Microscopy List {microscopy-at-aaem.amc.anl.gov} , Confocal Microscopy List {confocal-at-ubvm.bitnet} Message-Id: {Pine.3.05.1.9502011127.A7624-9100000-at-pauline.sdsc.edu} Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII
I am in need of access to a confocal scope in the San Diego area. Would anyone know of a lab that has one and who to contact? Thanks in advance for your help.
Marc
Marc C. Brande, M.S. SD3D Email List:3D Imaging San Diego 3D Imaging Group To subscribe/unsubscribe:listserv-at-bobcat.etsu.edu 3840 Camino Lindo To post a message:sd3d-at-bobcat.etsu.edu San Diego, CA 92122 Email:BRANDE-at-SDSC.EDU Voice:619-587-4830
Jan Coetzee asks about electronic vs. "dog-eared diary" for schedules. Dear Jan, We use a large desk calendar ourselves. It seems easier, since there are frequent changes, and everyone can check it instantly. I'm sure that a calendar page on a computer might do as well, but when a user calls, we are not always booted up, so it would take us more time than it does with paper and pencil. If your scopes are computer-controlled, it might save some time and/or effort to have schedules on the same computer--e.g. the computer could dial a user you need to contact re schedule changes--but we have not thought this to be worthwhile for us. Sometimes low-tech works very well! Yours, Bill Tivol
Jan Coetzee inquired about mfgrs of pltrs. Dear Jan, Melcor makes them. Their address, phone & fax are 1040 Spruce Street Trenton NJ 08648-4587 United States of America (our USA) (609) 393-4178 (phone) (609) 393-9461 (fax) One potential problem is that Peltiers have a limited delta-T, and can get very expensive. I asked Melcor about setting up a cold stage for epitax- ial deposition in a vacuum, and there was nothing which would get cold enough. Furthermore, if you wish to thermostat the device, you need to use a cooler which has a 100% duty cycle and output current proportional to the difference between target and actual temperatures. We found this out when we installed Peltiers in our darkroom as heaters/coolers to maintain developer temperature. The intermittant current put out by the usual commercially available devices blew the Peltiers out! Otherwise, Peltiers are marvelous devices. Good luck. Yours, Bill Tivol
I've received this request for help on locating micrographs of bacteria. I could not help this individual. So is there anyone out there that can?
Cheers....Nestor Z. Your Friendly Neighborhood SysOp =====================================
P.S.
I'm working on concept of adding an Image Library to the Software Library. When it's ready for contributions I'll post a message to the Listserver....
====================================
I need an electron microscope picture of Methanosarcina barkeri and Methanobacterium wolfei. I called the University of California at Berkeley and they gave me your address. They said that they don't keep a file of their past work and would charge $300 per picture to do the work again. I'm fairly certain that the pictures already exist somewhere. I believe that NASA may have some of them but I don't know who to ask there . Thank you for any help in getting any pictures of the two bacteria.
The following files have been delivered to you - please use the eXtract command in the browser to work with them:
MARSALL.ASC (format: Text) ---------------------------------------------------------------------- Thomas K. Wilson wilsontk-at-MUohio.edu Dept. of Botany Miami University ! Miami was a University when Oxford OH 45056 ! Florida belonged to Spain ! USA 513.529.4210 office 513.529.4243 fax
-------------- Enclosure number 1 ---------------- The following is posted as a favor to Marshall University, and to any interested applicants on the MSA-BBS. Please distribute this message to any and all interested people you may know.
I am not involved with this Supervisor search in any way whatsoever (Other than having promised to post this ASAP). Please contact Dr. W.B. Rhoten directly at the address below (his E-mail address is Rhoten-at-musom01.mu.wvnet.edu)
POSSITION OPEN
SUPERVISOR OF ELECTRON MICRSCOPY FACILITY
To supervise and perform day-to-day operations of the Electron Micrscopy Facility and assist students, research assistants, and investigators. Participate in graduate level EM Course.
Bachelor's degree including courses in biological and physical sciences and at least 3 years experience in EM, or a Masters degree and at least 3 years of relevant experience, or a Ph.D. degree. Qualifications include comprehensive knowledge of EM and ability to enhance educational and research activities. Experience in image processing and computers desired.
Qualified applicants should send a cover letter, resume, and list of at least 2 references to:
Dr. W. B. Rhoten Dept. of Anatomy, Cell & Neurobiology School of Medicine, Marshall University Huntington, WV 25704-9388
For full consideration submit application by March 1, 1995.
MARSHALL UNIVERSITY IS AN AFFERMATIVE ACTION/EQUAL OPPORTUNITY EMPLOYER
One of EM Unit users is studying developing Red deer. The samples we are examining are skull and pedicle (developing antler) taken from the deer at set time intervals over atwo year period. The samples are processed routinely, ie glutaraldehyde fixed,decalcified, OsO4 post fixed, uranyl acetate bloc stain, dehydrated and embedded in Agar 100 epoxy resin.
Our problem is that it now seems to be that the glycogen content is of some significance to the investigation. Unfortunately the above process is not ideal for showing the glycogen. Some recently processed samples using potassium ferrocyanide with the osmium are brilliant however we would like to enhance the glycogen in the previous blocks. Does anyone out there know a method to enhance "unstainable" glycogen in ultrathin sections? Thanks in anticipation.
Allan Mitchell South Campus EM Unit allan.mitchell-at-stonebow.otago.ac.nz
} I am looking for a way to convert my PostScript files into "regular" image } files. Does someone know of such siftwares on either Mac or Unix platforms? } Any information would be appreciated. } Do you mean postscript or encapsulated postscript? If postscript then one of the postscript interpreters available should do it. I use one an a PC called GoScript that outputs to TIFF files as well as printers. I am not sure, but you should take a look at the GNU Ghostscript interpreter that runs on virtually anything - certainly on unix systems. If you mean encapsulated postscript (EPS) then just import the files into a Mac word processor such as WordPerfect on Microsoft Word and then copy and paste to whereever you want to. +------------------------------------+ | Dr Douglas Arrell | | Mechanical Performance and Joining | | Institute for Advanced Materials | | 1755 ZG Petten | | Netherlands | | {ARRELL-at-JRC.NL} | | Tel. (+31) 2246 5287 | | Fax (+31) 2246 1917 | +------------------------------------+
Group - Responding to an inquiry from Nestor, Custom Medical Stock Photo is a company which purchases micrographs and then sells them to publications, etc. They have a considerable inventory - in microscopy, and a number "with" bacteria. Many readers may be interested in contacting them and explore the sale of their micrographs. I have, of course, no financial interest in the company.
Custom Medical Storck Photo, Inc. 3819 North Southport Ave Chicago, IL 60613 Tel.: 312-248-3200 Fax: 312-248-7427
Message-Id: {1995Feb03.100340.2874650620-at-dmcmail.ucsf.edu} To: Microscopy-at-aaem.amc.anl.gov (M)
Subject: Time: 9:45 AM 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
**************************************************************************** John A. Kloetzel, Ph.D. {kloetzel-at-umbc.edu} Department of Biological Sciences University of Maryland Baltimore County (UMBC) Catonsville, MD 21228-5329 USA Phone: (410) 455-2247 or -3913 (Lab) FAX: (410) 455-3875 ****************************************************************************
A few months ago a thread ran on fixation and embedding pepper contaminant artefact in biological ultrathin sections. A colleague of mine read the Pepper Summary I compiled and sent me the following fixation protocols that he has used successfully without pepper. Notice the first one in which glutaraldehyde, phosphate buffer AND OSMIUM are all mixed TOGETHER!
If anyone out there wants a copy of my Pepper Summary, contact me off-line at my e-mail and I will zip a copy out to you.
"1. For fixing cilia in mammalian trachea, I have used an "instant fixation" method using a combination of osmium, phosphate buffer, and glutaraldehyde - in the cold - for many years and have never seen the pepper described in the Pepper Summary you sent to me. Right - all that stuff in the same buffer dumped on the tissue. Works great, but may extract a bit of actin.
"The method I used was one described in a paper by Omnoto and Kung in the J. Cell Biol 87:33-46. I think it uses 50 mM NaPhosphate, pH 7.2, 2% OsO4, 2% glutaraldehyde. Add the Osmium just before you add the tissue and fix on ice for 10 min. If you want, you can remove the black fix after 10 min and add another slug of fix for another 10 min but that is optional.
"Omoto and Kung used it to fix Paramecium and their cilia. I have used it to fix mammalian trachea (with their cilia). The advantage is that it seems to "freeze" cilia in position, as opposed to glutaraldehyde, in which cells actualy swim for a dab before either being fixed or dying (we fix cells, we don't kill them, do we?). The osmium does not penetrate for more than a few cell layers but, with epithelial tissue or single cells, it does not make much difference.
"I have never tried that fix method on Chlamydomonas or Tetrahymena. Over the last year I have done a lot of embedding of Chlamy and have never seen pepper. For those beasts, I find that cacodylate gives the best preservation of the cytoplasm, although others find that phosphate buffer works fine too. I do fix with glutaraldehyde in culture medium (pretty much phosphate buffer), overnight in glut in cacodylate, rinse a few times in cacodylate, then into 0.5-1% OsO4 in cacodylate for 30-60 min on ice, rinse with a few changes of water and into uranyl acetate for a few hours prior to dehydrating in acetone and embedding in epon.
