particles collected in a beryllium processing facility for mineral forms of Be (BeO, Be, CuBe, BeF). Do you have TEM/EELS capabilities to run such analyzes. Work will commence in July.
Thank you for your interest. ------------------------------- Renee M Kalmes CIH Managing Scientist Environmental Group 149 Commonwealth Drive Menlo Park, CA 94025 (650) 688-1757phone (650) 688-1799 fax http://www.exponent.com
Dear listers, we have an ultratome III. When we get sections for TEM (thin sections), it produces sections with irregular thickness; moreover we have really difficult to get very thin sections. We suspect some of wrong at the level of the heating coil, but we do not have experience in that. Could someone help us?
Moreover, could someone indicate prizes and companies that sell ultramicrotome or are there some second hand ultramicrotome availbale?
Thanks a lot for your answers and time.
Cheers,
dr Enrico de Lillo Istituto di Entomologia agraria - Universit=E0 Bari - Italy tel. +39 080 5443105 fax +39 080 5442876 email: delillo-at-agr.uniba.it http://193.204.185.103/de_lillo.htm
Different lenses are corrected for different cover glass thicknesses, and there are lenses corrected for use with no cover glass. Your microscope manufacturer should be able to tell you more about details. I always thought that 0.17mm was the standard (which falls into the 1 1/2 thickness). No 1 cover glasses are better if you want to apply pressure more directly more locally, but they also break more easily. my 2p Best wishes
Stephan Helfer +44(0)131 248 2865; fax +44(0)131 248 2901 ------------------------------------------- To most people 'solutions' mean finding the answers. But to chemists solutions are things that are still all mixed up. (Science Explained)
Somehow I am missing part C of my Zeiss manual, especially part C 3.3 which explains how to mecahnically align the lenses to the Zeiss EM 10C/10CR electron microscope.
Does anyone know this procedure? Or is it possible for anyone to fax me this section of the manual that I am missing???
Thankyou.
Garry Burgess
Charge Technologist Electron Microscopy Laboratory Department of Pathology Health Sciences Centre Winnipeg, Canada Ph: 204-787-1508
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } An investigator is trying to look at some cross bridges in the germaria of a } mutant Drosophila. We have fixed the tissue in 2% Glut in 0.1M cacodylate } buffer, post fixed in 1% OSO4 in 0.lM cacodyalte, treated with 2% Tannic Acid } in the same buffer, and en bloc staining with 2% aqueous uranyl acetate, } followed by routine dehydration. The tissue is embedded in Spurrs's media. } Sections were cut in the 60 nm range. } Stained sections on formvar coated grids with 7% UA in absolute methanol, } followed by Lead Citrate. Also stained sections on plain grids with routine } UA and LC. We liked the plain grids better but the intensity of the stain was } not as great as we would like. The structures of interest are ER-like cisternae, } and are very hard to see. } } Does anyone have any ideas about how to improve the intensity of the stain } or suggestions on how to improve our techniques in order to see our tiny } structures in the germaria? } } } George Lawton } Chief Electron Microscopist } Microscopy and Imaging Service Center } UT Southwestern Medical Center at Dallas } 5323 Harry Hines Blvd. } Dallas, Tx 75235-9039 } Phone: 214-648-7291 } eMail: George.Lawton-at-email.swmed.edu } } Hi,
Your sections are too thin! And then they are on films! No wonder you cannot see anything. If you cannot use thicker sections and do away with films on grids, try the following (all tested successfully in our lab) Keep the tissue in osmium overnight in the refrigerator. Keep the tissue in the enbloc UA overnight in the refrigerator. Do not embed in Spurr's. Aside from its dangerous toxicity, it has the lowest contrast of the common epoxies. Use Luft (1961), perhaps medium hard (any textbook) and you will see an immediate increase in contrast. Spurr's is so highly crosslinked that poststains penetrate poorly. "Overexpose" your negatives in the TEM! That is, increase the negative density. This often works like charm. Then print on Grade 1 or 2 paper. If you need 3 paper, you need more negative density. Bye, Hildy
I was wondering if anyone uses a dilute solvent in the diamond knife boat = when collecting carbon coated grids? I suspect a strong concentration = would affect the glue holding the diamond knife in place, but wondered if = a dilute amount would even bother it? (Diatome knives are what I am using = if there are differences between the glue used in other brands). Reason = being, is that I am finding when I attempt to collect my sections, they = tend to scatter and most of them are found along the edges of the grid. = There is a hydrophobic effect happening with the carbon, and any quick = suggestions would be appreciated, to help alleviate this problem. Thanks, Susan
Susan Carbyn Electron Microscopy Technician Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada 32 Main Street, Kentville, Nova Scotia B4N 1J5 Canada
1) Try using Epon/Araldite embedding resins, old recipes. Those have unparalleled staining properties.
2) Do try it WITHOUT tannic acid, if you haven't yet. This should give notably better resolution.
3) How much time does the material spend in glutaraldehyde, OsO4? For high resolution of "tiny structures", it should not be left at any stage longer than necessary. Higher concentrations of glutaraldehyde (3-4%) may be worth trying, too.
4) Did you try using lower accelerating voltage?
5) Do stick with aqueous en bloc UA, and, as long as possible, naked grids.
Hope something helps. Sincerely, Vlad.
} } An investigator is trying to look at some cross bridges in the germaria of a } mutant Drosophila. We have fixed the tissue in 2% Glut in 0.1M cacodylate } buffer, post fixed in 1% OSO4 in 0.lM cacodyalte, treated with 2% Tannic Acid } in the same buffer, and en bloc staining with 2% aqueous uranyl acetate, } followed by routine dehydration. The tissue is embedded in Spurrs's media. } Sections were cut in the 60 nm range. } Stained sections on formvar coated grids with 7% UA in absolute methanol, } followed by Lead Citrate. Also stained sections on plain grids with routine } UA and LC. We liked the plain grids better but the intensity of the stain was } not as great as we would like. The structures of interest are ER-like cisternae, } and are very hard to see. } } Does anyone have any ideas about how to improve the intensity of the stain } or suggestions on how to improve our techniques in order to see our tiny } structures in the germaria? } } } George Lawton } Chief Electron Microscopist } Microscopy and Imaging Service Center } UT Southwestern Medical Center at Dallas } 5323 Harry Hines Blvd. } Dallas, Tx 75235-9039 } Phone: 214-648-7291 } eMail: George.Lawton-at-email.swmed.edu } } Vladislav V. Speransky School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Phone: 207 581 2998 Fax: 207 581 2969 Email: vladis-at-maine.maine.edu
Any tips, info., papers about the use of bacitracin when negative staining would be greatly appreciated.
Thanks, Kim
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Kimberly A. Riddle Florida State University tel: 850.644.6519 Biological Science Imaging Resource 119 Bio Unit I, 4370 fax: 850.644.0481 Tallahassee, FL 32306 riddle-at-bio.fsu.edu ~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 6 - October 14, 1999
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $2050 (Includes room and board, text, handouts, supplies)
Application Deadline: August 3, 1999
Admission application and information: Carol Hamel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu (Application forms available via Adobe Acrobat)
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical
measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, polarized light, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy, multiphoton excitation fluorescence microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratiometric-imaging laser tweezers and laser scissors
Applications to live cells will be emphasized; other specimens will be covered as well.
Students will have direct hands-on experience with state-of-the-art microscopes, video cameras, recorders and image processing equipment provided by major optical and electronics companies. Instruction will be provided by experienced staff from universities and industry.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual
I have used "Micro," a glassware cleaner, which contains EDTA to complex metals, to remove LaB6 residues. Use a fairly concentrated 25 to 50% solution in water and sonicate. Beware that this is only for stainless steels and will damage copper and brass components. Prof. Bigelow's suggestion about the hydrochloric acid sounds good and I'll have to try that next time.
regards,
Dave Audette OSRAM Sylvania Beverly, MA david.audette-at-sylvania.com
} -----Original Message----- } From: Keith Moulding [SMTP:mcmouldk-at-ust.hk] } Sent: Wednesday, June 16, 1999 11:56 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Cleaning Wehnelts } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } Any good chemical recipes for cleaning LaB6 oxide deposits? W seems to be } the flavour of the month. } } Keith. } }
Here in Australia the suppliers of coverglasses seem to promote No. 1 covers probably for the reason that adding a thin layer of mountant increases the thickness. Don't be fooled by what the manufacturers say their coverglass thicknesses are. A friend measured several boxes of them and found that they varied outside the limits stated. My friend was photographing Diatoms and needed the 'perfect' slide. He ended up measuring all coverglasses and separating out those that he wanted for photomicrography and used the others for routine work. All very time consuming and not really applicable to the busy lab.
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Dear Tom
Different lenses are corrected for different cover glass thicknesses, and there are lenses corrected for use with no cover glass. Your microscope manufacturer should be able to tell you more about details. I always thought that 0.17mm was the standard (which falls into the 1 1/2 thickness). No 1 cover glasses are better if you want to apply pressure more directly more locally, but they also break more easily. my 2p Best wishes
Stephan Helfer +44(0)131 248 2865; fax +44(0)131 248 2901 ------------------------------------------- To most people 'solutions' mean finding the answers. But to chemists solutions are things that are still all mixed up. (Science Explained)
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Could someone tell me the best way to prepare clay particles for TEM. I'm interested in small particles, micron to sub-micron in size in both the dry state and hydrated state. Thanks in advance. Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Is there an accepted minimum number of pixels that should be used in determining the linear distance between two resolved points (for example, in a resolution check) in the image of a sample. This would essentially identify the minimum magnification required for a given digital image size.
I'm thinking that the absolute minimum in an image with no noise would be three (e.g. dark - bright - dark), But more realistically should be something like 5 or 7.
Also, what is the minimum difference in contrast levels say for an 8-bit image to be able to say that the features are in fact resolved? Should the difference be greater than the square root of the pixel with the highest brightness?
} I was wondering if anyone uses a dilute solvent in the diamond knife boat } when collecting carbon coated grids? I suspect a strong concentration } would affect the glue holding the diamond knife in place, but wondered if } a dilute amount would even bother it? (Diatome knives are what I am using } if there are differences between the glue used in other brands). Reason } being, is that I am finding when I attempt to collect my sections, they } tend to scatter and most of them are found along the edges of the grid. } There is a hydrophobic effect happening with the carbon, and any quick } suggestions would be appreciated, to help alleviate this problem.
} Susan Carbyn
Susan -
The best approach is glow discharge. If your vacuum evaporator isn't equipped to do it, adding a Tesla coil to your system is inexpensive and should be simple.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
We have a Noran Voyager II EDXS system which runs on a Unix box under SunView. The data is stored in a proprietary format and can be converted to EMSA file format on the Voyager. A huge amount of data has been backed up on to floppy disk and reloading all the data on to the Voyager, converting it to EMSA format and the putting it all back on to floppies is not really an option we want to consider!!!
We were wondering if there were any programs around which could read the proprietary format on either a PC or Mac.
Thanks very much for any information you can provide.
Colin Veitch
Instrumentation Scientist Electron Microscopy Textile and Material Technology Group CSIRO Textile and Fibre Technology PO Box 21, BELMONT, Vic. 3216. Australia.
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
The question of when can you call a feature "resolved" is interesting. I was wondering if it could be described as resolved if its average pixel value and its standard deviation was significantly greater than the average pixel value of its surrounding nieghbors and their standard deviation? The test for significance may tell you the "n" needed or number of pixels needed to be valid. I dont know. What do you think?
Bob Derm Imaging Center University of Washington
On Wed, 23 Jun 1999, Walck. Scott D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Is there an accepted minimum number of pixels that should be used in } determining the linear distance between two resolved points (for example, in } a resolution check) in the image of a sample. This would essentially } identify the minimum magnification required for a given digital image size. } } I'm thinking that the absolute minimum in an image with no noise would be } three (e.g. dark - bright - dark), But more realistically should be } something like 5 or 7. } } Also, what is the minimum difference in contrast levels say for an 8-bit } image to be able to say that the features are in fact resolved? Should the } difference be greater than the square root of the pixel with the highest } brightness? } } Any ideas? } } -Scott } }
I am missing Part C (section C 3.3) of my original Zeiss manual which describes how to align the lenses of this microscope (mechanically), so I wondered if anyone could tell me the procedure, since I don't know how to do that. The microscope was moved from one room to another, and the lenses seem to have been thrown a little bit out of alignment.
Thankyou,
Garry Burgess Charge Technologist Department of Pathology } Health Sciences Centre
Scott - Nothing easier than dry clay preparation. Just tap against a small paint brush that has been "loaded" with some clay and let the particles settle onto a coated grid. Hold the grid with tweesers and tap these to release any excess and larger clumps. Carbon coat the grid.
Clay naturally includes water within its structure and dried clay is not "real clay". Years ago when I did some limited work on this, nothing useful was published. This may have changed, especially since suitable equipment is more common. Unfortunately ESEM is not likely to provide sufficient resolution. I am doubtful about "environmental" cells in TEM, since these too would lower possible resolution on account of window thickness. Freezing droplets of water and clay (mist) and using a cold stage would be the obvious technique. That equipment was not available years ago. I expect that its now possible to visulize hydrated clay, but the technique would be quite challenging. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Thursday, June 24, 1999 8:25 AM, Walck. Scott D. [SMTP:walck-at-ppg.com] wrote: } } } Could someone tell me the best way to prepare clay particles for TEM. I'm } interested in small particles, micron to sub-micron in size in both the dry } state and hydrated state. } Thanks in advance. } Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) }
Susan - No problem using a drop of alcohol (ethanol) added to the trough water. A lower water level gives a better placed light reflection and flatter surface. The lower water level also decreases the chance of wetting the block, but the lower surface tension increases that chance. D'oh. Usually no problem though.
It seems that the flatter trough water is largely unrelated to your pick-up problem. I suggest that if you have a nice ribbon, bring a grid up (rim may be bend to the most suitable angle for the tweesers) from underneath with the grid at about 45 degrees to the surface and the rim bisecting the first section. The ribbon is then held in place and centres nicely on the grid. If no nice ribbon is available, round up sections in the trough using a mounted hair or Teflon sliver. Use a loop and bring this up through the water surface centered on the sections. The loop picks up the last drop, which includes the sections. Place a coated grid on a filter paper and jiggle the loop minutely on the grid until the water goes into the paper and the sections will be well placed on the grid. Hydrophobic problems would be minimal if Butvar coated grids were used. Ultimately you can make carbon grids hydrophilic using a Glow Discharge Instrument. ProSciTech and most EM suppliers offer such units (PST only in Australasia) Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Thursday, June 24, 1999 2:56 AM, Susan Carbyn [SMTP:CarbynS-at-em.agr.ca] wrote:
} I was wondering if anyone uses a dilute solvent in the diamond knife boat when } collecting carbon coated grids? I suspect a strong concentration would } affect the glue holding the diamond knife in place, but wondered if a dilute } amount would even bother it? (Diatome knives are what I am using if there } are differences between the glue used in other brands). Reason being, is } that I am finding when I attempt to collect my sections, they tend to scatter } and most of them are found along the edges of the grid. There is a } hydrophobic effect happening with the carbon, and any quick suggestions would } be appreciated, to help alleviate this problem. } Thanks, } Susan } } Susan Carbyn } Electron Microscopy Technician } Atlantic Food and Horticulture Research Centre } Agriculture and Agri-Food Canada } 32 Main Street, Kentville, Nova Scotia B4N 1J5 } Canada } } Phone: (902) 679-5566 } Fax: (902) 679-2311 } } E-mail: carbyns-at-em.agr.ca }
Message text written by Garry Burgess } Hello knowledgeable folks,
I am missing Part C (section C 3.3) of my original Zeiss manual which describes how to align the lenses of this microscope (mechanically), so I wondered if anyone could tell me the procedure, since I don't know how to do that. The microscope was moved from one room to another, and the lenses seem to have been thrown a little bit out of alignment. {
Hi Garry ,
I do not know the mechanical alignment procedure for a Zeiss, but it is a=
TEM so basic alignment principles should do the job for you? Try this
ILLUMINATION LENSES 1. Large spot size with C1 (weak lens) bring C2 to crossover 2. Mechanically align the spot to the centre of the screen with C1 3. Smaller spot size with C1 (stronger lens) bring C2 to crossover 4. Mechanically align the spot to the centre of the screen with C2 5. Repeat 1 through 4 until you have a constant centre.
IMAGING LENSES 1. With a specimen in the microscope go to diffraction 2. Align the diffraction spot with the centre of the screen with the=
last lens (Could be called Projector 2, or just Projector?) 3. Slowly increase the magnification until around 30 to 60KX the image flips through diffraction (bright diffraction flash and the image flips over as another lens switches on). There will have been a similar action at a lower magnification but this is less important in most labs. 4. One step below this point centre a feature on the screen with the=
stage drives 5. One step above this point centre feature with the last but one le= ns mechanical alignments on the column (Could be called Projector 1 or Intermediate 2) 6. One step below this point centre the feature with the final lens mechanical alignments 7. Repeat 5 and 6 for a constant centre but with a flip over.
STANDARD PROCEDURES With any system the actions are always, or nearly always, the same. Cent= re one lens under a weak condition and then centre the other under a stronge= r condition. Similar procedures are used to bring gun and illumination deflection coil systems into a common alignment. My problem is how many imaging lenses do you have and how many of them may be aligned?
Please come back if you need more
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
-----Oorspronkelijk bericht----- Van: MichaelD {michaeld-at-amsg.austmus.gov.au} Aan: Tom Phillips {PhillipsT-at-missouri.edu} ; Stephan Helfer {S.Helfer-at-rbge.org.uk} CC: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com} Datum: jeudi 24 juin 1999 5:38 Onderwerp: Re[2]: coverglass thickness
The expression "cover glass thickness" as used by the manufacturers of microscope optics is misleading, as it means in reality:
the thickness of the layer of adhesive used + the thickness of the object + the thickness of the layer of mountant used + the thickness of the coverslip...
That's why it's usualy better to use thinner coverslips, especially for critical slides...
Mike Dingley and his friend are right: the number "1 1/2" only means that the majority of the coverslips is (about) 0.170 microns...
I've also measured some boxes of coverslips from a high-quality German Brand, using a precision micrometer. This was the result from some boxes of 22 * 22 mm coverslips chosen at random:
150 - 160: 21 p. 160-170: 56 p. 170 - 180: 23 p.
That's why it's sometimes advised in (amateur-)literature to measure the thickness of the coverslips. The thicker slips can than be used for large(r) whole mounts, the thinner for critical specimens. Of course, for amateurs this can be considered as "part of the fun", for pro's it's only time-consuming thus expensive!
I know from experience, that it's sometimes very difficult to obtain decent images from "routine slides", especially when using high-quality objectives. As an example: it's very difficult to make decent photomicrographs using, as I often do, an objective 45x/0.90 APO, due to the spherical abberation introduced as a result of incorrect cover glass thickness... These abb's become very noticiable when objectives with an N.A. higher than 0.50 are used... That's an interesting paradox as it means sometimes: "the better the optics, the worser the image"!
Possible remedies are:
* using cover slips of the correct thickness * using homogenious immersion objectives (if availlable) * correcting the tube length of the microscope (if possible)
(All the above is only applicable to microscopes/optics designed for a fixed tube length of 160/170mm: I don't have any experience with infinity corrected optics).
On Thu, 24 Jun 1999 14:06:00 +1000 jim {jim-at-proscitech.com.au} wrote: } I am doubtful about "environmental" cells in TEM, since } these too would lower possible resolution on account of } window thickness.
Dear Jim,
I must defend the "environmental" cells in TEM. We are one of several groups around the world (in USA, Japan and Europe)using this technique and we can obtain point resolution of 0.25nm. This will obviously depend on gas type and pressure but apertured cells do not suffer from window thickness effects.
We have used water vapour to study reactions but we have not to looked at hydrated clays.
Regards, Ron
Ron Doole. Department of Materials, phone +44 (0)1865 273701 University of Oxford, fax +44 (0)1865 283333 Parks Road. Oxford. OX1 3PH. ron.doole-at-materials.ox.ac.uk
Bob, Theoretically your definition sounds valid. In reality though, one complicating factor is noise. Then it comes down to reproducibility or validating a detection. Russ, Xerox
-----Original Message----- } From: Robert Underwood [mailto:underwoo-at-u.washington.edu] Sent: Wednesday, June 23, 1999 11:58 PM To: Walck. Scott D. Cc: Micro
Hi,
The question of when can you call a feature "resolved" is interesting. I was wondering if it could be described as resolved if its average pixel value and its standard deviation was significantly greater than the average pixel value of its surrounding nieghbors and their standard deviation? The test for significance may tell you the "n" needed or number of pixels needed to be valid. I dont know. What do you think?
Bob Derm Imaging Center University of Washington
On Wed, 23 Jun 1999, Walck. Scott D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Is there an accepted minimum number of pixels that should be used in } determining the linear distance between two resolved points (for example, in } a resolution check) in the image of a sample. This would essentially } identify the minimum magnification required for a given digital image size. } } I'm thinking that the absolute minimum in an image with no noise would be } three (e.g. dark - bright - dark), But more realistically should be } something like 5 or 7. } } Also, what is the minimum difference in contrast levels say for an 8-bit } image to be able to say that the features are in fact resolved? Should the } difference be greater than the square root of the pixel with the highest } brightness? } } Any ideas? } } -Scott } }
Nucip Guven in the Geosciences Department at Texas Tech University makes his living looking at clay with the TEM. Give Dr. Guven a call or look for some of his papers.
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Could someone tell me the best way to prepare clay particles for TEM. I'm interested in small particles, micron to sub-micron in size in both the dry state and hydrated state. Thanks in advance. Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
by moe.optics.rochester.edu (8.9.3/8.9.3) with ESMTP id IAA29867 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 24 Jun 1999 08:51:11 -0400 (EDT) Message-Id: {l03130303b397d7ea593b-at-[128.151.240.75]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
hello all-
i've noticed that the island size from a typical Au/Au-Pd sputtering system seems much bigger when viewed in a high resolution FESEM (meaning that it is easier to resolve the islands!). is there any trick to setting up a Cr system, or is it as simple as putting a Cr target in a high vacuum sputtering system? i have a turbo pumped vacuum station i'm thinking of building up into a Cr capable system and wonder if there are things to be aware of...
Brian McIntyre mailto:mcintyre-at-optics.rochester.edu Sr. Engineer lab: 716-275-3058/4875 River Campus EMLab fax: 716-244-4936 University of Rochester Rochester, NY 14620
"The most important thing a father can do for his children is to love their mother." - Unknown
We don't use any metals for charge collection coatings because of the visible-island-problem. We switched to carbon coating all SEM samples (that need coating) long ago. A little less contrast but no artifacts to 200kX. The carbon coating should be quite thin. About 10 nm or less--that's just a quick flash of carbon. More is definitely not better.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Noran now makes an EDS package based on a Windows NT workstation. All your data should be compatible with this new system. When we upgraded, we transferred all our old spectra onto Jazz discs using an FTP, and we were able to access them without a problem with the new software. The new system is called Vantage, and despite being on an NT machine, still retains the look, feel and functionality of the Voyager software. You may want to contact them and see if they'll offer to do the transfer for you.
Hope this helps,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
I just saw a demonstration of the Cresington High Resolution coater by Alan Berginc. This is a magnetron sputter unit. For Cr, he kept the shutter closed and watched the plasma until it changed color from a purple hue to this beautiful blue color. At that point, he said that the oxide on the target has been removed and that is when the deposition can start. I don't know what color-blind people will do. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Brian McIntyre To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
hello all-
i've noticed that the island size from a typical Au/Au-Pd sputtering system seems much bigger when viewed in a high resolution FESEM (meaning that it is easier to resolve the islands!). is there any trick to setting up a Cr system, or is it as simple as putting a Cr target in a high vacuum sputtering system? i have a turbo pumped vacuum station i'm thinking of building up into a Cr capable system and wonder if there are things to be aware of...
Brian McIntyre mailto:mcintyre-at-optics.rochester.edu Sr. Engineer lab: 716-275-3058/4875 River Campus EMLab fax: 716-244-4936 University of Rochester Rochester, NY 14620
"The most important thing a father can do for his children is to love their mother." - Unknown
I am starting to think about the 7th APEM, and was wondering if there will be a sufficient number of people interested in electron microscopy of polymers. What I have in mind is:
(i) an oral presentation on the deformation morphology of polyethylene;
(ii) a poster presentation on optimizing the morphology of polypropylene for use in crash barriers.
Nearly all my co-workers on these projects are from Asian countries. Of course, this is early days, and I would have some logistics to work out.
If you could let me know if there is a substantial polymer interesting, I would be glad to know.
You can see what polymers look like under the EM on:
http://www.reading.ac.uk/~spsolley/alien.html
Yours sincerely,
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Noran now makes an EDS package based on a Windows NT workstation. All your data should be compatible with this new system. When we upgraded, we transferred all our old spectra onto Jazz discs using an FTP, and we were able to access them without a problem with the new software. The new system is called Vantage, and despite being on an NT machine, still retains the look, feel and functionality of the Voyager software. You may want to contact them and see if they'll offer to do the transfer for you.
Hope this helps,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
} --- } } Is there an accepted minimum number of pixels that should be used in } determining the linear distance between two resolved points } (for example, in } a resolution check) in the image of a sample. This would essentially } identify the minimum magnification required for a given } digital image size. } } I'm thinking that the absolute minimum in an image with no } noise would be } three (e.g. dark - bright - dark), But more realistically should be } something like 5 or 7.
I'm not too sure what you're asking ... but if its in regard to how "resolution" is defined for digital images (pixels per inch) versus the historical definition (the distance between 2 resolvable points) ... a general working definition is pixel pairs, or the distance between the middle a first pixel and the middle a third with a discernable pixel between. For fine photography 7 line pairs is acceptable, but if you magnify an acceptable line pair, I don't think 5 pixels is needed ... a pixel pair should suffice. } } Also, what is the minimum difference in contrast levels say } for an 8-bit } image to be able to say that the features are in fact } resolved? Should the } difference be greater than the square root of the pixel with } the highest brightness? } That would seem an acceptable value ... I have only seen this issue addressed as the darker pixel being no brighter than the height of the lighter pixel (FWHM), but that wouldn't seem suitable for a pixel value. But again, resolution is generally defined by an average person's ability to see ... i.e, a photograph. It would be an interesting value to determine and standardize for the sake of quantitative instrumental measurements and image analysis.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I don't know where it was published ( I think in J. of Ultrastructure Research) but if you search by authors: Dr. Harry L. Malech & John P. Albert. "Negative Staining of Protein Macromolecules: A Simple Rapid Method" One of the slickest negative stains I've ever done. If all else fails I have a copy that I can send you.
Best,
Al Coritz Cryobiology Product Manager, Ventana-RMC
-----Original Message----- } From: Kim Riddle [mailto:riddle-at-bio.fsu.edu] Sent: Wednesday, June 23, 1999 11:57 AM To: microscopy-at-Sparc5.Microscopy.Com
To all,
Any tips, info., papers about the use of bacitracin when negative staining would be greatly appreciated.
Thanks, Kim
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Kimberly A. Riddle Florida State University tel: 850.644.6519 Biological Science Imaging Resource 119 Bio Unit I, 4370 fax: 850.644.0481 Tallahassee, FL 32306 riddle-at-bio.fsu.edu ~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I have been looking at niobium-rich precipitates in a AISI 430-like stainless steel. Using a QBSE detector and the atomic number contrast between the Nb-rich precipitates and the Fe matrix, I can acquire images of bright precipitates against a black background, clear enough for image analysis. I have been trying to use these images for volume fraction determinations, but am unsure whether this is reliable/feasible. Does anyone have any experience with a similar problem?
Thank you very much,
Adam Scott
******************************** Adam Scott MSc Student Department of Materials Engineering University of Cape Town 7701 South Africa Ph: 612929 (h) Ph: 650 3181 (w) Fax: 689 7571 ***********************************
Hello, I'm interested in using a resin for looking at a porous material's pore networks with a bright dye. Could anyone recommend some resins which are useful, or point me in the right direction to get a reference on different resins used for sample preparation and their properties?
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak 1 Cyclotron Road ICQ 23243541 http://nature.berkeley.edu/~gvrdolja MS90-1116 GAVrdoljak-at-lbl.gov Ernest Orlando phone (510) 495-2829 Lawrence Berkeley fax (510) 486-7797 National Laboratory cell (510) 290-6793 Berkeley CA 94720
Can anyone help a colleague to locate the following product or an equivalent?? Also, does someone have an address for a Histo listserver??
Thanks. Ann Lehman Trinity College Hartford, CT
--------------------
What I am seeking is not powdered albumen, but a commerical mixture of albumen, plus glycerine, and bacteriostatic somethings. I believe it is just called "Albumen Fixative" or "Albumen Solution". The word fixative refers to fixing (adhering) the paraffin sections to slides. We use it to coat slides so sections will stick. A large bottle sells for a nomimal price.
1) spatial resolution: I think, it is beneficial to go away from "what can we see", as that involves the observer, to "what can be transmitted". And the transmission is governed by the Nyquist and Shannon limits, which essentially say the same: To transmit a given frequency faithfully, it has to be sampled at twice that frequency. In other words, the theoretical resolution limit is given by half of your pixel frequency. In the real world, there is no abrupt cut-off, but higher frequencies are transmitted with decreasing accuracy. Of course, the resolution of the microscope and other factors play a role as well. If the microscope can resolve .3 nm, there is no point in sampling at 0.03 nm. The resolution will not increase.
2) Regarding significant changes in gray levels: This is strictly a matter of statistics. There are two sources of noise in the images: Noise that is produced by the camera and shot noise from the electron statistics. Let's say, the camera produces a noise of 2 gray values (1-sigma value) and you try to measure 100 electrons, and these electrons (100) produce a gray value of also 100 (to make it easier). The shot noise is then sqrt(100)=10. That is the 1-sigma value. The total 1-sigma noise is then 12 gray values. In other words, if pixel A has a value of 100 and pixel B has a value of 112, there is about a 68% chance that they are actually different. This chance increases to about 95% if Pixel B has a value of 124 and to about 99.7% if pixel B has a value of 136 (3-sigma). As you can see, the values change with the number of electrons, i.e., with exposure.
Hope that helps.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Exchange Administrator Sent: Thursday, June 24, 1999 1:57 AM To: Michael Bode
Hi,
The question of when can you call a feature "resolved" is interesting. I was wondering if it could be described as resolved if its average pixel value and its standard deviation was significantly greater than the average pixel value of its surrounding nieghbors and their standard deviation? The test for significance may tell you the "n" needed or number of pixels needed to be valid. I dont know. What do you think?
Bob Derm Imaging Center University of Washington
On Wed, 23 Jun 1999, Walck. Scott D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Is there an accepted minimum number of pixels that should be used in } determining the linear distance between two resolved points (for example, in } a resolution check) in the image of a sample. This would essentially } identify the minimum magnification required for a given digital image size. } } I'm thinking that the absolute minimum in an image with no noise would be } three (e.g. dark - bright - dark), But more realistically should be } something like 5 or 7. } } Also, what is the minimum difference in contrast levels say for an 8-bit } image to be able to say that the features are in fact resolved? Should the } difference be greater than the square root of the pixel with the highest } brightness? } } Any ideas? } } -Scott } }
If you are taking images of only one plane through your matrix, then I think there is no way you can measure volume fractions with certainty. You are only looking at a 2-dimensional cross section through a 3-dimensional object. Consider, for example, needle-like precipitates. If you cut them perpendicular to the long axis, you will see small circular pecipitates,if you cut them along the long axis, you'll see elongated pecipitates, and the area fraction on your images will change as well.
In other words: If you can assume, that there is no preferred orientation or other anisotropy in your precipitates, you should be able to use the numbers you get from the images. If you can't be sure of that, you probably need to cut the matrix in different directions and compare the results.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Adam[SMTP:ASCOTT1-at-ENGFAC.UCT.AC.ZA] } Sent: Wednesday, June 23, 1999 11:53:37 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: (Backscattered) - Ppts in Stainless Steel } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello
I have been looking at niobium-rich precipitates in a AISI 430-like stainless steel. Using a QBSE detector and the atomic number contrast between the Nb-rich precipitates and the Fe matrix, I can acquire images of bright precipitates against a black background, clear enough for image analysis. I have been trying to use these images for volume fraction determinations, but am unsure whether this is reliable/feasible. Does anyone have any experience with a similar problem?
Thank you very much,
Adam Scott
******************************** Adam Scott MSc Student Department of Materials Engineering University of Cape Town 7701 South Africa Ph: 612929 (h) Ph: 650 3181 (w) Fax: 689 7571 ***********************************
Now how to measure that practically for those of us who don't know how many whatsits are contributing to the signal we are digitizing?
May I suggest that an image be collected from a homogenous area, and maybe a defocused image at that, using the same conditions (signal strength) that would be used on the real image. Thus the only change in brightness from pixel to pixel should be due to random noise from whatever and all sources. A neighborhood of pixels could then be analyzed to determine the standard deviation of the measurements taken at that gray level. The exercise might need to be repeated at the mean gray level of the other feature. And I suppose the same info might be available from a gray level histogram, but the peak width would have to be translated into the standard deviation.
Then I suppose that if you were to have a triplet of pixels with the middle one darker than its neighbors by more than the threshold value, then you could say that you have resolved two features and the resolution of the image is eaual to the spacing of a couple of pixels. If it takes more than one pixel increment for the gray level to drop below the threshold, then would not your image be oversampled to that degree? your pixels are spaced closer than your microscope resolution warrants.
Then there will be the other question. What should the pixel spacing be to get good length measurements on a feature? If I measure a feature at 3 pixels, am I not implicitly saying that it is 3 +/- 1/2 pixels and that I have a 25% uncertainty? Seems that the answer will depend some on the size of the particles being measured. The resolution of the scope will be a limiting factor for small features, but how about larger ones?
Warren S.
Michael Bode wrote:
{snip} 2) Regarding significant changes in gray levels: This is strictly a matter of statistics. There are two sources of noise in the images: Noise that is produced by the camera and shot noise from the electron statistics. Let's say, the camera produces a noise of 2 gray values (1-sigma value) and you try to measure 100 electrons, and these electrons (100) produce a gray value of also 100 (to make it easier). The shot noise is then sqrt(100)=10. That is the 1-sigma value. The total 1-sigma noise is then 12 gray values. In other words, if pixel A has a value of 100 and pixel B has a value of 112, there is about a 68% chance that they are actually different. This chance increases to about 95% if Pixel B has a value of 124 and to about 99.7% if pixel B has a value of 136 (3-sigma). As you can see, the values change with the number of electrons, i.e., with exposure.
It should be a valid method if your surface represents a true cross section through your sample. If inclusions are plucked out during polishing, or if they are selectively etched or left behind during polishing or etching, then you would have errors. Your surface would not be a true sampling of the section.
Another thing to beware of is the choice of threshold level for detecting the particles. If your features are small in relation to your resolution, the measured fraction is very much a function of the choice of threshold level. Try multiple measurements on a single image and see how sensitive (or not) the measurements are to the threshold setting.
At 05:53 PM 6/23/1999 +0000, you wrote: } I have been looking at niobium-rich precipitates in a AISI 430-like } stainless steel. Using a QBSE detector and the atomic number contrast } between the Nb-rich precipitates and the Fe matrix, I can acquire } images of bright precipitates against a black background, clear } enough for image analysis. I have been trying to use these images } for volume fraction determinations, but am unsure whether this is } reliable/feasible. Does anyone have any experience with a similar } problem? } } Thank you very much, } } Adam Scott
} 2) Regarding significant changes in gray levels: } This is strictly a matter of statistics. There are two sources of noise } in the images: Noise that is produced by the camera and shot noise from } the electron statistics. Let's say, the camera produces a noise of 2 } gray values (1-sigma value) and you try to measure 100 electrons, and } these electrons (100) produce a gray value of also 100 (to make it } easier). The shot noise is then sqrt(100)=10. That is the 1-sigma value. } The total 1-sigma noise is then 12 gray values. In other words, if pixel } A has a value of 100 and pixel B has a value of 112, there is about a } 68% chance that they are actually different. This chance increases to } about 95% if Pixel B has a value of 124 and to about 99.7% if pixel B } has a value of 136 (3-sigma). As you can see, the values change with the } number of electrons, i.e., with exposure.
Dear Michael, Most of your two cents are OK, but there are a couple of ringers. First, assuming that the camera noise and shot noise are independent and both are nor- mally distributed, they do not add linearly, but rather as the square root of the sum of the squares. In your example, the total 1-sigma noise would be the square root of 104. Second, the error in the difference of two normally-distributed quantities is, again, the square root of the sum of the squares of the two errors. In your example, assume that the signal from the camera has been dark-current subtracted and that the error in the dark current (=camera noise) is 2, then if the two pixels have values of 100 and 112 with squared errors of 104 and 116, the square of the error of the difference is 220, or the error of the difference is about 15. Furthermore, there are systematic effects for adjacent pixels, so for the question of resolving features, these effects must be taken into account, and they can be complicated. Yours, Bill Tivol
I have appended some references for using bacitracin during negative staining. 'Hope you would find them useful. We use bactracin as a wetting agent routinely and get excellent results (see "Negative Staining" under "Gallery" in our web site (http://www.cimc.cornell.edu). Please get in touch with me if you have any questions.
-------------- 1
TI: MORPHOLOGICAL DESCRIPTION OF SURFACE STRUCTURES ON STRAIN B41 OF BOVINE ENTEROTOXIGENIC ESCHERICHIA-COLI BEARING BOTH K99 AND F41 ANTIGENS AU: DUCHET-SUCHAUX-M. BERTIN-A. DUBRAY-G. SO: J GEN MICROBIOL 134 (PART 4). 1988. 983-996.
AB: In order to describe morphologically the structures on the cell surface of bovine enterotoxigenic Escherichia coli, variants of reference strain B41 (K99+F41+) either negative for K99 and positive for F41 antigens (variants B41A, B41*C), or phenotypically negative for both antigens (variants B41B1, B41B2, B41*CB), and a transconjugant harbouring the K99 plasmid and expressing the K99 adhesin [transconjugant B41 .times. H510a:H510(2)] were examined by transmission electron microscopy using negative staining. Several negative staining procedures were tested for strain B41 and variant B41A: direct harvesting of strains into ammonium molybdate (2% w/v), with bacitracin (50 .mu.g ml-1) as wetting agent, gave the best results. Three morphologically distinct structures on the cell surface could be identified in cultures grown on Minca medium. Firstly, thin, filamentous, flexible fibrillar structures, presenting a helical structure and a mean diameter of approximately 3 nm, were recognized as K99 fimbriae, since they were present on strain B41 and on transconjugant H510(2), but not on K99-negative variants nor on the recipient strain H510a. Secondly, coil-like structures with a diameter of about 17-20 nm were observed on strain B41 and on variants B41A and B41*C. These structures appeared to consist of two more curled filaments (diameter 3 nm) joined to coil on themselves into dense spirals. They were very rare in variants B41B1 and B41B2 and were absent on variant B41*CB and on a transconjugate B41* .times. B41*CB, which had reacquired the K99 plasmid and which again exhibited K99 fimbriae. Strains B41 and variant B41A grown on 37.degree. C for 24 h on sheep-blood agar exhibited coiled structures like those seen on Minca medium. In contrast, after growth at 18.degree. C for 48 h (which inhibits the synthesis of F41 antigen), coiled structures were no longer expressed on the cell surface of strain B41 and of variants B41A and B41*C. Thus the presence of coiled structures correlated with the expression of F41 antigen in strains and variants, which suggests that F41 had a coiled morphology. Finally, straight fimbriae (diameter 6.5-7 nm) were observed on the cell surface of every strain and variant. Their expression on the cell surface was enhanced by several subcultures in static broth, and it was inhibited by subculture on agar, but not by culture at 18.degree. C after serial subcultures in static broth. These facts indicated that the straight fimbriae could be common fimbriae, and excluded their being F41 structures.
2
TI: NEGATIVE STAINING OF PROTEIN MACRO MOLECULES A SIMPLE RAPID METHOD AU: MALECH-H-L. ALBERT-J-P. SO: J ULTRASTRUCT RES.69 (2). 1979. * EN * 191-195.
AB: A simple negative stain technique is described which is suitable for high-resolution imaging of protein molecules in the range of 105-106 daltons. Protein solutions mixed with phosphotungstate stain containing bacitracin are applied to cleaned naked 400- or 500-mesh Cu grids resulting in the formation of thin films that are stable in the electron beam. Since no additional support film is present, the stain films are very thin and provide unusually high resolution images of protein molecules. The method is easy and relatively artifact-free compared to other high-resolution negative stain methods.
3
TI: NEGATIVE STAINING OF PROTEIN MACRO MOLECULES A SIMPLE RAPID METHOD AU: MALECH-H-L. ALBERT-J-P. SO: J ULTRASTRUCT RES.69 (2). 1979. * EN * 191-195.
AB: A simple negative stain technique is described which is suitable for high-resolution imaging of protein molecules in the range of 105-106 daltons. Protein solutions mixed with phosphotungstate stain containing bacitracin are applied to cleaned naked 400- or 500-mesh Cu grids resulting in the formation of thin films that are stable in the electron beam. Since no additional support film is present, the stain films are very thin and provide unusually high resolution images of protein molecules. The method is easy and relatively artifact-free compared to other high-resolution negative stain methods.
4
TI: WETTING AGENTS FOR BIOLOGICAL ELECTRON MICROSCOPY PART 1 GENERAL CONSIDERATIONS AND NEGATIVE STAINING AU: GREGORY-D-W. PIRIE-B-J-S. SO: J MICROSC (OXF).99 (3). 1973 (RECD 1974) 251-265.
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But the area fraction _should_ be the same regardless of anisotropy of particle shape as long as the particles are uniformly distributed and not concentrated one place or another. The area per particle will be smaller if cut across the long axis (instead of along the long axis), but there will be more particles in an image. Conversely for the elongated particle sections. But the area fractions will be the same barring polishing artifacts.
You are touching on the issue of particle size for which there would be a world of dependence on the the section orientation.
At 01:33 PM 6/24/1999 -0600, Michael Bode wrote: } Adam, } } If you are taking images of only one plane through your matrix, then I } think there is no way you can measure volume fractions with certainty. } You are only looking at a 2-dimensional cross section through a } 3-dimensional object. Consider, for example, needle-like precipitates. } If you cut them perpendicular to the long axis, you will see small } circular pecipitates,if you cut them along the long axis, you'll see } elongated pecipitates, and the area fraction on your images will change } as well. } } In other words: If you can assume, that there is no preferred } orientation or other anisotropy in your precipitates, you should be able } to use the numbers you get from the images. If you can't be sure of } that, you probably need to cut the matrix in different directions and } compare the results.
Hi Kim: The reference you want is: David W. Gregory and Brian J. S. Pirie "Wetting agents for electron microscopy of biological specimens" 234-235, Proc. Fifth European Congress on Electron Microscopy,(1972), Manchester,UK. They recommend as a minimum bacitracin concentration required to wet formvar coated grids of 7.5 ug/ml and 10ug for grids with carbon formvar or carbon substrates. The bacitracin can be mixed with distilled H20 and applied first to the grids and after removing all but a trace with filter paper, followed by applying your specimen and then stain solution. It also works well with simply adding to the negative stain solution.
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences Binghamton University Binghamton, NY 13902-6000
We've been doing some work on deposited tantalum, beta phase. When we do FIB sections and image them using ion channeling (ion beam generates secondary electrons for imaging), they look like "camouflage", i.e., NOT columnar or otherwise ordered in any way.
However, when we examine FIB'ed cross sections via TEM, the beta phase material DOES appear somewhat columnar. What gives? Which do we believe and why?
Thanks,
Mona St. Marie Failure Analysis Hewlett-Packard Inkjet
At 08:47 24/06/1999 -0400, you wrote: s!). is there any trick to setting up a Cr } system, or is it as simple as putting a Cr target in a high vacuum } sputtering system?
Cr sputtering demands a much more pure gas mixture than gold/pt/pd as Cr reacts with O2 and N2 and the oxides & nitrides are not as dense or conductive as the metal. For that reason the samples need to be looked at very soon after coating as the Cr oxidises.
i have a turbo pumped vacuum station i'm thinking of } building up into a Cr capable system and wonder if there are things to be } aware of... }
You need to finish up with a very pure low pressure atmosphere of your sputtering gas.
The Cr target has also to have oxide cleaned from it before sputtering with a short cleaning plasma before attempting the coating.
The Xenosput we use achieves all this very neatly. It admits very small quantities of pure xenon in a sealed off chamber then the final pump is carried out sputtering titanium in the coating chamber to scavenge all reactive gas (O2, N2, H2O etc.) and leaving just the xenon for the actual coating sputter.
We still need to take great care to dry all volatile materials from the specimen by warming to 60 deg. C (best overnight) and/or long turbo pumping before the sputter pump. Any traces of other gases (say evolved from the specimen or adhesives) and the sputtering just does not happen despite the creation of a plasma with the right current.
But if you are super careful with the purity of your argon you can approximate the same result in your system.
***************************************************** Mel Dickson, Deputy Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
I used a Noran Voyager EDS system fitted to Hitachi FE-TEM (HF-2000, 200kV) for composition analysis with nanometer scale. (When I analyzing, I aligned the beam spot size as about diameter of 2.5
nm)
I found an interface layer between substrate and epitaxial film on it. In order to identify the interface layer, accurate composition data is needed. I tried it using above EDS system of FE-TEM.