"I've also tried another method in which you fix with glut in phosphate buffer, rinse, then incubate overnight in uranyl acetate (in water) at 70 degrees C. It works on Chlamy and avoids osmium. It is supposed to eliminate the need for poststaining of sections, but I did not find this to be the case. I believe that I once read that the reason for staining sections is to put a dab of stain on structures at the last surface of the plastic that the beam sees before blasting through the objective lens. I don't know, but, for Chlamy, I usually use the more old fashioned method that I gave you above. Pepper does not seem to be one of my problems." ------------------------------------------------------------------------ Contributed to MICROSCOPY by:
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Dear Lucille, I just got a number from Kodak for technical questions, but they could probably direct you to a distributer. It is (800) 242-2424 x19. We usually use a local vendor, National Graphics, but I have not noticed a great range of prices with other vendors. Good luck. Yours, Bill Tivol
I am not aware that "cheap" is an a.k.a. for "reliable". Please be kind to the English language.
On Fri, 3 Feb 1995, Lucille A. Giannuzzi wrote:
} Can anyone recommend a reliable (a.k.a. cheap) U.S. vendor for TEM film? } } Thanks in advance, } Lucille Giannuzzi } } } ************************************************************************* } Lucille A. Giannuzzi, Ph.D. } } Dept. of Mechanical and Aerospace Eng. phone (407) 823-5770 } University of Central Florida fax (407) 823-0208 } 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu } Orlando, FL 32816-2450 USA } ************************************************************************* } } } }
DDoes anyone know about work done on immunolabelling of wood tissue of fruit trees. I'm interested in any kind of information I can get on fixation, embedding and the labelling procedure. I plan to be using ProteinA-colloidal gold to tag the antibody.I'll be using a confocal microscope for this study.If anyone knows any work done in this area could you please send me the references. Thank you in advance. my email address: komminen-at-student.msu.edu
Dear Fellow Microscopists, We have recently acquired a scanning electron microscope with a four crystal Wavelength Dispersive Spectrometer but the associated computer system is rather old and probably not worth re-activating. I know that there is software for the Macintosh, such as "D.T.S.A." and "Flame", for ENERGY Dispersive Spectroscopy but my question is "Is there any (Mac) software out there which can handle W(AVELENGTH)DS spectral acquisition and processing?". Any leads would be very much appreciated. Thanks.
Rick Hall Materials Science Program University of Delaware
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.
Sincerely,
Dr. George McNamara Universal Imaging Corporation George_M-at-Image1.com --------------------------------------
Subject: Time: 9:45 AM 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
The organizers of the 28-31 March 1995 Scanning 95 meeting want about 25 student volunteers to run the slide projectors for the sessions in exchange for full meeting registration ($275). Please contact: Mary K. Sullivan FAMS, Inc P.O. Box 832 Mahwah, New Jersy 07430,0832
or leave me a message. Debe Holmberg e-mail {dlholmberg-at-peseta.ucdavis.edu} Lab 916-752-9021 FAX 916-752-4604
I am going to be working on an TEM project that will be under "GLP" . GLP are guidlines for doing certain experiments for the FDA (I think the equivalent in Europe is ISO 9000). I was wondering if anyone out there is doing TEM under these guidlines? And if so what are they using for magnification calibration. I do not believe that there are any vendors who can supply a certified magnification standard for TEM, which, I think is required for GLP.
Thanks, David Leaffer Syntex Research david.leaffer-at-syntex.com
The organizers of the 28-31 March 1995 Scanning 95 meeting want about 25 student volunteers to run the slide projectors for the sessions in exchange for full meeting registration ($275). Please contact: Mrs. Mary K. Sullivan FAMS. Inc. P.O. Box 832 Mahwah, NJ 07430-0832
or leave me a message. Debe Holmberg Lab 916-752-9021 Fax 916-752-4604 dlholmberg-at-peseta.ucdavis.edu
Dear All, We are are multi-user electron microscope facility which has an extensive range of equipment and uses a wide range of techniques. Five technical staff from three contributing University departments are employed full-time to undertake for work coming from both inside and outside the University.
The role of the 'academic in charge' of the facility is shortly to come up for reassessment and so it is a pertinent time for us to reconsider what that role should entail. We are looking for feedback from other individuals as to what they see the contribution of an academic in this environment should be. Any opinions/suggestions?
} The role of the 'academic in charge' of the facility is shortly to come up } for reassessment and so it is a pertinent time for us to reconsider what } that role should entail. } We are looking for feedback from other individuals as to what they see the } contribution of an academic in this environment should be. Easy - 1. Keep abreast of all techniques which a) you have, b) exist, and c) potentially exist. 2. Be an expert practionioner of two or more of these. Publish a lot in your own name. 3. Give advice on the application of all techniques and on the high-level interpretation of all results. Publish jointly with others. 4. Raise funds to replace equipment and to buy new techniques as they become applicable. 5. Make sure all users publish, and tell you about it! 6. Establish a reputation as a scientist in some major subject area, not just as a microscopist. Publish a lot. 7. Keep friendly with the heads (or budget controllers) of all potential user departments. 8. Get to know lots of people in your institution by playing sport/drinking/etc with them. 9. In your spare time, publish some more.
I offer this advice after 20 years of running such a facility!
Peter Goodhew
---------------------------------------------------------------------------------------------------------- Professor Peter J Goodhew, Department of Materials Science & Engineering University of Liverpool LIVERPOOL Fax (44) (0)51 794 4675 L69 3BX, UK Tel (44) (0)51 794 4665 (secretary Debra) ---------------------------------------------------------------------------------------------------------- inter alia: Director of the MATTER project for educational software ----------------------------------------------------------------------------------------------------------
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Institute of Physics Publishing Techno House Tel: +44 272 297481 Redcliffe Way Fax: +44 272 294318 Bristol BS1 6NX England Telex: 449149 INSTP G
I am the Publisher of the journal Bioimaging which I hope you have heard of. I would like to place information about the journal, including tables of contents, on your bulletin board. Will this be possible and how should I do it?
Several years ago there appeared an article in Science discussing laboratory space design and a recent "New" model being implemented in England. Does anyone remember this article, and what has happened to the English experiment?
Microscopy List {microscopy-at-aaem.amc.anl.gov} , Functional Neuroimaging List {lat-at-po.cwru.edu} , Confocal Microscopy List {confocal-at-ubvm.bitnet} , Cell Bio List {cellbiol-at-net.bio.net} Message-Id: {Pine.3.05.1.9502081015.B14211-a100000-at-pauline.sdsc.edu} Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII
Can Anyone point me to sources (free to minimal cost) of analog (VCR tape) or digital timelapse movies of cells in culture? This is not for commercial use, only presentation demonstration. Of course I would credit each source in the presentation. Thanks in advance for any help you can give.
Marc
Marc C. Brande, M.S. SD3D Email List:3D Imaging San Diego 3D Imaging Group To subscribe/unsubscribe:listserv-at-bobcat.etsu.edu 3840 Camino Lindo To post a message:sd3d-at-bobcat.etsu.edu San Diego, CA 92122 Email:BRANDE-at-SDSC.EDU Voice:619-587-4830
Dr. Neal D. Evans Shared Research Equipment Program Oak Ridge National Laboratory Building 5500, MS 6376, Oak Ridge, TN 37831-6376 voice(615-576-4427) fax(615-574-0641) email(evansnd-at-ornl.gov
Is anyone aware of a company that sells reconditioned SEMs?
+---------------------------------------------------------+ | Stuart Wright Los Alamos National Lab, MST-6 | |.........................................................| | Mail Stop G770 phone: (505) 665-3647 | | Los Alamos, NM 87545 fax: (505) 667-5268 | +---------------------------------------------------------+
------ Forwarded message ends here ------
******************************************************************* Daniel E. Sampson dsampson-at-earthsci.ucsc.edu Instrumentation Specialist Phone: (408) 459-4992 Earth Sciences FAX: (408) 459-3074 University of California Santa Cruz, CA 95064 *******************************************************************
Stuart, Have you tried contacting the instrument manufacturers themslves? I believe that JEOL and Hitachi, for instance, may sell the reconditioned SEMs that comes in on trade-ins. Failing that, give us a call, we'll try to direct you to more sources.
Ellie Solit, Publisher of MICROSCOPE TECHNOLOGY & NEWS AND THE MICROSCOPE BOOK
On Wed, 8 Feb 1995, Stuart Wright wrote:
} Is anyone aware of a company that sells reconditioned SEMs? } } } +---------------------------------------------------------+ } | Stuart Wright Los Alamos National Lab, MST-6 | } |.........................................................| } | Mail Stop G770 phone: (505) 665-3647 | } | Los Alamos, NM 87545 fax: (505) 667-5268 | } +---------------------------------------------------------+ } }
Dear All, Further to my message of 8.2.95, thank you very much to those who have responded so candidly to my request for people's feelings and experiences on this topic. I appreciate that it can be a sensitive issue, not helped when one is replying to someone who doesn't even identify themselves properly. As I neglected to state who I am and where I work (I changed computers days ago and forgot to put the automatic signature on - the ramifications of this I am just becoming aware of...) that info now follows. Maybe now I'll get some more replies...
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
I have been asked to do particle sizing of a 10% fat immulsion (pH 8.0) and had hoped to be able to do a very quick negative staining procedure as this may need to be run frequently. I toyed with the idea of freeze fracture, but the time involved is not convenient for lots of runs. Has anyone tried this? What stains would you suggest? Is Osmium vaper fixation nessecary?
We have considerable experience with negative stain of phospholipid vesicles, lipoproteins, and triacylglyceride emulsions. In these circumstances fixation is not critical, and in fact can produce artefacts. PTA stain seems to work best. We concentrate our sample on grid by drying multiple drops under gentle nitrogen gas stream prior to negative stain. This avoids clumping in the suspension. Some sizing artefacts can occur as dryed particles of large size can deform slightly. See Ganz et al, 1991, J Lipid Res 31:163 for nice discussion of this. Good luck-
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
On Thu, 9 Feb 1995, Kathy Walters wrote:
} Dear Fellow Microscopists, } } I have been asked to do particle sizing of a 10% fat immulsion (pH 8.0) } and had hoped to be able to do a very quick negative staining procedure as } this may need to be run frequently. I toyed with the idea of freeze } fracture, but the time involved is not convenient for lots of runs. Has } anyone tried this? What stains would you suggest? Is Osmium } vaper fixation nessecary? } } Thanks for any help! } Kathy Walters } } }
Dear Kathy, Staining is not really my field, but I'd suggest a water-soluble heavy metal and NO osmium. The Os would only dissolve in the lipid and reduce con- trast. If possible, looking at a frozen, hydrated (or lyophyllized in-situ) specimen would be best. You don't specify either the matrix of the emulsion (I assume aqueous) nor the technique to be used (I assume TEM), but a possible protocol would be to add the stain to the emulsion and rapidly freeze, then cryo-section, examine on a Friday, raise the temp to ~-90C, come in Saturday and raise the temp to -80C, Sunday to -70C, and look at the freeze-dried spec- imen Monday. If you can do the particle sizing from the frozen-hydrated spec- imen, the last steps can be omitted. Keeping the stain strictly in the aqueous phase should prevent size changes, etc. in the lipid drops. Good luck. Yours, Bill Tivol
E.J. Fjeld Co, 3 Executive Park Drive North Billerica MA 01862 Phone 508-667-1416
Provides reconditioned AMRAY microscopes. They also make special stages and accessories. They've been around for a long time and are reliable.
Jacob Bastacky, MD 1-116 Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
Stuart, my attempts to reply to your question by email have resulted in 6 notices of undelivered mail. I left a voice message on your phone this morning. I think we can help you in your search.
Regards, Ellie Solit, Publisher/Executive Editor of Microscope Technology & News and The Microscope Book, a Smart catalog.
On Thu, 9 Feb 1995, Stuart Wright wrote:
} Is anyone aware of a company that sells reconditioned SEMs? } } } } +---------------------------------------------------------+ } | Stuart Wright Los Alamos National Lab, MST-6 | } |.........................................................| } | Mail Stop G770 phone: (505) 665-3647 | } | Los Alamos, NM 87545 fax: (505) 667-5268 | } +---------------------------------------------------------+ } }
Message-ID: {95020916094142E.RQDA-at-USCN.USCN.UGA.EDU} (UMass-Mailer 4.04) Neuroscience List {neur-sci-at-net.bio.net} , Microscopy List {microscopy-at-aaem.amc.anl.gov} , Digital Video List {digvid-l-at-ucdavis.edu} , Confocal Microscopy List {confocal-at-ubvm.bitnet} , Cell Bio List {cellbiol-at-net.bio.net} Message-Id: {Pine.3.05.1.9502091131.A22203-e100000-at-pauline.sdsc.edu} Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII
I thought this post should be quickly disseminated.
Marc C. Brande, M.S. SD3D Email List:3D Imaging San Diego 3D Imaging Group To subscribe/unsubscribe:listserv-at-bobcat.etsu.edu 3840 Camino Lindo To post a message:sd3d-at-bobcat.etsu.edu San Diego, CA 92122 Email:BRANDE-at-SDSC.EDU Voice:619-587-4830
---------- Forwarded message ----------
THE FOLLOWING RELEASE MOVED OVER PR NEWSWIRE ON TUESDAY, FEBRUARY 7, 1995 AT 11:42 AM, PST.
Apple Announces QuickTime Conferencing Open, Cross Platform Conferencing, Collaboration and Multimedia Communications Technology
SAN FRANCISCO, California--February 7, 1995--Apple Computer today announced a cross platform conferencing, collaboration and multimedia communications technology that allows personal computer users to share real-time information, images and sound anywhere in the world. Apple is currently making the technology, called QuickTime Conferencing, available to corporate allies who plan to create or have announced they are creating end user applications based on the technology. QuickTime Conferencing is a standards- based architecture that allows users to:
-- video conference and collaborate--to share and annotate text, images, screen capture, sound, video and virtual scenes real-time among fellow conference participants in a variety of locations worldwide. QuickTime Conferencing allows users to record conversations and transform those conversations into QuickTime movies. All of this can be done on a variety of networks such as an Integrated Services Digital Network (ISDN), the worldwide internet, local area and wide area networks and Asynchronous Transfer Mode (ATM) networks. QuickTime Conferencing can be used by a number of simultaneous users, the total number being only by available network bandwidth.
-- conduct cross platform video conferencing connectivity between Macintosh computers, PCs, UNIX systems and room-based conferencing systems through the use of the H.320 worldwide teleconferencing standard.
-- broadcast and view multimedia content--digital audio, music and video on a local or wide area network.
Through alliances QuickTime Conferencing technology is expected to yield product bundles such as: -- Apple Media Conference Kit--Consisting of the QuickTime Conferencing system extension, the Apple Media Conference application and a high quality, color video camera. -- Apple Media Conference Pro Kit--Consisting of the QuickTime Conferencing system extension, the Apple Media Conference application, a color video camera and an H.320 codec/ISDN adapter board. Being developed by Sagem/SAT, a leading international communications product company, the board is designed to allow interoperability between platforms (Power Macintosh to Macintosh, PC, UNIX and room systems) and full-screen image sharing. --Complete Media Conference System--Consisting of an Apple Media Conference Kit, a Power Macintosh 7100 AV, a 17 inch color monitor, external speakers and a keyboard.
Because QuickTime Conferencing is software-based, it is easily incorporated into new and existing third party products. As such, Apple believes that QuickTime-compatible products could yield extremely affordable prices: -- Apple Media Conference Kit--under $200 -- Apple Media Conference Pro Kit--under $1,750 -- Complete Media Conferencing System--under $6,000
Apple is working with a wide range of companies including telcos, network, software and hardware providers and developers to provide a range of solutions that take advantage of the benefits of QuickTime Conferencing (see associated releases). These allies have announced that they expect to make products available in the second quarter of 1995. From the home office to university campuses to the multinational enterprise network, QuickTime Conferencing will allow users to communicate with people across the country or across the world. Users won't have to worry about whether their hardware equipment, networking equipment and applications are compatible with the solutions being used on the other end of the network line. QuickTime Conferencing is designed to be fully operational with H.320 standards-based systems. "The introduction of QuickTime Conferencing will not only extend Apple's leadership in multimedia, but will make an important difference in the video conferencing and collaboration market," said Rick Shriner, vice president of Apple's Core Technologies Group. "Our goal in designing QuickTime Conferencing was to develop a solution that allowed people the opportunity to communicate and collaborate. By making it open in every sense of the word, our users can metaphorically break down the walls of their homes, schools and offices and expand the boundaries of their lives." QuickTime Conferencing users can have access to people, information, sights and sounds that could never be combined before. For example: -- An author in Tokyo, Japan and her publisher in San Francisco, California can view and discuss cover art for a new novel. They can each view the design at several different angles, change the visual perspective of the artwork, and annotate the image and accompanying text for the other to see. -- A sixth grade class in Dallas, Texas can discuss and view the effects of global warming with an environmental scientist at U.C. Berkeley's Lawrence Labs in California by using QuickTime Conferencing over the internet. -- A special effects producer in Hollywood, California can take a movie director on the East Coast through a virtual tour of a proposed set design. While the producer records their discussion as a QuickTime movie, the director can pan around the scene, zoom in to look at props and view the set design from a variety of angles. -- A breast cancer patient and her doctor in Fargo, North Dakota can consult with a leading oncologist in Boston, Massachusetts on her prognosis and course of treatment. The Boston physician can view her mammograms and annotate her medical chart as they converse. -- A CEO's company-wide address can be broadcast for easy viewing by all employees at their personal desktop.