However, I can not use any standard samples for the accurate composition
analysis. (I have no data of K factors which I concerned) So, I quantafy it by automatic calculation using the installed EDS software running on SUN workstation. In that software, it automatically corredted data by "Metallurgical and Biological Thin Section Correction"
So, I wonder about the accuracy. How much the error (percentage) when using that method and that system.
I also analyzed the composition of substrate and epitaxial film on it, at the same time and same analysis conditions. The results is quite well agreed to stochimetry of substrate and epitaxial film on it. (within 5 percentage error range of atomic composition).
Now I want to your kindful helps: 1. Normally, how much the error range by automatic calculation without standard sample.
2. Is there any reference literatures which mentioned the accuracy or error when analyzing the atomic composition using EDS fitted to FE-TEM without standard sample by automatic calculation?
I want to mention the error percentage and references literature for it in my manuscript.
Please help me.
Sincerely Yours.
Soon-Ku Hong. Institute for Materials Research Tohoku University, Sendai 980-8577, Japan Fax:+81-22-215-2074 Tel:+81-22-215-2073
Nestor Zaluzec wrote: ================================================ There is some work a few years ago that suggested that Osmium coating might be better than Cr in the long run.
Here is a reference.
Osmium Conductive Metal Coating for SEM Specimen Using Sublimed Osmium Tetroxide in Negative Glow Phase of DC Glow Discharge ,
A. Tanaka, J. Electron Microsc. 43: 177-182 (1994). ================================================= For those not with ready access to this publication, we have on our website, as part of the information provided for the OPC 40 Osmium Coater, some side by side comparisons, comparing osmium vs. other coating methods as well as other useful information about osmium coating. The URL is http://www.2spi.com/catalog/osmi-coat.html
There is also a link to the US Patent that describes the technology in quite explicit terms.
The advantages of osmium coating are several: a) layer is completely amorphous so there is zero grain size, b) it is a precious group metal and is therefore stable like gold, and does not oxidize or otherwise deteriorate as does chromium, and c) uses an ordinary rotary vane pump instead of a turbo so through-put is much faster. The amorphous nature of the coating is thought to be the reason why higher levels of conductivity are possible with thinner coatings than with other coating systems.
Disclaimer: SPI Supplies distributes the equipment for using this new method for applying conductive coatings so we have a vested interest in promoting this new technology.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi3spi-at-2spi.com
Look for us! ############################ WWW: www.spi.cc ############################ ==================================================
Ann - no opinion on that albumen material. But complete details about the busy Histology listserver can be found on our links page. Use control F and search for "Histonet". There is an internal link to give all info required. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Friday, June 25, 1999 2:10 AM, Lehman, Ann [SMTP:Ann.Lehman-at-exchange.cc.trincoll.edu] wrote: } } Can anyone help a colleague to locate the following product or an } equivalent?? } Also, does someone have an address for a Histo listserver?? } } Thanks. } Ann Lehman } Trinity College } Hartford, CT } } -------------------- } } What I am seeking is not powdered albumen, but a commerical mixture of } albumen, plus glycerine, and bacteriostatic somethings. I believe it is just } called "Albumen Fixative" or "Albumen Solution". The word fixative refers to } fixing (adhering) the paraffin sections to slides. We use it to coat slides } so sections will stick. A large bottle sells for a nomimal price.
Please unsubscribe until further notice, and thanks for all the information. Jerry ______________________ Jerome D. Schick, Ph.D. Semiconductor Devices and Electron Microscopy 26 Kuchler Drive LaGrangeville, NY 12540 Bus (914)223-7393 FAX (914)227-2743 jdschick-at-worldnet.att.net
the good news is that I use a product called Glycerin Albumen for sticking sections to slides - it carries the brand name of Gurr and was supplied by Searle Diagnostics of High Wycombe, England. The bad news is, of course, that I am in the UK.
I do notice that Agar Scientific supply Glycerin Albumen in 100ml bottles - catalog number L4185 price 14.65 UK pounds. If you can't source it in the US they may have a USA agent or their details are below: Agar Scientific Ltd 66A Cambridge Road Stansted Essex CM24 8DA England tel +44 (0) 1279 813519 Fax +44 (0)1279 815106
Good luck
Malcolm
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: jim-at-proscitech.com.au To: 'Lehman, Ann'; 'MSA Listserver'
Ann - no opinion on that albumen material. But complete details about the busy Histology listserver can be found on our
links page. Use control F and search for "Histonet". There is an internal link to give all info required. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Friday, June 25, 1999 2:10 AM, Lehman, Ann [SMTP:Ann.Lehman-at-exchange.cc.trincoll.edu] wrote: } } Can anyone help a colleague to locate the following product or an equivalent?? } Also, does someone have an address for a Histo listserver?? } } Thanks. } Ann Lehman } Trinity College } Hartford, CT } } -------------------- } } What I am seeking is not powdered albumen, but a commerical mixture of } albumen, plus glycerine, and bacteriostatic somethings. I believe it is just } called "Albumen Fixative" or "Albumen Solution". The word fixative refers to } fixing (adhering) the paraffin sections to slides. We use it to coat slides } so sections will stick. A large bottle sells for a nomimal price.
by bellevue.cnrs-bellevue.fr (8.9.1a/jtpda-5.3) with SMTP id PAA24807 for {microscopy-at-sparc5.microscopy.com} ; Fri, 25 Jun 1999 15:06:15 +0200 Message-Id: {2.2.32.19990625132141.00680278-at-bellevue.cnrs-bellevue.fr} X-Sender: riviere-at-bellevue.cnrs-bellevue.fr X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Please I wish to be unscribed for this E-mail: rommelue-at-bellevue.cnrs-bellevue.fr
We have a Philips 400 TEM that needs a new home as it is being replaced by a newer one this summer. The TEM is about 20 years old but is in very good working order and has been maintained by Philips since day one. I would like to see it go to a good home rather than dump it into a skip. We are not asking much for it - only a contribution to removing it carefully from the lab in one piece. If anyone is interested please contact me direct. Derek
******************************* Derek W. Penman Departmental Superintendent Preclinical Veterinary Sciences Royal (Dick) School of Veterinary Studies Summerhall Square Edinburgh EH9 1QH Tel: 0131 650 6087 Fax: 0131 650 6123 E-Mail: dpenman-at-ed.ac.uk
There is a great little book published by the Clay Minerals Society titled: Electron-Optical Methods in Clay Science. It is Volume 2 in their CMS Workshop Lectures series (1990). It is a compilation of workshop lectures ranging from electron microprobe to high resolution TEM. There are some good references to prep methods, their benefits and pitfalls, as well as a wealth of references. It is available for $21 from:
The CLay Minerals Society P.O.Box 4416 Boulder CO 80306
If it is helpful to you, one of the editors is Dr. Ian D.Mackinnon, Advanced Ceramics Development, University of Queensland who, I believe, directed the Electron Microscope center there. Contact me off-line and I'll find his e-mail and some others if you would like.
Tom Kremer Analytical Science & Technology Kimberly-Clark 920-721-4583 e-mail: tkremer-at-kcc.com
} ---------- } From: Walck. Scott D.[SMTP:walck-at-ppg.com] } Sent: Wednesday, June 23, 1999 5:25 PM } To: Micro } Subject: TEM of clay particles } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Could someone tell me the best way to prepare clay particles for TEM. I'm } interested in small particles, micron to sub-micron in size in both the } dry } state and hydrated state. } Thanks in advance. } Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } }
To microscopy list, I have a Sorvall MT-5000 in good working condition for sale. I'm asking $1,500, anyone interested please contact me at (410) 955-1365 work or (410) 889-8009 home
Oops, I think I goofed on that one. Thanks for pointing that out, Bill. Actually, what I wanted to do is to point out, that there are scientific methods to deal with these questions rather than trying to "eyeball" those numbers. I also agree, that there is "crosstalk" between resolution and gray level distinction, but I don't think, that is something that can be resolved in this forum.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: William Tivol[SMTP:TIVOL-at-WADSWORTH.ORG] } Sent: Thursday, June 24, 1999 2:21:13 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Image resolution checks with digital images. } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Michael Bode wrote:
} 2) Regarding significant changes in gray levels: } This is strictly a matter of statistics. There are two sources of noise } in the images: Noise that is produced by the camera and shot noise from } the electron statistics. Let's say, the camera produces a noise of 2 } gray values (1-sigma value) and you try to measure 100 electrons, and } these electrons (100) produce a gray value of also 100 (to make it } easier). The shot noise is then sqrt(100)=10. That is the 1-sigma value. } The total 1-sigma noise is then 12 gray values. In other words, if pixel } A has a value of 100 and pixel B has a value of 112, there is about a } 68% chance that they are actually different. This chance increases to } about 95% if Pixel B has a value of 124 and to about 99.7% if pixel B } has a value of 136 (3-sigma). As you can see, the values change with the } number of electrons, i.e., with exposure.
Dear Michael, Most of your two cents are OK, but there are a couple of ringers. First, assuming that the camera noise and shot noise are independent and both are nor- mally distributed, they do not add linearly, but rather as the square root of the sum of the squares. In your example, the total 1-sigma noise would be the square root of 104. Second, the error in the difference of two normally-distributed quantities is, again, the square root of the sum of the squares of the two errors. In your example, assume that the signal from the camera has been dark-current subtracted and that the error in the dark current (=camera noise) is 2, then if the two pixels have values of 100 and 112 with squared errors of 104 and 116, the square of the error of the difference is 220, or the error of the difference is about 15. Furthermore, there are systematic effects for adjacent pixels, so for the question of resolving features, these effects must be taken into account, and they can be complicated.
Does anyone have a complete reference analysis of U.S.N.M. #116725 Standard Andradite. This garnet was the subject of crystal structure analysis by Novak & Gibbs (1971, Amer. Mineral. 56, 791-825). I found a standard mount of one of these grains in the lab but have no other information except the structural formula given in the paper. Thanks in advance.
Dear Scott, When I used to teach the "Clays" lab and examine kaolinite, halloysite and brucite by TEM, SAED, SEM and EDS, I would prepare the clays by suspending a bit in ethanol and sonicating for 30 seconds, then letting one drop dry on a carbon-coated grid. For SEM the drop would dry on a polished graphite planchet. The kaolinite and halloysite are quite stable, but the brucite is a bit beam sensitive. I suspect the hydrated state is more determined by the vacuum of the EM than anything you do in preparation. At 06:25 PM 6/23/99 -0400, you wrote:
} Could someone tell me the best way to prepare clay particles for TEM. I'm } interested in small particles, micron to sub-micron in size in both the dry } state and hydrated state. } Thanks in advance. } Scott } } Scott D. Walck, Ph.D.
Regards, Mary
} Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
} } What I am seeking is not powdered albumen, but a commerical mixture of } } albumen, plus glycerine, and bacteriostatic somethings. I believe it is } just called "Albumen Fixative" or "Albumen Solution". The word fixative } refers } to } fixing (adhering) the paraffin sections to slides. We use it to coat } slides } so sections will stick. A large bottle sells for a nomimal price.
You can make your own from equal parts of glycerin and lightly beaten egg white (no yolk). Add a bit of thymol to prevent mold. "Animal Tissue Techniques" by Humason (any edition) has the recipe or e-mail me. Other texts on Microtechnique also have the recipe. Albumin-glycerin is less popular these days since egg white contains avidin and interfers with some immunostaining methods.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
We have a Penetron Swirling Shaker, and used to get vials for it from = John's Scientific. We were given a catalogue number from which we could = continue to order them from another company. It appears that they no = longer make them, and I was hoping that someone could tell me of a = supplier that makes comparable vials for this unit. Please reply to me directly. Thanks Susan
Susan Carbyn Electron Microscopy Technician Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada 32 Main Street, Kentville, Nova Scotia B4N 1J5 Canada
The Integrated Microscopy Core of Baylor College of Medicine in Houston = has a Philips 410 TEM for sale. Ancillary equipment includes a = rotating tilt holder, a multiple (3) grid specimen holder and a low dose = unit. This microscope has been on service contract since it was = purchased. We are asking $20,000. The buyer will be responsible for = moving it to its new location.
Hank Adams Technical Coordinator Integrated Microscopy Core Cell Biology Baylor College of Medicine Houston, TX 77030 713 7984952
This is a response to the reply Gary Gill posted after I posted a message questioning his suggestion (in Microscopy Today #99-4) that No.1 cover glasses were a more appropriate choice of thickness than No. 1 1/2 (the original two messages are included at the end of this message). My response, as I interpreted Gary's, should be viewed as a contribution to friendly scientific disagreement and not as a personal criticism of Gary. Furthermore, let me point out that Gary and I may disagree about which size cover glass to use because we have very different preps. He points out in his most recent posting that he is primarily looking at whole mounts of pap smears which vary in thickness. My laboratory uses mostly 0.5 um thick semi-thin plastic resin sections and some 8 um thick paraffin section.
I pulled out a micrometer and did some measurements on some batches of cover glasses sitting around. Fisher #1 1/2 (22 x 22) cover glasses (#12-541B) came in at 180 um (no variation in the 5 tested). Fisher #1 1/2 (22 x 50) cover glasses (#12-544B) ranged from 175 to 180 um. Corning #1 1/2 (22 x 22) cover glasses came in at 180 um (no variation in the 5 tested).
Corning #1 (22 x 22) measured 150 um (no variation in the 5 tested) while a batch of Corning #1 (22 x 60) ranged from 140 to 150 (avg 145).
I should point out that 2 years ago I measured the thickness of the Fisher brand coverslips and they were all about 170 and that is why I chose that brand. Obviously there is some inter-batch variation as well as intra-batch variation.
As an aside, I will point out the microscope slides sitting around my lab showed much more variation:
Fisher Superfrost 3 x 1" x 1 mm (12-550-12): 0.95 1.0, 1.01, 0.98, 0.975.
Fisher Frosted 25 x 75 x 1 mm (12-552): 0.99, 1.00, 0.99, 1.0, 1.0
Clay Adams Gold Seal Rite-on Micro Slides 25 x 75 mm - 0.97 to 1.07 mm thickness (#3050): 0.95. 0.96, 0.995, 1.04, 0.95.
More disturbingly, the flatness of the microscope slides also varied along their length.
Back to the question of the cover glass thickness:
I then took 5 Fisher Frosted slides that all measured 1.00 mm thick in the center of the slide (exact position marked with a diamond pen). These slides all had 0.5 um semi-thin plastic resin sections. They were all coverslipped with a set of cover glasses that all measured 180 um by placing a small drop of Permount (fresh - not overly viscous from sitting around for ages) and pressing the slide down on top of the cover glass with hand pressure for a few seconds and then heating on a slide warmer for a couple of hours. When I re-measured them at the center point, I got something very close to 1180 (my micrometer is marked at 10 um intervals so greater precision than 5 um is somewhat dicey). If I used glass slides that had 8 um paraffin sections (this was the thickness set on the microtome which I realize isn't precise) and coverslipped them using the same method, I got a number equal to the thickness of the glass + cover glass + 10 um (presumably the section thickness + mounting medium). Assuming the paraffin section was close to 8 um, it would imply the mounting medium added about 2 um. Although it would have been better if the No. 1 1/2 cover glasses had been closer to 170 um thick, the percent error in using ones that were 180 um would be less than starting with No. 1 cover glasses that were 150 um thick and hoping to get an even 20 um thick layer of mounting medium.
Finally, let me end with some published comments on cover glass thickness by notable authorities:
"It is therefore best to prepare a microslide with the No. 1 1/2 cover glass." John Gustav Daly in "Photography through the microscope" (1988) 9th edition, p. 20; Eastman Kodak Co.
"Standard coveslips are assumed to be 0.17 +/- 0.01 thick (with a refractive index of 1.515). Number 1 1/2 coverslips are nominally selected for this standard thickness." The author goes on to state that coverslips should be measured for the most critical work. Shinya Inoue (1986) "Video Microscopy" 1st edition. p. 134; Plenum Press, NY.
"No. 1 1/2 generally gives the greatest yield of usable cover glasses." G.P. Berlyn, J. Miksche (1976) Botanical Microtechnique and Cytochemistry. p. 8, Iowa State Univ. Press, Ames.
Original reply from Gary Gill:
} Correction: No. 1 cover glasses range 0.13-0.16 mm thickness; No. 1-1/2, } 0.17-0.19 mm (American Society for Testing Materials. Standard } Specification for Cover Glasses and Glass Slides for Use In Microscopy. } ASTM Designation E211-70, Effective 12.24.70). Or, No. 1 = 0.13-0.17 mm; } No. 1-1/2 = 0.16-0.19 mm (Interim Federal Specification Cover Glass, } Microscope. NNN-C-001434A, 01/08/71). No significant difference. } } Thickness of mounting medium for tissue sections, 3 sets of 4 slides broken } across the section, the broken edges trued up and polished and measured with } a micrometer microscope (Aumonier FJ, Setterington R. Some notes on the } mounting of histological sections. Proc Roy Micr Soc. 1967;2:428-9): } * Cover glass applied routinely (no pressure) = 10, 51, 63, and 76 } micrometers } * Cover glass weighted with 30 gm for 2 days = 18, 18, 20, and 30 } micrometers } * Cover glass with spring loaded clothespin for 72 hours = 5, 10, 10, } and 20 } micrometers } } Therefore, the thickness of mounting medium is substantial relative to the } difference between the range of thickness for No. 1 cover glass and the } tolerance of high dry achromat objectives to deviations from optimal } thickness of 0.17 mm (+/- 15 micrometers and more). Ergo, my recommendation } to use No. 1 thickness cover glasses. A modest bonus is getting more No. 1 } cover glasses per ounce for the same price as for No. 1-1/2. Fluorite and } apochromat objectives have higher NAs power for power than do achromats and } so are even more sensitive to cover glass (and mounting medium) thickness. } Objectives start to show intolerance to cover glass deviations at } approximately 0.6 NA (40X achromat). } } Cytologic preparations (e.g., conventional Pap smears, my field) are more } problematic than histologic sections. Pap smears can sometimes require up } to 12 or more drops of mounting medium to fill in all the valleys of thick } preparations. } } No. 1-1/2 cover glasses are suitable when there is little or no mounting } medium between the specimen and cover glass (e.g., cells grown in culture on } cover glasses, blood films spread on cover glass, cells on Nuclepore filters } dissolved on a cover glass). } } Gary W. Gill }
} } -----Original Message----- } } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] } } Sent: June 22, 1999 11:35 AM } } To: Microscopy-at-Sparc5.Microscopy.Com } } Subject: coverglass thickness } } } } In the May 1999 Microscopy Today (#99-4), Gary W. Gill of Diagnostic } } Cytology Laboratories, Inc., has a short article entitled "Cover Glass } } Perspectives." Although the article has a lot of interesting information, } } at one point the author states that one should not use No. 1 1/2 } } coverglasses even tho they have a nominal thickness of 0.16 to 0.19. He } } says to use No. 1 coverglass to make up for the "substantial and variable } } thickness of the mounting medium." This is counter to everything } } I was ever } } taught. I always use the minimum mounting media possible and press the } } slide down firmly on to the coverslip to ensure this is kept as thin as } } possible. } } } } Isn't standard to use #1 1/2 coverslips? Is there really a school of } } thought that one should use #1's?
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
The long arm of our EH&S division is reaching out to touch me. The goal is to establish a safe way to weigh the hazardous chemicals used in our EM lab.
In the olden days we just took common sense precautions, moved a balance into the hood if we had to, moved it back when we were done. But now that the official list of hazardous chemicals is getting longer and longer, it is now up to 3 pages, this is getting to be a hassle. We also have some users, often students, who do not have the common sense or confidence to know when to use the hood or how to move the balance.
The first shot would be to leave the balance in the hood and make everyone weigh everything there. Drawbacks to this approach are that we are not supposed to 'store' anything in the hoods, they are for doing work. Also the draft from the hood makes the balance reading unstable. We have though about draft shields, but our balance is so old we would have to fabricate one ourselves.
We have thought about a small desktop, HEPA filtered workstation, maybe like the kind some asbestos labs use. This would get the balance out of the hood, keep from contaminating the hood, and maybe do a better job of protecting against particulate dust from chemical powders.
Does anyone have a good plan for complying with modern regulations or do you have some ideas about where to look for free standing, ductless filter cabinets that don't cost a fortune. If necessary we will bite the bullet and get what's needed, but if we can do it with what we have already that would be great.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
We buy 20 ml glass scintillation vials with plastic caps (without aluminum foil liners - these vials are also great for fixing tissues in). We tare a vial with its cap on in our good scale. Then we go to our fume hood, add approximately the correct amount, cap it and re-weigh it. We then go back to the hood and generally add the appropriate number of mls of solvent to come up with a 1 mg/ml solution. We then use a pipetman to add the appropriate number of micro or milli grams to our solution. We have a small electronic scale in the hood for weighing out gram quantities of embedding resins, etc but it is not accurate enough for {100 mg measurements due to the air flow. The students sometimes use that for getting the approximate weight but a trained scientist can usually eyeball about the right amount and then go to the accurate scale for the precise amount. You waste a little but its simple to teach students. Tom
and cap} Hi: } } The long arm of our EH&S division is reaching out to touch me. The goal is } to establish a safe way to weigh the hazardous chemicals used in our EM } lab. } } In the olden days we just took common sense precautions, moved a balance } into the hood if we had to, moved it back when we were done. But now that } the official list of hazardous chemicals is getting longer and longer, it } is now up to 3 pages, this is getting to be a hassle. We also have some } users, often students, who do not have the common sense or confidence to } know when to use the hood or how to move the balance. } } The first shot would be to leave the balance in the hood and make everyone } weigh everything there. Drawbacks to this approach are that we are not } supposed to 'store' anything in the hoods, they are for doing work. Also } the draft from the hood makes the balance reading unstable. } We have though about draft shields, but our balance is so old we would have } to fabricate one ourselves. } } We have thought about a small desktop, HEPA filtered workstation, maybe } like the kind some asbestos labs use. This would get the balance out of the } hood, keep from contaminating the hood, and maybe do a better job of } protecting against particulate dust from chemical powders. } } Does anyone have a good plan for complying with modern regulations or do } you have some ideas about where to look for free standing, ductless filter } cabinets that don't cost a fortune. If necessary we will bite the bullet } and get what's needed, but if we can do it with what we have already that } would be great. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Hi! We use Epon\Aradlite sections to do immunogold technique. Specimen (pancreatic islets) was fixed in paraformaldehyde/low glut fixative. The procedure is three step: goat primary antibody, rabbit antigoat secondary Ab and protein A+gold. We have problems with sections. They do not stay on nickel grids. After procedure the grids are bluish/grinish in colour. It looks as if they were oxidized. We etch the sections with Na metaperiodate for 1H, block with BSA. Our buffer is phophate buffer. I have done this procedure before and I did not have this kind of problem. Is it possible that the grids are too old (they were purchased a few years ago)? Do any of you have an explanation why sections do not stay on the grids and what is causing the change in grids colour? Thank you Dorota
We have just decommissioned a Leica CLSM and are offering the parts for sale. A partial list includes: Omnichrome Ar/Kr laser with ~100 hours of use Omnichrome power supply TMC vibration damping table Leica Fluovert FU inverted microscope Leica microscope objectives (25x 0.75 na, 40x 1.3 na, 100x 1.2 na - all oil) a full set of dichros, filters, etc.