Because QuickTime Conferencing allows for sharing of multimedia data and reduces the time and expense of travel, it allows people to be more productive than ever before. "In the past people found video conferencing easy to resist because prices were high and the number of people they could communicate with was extremely limited," said Rick LeFaivre, senior vice president of the Apple Technology Group. "Now for what we expect to be very aggressive prices, people can conduct a media conference with virtually anyone, anywhere in the world. A Power Macintosh QuickTime Conferencing user can share QuickTime VR (virtual reality) images, annotate text documents and share digital music over networks from basic rate ISDN to the internet to ATM." Because QuickTime Conferencing is a software-based architecture, application developers, communications providers and hardware vendors can easily develop compatible solutions. For example, Crosswise Corporation, the maker of Face to Face, a cross-platform document conferencing application, developed a QuickTime Conferencing-compatible version of their software in just one month. A QuickTime Conferencing compatible application shares the interface of other QuickTime Conferencing-enabled third party applications, so customers can begin using applications quickly and easily. QuickTime Conferencing is based on Apple's award winning QuickTime technology. It is a conferencing architecture which allows support for both industry standards such as H.320, as well as proprietary architectures, and codecs such as Indeo by Intel Corporation. QuickTime Conferencing is transport, compression and media-device independent. Apple's built-in AV capabilities combined with the performance of the PowerPC RISC architecture, make it easy for users to make multimedia connections with others on the information superhighway almost as soon as they pull QuickTime Conferencing out of the box. "Having QuickTime Conferencing available in my home, office, and studio literally allows me to be in multiple locations at one time--it's the next best thing to having a Star Trek transporter," said Los Angeles-based screenwriter and multimedia special effects consultant Michael Backes, co-author of the screenplay for Jurassic Park and other motion pictures. "Within the next few months, I'll be counting on QuickTime Conferencing as the backbone for my business." "The short and sweet of QuickTime Conferencing is that it requires less network bandwidth and uses innovative technology," says Matt Ghourdjian, National Director of Technology at Howrey & Simon, a 300-lawyer law firm serving Fortune 50 clients. Howrey & Simon intends to use the product to send QuickTime movies of depositions and re-enactments for lawyers to use in court; for live document sharing; for consultation between partners; and to conduct tours of the firm's Washington, DC office from Los Angeles. "It's simply outstanding," says Chris Masten, Howrey & Simon's Technical Litigation Support Manager. To use the Apple Media Conference Kit on the Macintosh, users need at least 16 Megabytes of RAM, a 68040 or PowerPC-based Macintosh, System 7.5, a network interface such as Ethernet, ISDN, Token Ring, and optionally the ability to digitize audio and video using the built-in AV subsystem or a third party digitizer card. To use the Apple Media Conference Pro Kit on Macintosh, users need at least 16 Megabytes of RAM, an AV PowerPC-based Macintosh and an ISDN connection. To communicate with QuickTime Conferencing users from the PC and other platforms, users will need an H.320 compatible codec on their machine, available from a variety of vendors. QuickTime Conferencing technology is currently under development and products using the technology have not yet been completed. Apple will provide pricing and availability information when products are completed and ready for release. Apple Computer, Inc., a recognized pioneer and innovator in the information industry, creates powerful solutions based on easy to use personal computers, servers, peripherals, software, online services, and personal digital assistants. Headquartered in Cupertino, California, Apple (NASDAQ:AAPL) develops, manufactures, licenses and markets products, technologies and services for the business, education, consumer, scientific & engineering and government markets in over 140 countries.
-30-
Apple, the Apple logo, QuickTime and Macintosh are registered trademarks and Power Macintosh is a trademark of Apple Computer, Inc. Additional company and product names may be trademarks or registered trademarks of the individual companies and are respectfully acknowledged.
END
Applelink pathway: News Break Apple & Industry News PR Express Apple Press Releases 2/7/95
I know of a company that reconditions old AMRAY SEMs. The owner used to work at AMRAY before starting his own company. Company is E.J. Feld located in Massachusetts. I can give you the phone on Monday, 2/13 when I return to work. Dr. David A. Shifler Powell Labs Ltd. Baltimore, MD 31231 (410) 327-3500 (410) 327-7506 (FAX)
Thanks to all the replies to my recent TEM survey, I am well on my way to preparing the survey for publication. I need some information -- I'm a chemist, not a microscopist -- What do TEM users see as important trends in TEM? New technology? Important applications? Is there other technology that is replacing TEM in any way. How important are accessories like EDS and EELS? For what uses?
Any comments on today's TEM would be greatly appreciated -- I'll send a copy of the survey to any contributor.
Does anyone in the group have the phone number for PRESENTATION TECHNOLOGY? We are in the information gathering stage for the purchase of a 35mm slide maker and the phone number we have (408-774-3733) is not correct.
Thanks for your help.
Doug
Douglas W. Cromey, M.S. Cell Biology and Anatomy Arizona Health Sciences Center 1501 N. Campbell Ave. Tucson, AZ 85724 (602)626-2824 dcromey-at-ccit.arizona.edu
} Dear Microscopists, } } Does anyone know if there is any recent books about applications of } microbeam techniques to mineralogy and petrology? } } The one I have was published by Mineralogical Association of Canada } (Short Course in Microbeam Techniques) in May 1976. } } Thanks in advance. } } Long LIang } ARCO EPMA/SEM Lab } PLano, TX
Try: McLaren, A.C., 1991, TEM of minerals and rocks, Cambridge University Press, Cambridge
Boland, J.N. & FitzGerald, J.D. (eds), 1993, Defects and processes in the solid state: geoscience applications, Developments in Petrology, Vol 14, Elsevier, Amsterdam
Buseck, P.R. (ed), 1992, Minerals and reactions at the atomic scale: TEM, Mineralogical Society of America, Reviews in Mineralogy, vol 27.
Cheers Timon
------------------------------------------------ Timon Fliervoet, Timonf-at-earth.ruu.nl, Faculty of Earth Sciences, Department of Geology, Utrecht University, P.O. Box 80.021 3508 TA Utrecht, the Netherlands. tel: ++31 30 - 535054, fax: ++31 30 - 537725
Help please!! SiSTek is looking for TEM analytical services in the southwestern US, preferably in the Phoenix metropolitan area where we are located. We are a company that provides consultant services to a number of manufacturers of Si-related deposition systems and need *local* (turnaround time and iterative analysis is important for our clients) TEM support. We believe there is a company in the Phoenix area offering TEM services, but can't find the name. Does anyone know the name/contact?
Subject: Time: 9:13 AM OFFICE MEMO Ecomet Polisher for sale Date: 2/14/95
FOR SALE: ECOMET 1 8" Polisher/grinder, complete with various polishing discs, alumina and lapping oil. 115 volt, 5 amp, new price in 1988 = $1250.00 Best offer. Call Doug Davis of EM Lab at UC Berkeley at (510) 642-2085 or e-mail: doug_davis-at-maillink.berkeley.edu
Any tips on cutting wells out of 24 well cell culture plates would be greatly appreciated.
Joe Joseph A. DeMaro Washington University Medical School Department of Neurology 660 S. Euclid Rm 212 Biotech St. Louis, MO 63110 jad1-at-cec.wustl.edu 314-362-9448
LaserMaster Corporation - Imaging Division is the ONLY company that manufactures a 1800 dpi plain paper laser printers. I work for LaserMaster and have just completed printing the 100/300 dpi Round Robin Greyscale Test images. If you are interested in obtaining them, you may e:mail me at Gregb-at-Sales.LMT.com and I will mail you a printout of the test images from the LaserMaster printer. I can be reached at 1-800-950-6363 Ext: 3207 if you have questions about your specific situation and resolution needs.
Thank You all, Greg Begin - LaserMaster Corp. Scientific/Medical Imaging Div. /\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\\/\/\/\/\/\/\/\/\/
} Date sent: Thu, 09 Feb 95 07:42:30 -600 } From: Supratik_Guha-at-mail.mmmg.com (SG) } To: microscopy-at-aaem.amc.anl.gov } Subject: printers for gray scale prints
} I am looking around for a gray scale printer to attach with our Gatan } slow scan CCD image aquisition system and would appreciate any } suggestions. We would prefer not to get a dye-sub printer due to the } high costs of printing. I understand that there are 1800 dpi laser } printers available, could someone point out manufacturers for these } machines? } } Supratik Guha } Senior Materials Scientist } 3M Corporate Research Labs. }
When I want to process cells for SEM from 24 well culture dishes I use a Bunsen burner and a scoopula (the curved metal device for scooping out dry chemicals). First, the cells are fixed as usual with glutaraldehyde then washed in buffer and distilled water. Then, working within a fume hood and wearing a heatproof glove, the scoopula is heated until red then touched to the underside of the culture dish. The curved metal is about the right size for the 24 well dishes. It takes 2 or 3 times to heat and cut until the whole bottom is released. Once the initial cut is made you need to keep the cells wet and this is easily done using a squirt bottle of distilled water. This technique does not appear to cause damage to the cells but we normally look at the more centrally positioned cells to avoid any artefacts. Hope this helps.
Nancy Smith Cal State Hayward nsmith-at-csuhayward.edu
Hello, everyone: Does anyone know where I can find some reference on measurement of electron mean free path of different materials? The mean free path I mean here is the mean free path for measuring the thickness when doing EELS, so this includes electrons of all the energy losses rather than one particular energy loss. We are particularly interested in getting the right electron mean free path for Chrome Oxide and evaporated Carbon. Thanks a lot.
Eric Wang FB-10 Roberts Hall Univ. of Washington Seattle. Wa 98195 (206) 543-1514
} Any tips on cutting wells out of 24 well cell culture plates would be greatly } } appreciated. } Joe } Joseph A. DeMaro
Depending on what exactly it is you are doing, how about using Thermanox (Thermonox?) plastic slides on the bottom of the wells - they make them specially to fit into the wells of 12 well plates and possibly the 24 well ones too. They come sterilised and you just pop them into the well before you add medium and cells and remove later, fix, dry etc and mount on a stub. The slide surface properties are supposed to duplicate the ordinary well bottoms. Its easier than cutting wells out of the bottom of the trays. Regards R Easingwood
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
I work for an OEM of scanning electron microscopes and am interested in keeping up with the latest technology and issues. I would like to subscribe to the mailing list. Could you please let me know what I need to do.
We process a considerable number of cell cultures for both SEM and TEM, and have found what we consider to be the ideal system. For this purpose, we have our investigators culture their cells in Leighton tubes...these are cell culture tubes with a flat bottom which holds a long, narrow plastic coverslip. Once the cells are attached, the medium is replaced with fixative, then the coverslip (which is attached to a nifty little handle) is removed. The plastic on the coverslip is impervious to all solvents used in microscopy, and can be embedded and sectioned for TEM.
I wouldn't think of doing it any other way.
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
We have two Beseler enlarger needing adjustments. Any information on available service person in the south, please remit details directly to me. Number I called was disconnected!