If anyone is interested, please contact us for more information. +++++++++++++++++++++++++++ University of Pennsylvania Biomedical Imaging Core Laboratory Philadelphia, PA 19104
At 6:40 PM -0600 6/24/99, "MONA_STMARIE-at-HP-Corvallis-om3.om.hp.com"-at-Sparc5.Microscopy.Co wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mona,
The columnar structure that you see in cross-section TEM could be stacking faults (or other lattice defects) and not grains. We have observed similar microstructure in TiN films:
"Comparison of Sputtered Titanium Nitride on Silicon Dioxide and Aluminum-Alloy Thin Films," J.L. Drown, S.M. Merchant, M.E. Gross, D. Eaglesham, L.A. Giannuzzi, R.B. Irwin, Microscopy and Microanalysis, vol. 3 suppplement 2, (1997), 469.
Regards, Lucille
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-mail.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
I want to say thanks to all of the people who responded to my staining problem. I received over 25 responses, 5 within the=20 first 3 hours it was posted. Generally the responses fell into three groups: 1. Don't use Spurr's media 2. Cut thicker sections 3. Stain with LC, UA, and again with LC
The investigator has decided to redo the experiment and we will use one of the Epon media and cut slightly thicker sections without a film.
Thanks again. I am always amazed at how many responses one get and especially how fast.
George Lawton Chief Electron Microscopist Microscopy and Imaging Service Center UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, Tx 75235-9039 Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
----- Original Message ----- } From: David Henriks {Henriks-at-CompuServe.COM} To: Micro Listserver {microscopy-at-Sparc5.Microscopy.Com}
Project Engineer
Performs engineering studies, design (including drawings and = schematics), testing, and evaluation of intricate miniature mechanical = and electromechanical devices or systems involving vacuum, ion/electron = optics, and cryogenic technologies. This involves = development-engineering activities related to the commercialization of = new products, systems, and services. Activities may include = conceptualization of new products, feasibility studies, prototype = design and implementation of customer specifications, materials = selection, cost estimation, selection and design of equipment and = systems, production startup and performance verification, customer = training and follow-up services. Interfaces with marketing, sales, and = research in defining objectives and priorities for projects. Is = proficient in the use of geometric tolerancing, CAD systems, and = plotters in order to prepare detailed engineering documentation. = Supervises drafting staff and is responsible for the production of all = engineering documentation including test specifications and procedures. = Must be familiar with regulatory standards (CSA, SEMI, CE, VDE, PTB). = Provides existing product line support in design activities and/or = modifications of hardware for production. Typical assignments are = complex and require the use of initiative and judgment. Requires a = degree in mechanical engineering or engineering physics. An advanced = degree is preferred, combined with a minimum of 8 years = engineering/supervisory experience.
E.A. Fischione Instruments, Inc. is an equal opportunity employer.
Resumes and salary requirements should be sent to:
Human Resources Director, MLS E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 Phone (724) 325-5444 FAX (724) 325-5443 E-mail: info-at-fischione.com
Renee Kalmes rkalmes-at-exponent.com -----Original Message----- } From: Jerome D. Schick [mailto:JDSchick-at-worldnet.att.net] Sent: Friday, June 25, 1999 3:49 AM To: Microscopy-at-sparc5.microscopy.com
Please unsubscribe until further notice, and thanks for all the information. Jerry ______________________ Jerome D. Schick, Ph.D. Semiconductor Devices and Electron Microscopy 26 Kuchler Drive LaGrangeville, NY 12540 Bus (914)223-7393 FAX (914)227-2743 jdschick-at-worldnet.att.net
Accidentally delete files folders? =46disk or =46ormat the wrong drive= s? Lose data because of a virus? Are you or have you seen errors like, invalid drive specification, invalid media type or error reading drive c:? Hard drives are getting cheaper and cheaper, but the data on those drives can be invaluable.
However you have options. You can send the drive to a Data Recovery Professional and pay the price (thousands). You can buy an over the counter retail product which may due more harm than good, or you can use the software that the professionals use for this type of recovery . We are for a limited time, making the software we sell to the professionals available to the general public to handle the out cry for data recovery software we have experienced due to virus infection , and the simple fact that almost everyone now has a computer. Virus scanners are great, but the fact is they can not update virus signatures as fast as people are producing viruses.
More and more people are learning how to program. Unfortunately that means there are more programmers capable of producing viruses. Major companies spending millions of dollars on security and virus protection have lost data due to virus infection. It will get worse before it gets better. You don't have to spend thousands to recover your data if you have the right software. We are making this software available to you now, at a very reasonable cost! Act now and receive unlimited free technical support! This won't last long!
=46or more information please reply to: mailto:roon99-at-writemail.com?subject=3Dmore-info
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Accidentally delete files folders? =46disk or =46ormat the wrong drive= s? Lose data because of a virus? Are you or have you seen errors like, invalid drive specification, invalid media type or error reading drive c:? Hard drives are getting cheaper and cheaper, but the data on those drives can be invaluable.
However you have options. You can send the drive to a Data Recovery Professional and pay the price (thousands). You can buy an over the counter retail product which may due more harm than good, or you can use the software that the professionals use for this type of recovery . We are for a limited time, making the software we sell to the professionals available to the general public to handle the out cry for data recovery software we have experienced due to virus infection , and the simple fact that almost everyone now has a computer. Virus scanners are great, but the fact is they can not update virus signatures as fast as people are producing viruses.
More and more people are learning how to program. Unfortunately that means there are more programmers capable of producing viruses. Major companies spending millions of dollars on security and virus protection have lost data due to virus infection. It will get worse before it gets better. You don't have to spend thousands to recover your data if you have the right software. We are making this software available to you now, at a very reasonable cost! Act now and receive unlimited free technical support! This won't last long!
=46or more information please reply to: mailto:roon99-at-writemail.com?subject=3Dmore-info
********************************************************************* ** To be removed reply to mailto:petlm-at-hotbot.com?subject=3Dremove ********************************************************************* **
Accidentally delete files folders? =46disk or =46ormat the wrong drive= s? Lose data because of a virus? Are you or have you seen errors like, invalid drive specification, invalid media type or error reading drive c:? Hard drives are getting cheaper and cheaper, but the data on those drives can be invaluable.
However you have options. You can send the drive to a Data Recovery Professional and pay the price (thousands). You can buy an over the counter retail product which may due more harm than good, or you can use the software that the professionals use for this type of recovery . We are for a limited time, making the software we sell to the professionals available to the general public to handle the out cry for data recovery software we have experienced due to virus infection , and the simple fact that almost everyone now has a computer. Virus scanners are great, but the fact is they can not update virus signatures as fast as people are producing viruses.
More and more people are learning how to program. Unfortunately that means there are more programmers capable of producing viruses. Major companies spending millions of dollars on security and virus protection have lost data due to virus infection. It will get worse before it gets better. You don't have to spend thousands to recover your data if you have the right software. We are making this software available to you now, at a very reasonable cost! Act now and receive unlimited free technical support! This won't last long!
=46or more information please reply to: mailto:roon99-at-writemail.com?subject=3Dmore-info
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I have some problems with MT 7000 ultramicrotome and I need electronic circuit schematics. Can anyone help me?
Thank you!
Best regards
Sorin
********************************************* Sorin Lazar Dept. Electron Microscopy & Image Analysis National Institute for Biological Sciences Spl. Independentei, nr. 296, Bucharest Romania
We have a Philips 301 electron microscope that we would like to give to another non-profit who can use it. When last used it worked perfectly but no longer has a service contract.
Hoping for some assistance. I have been asked to research into costs and sources for sound proof material. To be more specific we want to reduce noise in a couple TEM rooms. We expect to acquire a high voltage FE TEM this fall. I have seen a foam product, usually black, attached to the walls in labs before. Everyone called it "eggshell" or "egg crate". Does anyone know of vendors or sources of same or similar products? Thanks in advance.
Joel McClintock EM Specialist U. of Kentucky (606)257-1242 jmcclin-at-pop.uky.edu
{fontfamily} {param} Times_New_Roman {/param} {bigger} RESEARCH ASSOCIATE- TECHNICIAN POSITION OPENING
UNIVERSITY OF ILLINOIS, URBANA-CHAMPAIGN
{/bigger} {/fontfamily} {bigger} {fontfamily} {param} Times {/param} {bigger} Qualif= ications:
Minimum requirement: Bachelor of Science degree. Applications from Masters or Ph.Ds are welcome. Postdoctoral fellowship appointments are possible. Previous lab and/or electron microscopy experience required.
Responsibilities:
To perform research in a laboratory of cell biology and structural biology. Work will combine electron microscopy and light microscopy while including tissue culture, immunostaining, and molecular biology. TEM will be done on a Phillips CM-200 equipped with a CCD camera. Work will include assisting with EM tomography and serial thin section 3-D reconstructions.
Closing Search Date: Applications will be accepted until the position is filled (as late as Oct. 1, 1999). Full consideration will be given to applications received prior to July 1, 1999 or until the position is filled.
=09
=09
The University of Illinois at Urbana-Champaign is an affirmative action, equal opportunity employer.
Can someone tell me the correlation between rpm's and g's when using centrifuges? We have a protocol that specifies 7.5 g's for a centrifugation step, but our readout is, of course, in rpm's. I assume there must be table somewhere, but I don't recall ever seeing one.
Thanks.
Randy Tindall Electron Microscope Specialist Electron Microscope Core Facility University of Missouri Columbia, MO 65211
Good Morning Everyone, Does anyone out there know how to assemble the carbon coating apparatus on a Polaron E6100 sputter coater (approx. 1985 model)? Although I know how to carbon coat, however, but not on this particular model. These parts are sitting in a bag and the instructions are very vague with no pictoral info. Help! Thank you all in advance for your help, Maria
Maria Fazio-Zanakis Bioimaging and Molecular Histology Hoechst Marion Roussel, Inc. 1-908-231-3357 Fax: 1-908-231-3962 Email: Maria.Fazio-Zanakis-at-hmrag.com
Good Morning Again Everyone, I need some assistance and/or advise on how to change the oil on a Polaron sputter coater E6100 that has not been changed for a long time. I can't seem to obtain a vacuum greater than 4 x 10 -4 torr. I don't mind changing it if this is the only recourse. Thanks again, Maria
Maria Fazio-Zanakis Bioimaging and Molecular Histology Hoechst Marion Roussel, Inc. 1-908-231-3357 Fax: 1-908-231-3962 Email: Maria.Fazio-Zanakis-at-hmrag.com
You are in luck. I needed this info and so searched for it. Nice guy, eh?
The formula is RCF = 0.00001118 x r x N squared
where RCF = relative centrifugal force r = rotating radius in centimeters N = rotating speed in revs per minute
I have a neat nomograph which I can fax to you. But you gotta give me a fax number that will work this time.
Also, remember to do the calculation from where the specimen will actually reside rather than the radius of the rotor. In a test tube which covers some distance remember that the forces will vary over the distance. It may be safest in this case to use the midpoint of the test tube. That's what I do.
Cheers,
JB
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
The foam panels we used are manufactured by Illbruck Acoustic Products, 3800 Washington Ave. N., Minneapolis, MN 55412, (612) 520-3620 or (800) 662-0032, fax (612)521-5639. They may have a dealer in your area. The local dealer we worked through was The Huff Co., Inc., 28915 Herky Dr., Suite 109, Lake Bluff, IL 60044, (847) 362-7440, fax (847) 362-0427.
More specifically, the panels are SonexOne, which have a ASTM E84 Class 1 flammability rating. They come in 2" and 3" thicknesses. We installed 2" panels for a JEOL JEM-4000EX and a Philips CM30T and 3" panels for an Hitachi H-9000. It can be ordered in the natural white or with a painted surface (we ordered a gray-painted variety for the 2" panels) or with a Hypalon polymer surface in different colors (we ordered a black Hypalon for the 3" panels). Our cost for the 2" painted panels was $290 per box (64 sq. ft. per box in 2'x4'panels).
In general, the entire room doesn't need to be covered with this foam. Indeed, only the walls of the separate utility rooms for our JEM-4000EX and Philips CM30T are covered, and the dealer thought that we didn't need to cover as much as we did. The utility rooms contain most of the noise-generating components (air-handling units, water chillers, pumps, etc.). The entire room for the H-9000 is covered.
You'll also need to buy some cartridges of latex multi-purpose construction adhesive which can be used for vinyl foams and plastics. We bought some from Grainger. You might be able to find some locally, or the foam dealer may have a recommendation. The Huff Co. said that "Liquid Nails" works ok, but we bought "OSI Pro-Series QB-350". You'll need about 1 cartridge per 32 sq. ft.
You must realize, however, that these panels are better at absorbing higher frequencies. For low frequencies, other strategies may be necessary: floating floors, high-density foam panels for cabinets, etc. } -----------------------------------------------------------------------. } } Hoping for some assistance. I have been asked to research into costs and } sources for sound proof material. To be more specific we want to reduce } noise in a couple TEM rooms. We expect to acquire a high voltage FE TEM this } fall. I have seen a foam product, usually black, attached to the walls in } labs before. Everyone called it "eggshell" or "egg crate". Does anyone know } of vendors or sources of same or similar products? Thanks in advance. } } Joel McClintock } EM Specialist } U. of Kentucky } (606)257-1242 } jmcclin-at-pop.uky.edu
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798 cook-at-aaem.amc.anl.gov
Randy: I think this is the correct info. You need to know the distance from the center of the rotor to the center of the tube to calculate the average g. It is much easier to use a table specific for the rotor. But if one is not available:
Relative Centrifugal Field (x g) = 1.12 r (RPM / 1000)(RPM /1000)
Note that I couldn't figure out how to write a superscript number to indicate you need to square the"(RPM /1000)" so I wrote it twice on purpose.
r = radius (usually used r (average) or the center point but also can use r (max) or r (min). This is for nonprecipitating solutions with a density less than 1.2 g/ml.
The angle of the rotor is not in this equation but it is important in the pelleting speed I believe. I think this is what the manufacturers call the "k factor".
Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I have a Link Analytical Lemas stage control system on an SEM. The control screen displays "status reset" and the joystick does not operate the stage system. I have tried to reset the system but have not been successful. If anyone has one of these systems and can help me diagnosis this problem, I would be very thankful.
Phoebe J. Doss Manager/Adjunct Instructor EM Lab Oklahoma State University
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hoping for some assistance. I have been asked to research into costs and } sources for sound proof material. To be more specific we want to reduce } noise in a couple TEM rooms. We expect to acquire a high voltage FE TEM this } fall. I have seen a foam product, usually black, attached to the walls in } labs before. Everyone called it "eggshell" or "egg crate". Does anyone know } of vendors or sources of same or similar products? Thanks in advance.
Try MarkerTek in NY. They have an 800# and a website.
I'm not clear if you want to reduce sound coming into the rooms or reduce reverberant sound generated within the room, in either case try IAC at "http://www.industrialacoustics.com/index.htm", they have solutions for most sound control problems.
Joel McClintock wrote:
} Hoping for some assistance. I have been asked to research into costs and } sources for sound proof material. To be more specific we want to reduce } noise in a couple TEM rooms. We expect to acquire a high voltage FE TEM this } fall. I have seen a foam product, usually black, attached to the walls in } labs before. Everyone called it "eggshell" or "egg crate". Does anyone know } of vendors or sources of same or similar products? Thanks in advance. } } Joel McClintock } EM Specialist } U. of Kentucky } (606)257-1242 } jmcclin-at-pop.uky.edu
--
Richard J. Mount Auditory Science Laboratory, Department of Otolaryngology & Brain and Behaviour Division/Research Institute The Hospital for Sick Children Toronto, Ontario, Canada (416) 813-6551; Fax (416) 813-8456 http://www.sickkids.on.ca/otolaryngology/Earhome1.asp
Hello, Just a quick question. I've got a metamorphic rock about 1"x3" I'd like to impregnate with a resin with flourescent dye to look at porosity. I've heard there is a flourescent dye you can add to LR White for just such a purpose. Does anyone know which one it is? Also, if there is an alternate resin/flourescent dye to use, could anyone let me know? Sincerely,
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak 1 Cyclotron Road ICQ 23243541 http://nature.berkeley.edu/~gvrdolja MS90-1116 GAVrdoljak-at-lbl.gov Ernest Orlando phone (510) 495-2829 Lawrence Berkeley fax (510) 486-7797 National Laboratory cell (510) 290-6793 Berkeley CA 94720
I am trying to locate calcium sites in fungal cells embedded in EPON resin. Has anybody any suggestions about the floating medium I could use during thin sectioning to avoid the dissolution and loss of the calcium from the cells?
We have had good results using Armstrong Soundsoak panels. The panels look nicer and have just about the same acoustic properties as the 2 inch eggcrate foam. Also, the vinyl coating is cleanable whereas most of the foam eggcrate is not. Obviously 4 inch or 6 inch eggcrate will do better, but there are diminishing returns (as well as diminishing space in your room). Low frequency is a problem no matter which type of panel you use.
Cheers, Henk
At 10:22 AM 6/28/99 -0400, Joel McClintock wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu An optimist believes that we live in the best of all possible worlds. A pessimist fears that this is true.
We need to have information on Thickness monitors that can be used in a Denton 502A. Basically we are looking for one which has the greatest range of motion since I understand that generally because of the water cooling they cannot move too much. Any and/or all information welcome. If I get enough I will catalog and make available to anyone who asks.
Thanks in advance.
Mei Lie Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
One source of the material you are looking for is a company called Illbruck and goes by the brand name of Sonex. There are a variety of grades with different types of coatings, colors, sound reduction levels, and also fire ratings. I use it extensively in the VG HB603z FEG AAEM room (http://tpm.amc.anl.gov). It's not cheap, but installation is simple. Watch out for your local fire codes as some brand of "foam-based" sound proofing are not adequately fire rated for general laboratory usage.
Nestor
Your Friendly Neighborhood SysOp
} } } Hoping for some assistance. I have been asked to research into costs and } sources for sound proof material. To be more specific we want to reduce } noise in a couple TEM rooms. We expect to acquire a high voltage FE TEM this } fall. I have seen a foam product, usually black, attached to the walls in } labs before. Everyone called it "eggshell" or "egg crate". Does anyone know } of vendors or sources of same or similar products? Thanks in advance. } } Joel McClintock } EM Specialist } U. of Kentucky } (606)257-1242 } jmcclin-at-pop.uky.edu
-West Marine has a foam/lead product that the use for engin covers that damps vibration well.
If the wall are not built yet build a wall with offset studs so that there in no physical connection between the surfaces of the faces of the two walls. then fill the space with fiberglass insulation.
Gordon
Gordon Couger gcouger-at-couger.com TDY Vancouver, B.C. 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Are you seeking calcium by x-ray microanalysis or EELS?
We have done XRMA since 1981. Our standard approach is to freeze-dry, otherwise you lose all the diffusible and maybe loosely bound elements with routine chemical processing, then infiltrate in resin (generally Spurr's) in some sort of vacuum situation. The tissue is not osmicated because of interfering peaks (for some elements).
We always cut thick sections, 1, 1.5 or 2 microns, on DRY glass knives. The grids, generally aluminium, formvar film coated, are held in reverse-action or clamped forceps (with a small O-ring?) and balanced on a match box or with BluTack so that the grid is just behind the knife edge. The sections are manipulated onto the grids with an eyelash. They are presed down with a polished metal rod although a technician we used to have could "patch-weld" them to the film with just the eyelash. Sometimes, if they stick to the rod, I sandwich them between two grids, press and separate. You can usually use the empty grid for the next sections. They are carbon coated prior to analysis (for conductance i.e. anti-charging) in the STEM.
The reason for thick sections is that you get extremely low count rates with ultra-thin sections. We can also use up to 200 kV for the really thick sections. Obviously you want to remove the objective aperture for the analysis. The images are nothing like conventional images!! But the chemical goodies should be present!!
You can do thin sections with the blocks by normal methods and contrasting first with osmium vapour from any osmium fixative if you don't want to dedicate some crystals to do this. It is possible that thin sectioning (with a water bath) could leach some elements from the surface layer of the block if it gets wetted.
If you are doing this for EELS, please post a synopsis of replies - I would be interested!
Good luck - Keith
Keith Ryan Marine Biological Association of the UK Plymouth, England
If you want to reduce the noise generated by the microscope and ancillaries in the TEM room then curtains around the walls are cheap and effective. Works well for our FEGTEM. Of course it will not cut out any bangs and crashes from adjacent rooms.
Ron
On Mon, 28 Jun 1999 10:22:22 -0400 Joel McClintock {jmcclin-at-pop.uky.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hoping for some assistance. I have been asked to research into costs and } sources for sound proof material. To be more specific we want to reduce } noise in a couple TEM rooms. We expect to acquire a high voltage FE TEM this } fall. I have seen a foam product, usually black, attached to the walls in } labs before. Everyone called it "eggshell" or "egg crate". Does anyone know } of vendors or sources of same or similar products? Thanks in advance. } } Joel McClintock } EM Specialist } U. of Kentucky } (606)257-1242 } jmcclin-at-pop.uky.edu } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Dear list, I'm in need of a formula for a staining technique. I've read that Orcein could be used to stain gums (i.e., gum arabic). If you have any information it would be greatly appreciated. Thanks.
John B. Crowe jcrowe-at-ora.fda.gov US FDA Forensic Chemistry Center 6751 Steger Dr. Cincinnati, OH 45237 phone (513)679-2700 fax (513)679-2761
"A PHILIPS EM-400T-FEG with ECON-EDAX system is available to anybody who can assume responsibility for all moving costs. It is currently not in use but was in use prior to shut-down. The microscope had an extra FEG gun and one FE tip. Comes with all baking equipment for the FEG. It is a good system for anybody who needs a second instrument and willing to set it up and get it running. The micrsocope is in the New Haven area in Connecticut. Anyone interested in the microscope should contact me by e-mail or phone. } } R. Ayer } {ayerr-at-aol.com} } Phone 203-389-6065 } Fax 203 876 8914
I use sodium chloride salt windows (optical crystals) for FT-IR microspectroscopy analysis of samples. Where would one look for information on cleaning the salt windows after analysis? I have not found a source that covers this topic on cleaning any type of crystals (KBr, KCl, NaCl, etc....).
Jeff Nicklaw
} } Email: nicklaj-at-basf.com } Name: Jeff Nicklaw } }
Cleaning the grids with acetic acid for 5-10 minutes (then rinse with water and dry them in an oven) will greatly improve your situation. The grids can be 'old', but the acid removes the oxidation that inhibits your section adhesion.
Gregg Sobocinski Parke-Davis Pharmaceuticals Ann Arbor, MI Gregg.Sobocinski-at-wl.com
-----Original Message----- } From: Dorota Wadowska [mailto:wadowska-at-upei.ca] Sent: Friday, June 25, 1999 4:21 PM To: microscopy-at-Sparc5.Microscopy.Com
Hi! We use Epon\Aradlite sections to do immunogold technique. Specimen (pancreatic islets) was fixed in paraformaldehyde/low glut fixative. The procedure is three step: goat primary antibody, rabbit antigoat secondary Ab and protein A+gold. We have problems with sections. They do not stay on nickel grids. After procedure the grids are bluish/grinish in colour. It looks as if they were oxidized. We etch the sections with Na metaperiodate for 1H, block with BSA. Our buffer is phophate buffer. I have done this procedure before and I did not have this kind of problem. Is it possible that the grids are too old (they were purchased a few years ago)? Do any of you have an explanation why sections do not stay on the grids and what is causing the change in grids colour? Thank you Dorota
Does anyone have a copy of Kessel and Kardon's "Tissues and Organs" (Freeman, 1979) they'd be willing to sell me?
Many thanks, Dee
Dee Breger Manager, SEM/EDX Facility Lamont-Doherty Earth Observatory Route 9W Palisades NY 10964 USA T: 914/365-8640 F: 914/365-8155 I: www.ldeo.columbia.edu/micro "Journeys in Microspace" (Columbia University Press, 1995) ____________________________________________________________________________ Automatic note: Sometimes I don't receive incoming emails (with no notice to the sender). If I don't respond to your message, please send it again! ____________________________________________________________________________ _
PhD in physics in search for a post doctoral. io will have finished my thesis on Sept 99. I'm actually working at Reims University (FRANCE) in the laboratory of analysis solid surfaces and interfaces. My research consists in exploration for new imaging technique X ray in total reflection (Application : topography observation of surface and interface solid/solid) and fluorescence X. In the team we use X ray microscopy in electrochimical systems. My competences : X ray microscopy : Projection, reflection and fluorescence Glancing X ray : Reletometry, TXRF, GIXF Experimentator data analysis preparation of thin layer by evaporation preparation of X-visible screen converter for image guide ........ Please send Email to receive my CV and details on my work. N.B : I will take part in XRM 99 conference to be held in Berkeley, CaliforniaAugust 1-6, 99. Sincerly
I've polished NaCl single crystals with an ethanol/water mix on a polishing cloth. Sorry, I can't remember the mixture ratio, but it had mostly ethanol to start with (90/10??). -Scott ---------- } From: Jeffrey M Nicklaw To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Colleagues
Anyone know the answer to this question? Please reply directly to the author as well as the Listserver.
I use sodium chloride salt windows (optical crystals) for FT-IR microspectroscopy analysis of samples. Where would one look for information on cleaning the salt windows after analysis? I have not found a source that covers this topic on cleaning any type of crystals (KBr, KCl, NaCl, etc....).
Jeff Nicklaw
} } Email: nicklaj-at-basf.com } Name: Jeff Nicklaw } }
The College of Natural Resources and Sciences at Humboldt State University is currently recruiting to hire an Equipment Technician III. Could you please tell me if it is possible to place a notice on your listserver? What is involved and what is the cost?
Thank you.
Barbara Westmoreland, AA/S College of Natural Resources and Sciences Humboldt State University Arcata, CA 95521-8299 (707) 826-5826 -- Phone (707) 826-3562 -- FAX westmore-at-laurel.humboldt.edu
It will be greatly be appreciated if anybody can give me information regarding
a. The type of detector the KE 4-quadrant BSE detector. b. The country and city where KE-4 quadrant BSE is made.
Thanks for the help. Andre Wong Faculty of Dentistry, UBC, 2199, Wesbrook Mall, Vancouver, BC V6T1Z3. tel. 604-822-2873 Fax 604-822-3562 e-mail: andywong-at-unixg.ubc.ca
Responding to the message of {v04011700b39d3b798264-at-[128.206.162.35]} from Tom Phillips {PhillipsT-at-missouri.edu} : } } Randy: I think this is the correct info. You need to know the distance } from the center of the rotor to the center of the tube to calculate the } average g. It is much easier to use a table specific for the rotor. But } if one is not available: } } Relative Centrifugal Field (x g) = 1.12 r (RPM / 1000)(RPM /1000) } } Note that I couldn't figure out how to write a superscript number to } indicate you need to square the"(RPM /1000)" so I wrote it twice on purpose. } } r = radius (usually used r (average) or the center point but also can use r } (max) or r (min). This is for nonprecipitating solutions with a density } less than 1.2 g/ml.
snip!
There seems to be a discrepancy of a factor of 10 in the numerical constant in the two equations for relateive centrifugal field presented by Bozzola - 11.18x10(-6) - and the above presented by Phillips - 1.12x10(-6).
Which is the correct one?
Thanks,
Gib
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu http://biosci.umn.edu/MIC/consortium.html
Harrick Scientific Corporation is a supplier of IR-VIS-UV accessories. According to a blurb sheet on an Internal Reflection Accessory we recently purchased "crystals are best cleaned in organic solvents such as toluene or MEK. Plasma discharge cleaning is also very useful for certain materials, but should not be used on KRS-5 cystals. Harrick's number is (914) 762-0020 should you have any other questions regarding the crystals they supply.
Regards, diana
Walck. Scott D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I've polished NaCl single crystals with an ethanol/water mix on a polishing } cloth. Sorry, I can't remember the mixture ratio, but it had mostly ethanol } to start with (90/10??). } -Scott } ---------- } } From: Jeffrey M Nicklaw } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Cleaning sodium chloride salt windows } Date: Tuesday, June 29, 1999 10:03AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Colleagues } } Anyone know the answer to this question? Please reply directly to the } author as well as the Listserver. } } Nestor } } ========================================================= } } Nestor } } I use sodium chloride salt windows (optical crystals) for FT-IR } microspectroscopy analysis of samples. Where would one look for information } on } cleaning the salt windows after analysis? I have not found a source that } covers } this topic on cleaning any type of crystals (KBr, KCl, NaCl, etc....). } } Jeff Nicklaw } } } } } Email: nicklaj-at-basf.com } } Name: Jeff Nicklaw } } } }
The difference is that my "r" was meant to be the radius in "mm" whereas John's was "cm". Therefore the equations are both correct. My original posting should have given the units. Sorry for the confusion. Tom
} } Responding to the message of {v04011700b39d3b798264-at-[128.206.162.35]} } from Tom Phillips {PhillipsT-at-missouri.edu} : } } } } Randy: I think this is the correct info. You need to know the distance } } from the center of the rotor to the center of the tube to calculate the } } average g. It is much easier to use a table specific for the rotor. But } } if one is not available: } } } } Relative Centrifugal Field (x g) = 1.12 r (RPM / 1000)(RPM /1000) } } } } Note that I couldn't figure out how to write a superscript number to } } indicate you need to square the"(RPM /1000)" so I wrote it twice on purpose. } } } } r = radius (usually used r (average) or the center point but also can use r } } (max) or r (min). This is for nonprecipitating solutions with a density } } less than 1.2 g/ml. } } snip! } } There seems to be a discrepancy of a factor of 10 in the numerical } constant in } the two equations for relateive centrifugal field presented by Bozzola - } 11.18x10(-6) - and the above presented by Phillips - 1.12x10(-6). } } Which is the correct one? } } Thanks, } } Gib } } Gib Ahlstrand } Electron Optical Facility, University of Minnesota, Dept. Plant Pathology } 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 } 612-625-9728 FAX, giba-at-puccini.crl.umn.edu } http://biosci.umn.edu/MIC/consortium.html
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
We have a Noran Voyager EDS system that has the capability to capture grey scale images from the SEM. We are interested in downloading those images. Does anyone know if it is possible to transfer those images (or any Noran file for that matter) from the EDS to a Zip Drive?
TIA
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (920) 42403404 wise-at-uwosh.edu http://www.uwosh.edu/departments/biology/wise/wise.html
HI I AM AN OLD JEOL SERVICEMAN OF TWENTY YEARS AND MY NAME IS MIKE SO I READ YOUR LETTER A LITTLE. YES YOUR PROBLEM WHICH IS COMMON IS IN THE ALPHA NUMERICS FACED IT MAY A TIME THE ANWER IS CORRECT CLEANING THE CONNTACTS USUALLY DOES IT, BUT ALSO RESET ALL THE BOARDS IN THE CPU AREA AND RIBBION CONNECTORS.
Good Morning Everyone, Does anyone out there know how to assemble the carbon coating apparatus on a Polaron E6100 sputter coater (approx. 1985 model)? Although I know how to carbon coat, however, but not on this particular model. These parts are sitting in a bag and the instructions are very vague with no pictorial info. Help!
and:
Good Morning Again Everyone, I need some assistance and/or advise on how to change the oil on a Polaron sputter coater E6100 that has not been changed for a long time. I can't seem to obtain a vacuum greater than 4 x 10 -4 torr. I don't mind changing it if this is the only recourse. Thanks again, Maria
Dear Maria,
I am contacting you as the original manufacturers of the E6100 Bench Top Evaporator. Although the E6100 has now been replaced with the E6300, we continue to support the E6100 and would be please to be of assistance.
Our local representative, Energy Beam Sciences Inc. (see below) will be in contact.
Energy Beam Science Inc. 11 Bowles Road, PO Box 468 Agawam MA 01001 USA Tel: 413 786 9322 Fax: 413 789 2786 ebs-at-ebsciences.com
Best regards Mike Wombwell Polaron range Business Manager V G Microtech The Birches Industrial Estate Imberhorne Lane East Grinstead West Sussex RH19 1UB UK Direct line: +44 (0)1342 310296 Switchboard: +44 (0)1342 327211 Fax: +44 (0)1342 315074 http://www.polaron-range.com E&OE
Here are some of the system parameters used in the design of our high resolution sputtering systems.
VACUUM: 250l/s dry turbo pump that produces vacuum levels {3X10-6 in 4 minutes. LN2 cold trap is optional but suggested to pump residual water vapor for refractory metal deposition. SPUTTERING MECHANISM: Ion beams are used to remove target material. The benefit to ion beam sputtering is that the sample is not exposed to plasma accelerating potential and the material evolves from targets at {30ev thereby eliminating radiation effects to samples. Ion beam sputtering is slow, approximately 10A/minute, but precisely controllable. Conductive thin films can be deposited at thickness selected by operator with proper mounting and tooling factor calibration of the quartz thickness microbalance. DRIVING GASES: 5 nine pure Ar gas is used in ion sources and dry N2 gas is admitted through the turbo when venting. TARGET MATERIALS: Iridium or platinum for FESEM imaging at {200kX. Thin films of Ir and Pt are indistinguishable. Ir films have the benefit over Pt of minimizing beam sensitive samples. When imaging in FESEM exceeds 200kX any of the refractory metals, Ta, Ti, W and Cr are suggested. Samples with refractory metal films as thin as 5A and thick as 125A do not exhibit structure when viewed at magnifications up to 500kX. Carbon can also be sputtered but conductive metal films are usually thin enough that x-ray production from the metal is in the noise. Au/Au-Pd or Pd targets are not suggested for high resolution imaging. THIN FILM THICKNESS MEASUREMENTS: Inherent accuracy of QTMB is {1A when sputtered material is at low energy and hot blobs of material do not impinge on the quartz sensor. These conditions are met by sputtering mechanism, mounting and tooling factor calibration procedures of the IBS and IBSE. Water cooling the sensor typically required to stabilize the QCMB is unnecessary when ion beam sputtering. MOVING SPECIMEN: Since material evolves from the target normal to the target plane it is necessary to tilt and rotate the specimen. Different tilt angles and RPMs are necessary to insure uniform and complete coverage of various specimen topography. OPERATION: Operation of ion beam sputtering systems requires some understanding of the ion sources and vacuum conditions (purity). There is no black magic to operate these systems. Parameters are repeatable and produce repeatable results: high contrast conductive, continuous thin films without grain structure or other artifacts.
Technical References and brochures on the VCR IBS & IBSE systems now manufactured by South Bay Technology are available.
The annual Golf tournament at the MAS/MSA meeting in Portland will be on Sunday, Aug. 1, 1999. To sign up please contact the Rebedeau Group -at- mbrebedeau-at-aol.com
See you all there,
John Arnott -- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
I am having some problems during ultrathin sectioning with my block face getting wet as the diamond knife makes the first contact with the block and it happens with every stroke thereafter. I have dried the block face with filter paper or kimwipe before the next cutting stroke but it is not helping at all. Sometimes it happens during sectioning too. All my blocks are embedded in Spurr's and these are primarily plant tissue samples. I have switched knives, cleaned them before sectioning, lowered the water level in the boat, changed the clearance angle, but nothing has been working so far. I am not sure whether it is a lack of humidity problem or something else.
I will appreciate some help in this matter.
Thanks,
Soumitra
Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
I would like to thank all of you who responded to my original inquiry about scanners. I received several detailed answers, as well as requests for posting a summary of the answers as I promised, so here goes.
In my original message, I had inquired about whether members of the listserv had direct experience with either the UMAX Powerlook III or the Scan Maker 5, and could give me advice as to which would be the best scanner for TEM negatives. The UMAX costs $1200 and the ScanMaker costs $2400.
The bottom line is that there appears to be some reason for the price difference, (beyond the issue of glass in the optical path I mentioned). However, both scanners are of high quality and there are several users who are very satisfied with the UMAX. I also received good comments about the Agfa DuoScan. Apparently the UMAX geometry does eliminate the Newton rings, although some questions were raised as to the mechanics, durability, availability of drivers for NT, and customer support (which was also raised for the Scanmaker). I also got several recommendations to purchase a full-featured version of Photoshop or similar software.
There were two very detailed answers from Larry Thomas and Richard Edelman, which I reproduce below, and in which some of these points are amplified.
Several people commented that the quality of current scanners liberated them from spending time in the darkroom, so I will be buying one soon, and the comments here will make me ask some tough questions of the manufacturers.
I'm replying directly rather than to the listserver group.
My experience with using flatbed scanners for TEM negatives includes both of the units you're considering, as well as the considerably cheaper Microtek 4. These certainly aren't the only scanners around, but I would rate them highly in their respective 'middle' price ranges. Considering that they all produce high-quality scans, the criteria I consider most important are:
1. Price (of course). 2. Optical resolution: Given the choice, I would get a higher resolution (1K or 1.2K dpi ) unit rather than a 600 dpi one. Having said that, I only use the full 1K /1.2K resolution for scanning lattice images taken at 1MX or so. My main purpose in this case is to aid in matching the sampling resolution for Fourier transform (diffractogram) analysis. For routine scanning of TEM negatives, I normally use 600 dpi scanning to get reasonable print resolutions (200 dpi final print resolution at final print magnifications is adequate for publication printing) . 3. Dmax: Get the highest Dmax you can for the $$. TEM negatives of crystalline materials have relatively large density ranges and often must be taken with estimated exposures; A large Dmax can be used to compensate for overexposures in many cases. 4. Bit depth: All else being equal and if I could find one I could afford, for really serious scanning I'd consider a unit with full 16-bit grayscale (48-bit RGB). A high bit depth is important for [our] TEM negatives because they tend to have large density ranges. All the units we're considering here are 36-bit color scanners (12-bit grayscale), although the Umax as I recall has some claim of 14-bit 'effective' scans. Not sure what this means or if I believe it. 5. Practical scanning speed: By this I mean how long it really takes to do a scan, including overhead times for preview scanning, warmup, self-calibration. My impression is the Powerlook III is much slower by this criterion. The difference in work throughput can be significant. Don't take my word for this --check it for yourself if possible. 6. Scanner control software: Both Microtek and Umax leave much to be desired in the category of basic scanner controls. One semi-hidden feature I like in the Microtek program (ScanWizard) is an exposure control. Although limited in range, it allows some compensation for misjudged exposures in the TEM. Umax seems to lack a comparable feature. I find the histogram controls in both programs especially wanting for scanning high-bit images. After experimenting with software controls, I've gotten best results setting the scan parameters manually but will admit that the automatic controls do a fairly good job. I've also compared outputting 8-bit grayscale images vs outputting raw high-bit (12-bit in this case) images from the scanner to an image processing program (Photoshop). I've gotten the best results (least data loss) outputting the raw high-bit images and processing them in Photoshop before reducing them to 8 bits for printing. A third-party vendor has effective plug-in filters for unsharp masking and similar operations on high-bit (16-bit) images in Photoshop. One additional 'tip' if you missed my previous comments on this to the listserver is that it's important to scan TEM negatives with the positive transparency setting. You can invert the image contrast separately. Both Microtek and Umax assume that a negative transparency scan involves low-contrast 35 mm photographic film, and they automatically apply lookup tables that boost the output contrast of the scanned images. The result for TEM negatives is severe posterization (loss of grayscales).
7. Operating convenience: MIcrotek's system of a sliding drawer for negatives is highly preferable to placing the film on a glass platen. Not just because of the dust and Newton's ring problem, but because the films are easier to place, precisely position, and remove from the drawer cutouts without getting one's fingers all over them. The Microtek scanners don't come with a 3-1/4 x 4 " insert, unfortunately, but it's simple to make one from cardboard.
If I were buying today for routine TEM support, I would get the Scanmaker 5. An easy choice. However, prices have tumbled and the new scanners seem to keep getting better. I'd keep an open mind.
} I am considering Microtek ScanMaker5, and the UMAX Powerlook III. } Both scanners work in transparency and reflection mode, and come } with a SCSI interface and lots of software, and can handle various sizes. } The Scanmaker5 is listed at 1000 x 2000 dpi optical resolution, and } 3.6 maximum optical density (Dmax). It costs about $2400 } The UMAX Powerlook III is listed as 1200 x 2400 dpi optical resolution and } 3.4 maximum optical density. The price is about $1200.
I was looking at the same question two years ago ($1,300 vs $2,500 and went with the Umax based on cost), and loking to do the same things - scan EM negatives in a central facility. SO here are some of my thoughts.
I haven't worked with the Scanmaker but I have worked with Umax scanners and also have an Agfa Duoscan. My honest conclusion after having worked with several Umax scanners and talked with other people is that with the Umax you get what you pay for; i.e. there are reasons why the others cost twice as much. This will be intuitivly obvious to you if you have the oportunity to simply pick up both scanners. The Umax scanners are very cheaply designed and assmebled - and they are looking for the low cost market. Two years ago I bought a Umax Gemini 16D (the powerlook II replaced it the next month, so other than 600/1200DPI it was basically the same as the power look series - the higher resolution was only optically attainable via a lens shift which meant it was only applicable in the central 4" not the full 8" width of the of the scanning bed). There are two different light sources for reflected vs transmitted. The transmitted light travels via a separately motorized cariage system over the flatbed - so there are two separate mechanical motion systems to have problems. Since the upper (transmitted light) carriage is above the glass the metal and lubricant that is rubbed off (scapped off since there are no bearings on the travel rails) it falls on to the inside surface of the top glass. You need to remove the top glass periodically and clean off the debris.
O.k., next issue within six months of purchase Umax had basically stopped support of the scanner (well they had replaced it with the Powerlook right?) driver support stopped - and they didn't work really well under NT any way. But Umax Tech support didn't know anything about the Gemini's any longer either - I could only get answers from one of the tech support managers who'd been around for awhile and had one on his desk. This was with a 12-month warranty. At 13.5 months of age, $1,300 original purchase price, the transmitted light unit died. The low mag lens no longer worked properly (so we were limted to a 4 x 12" reflected light scanning area). Had all sorts of problems with the driver versus an Adaptec SCSI card (The Umax supplied SCSI card basically never worked right). 14-months after I bought the Umax I bought an Agfa Duoscan for $2,400, and (knock-on-wood) I haven't had a lick of trouble with it in the last 7 months - I count it as an expensive leanring experience. Having experienced other Umax scanner problems (though not with a Powerlook III) with other people - some of whom just wound up returning them - I would not recommend anyone buying a Umax scanner. (I think they are still listed as one of the top sellers and that is simply due to price not quality).
There's my $0.02 worth. Oh, BTW I have also found out that the Linocolor scanners (along standing and recommended scanner maker) are actually manufactured either by Umax or from Umax components - so I didn't buy on of them either.
(I do not have any financial ties with any scanner company - that I know of any way...) Good luck!
Richard E. Edelmann, Ph.D.
} Thanks very much for your detailed assessment. I was } considering going with the UMAX, but what you say gives me } real pause. Sounds like you had a really painful experience. } Other people have also commented that the UMAX is } flimsy, but appear reasonably happy with it, although those } that have used both think the scanmaker is superior. One concern } you raise, of not working well under NT (which is what I will run) } is especially troublesome. What problems did you run into?
As you have learned by now under NT hardware drivers are everything, and alot of hardware out there has drivers for 95 & 98 but Nt drivers either don't exist or are "soon to be released" (yeah right). The number one rule I have found regarding Nt and hardware is: Download the NT drivers from the manufacturers web site BEFORE placing any purchase orders - no drivers available no purchase order, period (I have waited upto 1.5 years for "soon to be released" drivers).
Umax vs NT:
(1) As with most scanners, the scanners are slower than average and higher SCSI cards (I haven't seen a true SCSI-Wide [SCSI-2] let alone a SCSI-3 scanner yet) and you will run into SCSI bus time out errors. If you are using an Adaptec SCSI card you need to set the "Maximum Sync Transfer Rate" in the Adaptec BIOS for the scanner as low as it will go. (Never use an NCR SCSI card - they are nothing but trouble)
(2) Why not use the scanner supplied SCSI Card? These cards are (Generally) the cheapest card available, usually ISA with fixed (i.e. non-changable) interupts or antiquated PCI cards (again requiring Legacy interupt support). If you have a real SCSI card (i.e. you're using SCSI HD's or a CDR, etc.) in the system then chances are the scanner SCSI card will conflict. If you run the scanner through the real SCSI card then Umax's solution was - "hey its not our card, use the twain drivers and seek help else where." (I hate to imagine what a cheap scanner scci card and a iomega scsi zip card would do with each other - although Iomega may have switched to the bottom end Adaptec cards).
(3) Due to intermittent scanner problems the UMAX NT drivers got corrupted (I know, sounds odd but true). Had to uninstall the umax drivers / software and reinstall - only the umax uninstall didn't uninstall much of what it installs! So I routinely had to manually uninstall the software, which happened to get installed in four different subdirectories!