************************************************************ *Cesar D. Fermin, Ph.D \|*|/ Fax (504) 587-7389 * *Tulane Medical School /|*|\ Answ. Mach.(504) 584-2618 * *Pathology/SL79 \|*|/ Secretary (504) 584-2436 * *New Orleans, La 70 112 /|*|\ Lab (504) 5841 * *Fermin-at-TMC.Tulane.edu -} Director of Morphological Services* ************************************************************
Back around '82 or '83 I found that I had a problem with my Besseler's constantly slipping focus just a tiny bit. So I put pipe clamps (those metal rings that can be tightened of loosened with the turn a a screw) on the metal guides for the focus which I would tighten when focusing the bellows particulalry tight. -mc
On 15 Feb 1995, Fermin, Cesar wrote: } ref.: Beseler enlarger tune-up.
For those interested, Leighton tubes for cell culture are manufactured by Corning Costar and are available from Fisher Scientific (Cat.# 07-200- 367). They appear in the 95-96 catalog on page 721.
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
Dear Eric, If no expert in the field has a table of the mfp's, I can fax you tables of stopping powers for electrons in carbon, oxygen, iron & titanium-- you have to interpolate for chromium and calculate the mpf from dE/dx. Good luck. Yours, Bill Tivol
Many thanks to all who responded for my call for help with locating TEM service near us in the Phoenix metro area. As we thought, there is an established group in Phoenix who have been around for a few years and who provide TEM services.
For anyone else who might be interested, the company is called NanoTEM, Inc, 7620 E. McKellips Rd., Suite 4109, Scottsdale, Arizona 85257, phone 602 759 2808, fax 602 947 7615.
Greetings, Is there something out there that will make a thin film that isn't formvar? I have been using wire loops coated with a film of 1.2% formvar to support my small samples during rapid freezing and substitution. This works fine for acetone substitution but we would like to try Tetrahydrafuran (THF) as a substitution medium and, alas, THF eats the formvar.
We get a formvar film on the wire loop by casting small rectangles of formvar on water and then trasfering one to a loop.
Are there other compounds that could be used to make a film across the loop and that might survive THF??
I am trying to prepare polished sections from steel samples for EPMA analysis. Are there any recommended abrasive/size for rough grinding, fine grinding, rough polishing, and final polishing?
Your help is high appreciated.
Long Liang ARCO EPMA/SEM Lab Plano, TX LLIANG-at-is.arco.com
} } I am trying to prepare polished sections from steel samples for EPMA } analysis. Are there any recommended abrasive/size for rough grinding, } fine grinding, rough polishing, and final polishing?
I have always stuck to the simple silicon carbide paper (in steps from 120 to 1200 grade) and then diamond (6,3,1um) route, and found no problems with that.
Doug
+------------------------------------+ | Dr Douglas Arrell | | Mechanical Performance and Joining | | Institute for Advanced Materials | | 1755 ZG Petten | | Netherlands | | {ARRELL-at-JRC.NL} | | Tel. (+31) 2246 5287 | | Fax (+31) 2246 1917 | +------------------------------------+
CALL FOR PAPERS STUDENT COMPETITION TO BE HELD AT THE UNIVERSITY OF WISCONSIN WHITEWATER, WISCONSIN Friday, March 24, 1994 AWARDS: FIRST PLACE $100 SECOND PLACE $75 THIRD PLACE $25 Abstracts will be published in Midwest Microscopy. Microsgraphs from first place winner will be on the cover of Midwest Microscopy. Students will be judged on written abstract, presentation, and quality of study. Student competition is open to undergraduate and graduate students and may involve any type of microscopy. Student and sponsoring faculty member must be members of MSEM. Abstracts should be submitted before March 15 to : Dr. Lance Urven University of Wisconsin- Whitewater 800 West Main,Whitewater, WI 53190
John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu Time: 11:59 AM
Date:2/20/95 URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html NC EMAL
I have some colleagues that want to electropolish some fairly heavily deformed steel. Composition is: 0.4% C 0.6-0.9%Si 22-24%Cr 7-9%Ni Trace N Balance Fe. Does anyone have a good starting solution and condidtions for this alloy? Many Thanks. John Mansfield.
} I am trying to prepare polished sections from steel samples for EPMA } analysis. Are there any recommended abrasive/size for rough grinding, } fine grinding, rough polishing, and final polishing?
A step that may be advantageous is the final polish of the steel by use of a colloidal silica type solution [0.05 micron, with 9.8pH]. Dampen the cloth [such as a Buehler Mastertex] with distilled water; apply liberal amount of the solution [such as Buehler Mastermet] to the cloth, and apply firm pressure to soft pressure over a period of ~45 seconds, rotating the sample quite abit. A final word of caution: this solution [Mastermet], besides having the high pH [rough on skin], will crystallize into small, -very hard- particles. Is therefore highly advised to filter the solution a few times into a smaller bottle before each use. Have found this stuff to be -very- effective tho'. LECO also has a similar solution, but have not used it enough to get comfortable with it as the Mastermet.
In the previous steps I have used the 120 to 800 grit SiC papers, then a 9, 6, 1, & sometimes 1/4 micron diamond paste, on Texmet for the rough polish (9 & 6), and then Mastertex for the 1/4u. All depends on the grade and condition of the steel tho'... [all these recommendations are based on me hand polishing individual samples, and 3-6 samples in a ring held in hand]
Hope this helps, -Rob PS: I have no ties with Buehler, just a satisfied customer for +10 years. ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, | Rob Tayloe | MSM Spelunkers Club /\v/\ | | Metallographic Lab. | Missouri Speleological Survey /\v/\ | | Rolla Research Center | Bat Conservation International /\v/\ | | U.S. Bureau of Mines | Missouri Cave & Karst Conservancy | | tayloe-at-rorc.usbm.gov | National Speleological Society #32993 /\v/\ | | (314) 364-3169 x247 | American Cave Conservation Association | ''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''
The techniques described by Rob Tayloe are certainly in agreement with the references I have for polishing steel and will work quite well. The only note I would make is that we do supply a Colloidal Silica which DOES NOT CRYSTALLIZE. This is an important feature as Rob has mentioned because the crystallized particles can be a real problem if you do not realize they are there.
Our Non-Crystallizing Colloidal Silica is available as follows:
Part No. Description Price CS1-16 Non-Crystallizing Colloidal Silica 16 oz bottle $16.00 CS1-128 " " " 1 gallon bottle 78.00
If you'd like more information (MSDS etc) on this prodcut or any of our other products, please let me know.
Best regards-
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
one of my users is looking at TEM of insect heart. Does anyone have some good references on insect heart ultrastructure? Some of the cells associated with the heart appear highly vesiculated at the cell periphery. The morphology of these cells looks good, so we don't feel these stuctures are an artifact, but we have so far been unable to identify what kind of cell or cell type it might be. Can anyone suggest a reference or an investigator we could contact in this regard?
---------------------------------------------------------- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University San Diego CA 92182-0057 phone: (619) 594-4523 fax: (619) 594-5676 email to sbarlow-at-sunstroke.sdsu.edu
} SUSAN R. SESACK wrote: } } Would someone please provide me with instructions for enrolling in } the Internet bulletin board for histology and microscopy? Much } obliged.
To subscribe, please send a mail to "Listserver-at-AAEM.AMC.ANL.GOV" with "Subscribe Microscopy your_id-at-e-mail" in the body. Enjoy it.
Holland Cheng -------------------- Structural Biology Department of Biological Sciences Purdue University W. Lafayette, IN 47907-1101
} SUSAN R. SESACK wrote: } } Would someone please provide me with instructions for enrolling in } the Internet bulletin board for histology and microscopy? Much } obliged.
To subscribe, please send a mail to "Listserver-at-AAEM.AMC.ANL.GOV" with "Subscribe Microscopy your_id-at-e-mail" in the body. Enjoy it.
Btw, can someone in the server fix the returning route so that the reply can a global one? I would like to be on a list that I can see discussions (questions and answers) rather than a collection of of questions. Thanks in advance!
Holland Cheng -------------------- Structural Biology Department of Biological Sciences Purdue University W. Lafayette, IN 47907-1101
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ ++++I would like to add a small point about polishing specimens for analysis in the electron probe microanalyzer (EPMA). I prefer to leave the scratches in from the 1/4micron diamond polishing step in order to have an image feature on which to focus with the light optics. Since quantitative x-ray microanalysis samples should be flat-polished but unetched, it is hard to find a suitable surface feature to use for focusing without these fine scratches. For some reading on this, try Chapter 11 of Goldstein et al., Scanning Electron Microscopy and X-ray Microanalysis, 2nd edition, Plenum Press, 1992.