(4) Umax scanner errors are either error code numbers - which are rarely listed in the error code list at Umax's site - or are blinking light sequences on the scanner - which aren't defined in the manual or on the web site. (Took me fours days to find out that the error was that one of the lamps had burned out - oh the lamps stay lit for 45 minutes of inactivity, but still take 10-15 seconds on each scan " Please wait - Warming up the lamps")
(5) In addition to the actual NT Drivers, each scanner comes with software which actually allows you to set the operating parameters for the scanner (When you go File} inport } scanner in something like Adobe Photshop, photoshop initalizes the scanner software - Oh, if you use Adobe PS to scan, if you close the scanning window you must then exit adobe PS before re-openning the scanner software or the system will hang). The last version of the Umax software I worked with wasn't all that user friendly, and was a little problematic to find the settings you wanted to get to - and in the last couple months before I ditched the Umax the "Pre-view" image really had nothing to do with the scanned image interms of brightness, contrast, colors, or cropping placement.
Again I hope this info helps. I'm sorry to appear so negative with regards to Umax.
Richard E. Edelmann, Ph.D.
*************************************************************************** Department of Nuclear Engineering 814-865-0036 The Pennsylvania State University fax: 814-865-8499 231 Sackett Bldg, University Park, PA 16802-1408 http://www.nuce.psu.edu
Two Diatome ultramicrotomy knives for ultrathin, semithin and cryo sectioning for sale.
Knife number 1 is 4.00mm, brand new, never used, and in the original factory sealed Diatome case. Best offer over $2,720.00
Knife number 2 is 2.70mm, brand new, never used, and in the original factory sealed Diatome case. Best offer over $1,920.00
Both knives are in the earlier original, oval shaped boat and both have the original unbroken wax sealed insignia protecting the unopened case. E mail your bid to cshear-at-rivnet.net or telephone (804) 462-0912.
This is installed on my Cambridge S250 M3, the manual was published in 1985.
Bill Giles TIMET William.Giles-at-TIMET.com
} -----Original Message----- } From: Andre Wong [SMTP:andywong-at-interchange.ubc.ca] } Sent: Tuesday, June 29, 1999 11:43 AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: KE 4-quadrant dector } } ------------------------------------------------------------------------ } } } It will be greatly be appreciated if anybody can give me information } regarding } } a. The type of detector the KE 4-quadrant BSE detector. } b. The country and city where KE-4 quadrant BSE is made. } } Thanks for the help. } Andre Wong }
I have had success avoiding the wetted-block problem when I microtome under humid conditions with the diamond-bonding material painted with Kodak Photo-Flo (a suggestion I saw in a Microstar knife book). I think that the Photo-Flo enables the knife edge to stay wetted from edge-to-edge while the water level can be lower than usual. } } Dear all microscopists, } } I am having some problems during ultrathin sectioning with my block face } getting wet as the diamond knife makes the first contact with the block and } it happens with every stroke thereafter. I have dried the block face with } filter paper or kimwipe before the next cutting stroke but it is not } helping at all. Sometimes it happens during sectioning too. All my blocks } are embedded in Spurr's and these are primarily plant tissue samples. I } have switched knives, cleaned them before sectioning, lowered the water } level in the boat, changed the clearance angle, but nothing has been } working so far. I am not sure whether it is a lack of humidity problem or } something else. } } I will appreciate some help in this matter. } } Thanks, } } Soumitra } } } } Soumitra Ghoshroy Ph.D. } Electron Microscopy Lab } Box 3EML } New Mexico State University } Las Cruces, NM 88003-8001 } Tel: 505-646-1531/3600 } Fax: 505-646-5665 } e-mail: ghoshroy-at-nmsu.edu
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798 cook-at-aaem.amc.anl.gov
PhD in physics in search for a post doctoral. I will have finished my thesis on Sept 99. I'm actually working at Reims University (FRANCE) in the laboratory of analysis solid surfaces and interfaces. My research consists in exploration for new imaging technique X ray in total reflection (Application : topography observation of surface and interface solid/solid) and fluorescence X. In the team we use X ray microscopy in electrochimical systems. My competences : X ray microscopy : Projection, reflection and fluorescence Glancing X ray : Reletometry, TXRF, GIXF Experimentator data analysis preparation of thin layer by evaporation preparation of X-visible screen converter for image guide ........ Please send Email to receive my CV and details on my work. N.B : I will take part in XRM 99 conference to be held in Berkeley, California August 1-6, 99. Sincerly
For those interested in playing in the M&M Annual Golf outting to be held on Sunday August 1 (just prior to the start of this years annual meeting in Portland, Oregon), there are still plenty of tee times available. Regardless of your handicap, we invite everyone to play. It promises to be a healthy, fun filled event with prizes for everyone! The tournament will be held at the Skamania course in the heart of the Columbia River Gorge National Scenic area. Consult the "Registration Bulletin" for further details. Please be sure to register for this event via the Meeting Manager. If you do not have a form, please contact Doug Keene (DRK-at-SHCC.ORG or 503-221-3434) prior to July 18 for copy. Include a FAX number with your request or if you prefer you will be sent a PDF or JPG file as an attachment to an e-mail. The registration deadline of July 18 is fast approaching! ---------------------- Douglas R. Keene Shriners Hospital Microscopy Facility 3101 S.W. Sam Jackson Park Road Portland, Oregon 97219 503-221-3434 DRK-at-shcc.org
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On Mon, 28 Jun 1999, Joel McClintock wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hoping for some assistance. I have been asked to research into costs and } sources for sound proof material. To be more specific we want to reduce } noise in a couple TEM rooms. We expect to acquire a high voltage FE TEM this } fall. I have seen a foam product, usually black, attached to the walls in } labs before. Everyone called it "eggshell" or "egg crate". Does anyone know } of vendors or sources of same or similar products? Thanks in advance.
We used Sonnex Acoustical Material for our 200kV FE room, from
Illbruck Inc. 3800 Washington Avenue No. Minneapolis MN 55417
Tel: 612-521-3555
Alan
Alan W Nicholls, PhD Manager - Electron Microscopy Service Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
There may be other ways to elinate the problem of block wetting, but our solution is to use a "staticmaster" ionizing unit, model 2U500. It is mounted on a flex-neck holder, also available thru NRD, Inc (2937 Alt Blve., N, Grand Island, NY 14072-1292). The head of the unit is directed to the block face / knife edge and ionizes the air and by some magic I don't understand it effectively controls block wetting. Of course it is also important to keep your water level at a reasonable level and not to have any gap between the block and knife edge as you begin sectioning or you water will fill this gap.
I hope this helps, ---------------------- Douglas R. Keene Shriners Hospital Microscopy Facility 3101 S.W. Sam Jackson Park Road Portland, Oregon 97219 503-221-3434 DRK-at-shcc.org
Actually, Tom, your equation was a little more correct, with the exception of telling us about the measurement in millimeters. The proper constant is 1.12 E-05 for speed in rpm and a radius in cm.
RCF = 1.12 E-05 x r (cm) x [N (rpm)]^2
The acceleration is equal to rw^2 where r is the radius and w (omega) is the rotational speed in radians per second. When you measure the radius in centimeters and calculate the acceleration relative to gravity (9.8 m/s^2) and convert from radians to revolutions and from per seconds to per minutes, you arive at the 1.12 E-05 factor.
So much for my engineering background creeping to the surface. Warren Straszheim
At 03:44 PM 6/29/1999 -0500, Tom Phillips wrote: } } The difference is that my "r" was meant to be the radius in "mm" whereas } John's was "cm". Therefore the equations are both correct. My original } posting should have given the units. Sorry for the confusion. Tom
We don't deal with biological materials but we have encountered the same problem you are describing with some polymers containing particles of layered minerals. Eventually we solved the problem by doing cryo and collecting dry instead of room temperature sectioning.
I hope this helps.
Jordi Marti
} } I am having some problems during ultrathin sectioning with my block face } getting wet as the diamond knife makes the first contact with the block and } it happens with every stroke thereafter. I have dried the block face with } filter paper or kimwipe before the next cutting stroke but it is not } helping at all. Sometimes it happens during sectioning too. All my blocks } are embedded in Spurr's and these are primarily plant tissue samples. I } have switched knives, cleaned them before sectioning, lowered the water } level in the boat, changed the clearance angle, but nothing has been } working so far. I am not sure whether it is a lack of humidity problem or } something else. } } I will appreciate some help in this matter. } } Thanks, } } Soumitra } } } } Soumitra Ghoshroy Ph.D. } Electron Microscopy Lab } Box 3EML } New Mexico State University } Las Cruces, NM 88003-8001 } Tel: 505-646-1531/3600 } Fax: 505-646-5665 } e-mail: ghoshroy-at-nmsu.edu
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798 cook-at-aaem.amc.anl.gov
Hi Soumitra, Sorry your having problems. One thing you shouldn't do is dry your mount with anything, especially filter paper as there are some extractables which encourage further wetting. When your block gets wet from the boat just let it dry by evaporation. If you think the knife or block may have some contamination, wash them with a mild detergent and rince with hgih purity water and allow to air dry. Also never stop the block face near the knife edge as a static knife will likely wet. Keep the block face moving while it near the knife edge. Check to make sure water has not collected on the back edge of the knife as this can be invisible to you but will rewet your block face even though it appears dry Also make sure the water you are using is pure with no residuals. Good luck, Russ, Xerox
-----Original Message----- } From: Soumitra Ghoshroy [mailto:ghoshroy-at-nmsu.edu] Sent: Wednesday, June 30, 1999 10:42 AM To: Microscopy-at-sparc5.microscopy.com
Dear all microscopists,
I am having some problems during ultrathin sectioning with my block face getting wet as the diamond knife makes the first contact with the block and it happens with every stroke thereafter. I have dried the block face with filter paper or kimwipe before the next cutting stroke but it is not helping at all. Sometimes it happens during sectioning too. All my blocks are embedded in Spurr's and these are primarily plant tissue samples. I have switched knives, cleaned them before sectioning, lowered the water level in the boat, changed the clearance angle, but nothing has been working so far. I am not sure whether it is a lack of humidity problem or something else.
I will appreciate some help in this matter.
Thanks,
Soumitra
Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
} I am having some problems during ultrathin sectioning with my block face } getting wet as the diamond knife makes the first contact with the block and } it happens with every stroke thereafter. I have dried the block face with } filter paper or kimwipe before the next cutting stroke but it is not } helping at all. Sometimes it happens during sectioning too. All my blocks } are embedded in Spurr's and these are primarily plant tissue samples. I } have switched knives, cleaned them before sectioning, lowered the water } level in the boat, changed the clearance angle, but nothing has been } working so far. I am not sure whether it is a lack of humidity problem or } something else.
In addition to the other insights and solutions offered by others, let me offer this. Not always, but very frequently I find that water jumps up onto the block face if there are *any* holes, tiny or large. For plant material there may be microscopic areas that are incompletely infiltrated, thus attracting the water. Sometimes with a good diamond knife and lots of patience you can get the block face sort of polished enough to avoid this problem. Otherwise you may have to resort to the kinds of tips posted here. I keep squares of lens tissue handy and sometimes have to swipe the block face between each cut. As long as the back of the knife remains dry, this works pretty well.
If you can trim the block closer to the area of interest, the smaller block face may have fewer holes and present fewer problems.
Good luck!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/mcroangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Those of you that are aquiring either TEM or LM digital images as a core facility, how many are storing these images as archival for the investigator (especially if the investigator has been given the a copy of the original file? Secondly, how long do you feel these images need to be stored (research based images ,not clinical)?
The answers to these type of questions is going to formulate our decision on the type of storage media we use. We were using a 1.3 GB magnetic optical and copying on redundant discs. We chose an MO due to their archival capabilities (30 years) but the drive went bad (4 y old) and now I'm wondering if I need to spend the $1,000 plus cost of another MO drive when things like Jazz and some SCSSI tape backups are just as fast and large. At this time, we do not have more than 2 GB of data to store and some of that is 3 or 4 years old and the projects published. I guess it all comes down to who should be in charge of the data. I appreciate any insights in these matters.
I have that same problem at times and one thing that seems to stop it is the use of an anti static gun. I use a product called Zero Stat 3 which looks like a small pistol that emits ?? charges in the direction it is pointed, in this case the block face and diamond just as the block starts it's downward stroke. It seems to neutralize the water and block. If I start seeing the water rise up as the block approches the knife edge the water will usually jump to the face. The next pass I dry the block and back of the knife and start using the zero stat and I get beutifull sections ther after. I have seen them in several of the EM distributers catalogs. Someone told me you used to get them for static on record players??..... what ever those are? Ha Ha Ha, oops I'm dating myself.
Soumitra, when all else fails there is another possibility - the microtome's retraction mechanism. Ultrmicrotomes without a specimen bypass on the upstroke, relay on a retraction movement. In the Ultratomes that was achieved through a large electro magnet flexing the quite solid substage of the knife-holder down and so pivoting the knife. This results in an approximate 25um retraction movement at the knife edge during the block's upstroke. It is a pretty reliable mechanism, but I had this fail some years ago. The upstroke retraction was reduced to less than 5um and block wetting was inevitable. A quite maddening problem. If you use such a microtome, just check that the retraction movement is obvious during the cutting cycle. You could mount a scale and try and measure the movement using a crosshair in the eyepieces; I would be concerned if its much under 20 um. A mechanical workshop would have a clamping micrometer with a probe which can measure the retraction. Ah, for the joys of ultramicrotomy. Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Thursday, July 01, 1999 12:42 AM, Soumitra Ghoshroy [SMTP:ghoshroy-at-nmsu.edu] wrote: } } } Dear all microscopists, } } I am having some problems during ultrathin sectioning with my block face } getting wet as the diamond knife makes the first contact with the block and } it happens with every stroke thereafter. I have dried the block face with } filter paper or kimwipe before the next cutting stroke but it is not } helping at all. Sometimes it happens during sectioning too. All my blocks } are embedded in Spurr's and these are primarily plant tissue samples. I } have switched knives, cleaned them before sectioning, lowered the water } level in the boat, changed the clearance angle, but nothing has been } working so far. I am not sure whether it is a lack of humidity problem or } something else. } } I will appreciate some help in this matter. } } Thanks, } } Soumitra } } } } Soumitra Ghoshroy Ph.D. } Electron Microscopy Lab } Box 3EML } New Mexico State University } Las Cruces, NM 88003-8001 } Tel: 505-646-1531/3600 } Fax: 505-646-5665 } e-mail: ghoshroy-at-nmsu.edu
G'day, Would any of you kind microscopists out there be able to advise me please? Have any of you done any kind of microscopy on seal teeth, tooth structure etc especially in relation to the animal's age? Thank you Gerry
Geraldine Nash Electron Microscopist
EM Unit Australian Antarctic Division Channel Highway Kingston Tasmania 7050 Australia Ph: + 61 03 62 323 322 Fax: + 61 03 62 323 351
At 02:40 PM 6/30/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Why not just store it on CD-R in ISO-9660 format? Make 2 copies as insurance. Each copy would cost about $2.
Is there any standard data format for spectra of filters or fluorescent dyes for storage of transmission or absorption / emission values related to wavelength?
I would like to have spectra in a standard format which I can overlay for comparison of crosstalk, for checking suitability of combinations of filters or dichroics, fluorochromes, laser or arc lines etc.. Molecular probes as well as BioRad have dye or filter spectra available on their web sites, but only as GIF images etc. I am sure that manufacturers have general information at hand, but I think they do not want to distribute this information freely (at least not as tables).
What I imagine is a standard format as simple as possible (like ASCII tables) relating, e.g. wavelenght (in nm) to transmission, excitation or emission strength etc., and that such files can be imported and displayed by standard plotting software. Links to websites with standard spectra and related software would be helpful.
Thanks!
Guenter
---------------------------------- Dr. Guenter Giese MPI fuer Medizinische Forschung Jahnstr. 29 D-69120 Heidelberg Phone (Germany or 0-)6221-486-320 Fax (Germany or 0-)6221-486-325 e-mail: ggiese-at-mzf.mpimf-heidelberg.mpg.de ------------------------------------------
The correct adress is : hussein.jibaoui-at-univ-reims.fr
PhD in physics in search for a post doctoral. I will have finished my thesis on Sept 99. I'm actually working at Reims University (FRANCE) in the laboratory of analysis solid surfaces and interfaces. My research consists in exploration for new imaging technique X ray in total reflection (Application : topography observation of surface and interface solid/solid) and fluorescence X. In the team we use X ray microscopy in electrochimical systems. My competences : X ray microscopy : Projection, reflection and fluorescence Glancing X ray : Reletometry, TXRF, GIXF Experimentator data analysis preparation of thin layer by evaporation preparation of X-visible screen converter for image guide ........ Please send Email to receive my CV and details on my work. N.B : I will take part in XRM 99 conference to be held in Berkeley, California August 1-6, 99. Sincerly
I received the following inquiry today and wondered if any of you could help. Please reply directly to: Catrinel Stanciu [SMTP:stanciu-at-mbi-berlin.de] {mailto:[SMTP:stanciu-at-mbi-berlin.de]} as this person is not on the listserver.
-----Original Message----- } From: Catrinel Stanciu [SMTP:stanciu-at-mbi-berlin.de] {mailto:[SMTP:stanciu-at-mbi-berlin.de]} Sent: Wednesday, June 30, 1999 12:18 PM To: ebs-at-ebsciences.com {mailto:ebs-at-ebsciences.com}
Dear Microscopist,
I have an Indium Wire and is not aware of its use. It was come along the em kit. Could any one experience the use to it in electron microscopy.
I am also search for an e-mail address of Professor Willeur C. Bigelow.
Thanks in advance.
Rajdeep Dongre Electron Microscopy Laboratory Agharkar Research Institute G.G. Agarkar Road Pune- 411 004, India
Dear Catrinel, What is the wavelength of the laser emission? The choice of substance for the film will depend on the match between its absorption spectrum and the emission spectrum of the laser. Ideally, these should not overlap. An additional consideration would be that energy absorbed by the specimen could be transferred to (and thereby damage) the film, but this is likely to depend only on the mechanical strength of the film regardless of what it is made of, and anything suitable for HRTEM has to stand up under a similar process for energy absorbed from the elec- tron beam. Good luck. Yours, Bill Tivol
Boy I got a lot of responce on this one but........... It seems most people are using CD's. Our computer system people suggested NOT using cd r, or cd rw's stating that the hardware / software is still not standardised. Our falculty computer cluster has been using Zip (too small ?) and Jazz drives for years with no problems, and everyone seems to have one or both, and yes everyone has a CD too.
I will collect all the data and submit it to the server...and our computer "experts".
But I quess I still ask why are we storing their data, other than to have a repository of images for other uses? Can we use their images without their permission since it is the investigators research or do you (that allow outside use) have the investigator sign off rights on their photos?
Can we bring this topic up at the upcoming meetings? Thanks
You can buy polypropylene film from Goodfellow Corporation as thin as 4 microns. Chemplex sells it, too, but somene seems to have stolen my Chmeplex catalog.
Alternatively, my company MOXTEK has for years offered a service to refurbish CAMECA column seperation windows with our AP1 film, which is stronger, less permiable, and has higher nitrogen x-ray transmission than polypropylene.
best regards mark
Mark W. Lund, PhD VP Engineering } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"This is a YOUNG business...How can I tell you what YOUR job is when I don't know what MINE is?" --Pogo
Yes, I would suggest the same storage medium, CD-R.
Remember, that CR-R can be sensitive to exposure to light for long times. So, in order to preserve them, you should not expose them to bright light (direct sunlight) for too long, or to too high a temperature. Storing them in a cabinet with doors should be adequate. Unlike magnetic media, CD-Rs don't care about magnetic fields.
Another option with higher capacity would be DVD. But I guess for writable DVDs you'll have to wait for a while.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Exchange Administrator Sent: Thursday, July 01, 1999 1:19 AM To: Michael Bode
At 02:40 PM 6/30/99 , you wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am having some trouble getting thin sections of some seeds I want to do EM immunolabeling on. I have ttried embedding some seeds in LR White (osmicated) or LR Gold (unosmicated). I took a week slowly infiltrating them with a dilution series of plastic resin but when I went to section them, they just popped out like stainless steel bb's. There is no hint at all that the embedding medium penetrated into them. Any experts out there with hints on fixing seeds?
TIA, Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
} I have an Indium Wire and is not aware of its use. It was come along the } em kit. Could any one experience the use to it in electron microscopy. }
Dear Rajdeep, Indium is used to make a vacuum seal for ultra-high vacuum applications. There should be a place where it would be used instead of an o-ring. This allows a vacuum seal without the use of grease or rubber, which can volatilize or outgas and degrade the vacuum. When you find the place for which the indium is intended to be a seal, place the indium so that it completely fills the seal volume and then some, so that when you tighten the seal the indium will be com- pressed. Indium is a soft metal and it will flow into the irregularities in the seal volume. Yours, Bill Tivol
Just a quick question to the wealth of knowledge on the microscopy list server,
How important is the WPE number on the Eponate 12 Resin bottle from Ted Pella when calculating the mixture of epon? i.e. should I recalculate the mixture every time the Eponate 12 Resin has a different WPE number???
Any help would be appreciated...
Eric A. Rosen Dept. Pathology UCLA Medical Center Electron Microscopy Lab...
} } I am having some trouble getting thin sections of some seeds I want to do } EM immunolabeling on. I have ttried embedding some seeds in LR White } (osmicated) or LR Gold (unosmicated). I took a week slowly infiltrating } them with a dilution series of plastic resin but when I went to section } them, they just popped out like stainless steel bb's. There is no hint at } all that the embedding medium penetrated into them. Any experts out there } with hints on fixing seeds? }
Dear Tom, We had similar problems with pollen grains. The trouble was with the difference between the hardness of the pollen and that of the resin. By varying the composition of the resin, we were able to make a sufficient match so that
sections could be cut--in our case, thick sections--with many--but not all-- of the pollen grains still in the sections. Good luck. Yours, Bill Tivol
The Tips & Tricks site has a few "difficult embedding" discussions archived. Go to:
http://www.biotech.ufl.edu/~emcl/index.html
follow the tips link and either run a search or manually pick through the TEM section. I am a year behind in archiving and will forward anything else I have to you but not to the list as a whole. If there is interest I will post the rest to the archive.
At 09:44 AM 7/1/1999 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
I am interested in learning about sample preparation and previous work performed on dye adsorbed onto mica investigated via fluorescence microscopy.
Is it possible to demonstrate using fluorescence microscopy the charge anisotropy associated with the edges and faces of mica? How? Is there a pH dependence?
What literature is available that discusses fluorescence microscopy with
mica as a substrate?
Thank You for any tips or leads!
Christian Honeker Max Planck Institute for Polymer Research
I am trying to do immunolabeling of semithin sections of LR White (medium) blocks along with TEM of ultrathin sections.
I'm using the protocol for infiltration and embedding given by Brorson (Micron 1997, 1998): from 70% EtOH, 2 x 96% EtOH for 20 min., LR White/96% EtOH (1:1) for 2 hr, complete resin for 5 hr, embed in gelatin capsule, polymerize -at- 56 deg. C. ~40 hr
My questions are:
(1) I'm also using Brorson's protocol for immunolabeling. Brorson was immunolabeling TEM sections with gold-conjugated probes. I'm doing labeling of LM sections using peroxidase/DAB. My labeling has been weak at best. I'm going to try antigen retrieval with hot citrate buffer. Any other suggestions?
(2)My TEM sections have little holes. I can't think why; my tissue is fish heart muscle. This should be easy. My oven temp. is sometimes not very stable, but I can't see how this would cause holes. Should I try vacuum pressure during infiltration?
Thanks,
-- Sonia Cawsey McGowan Copley Library, University of San Diego email: scawsey-at-acusd.edu home page: http://www.acusd.edu/~scawsey
} -----Original Message----- } Boy I got a lot of responce on this one but........... } It seems most people are using CD's. Our computer system people } suggested NOT using cd r, or cd rw's stating that the hardware } / software is still not standardised. ...
I don't understand your "computer system people" ... that is, if the CDRW software offers you the option "write a disk which can be read by any platform" (?!?!?) ... even if they change the standard, new CD readers will always be able to read old standards ... they know if they do not, their drive will be outsold by another which does. Four years ago, we went with Fujitsu MOs ... they work beautifully, and we have 4 years of worked archived on them ... be we now have to admit they didn't catch on universally. We have since changed to CDs, but we'll not move the MO archives until we are down to our last MO drive, but all four of them are working as good as new. Regarding "zip drives" ... handy? yes ... an archive medium? NO! ... please do not trust magnetic media ... always go with optical or magneto-optical ..
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
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Hi Soumitra, Sorry your having problems. One thing you shouldn't do is dry your mount with anything, especially filter paper as there are some extractables which encourage further wetting. When your block gets wet from the boat just let it dry by evaporation. If you think the knife or block may have some contamination, wash them with a mild detergent and rince with hgih purity water and allow to air dry. Also never stop the block face near the knife edge as a static knife will likely wet. Keep the block face moving while it near the knife edge. Check to make sure water has not collected on the back edge of the knife as this can be invisible to you but will rewet your block face even though it appears dry Also make sure the water you are using is pure with no residuals. Good luck, Russ, Xerox
-----Original Message----- } From: Soumitra Ghoshroy [mailto:ghoshroy-at-nmsu.edu] Sent: Wednesday, June 30, 1999 10:42 AM To: Microscopy-at-sparc5.microscopy.com
Dear all microscopists,
I am having some problems during ultrathin sectioning with my block face getting wet as the diamond knife makes the first contact with the block and it happens with every stroke thereafter. I have dried the block face with filter paper or kimwipe before the next cutting stroke but it is not helping at all. Sometimes it happens during sectioning too. All my blocks are embedded in Spurr's and these are primarily plant tissue samples. I have switched knives, cleaned them before sectioning, lowered the water level in the boat, changed the clearance angle, but nothing has been working so far. I am not sure whether it is a lack of humidity problem or something else.
I will appreciate some help in this matter.
Thanks,
Soumitra
Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
I received the following inquiry today and wondered if any of you could help. Please reply directly to: Catrinel Stanciu [SMTP:stanciu-at-mbi-berlin.de] {mailto:[SMTP:stanciu-at-mbi-berlin.de]} as this person is not on the listserver.
-----Original Message----- } From: Catrinel Stanciu [SMTP:stanciu-at-mbi-berlin.de] {mailto:[SMTP:stanciu-at-mbi-berlin.de]} Sent: Wednesday, June 30, 1999 12:18 PM To: ebs-at-ebsciences.com {mailto:ebs-at-ebsciences.com}
Dear Microscopist,
I have an Indium Wire and is not aware of its use. It was come along the em kit. Could any one experience the use to it in electron microscopy.
I am also search for an e-mail address of Professor Willeur C. Bigelow.
Thanks in advance.
Rajdeep Dongre Electron Microscopy Laboratory Agharkar Research Institute G.G. Agarkar Road Pune- 411 004, India
I need to prepare TEM samples from a Fe-Co bulk material. Can someone suggest me an electrolyte for twin-jet electropolishing and the corresponding conditions?
Dear Tom, I've used a procedure covered by VA Lindley in Microscopy Research & Techniques 21:355-360 (1992) "A New Procedure for Handling Impervious Biological Specimens" with great success after I struck a similar problem with weevils (never had such a problem with Spurrs resin completely failing to stick to a sample). The paper recommends a pre treatment with gamma-glycidoxypropyl trimethoxysilane and subsequent embedding with LR White. Its simple & worked amazingly well with re-embedded weevils (they were irreplaceable samples so I stripped off the Spurrs). The paper mentions several samples which they tested the method on, including Salicornia oil seeds. I like it.
Good luck.
Regards,
Richard } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
Re: TEM, Phillips EM400 Good Morning Microscopy List Members With the kind help of the group, perhaps we will find certain salvaged parts for the Phillips EM400 TEM that we are in need of.... specifically: the Emission Chamber, the Electron Gun (Insulator and Wehnelt Assembly), the High Voltage Cable and the Vacuum Block (Upper Column). If anyone can help, we can be reached at T. (305) 669-0233 or email. Thanks. Stephen Quinto natural-immunogenics corp.
We are looking for Philips EM 400 series TEM in any condition, or at least for the following parts: emission chamber and upper column vacuum block, electron gun (insulator and Wehnelt assembly), and high voltage cable.
Indium wire is used as a vacuum seal in the column of Philips EM400 = (near the top of the liner tube, if my memory serves me right). This is = a one-use seal and has to be scraped out with a special tool supplied = for the purpose and replaced with new wire each time the vacuum seal is = broken.
-----Original Message----- } From: Rajdeep [SMTP:rajdeep-at-aripune.ernet.in] Sent: Friday, July 02, 1999 10:00 AM To: Microscopy-at-Sparc5.Microscopy.Com
Dear Microscopist,
I have an Indium Wire and is not aware of its use. It was come along = the em kit. Could any one experience the use to it in electron microscopy.
I am also search for an e-mail address of Professor Willeur C. Bigelow.
Thanks in advance.
Rajdeep Dongre Electron Microscopy Laboratory Agharkar Research Institute G.G. Agarkar Road Pune- 411 004, India
Shaf makes a good point in that CD's are platform independent. Most storage media suffer because the drive will eventually breakdown, at which point you will have to try to find an outdated drive to be able to access your archive.
We have been archiving on CD-R's for 5 years now. We started with Win 3.11, moved to Win 95, and are now using Win 98 & NT. All of the original discs are still readable, even ones written with the early CD-R software. We frequently give out data on the CD's and have had no problems with there accessibility on other computers.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: shAf {mshaf-at-darkwing.uoregon.edu} To: RLVAUGHN-at-UNMC.EDU {RLVAUGHN-at-UNMC.EDU} ; Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
I have worked on EM equipment that uses indium wire as a vacuum seal. Indium allows you to make a vacuum seal between two flat surfaces. In the ADEM microscope we used it as a vacuum seal between the ceramic and stainless steel in the gun. If it was in your EM service kit, I would hang on to it.
Bill Carmichael Electron Microscopy Faculty Madison Area Technical College Madison, WI
http://electron-microscopy.madison.tec.wi.us
billc-at-jvlnet.com
Rajdeep wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Microscopist, } } I have an Indium Wire and is not aware of its use. It was come along the } em kit. Could any one experience the use to it in electron microscopy. } } I am also search for an e-mail address of Professor Willeur C. Bigelow. } } Thanks in advance. } } Rajdeep Dongre } Electron Microscopy Laboratory } Agharkar Research Institute } G.G. Agarkar Road } Pune- 411 004, India
I'd like to get the information about supliers of the systems of cathodoluminescence detectors for SEM wich can include a monochromator to get a spectra of the radiation in the NIR range (400-1200nm). We know about Oxford system but it's a high end priced. Any suggestions will be appreciated. kind regards to all...
Krzysztof Herman * LABSOFT * Biuro Techniczno-Handlowe ul.Ba=BFancia 45A, 02-892 Warszawa PL tel/fax: (+48 22)6446233, tel: 6449750, 6449753 mobile: (+48 601)307456, (+48 501)213438 E-mail: kherman-at-labsoft.com.pl http://www.labsoft.com.pl
Hi Krzysztof - If you dont need a defined monochromatic image, a cheaper pan-chromatic imaging system used in conjuction with a fibre optic light guide as close as possible to the specimen, a vacuum feedthrough and an Ocean Optics SD2000 spectrometer does quite well. You can pick from a number of gratings for the appropriate wavelength range, but its probably a good idea to include the most sensitive. Ocean Optics have a good website, all the information is there. The only tricky bit is aligning the fibre optic - it has to be very precise.
Good luck
Sally Stowe ( no connection with any company, etc....)
Dr Sally Stowe Facility Coordinator Australian National University EM Unit Research School of Biological Sciences Box 475, ACT 2601, Canberra, Australia FAX 06 (0)2 6279 8525 http://www.anu.edu.au/EMU
I'd like to get the information about supliers of the systems of cathodoluminescence detectors for SEM wich can include a monochromator to get a spectra of the radiation in the NIR range (400-1200nm). We know about Oxford system but it's a high end priced. Any suggestions will be appreciated. kind regards to all...
Krzysztof Herman * LABSOFT * Biuro Techniczno-Handlowe ul.Ba=BFancia 45A, 02-892 Warszawa PL tel/fax: (+48 22)6446233, tel: 6449750, 6449753 mobile: (+48 601)307456, (+48 501)213438 E-mail: kherman-at-labsoft.com.pl http://www.labsoft.com.pl
I have received a request for TEM characterization of antiferromagnetic = nanocrystalline MnF2 powder. Is it okay to load the nanocrystalline = powder on to C coated copper grids / holey carbon film on copper grids? = I have used this method earlier for non-magnetic nanocrystalline oxide = ceramic powders. What will be the effect of the magnetic field of the = objective lens on the material? Is there a risk of contaminating the = objective lens pole pieces? What precautions should I take? I use a JEOL = 2000 EX II (T) and a Philips CM200.
Ideas / suggestions / advise requested, please.
Best wishes. --- R Divakar PMS, IGCAR, Kalpakkam 603102, India ----
I would like to know if any of you has had experience in TEM of block copolymers, specifically poly-(propylene-ethylene) impact copolymers? I use RuO4 to enhance the contrast between the rubber phase and the matrix. The RuO4 is prepared from the oxidation of hydrated RuO2 with NaIO4 (sodium periodate). I've tried using chlorosulphonic acid to harden the blocks before staining but realised that the acid was destroying both the particles and the matrix. I've sectioned both at RT and at cryo- temperatures but there is not much improvement. One of the main problems I'm facing is the poor penetration of the stain.
The literature that I've come across shows the structure to nice round particles in a semicrystalline matrix but my experience has shown me that there are more than two constituents and it's very rare that I come across rubber particles that are spherical with some crystallinity inside.
I would appreciate it if you could give me information as to the preparation of the solution, the specimen, the staining procedure, the ideal sectioning conditions, etc.
Sincerely Siyabonga Siyabonga Mange BSc (Pure and Applied Chemistry;UCT) MSc (Applied Science) Final year Materials Engineering Department Menzies Building, Level 2 UCT
Having spoken to someone who has used an In seal before, are there not problems with baking? In has a low melting point, so if you use it as a seal in any U= HV system that is baked, would you not be depositing In throughout the system?
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The =C5ngstr=F6m Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
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Sonia, A couple of things to think about: 1. What type of fixation are you using? That has great bearing on number of antigenic sites available for ICC. 2.Does your oven cycle over 60oC when in the heating phase? High temperatures can really effect the amount of labeling. It might be a good idea to lower the average temperature to 50oC so your chance of the oven overshooting is less. 3. 40 hours in the oven is fairly long....usually 24hrs is sufficient with LRW. 4. If possible, polymerize in a vacuum oven that allows you to back-fill with nitrogen. Keep vacuum low but on slightly to help eliminate any gas from within the sample. You can often find old paraffin ovens that can be used for this purpose. 5. How good is your antibody? Do you know that it will give a strong label? Not all antibodies are equal. What concentration are you using? 6. What size gold are you using? The larger the gold, the less actual particles will bind. 7. Do you have another antibody which you know labels well that can be used for a method control? Before you blame the prep (rather than the antibody) you should try to react your material with another antibody that you know is powerful and has a strong reaction and well defined localization pattern. 8. How about your background? If you have no background, even a weak label can be informative. 9. Pinholes in the resin may mean that too much water is being retained in the tissue. Try dehydrating through 100% ETOH and then make sure to keep samples in closed containers to minimize moisture uptake from the air.
ICC is not an exact science and there are often problems. Don't give up because when a reaction works, it can reveal really significant information. I have had numerous examples of an ICC reaction that didn't make sense (i.e. was different than our preconceived result expectations) which later led to months of intensive research using other techniques and important new conclusions.
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
Sonia Cawsey McGowan wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
by hermes.Teknikum.UU.SE (8.9.1a/8.9.1aa) with SMTP id QAA06354 for {microscopy-at-MSA.Microscopy.Com} ; Fri, 2 Jul 1999 16:56:07 +0200 (MET DST) Message-Id: {3.0.6.32.19990702163630.008f9e10-at-pop.teknikum.UU.SE} X-Sender: joba-at-pop.teknikum.UU.SE X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32)
Having spoken to someone who has used an In seal before, are there not problems with baking? In has a low melting point, so if you use it as a seal in any UHV system that is baked, would you not be depositing In throughout the system?
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The =C5ngstr=F6m Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
Hi Soumitra, sectioning can be very frustrating sometimes. Here are some solutions I have found to block wetting, in addition to the helpful suggestions of Rus Gillmeister: Are there any tiny nicks in the edges of your block face? They tend to draw water by capillary action. Make the edges of your pyramid very clean. Lowering the surface tention of the water is helpful, so you get a concave surface and can lead the water right up to the knife edge. You can do that with a drop of highly diluted Photoflo, but I have better luck with saliva. I use a toothpick that has been in my mouth to lead the water up to the knife edge, and it is just enough. The only problem is epithelial cells that sometimes show up in the 1-micron sections. However, they are no problem on thin sections, as they are not fixed, so they just disappear. During dry weather, a hot cup of coffee next to the ultramicrotome may be enough to humidify the air. I have even known people who put wet towels around their microtome. Anyway, good luck, and keep at it and it will get better. Honest. Joyce
Interesting. We have had occasions, where people with older CD-ROMs could not read CD-Rs created with a CD burner. Whether that is particular to a specific burner or a general problem I can't say. On the other hand it's a problem that can be solved by purchasing a new $50 CD-ROM.
But back to the original question of whether the facility has to keep images: Unless there are any laws or regulations requiring you to keep images as records, I would think you are free to set your own policy. I would set up a reasonable policy (like: we keep electronic copies for a year or two on CD. After that the CDs are discarded or sent to the user), let everybody know about this policy and deal with special circumstances one by one. I am pretty sure, that if you have a reasonable policy, 95% of your users will simply accept them.
When I worked with TEMs, we would give the users prints of the negatives but keep the negatives. of course once in a while somebody wanted to make a print of an old negative, but that was not too frequent.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Colin Reid[SMTP:CREID-at-TCD.IE] } Sent: Thursday, July 01, 1999 11:36:19 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Digital Archiving continued } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Shaf makes a good point in that CD's are platform independent. Most storage media suffer because the drive will eventually breakdown, at which point you will have to try to find an outdated drive to be able to access your archive.
We have been archiving on CD-R's for 5 years now. We started with Win 3.11, moved to Win 95, and are now using Win 98 & NT. All of the original discs are still readable, even ones written with the early CD-R software. We frequently give out data on the CD's and have had no problems with there accessibility on other computers.
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: shAf {mshaf-at-darkwing.uoregon.edu} To: RLVAUGHN-at-UNMC.EDU {RLVAUGHN-at-UNMC.EDU} ; Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
For the past 16 years I've used an adhesion promotion step to embed difficult materials for diamond knife cross-sectioning. Typically hard, non porous materials such as glass, diamond, silicon, minerals, ceramics, semiconductors, metals, optical coatings and insect parts.
Prepare a 1% solution of Dow Corning's silane co-polymer Z-6040 (3-glycidoxy-propyltrimethoxy silane) in water and alcohol (50/50). Treat your samples in the solution for approximately an hour. Transfer them to filter paper to remove liquid and embed as normal in Spurrs or LR White. These coupling agents are the adhesion promoters used to bind fibers to polymer in fiberglass. Dow has a variety of other silane coupling agents with specificities for epoxies, acrylics, polyesters, phenolics, urethanes, etc. These are typically sold in 55 gal drums but smaller quantities are available.
Phil Swab Deposition Sciences Inc. Santa Rosa, CA 707-566-3718
-----Original Message----- } From: Tom Phillips [SMTP:PhillipsT-at-missouri.edu] Sent: Thursday, July 01, 1999 7:45 AM To: Microscopy-at-Sparc5.Microscopy.Com
I am having some trouble getting thin sections of some seeds I want to do EM immunolabeling on. I have ttried embedding some seeds in LR White (osmicated) or LR Gold (unosmicated). I took a week slowly infiltrating them with a dilution series of plastic resin but when I went to section them, they just popped out like stainless steel bb's. There is no hint at all that the embedding medium penetrated into them. Any experts out there with hints on fixing seeds?
TIA, Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Dear Greg Strout, John Warrack, Bill Tivol,Markus Meyenhofer,Mike Wombwell Mike Whittlesey and Jim Fotinopoulos, My sincerest thank you's to you all for your most invaluable help and for going out of your way to provide me the help I needed to resurrect this Polaron. Greg: a gracious thank you for going the "extra mile" to the point of sending me those beautiful images. What a help they are! It has been a pleasure conversing with you. John: those directions were extremely critical in assisting me with the carbon evaporating. I may contact you soon if I need further assistance. You are very kind. Bill: thank you for taking the time to provide such detailed and thorough oil changing instructions. You have this down to a science! Markus: It has been a pleasure speaking with you. Your help has truly been a benefit. Keep in touch. Mike W. and Mike W.: Thank you both for your valuable advise. Now I should be ready for the "maiden voyage". I will call you to further follow up. Jim: It was a pleasure to encounter your services. Your help was greatly appreciated. See you soon.
What a blessing this has been. Microscopists are truly a unique breed. I'm proud to be among the best.
Most Sincerely,
Maria
Maria Fazio-Zanakis Bioimaging and Molecular Histology Hoechst Marion Roussel, Inc. 1-908-231-3357 Fax: 1-908-231-3962 Email: Maria.Fazio-Zanakis-at-hmrag.com
} } Re: TEM, Phillips EM400 Good Morning Microscopy List } Members With the kind help of the group, perhaps we will find certain } salvaged parts for the Phillips EM400 TEM that we are in need } of.... specifically: the Emission Chamber, the Electron } Gun (Insulator and Wehnelt Assembly), the High Voltage Cable and the } Vacuum Block (Upper Column). If anyone can help, we can be reached } at T. (305) 669-0233 or email. Thanks. Stephen Quinto } natural-immunogenics corp. } }
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax raharris-at-ucdavis.edu
Yes, underfill boat (concave surface), making sure knife edge is wet, but also decrease clearance angle and cut faster. When aligning, don't ever stop face even with the knife edge when it's really close.
Most important if the face is getting water on it is to trim the block with the glass knife so that scratches from trimming go parallel with the block bottom edge, rather than vertical. Trimming with a razor blade creates little troughs for the water to run right up onto the face. If you don't know how to do this, and want instructions, email back.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} -----Original Message----- } } ... } It seems most people are using CD's. Our computer system } people suggested NOT using cd r, or cd rw's ...
I thought I better post a caveat to my last message which would have implied CD/RW the only way to go. I've been spending all of this Friday trying to retrieve archived images from a CD written here. Bad writes ... not because there isn't any one standard format for acrchiving files to CDs across platforms, but because, either (1) I was cheap and bought inexpensive CDs, OR (2) because my CD writer needed its BIOS flashed for better support for silver CDs. A lesson learned ... buy best quality, "tried and true" gold CDs for your archiving purposes (!!!)
have a *happy* 4th ... shAf {} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Please respond directly, offline, to sbarlow-at-sunstroke.sdsu.edu
I am interested in hearing from vendors that specialize in converting older, analog Scanning Electron Microscopes to digital computer controlled microscopes.
I am also interested in vendors that specialize in SEM digital image capture
thanks.
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Chairman, Educational Outreach subcommittee promoting access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
All: I'm looking for some information on ultramicrotomy of materials, specifically semiconductors. I'm totally in the dark about it, except that I have heard the words. Book titles, articles, web sites, helpful hints, any information would be greatly appreciated.
Hi Eric - Put the blocks back into the oven; overnight at 80 C should harden them at bit better. Remember when the blocks come out of the oven they will be quite soft for a few minutes and take a couple of hours to reach maximum hardness. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Saturday, July 03, 1999 6:18 AM, ERIC [SMTP:biology-at-ucla.edu] wrote: } } } } Today over here at UCLA we are having a slight problem with soft blocks } trying to cut 1 micron sections first... } } any suggestions as to what to do to get the sections off the glass knife } and onto a glass slide?? } } The sections seem to get stuck on the edge of the knife after they have } been cut and are floating in the water but stuck to the knife edge?? } } Any help would be appreciated... } } Eric A. Rosen } Dept Pathology } UCLA Medical Center }
I would appreciate it if someone out there in the scientific community has information to offer on how the following can be used to make water safe for drinking (in terms of killing biological contaminants).
[1] sodium hypochlorite: Can one use household bleach (5.25% sod hypoch)? What dilution is effective and safe to use?
[2] potassium permanganate: What concentration to use?
Thank you for any help or suggestions where to find the info.
Hi Jonathan: A complex subject. The melting point of Indium is 156.61 degrees C. I expect none of the TEM lens parts get that hot during a bake out. Some manufacturers restrict In seals to a couple of positions, like the gun chamber. I do not have the figures, but believe that Indium, even near its melting point, when compared with good vacuum greases would outgas at much slower rate. Furthermore, a few indium metal atoms would not be contaminants which would cause problems similar to greases and oils and their degraded products. I believe to revert manufacturers In seals to polymers seals would be unwise. Comparing current instruments with those from the 70th, arguably cleaner vacuum systems have improved TEM performance most and In seals are part of that advance. The discussion should be if in turbo and ion getter pumped systems more In seals should be used in place of polymer seals. Disclaimer: PST supplies In seals - but also vac greases. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com.au Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
On Saturday, July 03, 1999 12:37 AM, Jonathan Barnard [SMTP:Jonathan.Barnard-at-Angstrom.UU.SE] wrote: } } Having spoken to someone who has used an In seal before, are there not } problems } with baking? In has a low melting point, so if you use it as a seal in any } UHV } system that is baked, would you not be depositing In throughout the system? } } ******************************************************** } Dr Jonathan Barnard } } Analytical Materials Physics } The Angstrom Laboratory, Uppsala University } P O Box 534, SE-751 21 Uppsala, Sweden } Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 } } ******************************************************** } }
You don't have to use staining: our permanganic etching technique is ideal for PP-PE "impact" copolymers. You can see a picture of the result on:
http://www.reading.ac.uk/~spsolley/pp_impact.html
If you want any further information I can send it to you.