Good luck, Prof. C E Lyman
Electron Optics Lab Lehigh University 5 East Packer Avenue Bethlehem, PA 18015 ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
} } I am trying to prepare polished sections from steel samples for EPMA } } analysis. Are there any recommended abrasive/size for rough grinding, } } fine grinding, rough polishing, and final polishing? } } A step that may be advantageous is the final polish of the steel by } use of a colloidal silica type solution [0.05 micron, with 9.8pH]. } Dampen the cloth [such as a Buehler Mastertex] with distilled water; } apply liberal amount of the solution [such as Buehler Mastermet] to the } cloth, and apply firm pressure to soft pressure over a period of ~45 } seconds, rotating the sample quite abit. A final word of caution: } this solution [Mastermet], besides having the high pH [rough on skin], } will crystallize into small, -very hard- particles. Is therefore highly } advised to filter the solution a few times into a smaller bottle before } each use. Have found this stuff to be -very- effective tho'. LECO also } has a similar solution, but have not used it enough to get comfortable } with it as the Mastermet. } } In the previous steps I have used the 120 to 800 grit SiC papers, then } a 9, 6, 1, & sometimes 1/4 micron diamond paste, on Texmet for the } rough polish (9 & 6), and then Mastertex for the 1/4u. All depends on } the grade and condition of the steel tho'... [all these recommendations } are based on me hand polishing individual samples, and 3-6 samples in a } ring held in hand]
Dear Tobias, A carbon film--evaporated onto freshly-cleaved mica--should do the trick. After evaporation, float the film onto water and pick it up with the loop. You may have to experiment with thickness etc. to get the proper mech- anical strength, but it should certainly survive the THF. You may also want to try a mesh grid instead of the loop if the carbon film is not strong enough. Good luck. Yours, Bill Tivol
Message-Id: {9502212303.AA26486-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Re: TEM/formvar substitute/thin films Orig-Author: {tivol-at-tethys.ph.albany.edu}:ddn:wpafb ----------------------------------------------------------- Dear Tobias, A carbon film--evaporated onto freshly-cleaved mica--should do the trick. After evaporation, float the film onto water and pick it up with the loop. You may have to experiment with thickness etc. to get the proper mech- anical strength, but it should certainly survive the THF. You may also want to try a mesh grid instead of the loop if the carbon film is not strong enough. Good luck. Yours, Bill Tiv
I've done this quickly with "Image" by measuring the distance from the graph axis (x or y) to the data points in question. To calibrate (pixels to data units), just measure the distance between axis values.
On Wed, 22 H Feb 1995 Noonan_Eddie/perth-at-perth.atd.cra.com.au wrote:
} I have been asked by a colleague of mine who at present does not have } access to the Microscopy Listserver and who presently is sourcing a } new SEM for his lab to put out the following question. } } "In the next few weeks I shall be ordering a new analytical SEM. We } will use this SEM almost exclusively for EDS analysis. A motorised } stage will also be required for the SEM to accommodate overnight runs. } Therefore I require an ultra stable, reliable instrument. If any one } with experience in this area has any thoughts I would be grateful to } hear from you." } } Thanks in advance } } Mark Stewart } } Replies to: ejn-at-perth.atd.cra.com.au }
Jan Coetzee and Philip Oshel have posted about blood fixation but I havn't seen any replies on this topic. We have a project that will try to tie distortion of red cells to heat stress so fixation needs to be as artefact free as we can get it. Please Jan or Philip can you mail me with the best method you can recommend?
1) In the February '95 Biotechniques, New Products section, there's a blurb for a "Probe Clip" single slide incubation chamber sold by Grace Bio-Labs. Anyone have experience with these and/or a contact phone/fax # for Grace Bio-Labs?
2) A grad student in our lab has been doing ImmunoGold Silver (LM) immunostaining followed by BrDU - HRP - DAB for a second antigen. The silver was originally present, but faded out in the second reaction. Could have resulted from a number of factors, but what we were wondering is - is it possible to stabilize the silver with a sodium thiosulfate "fixer" step after the IGSS to protect it in subsequent immunoreactions, dehydrating and coverslipping? Any practical suggestions would be appre- ciated.
3) A colleague in Australia wants to purchase an antiserum to met- enkephalin. We have been buying from Incstar and getting good results, but the Australian distributor for Incstar charges outrageous prices. Does anyone know of an alternate antibody supplier that may be less ex- pensive for Australian customers?
------------------------------------------------------------------ |Glenn Holm *mime mail ok* Internet:karuzis-at-wccf.mit.edu | |M.I.T Dept. of Brain + Cog. Sci. This VAX doesn't do NeXTmail | |Cambridge, MA 02139 "Real Neuroscientists don't do gels!" | ------------------------------------------------------------------
Recently I posted a request for methods for lipid sizing. I have put together a summary of that request. If anyone would like a copy I will be happy to send you one, but it is rather lengthy for the listserver.
Marc C. Brande, M.S. SD3D Email List:3D Imaging San Diego 3D Imaging Group To subscribe/unsubscribe:listserv-at-bobcat.etsu.edu 3840 Camino Lindo To post a message:sd3d-at-bobcat.etsu.edu San Diego, CA 92122 Email:BRANDE-at-SDSC.EDU Voice:619-587-4830
On Thu, 23 Feb 1995, Glenn Holm wrote:
} 3 separate requests for information: } } 1) In the February '95 Biotechniques, New Products section, there's } a blurb for a "Probe Clip" single slide incubation chamber sold by } Grace Bio-Labs. Anyone have experience with these and/or a contact } phone/fax # for Grace Bio-Labs? } } 2) A grad student in our lab has been doing ImmunoGold Silver (LM) } immunostaining followed by BrDU - HRP - DAB for a second antigen. } The silver was originally present, but faded out in the second reaction. } Could have resulted from a number of factors, but what we were wondering } is - is it possible to stabilize the silver with a sodium thiosulfate } "fixer" step after the IGSS to protect it in subsequent immunoreactions, } dehydrating and coverslipping? Any practical suggestions would be appre- } ciated. } } 3) A colleague in Australia wants to purchase an antiserum to met- } enkephalin. We have been buying from Incstar and getting good results, } but the Australian distributor for Incstar charges outrageous prices. } Does anyone know of an alternate antibody supplier that may be less ex- } pensive for Australian customers? } } ------------------------------------------------------------------ } |Glenn Holm *mime mail ok* Internet:karuzis-at-wccf.mit.edu | } |M.I.T Dept. of Brain + Cog. Sci. This VAX doesn't do NeXTmail | } |Cambridge, MA 02139 "Real Neuroscientists don't do gels!" | } ------------------------------------------------------------------ }
I believe this question was asked on this list before, so I am sorry if I am repeating. A colleague here is having problems with negatives being ruined by static discharge in a JEOL 2010 TEM. JEOL suggests drying the negatives inside the TEM vacuum which works but is really not good and very inefficient. The problem began only recently when atmospheric humidity is increasing here. Does anyone know the cause of the static discharge problem and/or have a practical solution.
Norman Elliott | E-mail: nee-at-lanl.gov Los Alamos National Lab | Fax: 505-665-2104 MST-7 MS E549 | Voice: 505-667-1587 Los Alamos, NM 87545 |
I would like to announce the establishment of an FTP site for the Macintosh/Power PC based digital imaging microscopy software IPLabspectrum from Signal Analytics Corporation. There are several reasons for development of this site: 1. To provide individuals interested in developing a digital imaging microscopy system a chance to access and download demonstration versions of this particular Macintosh/PowerPC based digital imaging software for evaluation purposes.
2. To provide users of IPLabspectrum software for a site to download and upload user-generated system extensions and scripts .
Although the software is from a commercial source, the establishment of the FTP site was a decision by made by myself and other users of the Digital Imaging Microscopy Facility here at the University of Alabama at Birmingham as one way of promoting the digital imaging microscopy as a research tool. Signal Analytics Corporation, the developers of the software, has generously made available demonstration versions of their entire software line for FTP download purposes. If any other developers of imaging software wish to use this site as a repository for demonstration versions of their software I would be more than happy to accomodate them.
The demonstration versions of this software are organized into several folders, and separated into Macintosh and PowerPC versions of that software. There are versions of the software that support specific imaging boards (e.g. Scion LG-3) in the demo folders. Also in the site are demo versions of calcium ratio (IPLab Ratio), Three Dimension Reconstruction, and Multiprobe (FISH) extensions. In addition is a separate program for reading scanned images of electrophoretic gels. (IP Lab Gel)
There is a folder (directory) for Uploading of Scripts, Extensions, Messages, ect. provided-however the caveat there is that downloading the extensions from that directory is at your own risk, since this is a public access directory. If you choose to download from this directory, please take the time to scan the files for viruses or other nasties. Our aim is to monitor the upload directory as the usage increases, test the material in the directory for problems (bugs, viruses, and other annoying creatures) and then shift those programs that we consider problem free to a specific download-only directory. At present this download-only directory for user-uploaded files does not exist.
To access the site use the address FTP.BMG.UAB.EDU, logon as ANONYMOUS to enter the public directory. In the directory is a subdirectory (folder) IP_Labspectrum. Within this subdirectory are other directories containing the various demos of the software. I also included the shareware programs FETCH and BinHex 5.0 in a directory named FTP_SITE_Utilities for individuals (like myself) who rather use FETCH as a front end for FTP browsing and transfer.
This site is still undergoing development, so please forgive the style of organization. Any comments, suggestions, ect. about the site, digital imaging microscopy would be greatly appreciated. My e-mail address is KJMcCarthy-at-CellBio.BMG.UAB.EDU
Best Regards Kevin McCarthy Assistant Professor Department of Cell Biology University of Alabama at Birmingham Birmingham, Alabama 35294 Phone 205-934-9923/9924 Fax 205-934-7029
Return-Receipt-To: FRAWLEY-at-a1.resmel.bhp.com.au Registered-Mail-Reply-Requested-By: FRAWLEY-at-a1.resmel.bhp.com.au Mr-Received: by mta VULCAN.MUAS; Relayed; Fri, 24 Feb 1995 11:44:58 +1100 Mr-Received: by mta VULCAN; Relayed; Fri, 24 Feb 1995 11:59:09 +1100 Disclose-Recipients: prohibited
I am interested in using an extraction replica technique to determine MnS precipitate size distributions in a TEM.
The technique being considered is as follows: -Polish sample -Etch to give some surface relief -Carbon coat -Using blade cut 1mm size grid on surface -Release carbon film containing ppt's. from surface using etchant -Wash carbon replica in alcohol and water -Position on TEM grid.
My problem is finding an etchant that will not attack the very small MnS ppts. It has been reported that some etchants attack the Mn in these ppt's.
Any suggestions on the technique or the etchant would be appreciated.
Regards Leo D Frawley frawley-at-a1.resmel.bhp.com.au
} Date sent: Thu, 23 Feb 1995 14:09:10 -0700 } To: microscopy-at-aaem.amc.anl.gov } From: nee-at-beta.lanl.gov (Norman Elliott) } Subject: TEM negatives
} Dear list, } } I believe this question was asked on this list before, so I am } sorry if I am repeating. A colleague here is having problems with } negatives being ruined by static discharge in a JEOL 2010 TEM. JEOL } suggests drying the negatives inside the TEM vacuum which works but is } really not good and very inefficient. The problem began only recently when } atmospheric humidity is increasing here. Does anyone know the cause of the } static discharge problem and/or have a practical solution. } } We had this static discharge problem when first we bought our 2010. It was solved when JEOL replaced the Teflon drive gear (large) in the camera with a conducting (metal) one. Coating the gear with a conducting spray didn't work! We also installed a separate vacuum desiccator for the plates (diffusion pump and liquid nitrogen trap).
Peter Smith ps-at-bunyip.ph.rmit.edu.au RMIT App Physics Ph: (03) 660 3390 Office GPO Box 2476V (03) 660 2205 Lab Melbourne Fax: (03) 660 3837 Vic 3001 AUSTRALIA
About your discharge problem, we believe this is indeed strongly linked with the vacuum. Therefore we always use one or more desicators before the plates enter the microscope. This way also the vacuum in the microscope restores faster. The problem also depends on the plates: we had the experience that Ilford plates always gave discharge, while Kodak and Agfa did not in the same environment. Also handling of the plates can be important: sliding plates over one another can cause discharges before and after use in the microscope. Hope this can be of help, Nick Schryvers
I had some static discharge problems with my JEOL 1200exII some time ago. Every 8th to 10th neg had streaks on it. they were thin and sharp with two or three finger like projections coming fron the same point. The service tech. cleaned the camera (several times) and that cleared the problem up. It did take several tries and return trips to get the dirt (thought to be a metal fragment) cleaned up.
I hope this helps. Larry Hawkey hawkey-at-neuro.duke.edu 919-641-6425
Norman, Although I am not familiar w/ this particular model of JEOL, could it be possible that the static discharge is occuring before or (more likely) after the film has gone into/come out of the scope? We had an individual in the lab I was in before who tended to wear synthetic fabrics and on particularly dry Arizona days they could really ZAP the film. One solution is to wear a wrist grounding strap when handling the film (like the kind that people are supposed to wear when working inside their computers). Hope this helps. Doug
Douglas W. Cromey, M.S. Cell Biology and Anatomy Arizona Health Sciences Center 1501 N. Campbell Ave. Tucson, AZ 85724 (602)626-2824 dcromey-at-ccit.arizona.edu
Dear Norman, We, too, have had occasional problems with static discharges on our films--especially with LoDose, which is very sensitive to light. Our worst problems occur when the humidity is low. You must be very careful when sepa- rating films from the stack and when removing the stack from the package. You might also check if your lab coats produce static electricity--we just got new polyester coats which are terrific generators. If you are completely dark-ad- apted and loading the folm in total darkness, you can see the discharges as they occur, so at least you will know what is going on. [oops, I meant film] If your desiccant is too efficient, the discharge problem will be worse. Good luck, sometimes the procedures necessary to prevent these discharges are te- dious. If all else fails, try using 4489 film--it's not nearly as sensitive as SO163, but it is much finer grain and gives excellent images. We've never had discharge problems with it, probably because discharges are not intense enough to show up. Yours, Bill Tivol
In the past we too have had problems with static discharge ruining negatives (the film was SO163 being used in a JEOL 4000EX). We found that the discharges were coming from our vinyl gloves; when we switched to cotton gloves the problem went away. Additionally, when loading the camera we never use pre-dessicated film, but instead load the camera with film that has been at atmosphere (and warmed up to room temperature if it's been in the refrigerator). We then dessicate the camera box prior to putting it in the microscope.
On a related topic, on occasion we've observed a fine network of cracks in the emulsion of our SO163 film. Under an optical microscope the film looks similar to a dried out mud flat with cracks spaced rougly 0.05 mm apart. These cracks would appear regardless of whether we dried the film at room temperature overnight or in a heated film drier. The cracking also appeared with both new and old chemicals (D19 diluted 2:1 4 minutes; kodak rapid fixer 5 minutes). Thinking we had a bad batch of film we sent some of the material to Kodak, but they were unable to duplicate the effect. Finally, it was suggested that we reduce the concentration of hardener in the fixer to half of what Kodak recommends. This seems to have reduced the cracking quite a bit, although not entirely. Has anyone had similar problems?
+---------------------------------------------+ ! Douglas L. Medlin ! ! Physical Properties of Materials Department ! ! Mail Stop 9402 ! ! Sandia National Laboratories ! ! Livermore, California 94551 ! ! ! ! (510) 294-2825 ! ! dlmedli-at-california.sandia.gov ! +---------------------------------------------+ .\
Is there anyone out there who has any tips/techniques on polishing hygroscopic material for microanalysis. Specifically, I need to polish some CaO particles with CaC rinds. I could probably polish them with silicone oil, but how do I get the oil off the samples before I put them in the microprobe? Any comments or suggestions would be appreciated.
Thanks in advance
Glenn ********************************************************************** * Glenn Poirier glenn_p-at-geosci.lan.mcgill.ca * * Electron Microprobe Lab Phone: (514) 398 6774 * * Earth and Planetary Sciences Fax: (514) 398 4680 * * McGill University THERE ARE THREE SIDES TO EVERY STORY; * * Montreal, Quebec YOUR SIDE MY SIDE AND THE TRUTH * **********************************************************************
I am a graduate student currently working on a project utilizing EDS to get quantitative elemental information of atmospheric particles. In past few monthes, I have been trying to get hold of a article cited below through the interlibrary service and without any success. It will be greatly appreciated if anyone out there can direct me where and how to get this article.
Heinrich, K. F. J. (1987) "Mass Absorption Coefficients for Electron Probe Microanalysis." in "Proc. 11th Int. Cong. on X-ray Optics and Microanalysis." edited by J. Brown and R. Packwood, published by J. D. Brown, University of Western, Ontario, pp67-119.
Po-Fu Huang Particle Technology Laboratory Department of Mechanical Engineering University of Minnesota (Office) (612) 626-7227 (Lab) (612) 625-7307 (Fax) (612) 625-6069
We pre-pump negatives in a separate vacuum chamber with phosphorus pentoxide powder placed in the bottom of chamber. This film is loaded into a spare cassette and has always been suitable for use after being pumped for 1-2 hours or left under vacuum. After about a week, the powder absorbs moisture and must be replaced. The old powder can be neutralized with NaOH pellets and when NaOH pellets have dissolved, can be discarded by flushing with water.
Marge Hukee EM Core Facility hukee.margaret-at-mayo.edu Mayo Foundation (507) 284-3148 ---------------------------------------------------------------------------- --
I am currently writing a grant proposal, and I have been knocking my head against the wall looking for some sort of data on the optical transmission properties of nervous tissue (the brain). Does anyone have a good reference, or, from a practical standpoint, how thick do brain tissue slices have to be to obtain good optical transmission (esp. relative to other fatty tissue slices like those from the earlobe, whose transmission characteristics I can find since people have been doing pulse oximetry.).
Thanks in advance,
John Conroy University of Utah Dept. Bioengineering
We had some static discharge on our 35 mm film in the past. It occurred when pre-desiccated bulk was being rolled onto cassettes. The problem disappeared after bulk film was left in atmosphere. We never had problem with plates because we did not pre-desiccate plates. One may try leaving the exposed plates in a humid chamber for a while before developing them, if the problem persisted.
I have an LKB Ultratome III that is surplus to our needs. Purchased in 1978, almost never used. If you might be interested in making an offer or know of a worthwhile place for a donation, please let me know.
Thanks,
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95060 emlab-at-ucsco.ucsc.edu (408) 459-2477
Glenn (and others) I have polished B2O3 with the 3M lapping film and no water. It doesn't produce a great polish, but you can usually find a few spots where you can squeeze a beam in. If you don't use the 3M film, let me know and I'll tell you where you can get some or even send you a scrap. Maggy Piranian M.U.N.
On Fri, 24 Feb 1995, Glenn Poirier wrote:
} Greetings Microscopists } } Is there anyone out there who has any tips/techniques on polishing hygroscopic } material for microanalysis. Specifically, I need to polish some CaO } particles with CaC rinds. I could probably polish them with silicone } oil, but how do I get the oil off the samples before I put them in } the microprobe? Any comments or suggestions would be appreciated. } } Thanks in advance } } Glenn } ********************************************************************** } * Glenn Poirier glenn_p-at-geosci.lan.mcgill.ca * } * Electron Microprobe Lab Phone: (514) 398 6774 * } * Earth and Planetary Sciences Fax: (514) 398 4680 * } * McGill University THERE ARE THREE SIDES TO EVERY STORY; * } * Montreal, Quebec YOUR SIDE MY SIDE AND THE TRUTH * } ********************************************************************** }
EM folk: I'm trying to use T-max 100 in the 35mm camera on my Zeiss EM 10C. Everything works well (you get nice short exposures, and this particular camera can be handled in the light, so there's no problem with using a panchromatic film). But the film gets *BRITTLE* in high vacuum. I have snapped the film in half simply by bending it as I unloaded the camera. Any hints, or do I just need to switch to one of the orthochromatic films? If so, which one(s) have you tried and liked? Julian Smith III Biology Winthrop University Rock Hill, SC 803-323-2111 (vox)
Message-ID: {MAILQUEUE-101.950227084552.256-at-physics.uwc.ac.za} To: microscopy-at-aaem.amc.anl.gov
Douglas Medlin wrote about fine cracks appearing on some of their TEM negatives. I believe you are experiencing what photographers called :crazing:. This is an effect that occurs when the temperature of the developer and fixer is not the same, or at least the difference is not small enough. It is exactly as you describe it, a fine line structure on the film that looks that mud cracks.