On Fri, 2 Jul 1999, Siyabonga wrote:
} I would like to know if any of you has had experience in TEM of block } copolymers, specifically poly-(propylene-ethylene) impact copolymers? } I use RuO4 to enhance the contrast between the rubber phase and the } matrix. The RuO4 is prepared from the oxidation of hydrated RuO2 } with NaIO4 (sodium periodate). I've tried using chlorosulphonic acid } to harden the blocks before staining but realised that the acid was } destroying both the particles and the matrix. I've sectioned both at } RT and at cryo- temperatures but there is not much improvement. One } of the main problems I'm facing is the poor penetration of the stain.
Whichever method you use, here's wishing you success,
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
we get good TEM specimens of PS/PE block copolymers if we first cut the material with the cryo-ultramicrotome (knife at -50degC, specimen at -140degC, 6deg cutting angle). Afterwards we stain the thin sections (on 1000mesh copper grids) with RuO4. Therefor we prepare a fresh mixture of about 100mg RuCl3 hydrate with ca. 5ml of 10wt% NaClO solution (in a vented hood of course!). Then we place the specimens over the mixture and keep them in the fumes for about two hours. Afterwards we wash them first with water and then with ethanol.
Good luck,
Petra
} Dear Colleagues, } } I would like to know if any of you has had experience in TEM of block } copolymers, specifically poly-(propylene-ethylene) impact copolymers? } I use RuO4 to enhance the contrast between the rubber phase and the } matrix. The RuO4 is prepared from the oxidation of hydrated RuO2 } with NaIO4 (sodium periodate). I've tried using chlorosulphonic acid } to harden the blocks before staining but realised that the acid was } destroying both the particles and the matrix. I've sectioned both at } RT and at cryo- temperatures but there is not much improvement. One } of the main problems I'm facing is the poor penetration of the stain. } } The literature that I've come across shows the structure to nice } round particles in a semicrystalline matrix but my experience has } shown me that there are more than two constituents and it's very rare } that I come across rubber particles that are spherical with some } crystallinity inside. } } I would appreciate it if you could give me information as to the } preparation of the solution, the specimen, the staining procedure, } the ideal sectioning conditions, etc. } } Sincerely } Siyabonga } Siyabonga Mange } BSc (Pure and Applied Chemistry;UCT) } MSc (Applied Science) Final year } Materials Engineering Department } Menzies Building, Level 2 } UCT } } Tel.:(021) 650 3181 (o/h) } (021) 387 4117 (a/h) } 083 583 8182 } Internet:SMANGE-at-ENGFAC.UCT.AC.ZA -------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu Visit our WWW site! http://www.crpcu.lu/~wahlbrin
I'm looking for some information on ultramicrotomy of materials, specifically semiconductors. I'm totally in the dark about it, except that I have heard the words. Book titles, articles, web sites, helpful hints, any information would be greatly appreciated.
Hi Becky,
Volume 31, No. 4 (July 1, 1995) of Microscopy Research and Technique was dedicated to ultramicrotomy of hard materials. In particular, there is a paper by S.R. Glanvill (page 275) on "Ultramicrotomy of semiconductors and related materials".
Best regards, Joergen
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
Just to add my own tricks to the excellent comments that you've received here, I would say that once you get the block face wet, drying it with a kimwipe never seems dry it enough for me. And one it's been wet, it has a tendency to get wet again much easier. So if I ever manage to get a wet block face (which happens less and less as you gain experience), I not only dry it with a kimwipe, but I blow dry the whole block face and knife edge with an air gun. (from one of those cans of compressed air) The diamond knife can stand drying from the air gun with no problem, and it's especially important to dry the back of the knife, and the block face.
Because after the block face gets wet once, then unless it is more than 100% dry after that, it will immediately get wet again and again and again....and slowly increase your frustrations until you require psychiatric treatment.
I want to do elemental mapping on catalyst particles with EELS. We have a Philips CM200 microscope equipped with a GATAN Image Filter. How does one prepare catalyst powders for EELS elemental mapping ? I suppose the powder has to be set in some kind of resin and then thinned by microtoming or some thinning technique to sufficient thickness. What kind of resin works best ? How thick should the sections be ?
Thanks
Willem Erasmus Scientist, Basic Catalysis Research Sasol Technology Tel : +27 +16 9604211 Fax : +27 +16 9602826
Re: Ted Dunn's request for info on using bleach or potassium permanganate for purifying drinking water:
Household bleach (5.25% sodium hypochlorite) can be to disinfect drinking water for bacterial contamination. The amount you would use depends on the amount you are trying to disinfect. You should test the water 20 to 30 minutes after adding the bleach to check for the amount of free chlorine that is present in the water. You should have a free chlorine residule if 0.2 to 0.5 mg/L ( the greater the amount of contaminates in the water, the more chlorine it will tahe to reach this amount of residule chlorine). Kits can be purchased from various sources to measure the amount of chlorine present (the one I am familar with is made by Hach Chemical Co. but I am sure there are others available). These are similar to the kits used for measuring chlorene levels in swimming pools but test at lower ranges. Please be aware that this will not kill all pathogens. Those that are capable for forming cysts are an example and they can only be removed by filtering.
I do not know what the acceptable method is for using potassium permanganate.
If you are only wanting to purify small amount of water such as for use when backpacking, I would suggest you consider purchasing a hand held filter. These can remove all pathogens and are available at most local stores that carry camping and backpacking supplies.
Your best source of information is your local Boy Scout or camping store. We have used both sodium hypochlorite, iodine, and filtration. They each have their advantages & drawbacks. For cleaning biological out of water lines in a manufacturing setting (i.e., DI water), sodium peroxide is often used. For cleaning wells, dry sodium hypochlorite is used. The potassium permanganate is usually reserved for cuts and treating tropical fish.
Roy Nelson Material Testing Laboratory
DUNNTEM-at-aol.com"-at-sparc5.microscopy.com wrote: I would appreciate it if someone out there in the scientific community has information to offer on how the following can be used to make water safe for drinking (in terms of killing biological contaminants). [1] sodium hypochlorite: Can one use household bleach (5.25% sod hypoch)? What dilution is effective and safe to use? [2] potassium permanganate: What concentration to use?
Thank you for any help or suggestions where to find the info.
"When I were a lad", the British Army used to use a two tablet system: first one would add a "sterilizing tablet" to one's water bottle for a time, followed by a "thio tablet".
As regards the sterilizing tablet, to the best of my knowledge it was not hypochlorite as such, but something like Chloramine-T, the common name for
N-chloro p-toluene sulfonamide
which releases hypochlorite when dissolved in water. It is made from a by-product of saccharin manufacture. The "thio tablet" SOUNDS LIKE sodium thiosulfate, which would reduce any unreacted hypochlorite to chloride.
I would guess that these things might be on sale in a camping equipment shop.
On Sat, 3 Jul 1999 DUNNTEM-at-aol.com-at-Sparc5.Microscopy.Com wrote:
} I would appreciate it if someone out there in the scientific community has } information to offer on how the following can be used to make water safe for } drinking (in terms of killing biological contaminants). } } [1] sodium hypochlorite: Can one use household bleach (5.25% sod hypoch)? } What dilution is effective and safe to use? } } [2] potassium permanganate: What concentration to use?
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Willem Erasmus wrote: ============================================== I want to do elemental mapping on catalyst particles with EELS. We have a Philips CM200 microscope equipped with a GATAN Image Filter. How does one prepare catalyst powders for EELS elemental mapping ? I suppose the powder has to be set in some kind of resin and then thinned by microtoming or some thinning technique to sufficient thickness. What kind of resin works best ? How thick should the sections be ? ================================================= The ultimate goal is to have the sectioned sample "supported" in a way in which there are locations where the sample can be viewed without any support film. Also the section must itself be cut so it is absolutely of minimum thickness. For longer exposure times, drift can be minimized further through the use of higher grid mesh sizes (smaller hole sizes).
We have found that "supporting" the section on a holely carbon or Formvar ® film provides a stable section for viewing, with data being taken only from parts of the section that are suspended over a "hole". Hence the data is taken "neat" and without contribution (absorption) from a support film.
What resin to use? Vacuum embedding of at least some of the "Epon substitutes" (we have a natural preference for our own SPI-Pon™ 812 resin) gives quite an acceptable result. We use our own SPI "materials science" diamond knives but with 45° angles, not 55°. We have ourselves never been able to make thin enough sections with the 55° knives. 35° in theory would be even better but the knife will wear out much more quickly, perhaps too quickly for most budgets. Clearly other brands of diamond knives should work as well.
Just remember that section thickness is the critical issue and everything done must be from the perspective of minimizing that thickness (in order to minimize back ground effects).
Disclaimer: Our firms provides diamond knives and embedding resins and also performs sample preparation of this type as a service to others.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
A colleague at a local university is looking for a TEM of a centrisome for use in an in-house, intro biology lab manual. Does anyone have a photo they could donate (JPEG file is preferred)?
TIA
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (920) 42403404 wise-at-uwosh.edu http://www.uwosh.edu/departments/biology/wise/wise.html
A teacher has just contacted me with a request for help. Does anyone want to have an enjoyable, rewarding experience this fall? If you respond, please copy to me, so that I know that she's getting help.
} I currently teach the Talented and Gifted at my school, Findley Oaks Elem in } Duluth, GA (Atlanta Metro). When I ordered Microscopic Explorations: A GEMS } Festival Guide, I noticed the assistance your society provides as it relates } to volunteers, and obtaining microscopes. } } Our Talented and Gifted program focus is science, although we interweave all } the other disciplines. I am very much interested in meeting with one of } your members this summer to put some plans in place for the fall. Also, I } will be attending one of the GEMS workshop at the University of California, } Berkeley in August 1999. } } Any assistance you will provide is appreciated. } } Wanda Pennywell Rachal "MSPENNY" {MSPENNY-at-email.msn.com}
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Does anybody have experience (positive) with processing yeast for TEM, both conventional and rapid freezing - freeze-substitution? Favorite protocols, comments and tips would be much appreciated.
(I do thank greatly Scott Whittaker who has already directed me to some really good information on his tips website).
Sincerely, Vladislav V. Speransky School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Phone: 207 581 2998 Fax: 207 581 2969 Email: vladis-at-maine.maine.edu
Dear Willem, probably the simplest method to mount your partciles is to disperse them on a carbon holey grid using an organic solvent. I have seen others do this very successfully with ethanol for example. Put some of the catalyst in a test tube, add some ethanol shake to mix/disperse. Using a plastic pipette drop some of the mixture onto a carbon holey grid and let it dry. It should now be ready.
I hope this helps, Jonathan
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131
I would like you to suggest the following problem:
I want to observe the arrangement of 2 proteins forming a ring shape- pore on Erythrocyte membrane (the ring shape is already observed by using TEM). The previous data suggests that they may form in to 6 sub unit -pore . I intend to use immunogold labelling 1 protein in order to adjust the interaction of two. But this pore is very small just about 3 and 7 nm inner and outer diameter respectively. So I am afraid of a steric hindrance and epitop exposure problem.
Could you please recommend which method and what kind of Microscope I should apply for my work?
Thank you in advance.
Nguyen Anh thi Van Lab. of Applied Microbiology Dep. of Biochemistry Faculty of Agriculture Tohoku University Sendai Japan
Caution re: If you are only wanting to purify small amount of water such as for use when backpacking, I would suggest you consider purchasing a hand held filter. These can remove all pathogens and are available at most local stores that carry camping and backpacking supplies.
FILTERS WON'T REMOVE VIRUSES, E.G., HEPATITIS A VIRUS.
On Mon, 5 Jul 1999 msteglic-at-notes.mdacc.tmc.edu-at-sparc5.microscopy.com wrote:
} Date: Mon, 5 Jul 1999 09:35:28 -0500 } From: msteglic-at-notes.mdacc.tmc.edu-at-sparc5.microscopy.com } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: potable water } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Re: Ted Dunn's request for info on using bleach or potassium permanganate for } purifying drinking water: } } Household bleach (5.25% sodium hypochlorite) can be to disinfect drinking water } for bacterial contamination. The amount you would use depends on the amount you } are trying to disinfect. You should test the water 20 to 30 minutes after adding } the bleach to check for the amount of free chlorine that is present in the } water. You should have a free chlorine residule if 0.2 to 0.5 mg/L ( the greater } the amount of contaminates in the water, the more chlorine it will tahe to reach } this amount of residule chlorine). Kits can be purchased from various sources } to measure the amount of chlorine present (the one I am familar with is made by } Hach Chemical Co. but I am sure there are others available). These are similar } to the kits used for measuring chlorene levels in swimming pools but test at } lower ranges. } Please be aware that this will not kill all pathogens. Those that are capable } for forming cysts are an example and they can only be removed by filtering. } } I do not know what the acceptable method is for using potassium permanganate. } } If you are only wanting to purify small amount of water such as for use when } backpacking, I would suggest you consider purchasing a hand held filter. These } can remove all pathogens and are available at most local stores that carry } camping and backpacking supplies. } } Mannie Steglich } Houston, TX } } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I would like you to suggest the following problem:
I want to observe the arrangement of 2 proteins forming a ring shape- pore on Erythrocyte membrane (the ring shape is already observed by using TEM). The previous data suggests that they may form in to 6 sub unit -pore . I intend to use immunogold labelling 1 protein in order to adjust the interaction of two. But this pore is very small just about 3 and 7 nm inner and outer diameter respectively. So I am afraid of a steric hindrance and epitop exposure problem.
Could you please recommend which method and what kind of Microscope I should apply for my work?
ELECTRON MICROSCOPY TECHNICIAN, SENIOR Duties include all aspects of TEM, including negative staining and thin sectioning. It's an exciting job working with many different kinds of clinical and research specimens, physicians and researchers. It is a large lab with 4 other amiable EM techs who share duties. The salary is in the $27K range, plus 24% benefits (total--~$34.3K) which is reasonable for the cost of living here. Durham is a cosmopolitan city with 3 major universities nearby, and hence, all kinds of cultural entertainment; it is situated halfway between the beach and the mountains (about 3.5 hrs to each). Called the "City of Medicine," for it's high doctor/citizen ratio because of numerous hospitals, Durham also claims the team that "almost won" the NCAA tournament and that sent 3 players this year to the NBA (heaven help us next year). It is home to the Durham Bulls (baseball) and a half hour drive from the Carolina Hurricanes (hockey). With all this, it is still a relatively small city (~150K) where you can live in the country with only a 20 min drive to work, if you choose. Requisite job qualifications include experience in TEM, either through college courses or on-the-job training. Call or reply by email if interested.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Thanks to all of you who took the time to respond to my posting regarding block face getting wet. I received lots of very helpful suggestions.
This is truly an excellent resource for microscopists.
Cheers,
Soumitra
Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
Sample preparation required to perform EELS on catalyst particles varies, depending on the composition and hardness of the catalyst. We have good luck with many alumino-silicate and similar catalysts by embedding the finely ground powders in LR White resin (acrylic), and then microtoming to acquire very thin sections (references available). Many other supported catalysts do much better with an epoxy system such as Spurr's Low Viscosity resin. Some materials are easier examined by dispersing fine particles on a holey carbon support.
We'll need more info before we can give you any specific sample prep to follow. What is your catalyst material?
Good Luck, Brad ____________________________
I want to do elemental mapping on catalyst particles with EELS. We have a Philips CM200 microscope equipped with a GATAN Image Filter. How does one prepare catalyst powders for EELS elemental mapping ? I suppose the powder has to be set in some kind of resin and then thinned by microtoming or some thinning technique to sufficient thickness. What kind of resin works best ? How thick should the sections be ?
Thanks
Willem Erasmus Scientist, Basic Catalysis Research Sasol Technology Tel : +27 +16 9604211 Fax : +27 +16 9602826
} Rick Vaughn wrote ... } } } It seems most people are using CD's. Our computer system } } people suggested NOT using cd r, or cd rw's ... } } I thought I better post a caveat to my last message which } would have implied CD/RW the only way to go. I've been spending } all of this Friday trying to retrieve archived images from a } CD written here. Bad writes ... not because there isn't any one } standard format for acrchiving files to CDs across platforms, } but because, either (1) I was cheap and bought inexpensive CDs, } OR (2) because my CD writer needed its BIOS flashed for better } support for silver CDs. A lesson learned ... buy best quality, } "tried and true" gold CDs for your archiving purposes (!!!)
Dear List:
All of this talk about digital archiving reminds me that film, that semi-transparent material with the silver halide coating, has a storage life of 100+ years when properly processed.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
ELECTRON MICROSCOPY TECHNICIAN, SENIOR Duties include all aspects of TEM, including negative staining and thin sectioning. It's an exciting job working with many different kinds of clinical and research specimens, physicians and researchers. It is a large lab with 4 other amiable EM techs who share duties. The salary is in the $27K range, plus 24% benefits (total--~$34.3K) which is reasonable for the cost of living here. Durham is a cosmopolitan city with 3 major universities nearby, and hence, all kinds of cultural entertainment; it is situated halfway between the beach and the mountains (about 3.5 hrs to each). Called the "City of Medicine," for it's high doctor/citizen ratio because of numerous hospitals, Durham also claims the team that "almost won" the NCAA tournament and that sent 3 players this year to the NBA (heaven help us next year). It is home to the Durham Bulls (baseball) and a half hour drive from the Carolina Hurricanes (hockey). With all this, it is still a relatively small city (~150K) where you can live in the country with only a 20 min drive to work, if you choose. Requisite job qualifications include experience in TEM, either through college courses or on-the-job training. Call or reply by email if interested.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Thanks to all of you who took the time to respond to my posting regarding block face getting wet. I received lots of very helpful suggestions.
This is truly an excellent resource for microscopists.
Cheers,
Soumitra
Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
Our resident entomologist/electrophysiologist would like to delve into the fascinating world of microscopy and imaging, and wants to take me along with her.
My microscopy background has not included insects, and I am not well versed in their pecularities in preparation (such as how to deal with penetration of fixatives and the like through exoskeleton), so I am asking for your help with this.
At this point, I would very much appreciate it if people might point me in the direction of some good references and/or books on the subject of preparing cleared whole mounts of insects as well as techniques for fixing and embedding insect parts (especially antennae) for LM and EM.
Please reply off line. Thanks in advance.
Paula Allan-Wojtas Research Scientist, Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
} ---------- } From: Geoff McAuliffe[SMTP:MCAULIFF-at-UMDNJ.EDU] } Sent: Tuesday, July 06, 1999 9:32:49 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Digital Archiving continued
..
} Dear List:
} All of this talk about digital archiving reminds me that film, that } semi-transparent material with the silver halide coating, has a storage life } of 100+ years when properly processed.
.. and stored!! Exposure to light, humidity, chemical vapors, etc., or scratches due to handling can destroy negatives as well. I agree, that some negatives have a proven record of 100+ years, but how long do you relly need access to the negatives? I have folders and folders of TEM negatives from before the "digital age" in my shelves, and frankly, they will probably stay there until I die and then go to the trash without anybody ever looking at them again. And my filing system is not perfect either. If somebody asked me for a certain negative I took, say 10 years ago, I would probably have to look for a long, long time to find it. Some of the images went into publications, so there is another record of these. Others that I need I have on my PC and they will get transferred to other media until they become yesterday's news. Most of them are redundant and not needed, but since I had no way of making sure I recorded the right pictures I took many more than I needed.
While I agree, that negatives can survive at least a century, it is other factors like databasing, ease of transfer, and less work with chemicals that I would consider as well. The problem we have with recalling old data is usually not that the data is gone, but that the technology has changed and there is no longer access to the media (remember 8 inch disks :) ? I am convinced, that even in 50 years you can find somebody who can scrape the data from a CD and put it on a then current medium (holographic storage??).
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
I'm trying to get some information for a client who wants to try making his own glass Ralph (histo) knives. He says he heard something about a technique for breaking these by hand, without the special knifemaker. Does anyone out there in list-land know anything about this?
Thanks much.
Randy Tindall Electron Microscope Specialist Electron Microscope Core Facility University of Missouri Columbia, MO 65211
I'm trying to get some information for a client who wants to try making his own glass Ralph (histo) knives. He says he heard something about a technique for breaking these by hand, without the special knifemaker. Does anyone out there in list-land know anything about this?
Thanks much.
Randy Tindall Electron Microscope Specialist Electron Microscope Core Facility University of Missouri Columbia, MO 65211
Responding to the message of {v03007801b3a834474a66-at-[206.69.208.21]} from JoAnn Buchanan {redhair-at-leland.Stanford.EDU} : } } } Dear readers, I have a problem with very pale staining of my embed 812 } resin thin sections. I often need to use formvar and I cut 50nm thin } sections of cultured cells and fish embryos. I have tried numerous recipes } for both UA (including methanolic) and Lead (Sato's, lead nitrate, lead } citrate). It seems like the stain works well for a couple of times, then } flakes out. I have tried making the lead up fresh each time but even that } fails. The tissue is microwave processed including an en bloc uranyl } acetate step. I take great pains not to get any precipitate by using an } NaOH rinse. I stain 10-15 mins in UA and 5 min. in lead. Does any one have } any suggestions, ideas? What is the best lead stain?? Thanks in advance.
I, too, cut 50-55 nm sections of Embed 812, tho of plant materials, and also get slightly fainter staining, and I attribute it to just the fact that the sections are quite thin and there isn't a lot of material to stain - no Formvar on these grids. If I cut 60-75 nm sections, they do stain better. I'm cutting thin for better resolution.
Try cutting some thicker sections and look at those just to check your stains. I use 3-4% UA for 20-30 minutes, followed by Sato lead for 3-4 minutes. Hope this helps.
Gib
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu http://biosci.umn.edu/MIC/consortium.html
} Dear readers, I have a problem with very pale staining of my embed 812 } resin thin sections. I often need to use formvar and I cut 50nm thin } sections of cultured cells and fish embryos. I have tried numerous recipes } for both UA (including methanolic) and Lead (Sato's, lead nitrate, lead } citrate). It seems like the stain works well for a couple of times, then } flakes out. I have tried making the lead up fresh each time but even that } fails. The tissue is microwave processed including an en bloc uranyl } acetate step. I take great pains not to get any precipitate by using an } NaOH rinse. I stain 10-15 mins in UA and 5 min. in lead. Does any one have } any suggestions, ideas? What is the best lead stain?? Thanks in advance. } } JoAnn Buchanan } Molecular and Cellular Physiology } Stanford University Scholl of Medicine } Stanford, CA 94305