Dirk Knoesen Department of Physics, University Western Cape, Bellville, 7530 South Africa e-mail: dirk-at-physics.uwc.ac.za Phone: (+21) 959 2236 Fax: (+21) 959 3474
We use Eastman motion picture film (5302 fine grain release positive film) in Philips EM300 in the past and currently in Zeiss EM902 without any problem.
One suggestion I would make would be polishing with abrasive lapping films. These are abrasive particles which are bonded to a polyester film. It is a higher grade of traditional SiC grinding paper and it comes in micronized particle sizes. The material is designed to be used either with or without a lubricant which should take care of your problem.
While it is always preferable to polish with some sort of lubricant, this is a viable alternative for anhydrous materials. I must also add that the lapping film is one of our products so I do have a vested interest in this suggestion. If you would like a sample to try out, please contact me. It is avaialable in Aluminum oxide down to .05 micron and in Diamond down to 0.1 micron. Typical price for Al2O3 film is about $1.39 per 8" PSA Disc and for Diamond about $24 per 8" PSA disc. 8" Plain Back Diamond Film is more typically used and is priced at about $20 per 8" Disc. Prices, of course, will decrease in quantity. The diamond plain back films are used extensively in Tripo Polishing for SEM and TEM applications.
Best regards-
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
I have used Kodak Fine Grain Release Positive (5302) in Philips microscopes for 30 years. Never gave a problem with cracking with the unperforated (no longer available) or perforated stock. Gave some problems with "lightning strikes" on the final 4-5 frames out of 40. Philips have special cameras with large diameter hubs so the film is never bent much and used unperforated film so there was no tendency to crack at perforations. A 35 mm camera in our Hitachi H-7000 gives very fine cracking across the film between the edges of the perforations even when using the film Hitachi recommends. This can only be avoided by using film which is only JUST dry enough, shooting the whole roll and processing it in about 4 hours.
For your information, Materials Microscopy is a new newsletter dedicated to the professional interests of the working materials microscopist. Those who wish to receive a free subscription should provide us with their full mailing address via FAX -at- (602) 947-7615 or E-mail: MatlsMicrs-at-aol.com. Contributed articles should be mailed to: Materials Microscopy P.O. Box 2014 Scottsdale, AZ 85252 Please contact me if you need any further information. Thank you.
Rene E. Nicholas Circulation Manager Materials Microscopy
Hi Barbara! I tried to no avail to send you the present message at your email address: it stubbornly bounced back, so I eventually post it to the microscopy list. I apologize to the members of this list if they should feel this is wasting bandwidth: just delete and go on to the next message ;-)
Barbara! I am afraid I did not make my question clear enough ;-) What I wanted to know was: 1) the testes you wish to fix are from what species ?
2) for what kind of microscopy are they to be processed ? I.e. T.E.M., S.E.M., or light microscopy ?
3) What's the embedding medium?
4) in the case of light microscopy, do you contemplate using some special technique (besides for recognizing the different stages of the seminiferous epithelium) which would require that some specific chemical would be either required or, on the contrary, avoided?
In my personal experience with rat testes and light microscopy (this is rather very long ago: more than 30 years! :-( ), Bouin and other picric acid containing variations thereof are not optimal fixatives for preservation of the acrosomial apparatus of spermatids, which is one of the features used by most classification methods (Clermont's among others...). The best fixatives for light microscopy of paraffin embedded tissue are those containing potassium dichromate such as, for example, Helly's fluid (Zenker + formaldehyde), but they also are followed, after rinsing and dehydration, by shrinkage which is still more important than that observed after Bouin's or Bouin-Hollande's (somewhat better) fluids.
BTW, what's wrong with picric acid? It is always shipped in 'moist condition' in order to minimize hazards. I work in our lab since 1953, with Bouin, Bouin-Allen, Bouin-Hollande, Dubosq-Brazil etc., every day, and we never experienced any problem. We store the picric acid for our needs in the form of a saturated aqueous solution which keeps for as long as you will. This stock solution is then used for preparing all picric acid containing fixatives.
There is no point in trying to eliminate the picric acid from the fixed pieces of tissue before sectioning. If the presence of picric acid in the slides should hamper subsequent staining (a rare occurrence, btw), then the easiest and expeditious way to get rid of it is to pretend your tissue was fixed with a sublimate containing fluid: at the rehydration step, your slides should undergo treatment by alcoholic iodine (or lugol solution) and sodium thiosulfate in the usual way and 'voila': no more picric acid on the slides within 2 minutes! Under no circumstances should the pieces of tissue fixed with a picric acid containing fixative undergo rinsing in water! This is sheer heresy ;-) because it induces a lot of swelling in the tissue before shrinking even more during dehydration (I never found out which histologist ever advocated such method, but I know quite a number of renowned ones who firmly condemned it, and with good reason). If you insist on rinsing the pieces of testes after fixation, then merely increasing the number of steps through 90x alcohol should do the trick; you may even add a thin layer (1-2 mm thick) of lithium carbonate on the bottom of the alcohol containing flasks: this helps dissolving part of the picric acid away.
Possibly the method which should yield the best pictures has been documented in a book entitled 'Histological and histopathological Evaluation of the Testis', by Russell,Ettlin, Sinha Hikim and Clegg: Cache River Press 1990, (15777 Bolesta, Box 129, Clearwater, Fl 34620): ISBN 09627422-0-1.
At the end of the book, there is an appendix about the materials and methods which you might find useful (although perfusion fixation of rat testes is among the most frustrating experiences I ever had, even with a lot of heparin ). This would also require embedding in epoxy resin and making large semi-thin sections, which I don't know whether you are prepared to do ;). Glutaraldehyde certainly seems to me the best choice, but then paraffin embedding would be ruled out (too hard and too brittle after paraffin embedding). HTH, and good luck. Let me know about the outcome (good, I hope) John (correction: alcohol above should read 90 percent in volume)
*********************************************************** * Jean C. Desclin (John), Associate Prof. of Histology * * Laboratory of Histology - Faculty of Medicine * * Brussels Free University (U.L.B.) * * e-mail: desclinj-at-ulb.ac.be (internet) * * snail mail: route de Lennik 808 * * B - 1070 Brussels Belgium * ***********************************************************
To whomever, Would someone please repost the recent info on lipid sizing (K.Walters?) and the three responses on lipid negative staining. I had another crash and lost these responses. Thanks Mike D.
I am currently writing a grant proposal, and I have been knocking my head against the wall looking for some sort of data on the optical transmission properties of nervous tissue (the brain). Does anyone have a good reference, or, from a practical standpoint, how thick do brain tissue slices have to be to obtain good optical transmission (esp. relative to other fatty tissue slices like those from the earlobe, whose transmission characteristics I can find since people have been doing pulse oximetry.).
Thanks in advance,
John Conroy University of Utah Dept. Bioengineering
Dear John,
It has been years since I worked in this area, but I suggest you might look at the literature for the optical transmission properties of retina. As a nervous tissue, its optical properties should be quite similar to the CNS as long as the measurements are not made in the vicinity of the fovea. And, of course, due the importance of retinal transparency for vision, its optical properties must be well known. For a place to start I'd have a look at the "Handbook of sensory physiology" Springer-Verlag.
Just my two cents worth, Steven ----------------------------------------------------------------------------- Steven L. Goodman, Ph.D. Dept. Animal Health and Biomedical Sciences 608-262-0816 (office) University of Wisconsin 608-262-7420 (FAX) 1655 Linden Drive Madison, WI 53706 SLG-at-AHABS.WISC.EDU --------------------------------------------------------------------------
} Return-Path: {delannoy-at-welchlink.welch.jhu.edu} } Received: from AAEM.AMC.ANL.GOV by welchlink.welch.jhu.edu (5.0/SMI-SVR4) } id AA04845; Tue, 28 Feb 1995 09:52:41 -0500 } Date: Tue, 28 Feb 1995 09:38:53 -0500 } Message-Id: {9502281438.AA02618-at-welchlink.welch.jhu.edu} } X-Sender: delannoy-at-welchlink.welch.jhu.edu } Mime-Version: 1.0 } To: microscopy-at-aaem.amc.anl.gov } From: delannoy-at-welchlink.welch.jhu.edu (Michael Delannoy) } Subject: lipid sizing and neg. stain } X-Mailer: {Windows Eudora Version 1.4.2b16} } Content-Type: text/plain; charset="us-ascii" } } To whomever, } Would someone please repost the recent info on lipid sizing (K.Walters?) } and the three responses on lipid negative staining. I had another crash and } lost } these responses. } Thanks } Mike D. } }
Just a note to let you guys know that our link from this mailing list to the Usenet newsgroup sci.techniques.microscopy has been broken. The gateway at sci.techniques.microscopy.usenet-at-decwrl.dec.com has been closed down. Dec no longer support it. If you want as much coverage for you questions and posts as possible I encourage you to post both to this list and also to the Newssgroup.
If anyone knows how to set up a mail gateway to the Newsgroup, and also from the Newsgroup to the mailing list, I would be very interested to hear how to do it so we could reestablish the link and possibly make it bi-directional this time!
Many thanks Jfm.
______________ John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
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