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We have recommended Ross Ultra Super Glue to our customers as the best glue to use for Tripod Polishing applications. Recently, Ross has "improved" their formula and it is no longer satisfactory.
This note serves 2 purposes:
1) As a warning to Tripodders to beware of the "new and improved" Ross Ultra Super Glue (Blue label)
2) To ask if anyone out there has found another suitable glue to use. The main concern is for wedge polishing applications where the sample will be reduced to only a few microns in thickness.
There are super glues out there that work fine, but we are on a quest to find the best. Any help you can provide would be great!
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } What are some effective solvents for removing adhesive tape, spots, etc. } } from aluminum SEM mounts. Acetone is marginal as are other ketones. } } I've tried several alcohols without success. Caustics are out because } } they eat the aluminum. Pure and mixed acids don't work either. I've } } even tried WD-40 and liquid dish soap. } } } } The ideal solvent would have low toxicity, low cost, etc... Water } } solubility would be a bonus. I'd really like a chlorinated solvent } } vapor degreaser! Fat chance. } } } } Any suggestions? Terpenes, perhaps? } }
I have been using petroleum ether with considerable success. Used in a fumehood it is quite safe on aluminium, most plastics and other surfaces such as glass, wood and even paper.
-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.- Glynis de Silveira University of Cambridge Department of Materials Science and Metallurgy E-m:gds1002-at-cam.ac.uk Pembroke Street, UK Tel:+44(0)1223 334434 CB2 3QZ Fax:+44(0)1223 334567
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I have bought a AGFA DUOSCAN scanner recently, it has a tray for positives and one for negatives up to A4 size (~8"X11")
Resolution is 2000x1000dpi optical (4Kx4K interpolated but who would use that!).
Works fine for me but I find it rather slow!
Usual disclaimer apply, I have no connection with AGFA I am only a customer...
Jean-Marc Boichat email: jean-marc.boechat-at-chma.mhs.ciba.com EM LABS FO 5.1 phone:+4126 435 6979 fax: +4126 435 6907 Ciba research Center P.O. Box 64 CH- 1723 Marly 1 When things go wrong, don't go with them! Switzerland
Disclaimer: "nobody in this company ever cared for what I said, why would they start now".
X400-Received: by host241.abbott.com (Internal Mail Agent-1); Thu, 1 May 1997 08:27:29 -0500 Alternate-Recipient: prohibited
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On Wed, 30 Apr 1997, Crossman, Harold wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } What are some effective solvents for removing adhesive tape, spots, etc. } from aluminum SEM mounts. Acetone is marginal as are other ketones. } I've tried several alcohols without success. Caustics are out because } they eat the aluminum. Pure and mixed acids don't work either. I've } even tried WD-40 and liquid dish soap. } } The ideal solvent would have low toxicity, low cost, etc... Water } solubility would be a bonus. I'd really like a chlorinated solvent } vapor degreaser! Fat chance. } } Any suggestions? Terpenes, perhaps? } } } ------------------------------------------------ } Opinions or statements expressed herein, rational or otherwise, do not } necessarily reflect those of my employer. } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com} } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com} } Harold, have you tried soaking them in a 10% solution of Microsolution? This has worked for me several times I've wanted to reuse stubs. Use hot water (the hotter the better) when starting then add the Microsolution. It works best with hot water but letting things soak in cold water overnight works O.K. too. Hope this helps.
Peace through Christ,
Phil Rutledge, Director e-mail: prutle1-at-gl.umbc.edu Center for Microscopy voice: (410) 455-3582 UMBC Dept. of Biology fax: (410) 455-3875 Catonsville, MD 21228 ///// / / / / /////// /////// / / /////// /////// / / / / / / / / / / / / /////
If you are not concerned with a smooth surface Al stub, I just grind off the samples on a piece of fine sandpaper, dump these into acetone and let it sit for several hours. Pour off dirty acetone, add clean acetone and then pull out stubs and let dry on paper towels. I normally process about 30 or so stubs at a time. Most of my users take the stubs with them, so I don't have to do this often. Bruce Cutler, Microscopy Lab, Univ. Kansas, Lawrence
} Harold, } have you tried soaking them in a 10% solution of Microsolution? } This has worked for me several times I've wanted to reuse stubs. Use hot } water (the hotter the better) when starting then add the Microsolution. } It works best with hot water but letting things soak in cold water } overnight works O.K. too. Hope this helps. }
I have used microsolution to clean vacuum parts, mainly stainless steel. I have found that it severly attacks aluminum parts, therefore, I no longer use it for aluminum.
Michael T. Marshall Research Engineer, Electron Microscopy University of Illinois at Urbana-Champaign Frederick Seitz Materials Research Laboratory 104 South Goodwin avenue Urbana, IL 61801-2985 (217) 244-8193 fax: (217) 244-2278
Harold, The adhesives we usually encounter in microscopy are probably at least 60% cross-linked so there is no magic, modest hazard, solvent that will fully dissolve them. - Heptane will swell some adhesives and then a heptane wetted paper towel will wipe the adhesive off the mount. - THF will dissolve the non cross-linked component of some adhesives and that could make the adhesive easier to remove. - We use acetone, ethanol and muscle.
As always, read the MSDS for the solvent and obey the numerous, frequently changing, local, state and federal regulations for transportation and disposal.
...the usual disclaimers...At 11:42 AM 4/30/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes, one more request for info on the best camera to use with our microscope-computer set up. I'm sorry for not keeping all the info that has been out on this subject a few months back, but I thought there was no way the money would show up for one any time soon. Wrong. We have an Olympus BH microscope and a Power Mac computer. We are interested in using the NIH image software with this set up, but we need a CCD camera with at least good resolution and any software/hardware to link the camera and computer. Notes on personal experience with these systems would be appreciated. I will need to know how much the camera and interface will cost too. Once again, sorry for not paying closer attention the first time.
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In response to Mr. Henriks 'quest' for better Super-Glues for Wedge Polishing of TEM samples:
Through a bit of research on the subject, it has become apparent that the cyanoacrylate adhesives used for Wedge Polishing in general are not capable of forming a strong chemical bond to the surface of glass. This is due mainly to their affinity for materials to which water will adsorb. (Thus the problem with 'Super-Gluing' one's fingers together.) Since the majority of vendors selling 'Tripod'-like polishing fixtures for wedge polishing also supply only glass or PYREX stubs for mounting the sample, it is no surprise that the samples are often lost after hours of effort has been expended on thinning the sample.
For years, wedge polishers have been switching from one cyanoacrylate product to the next in order to find one that will adhere better to glass. Two years ago, after looking into the problem, it became clear to me that using a transparent polymer instead of glass for the mounting stub would solve the problem. The trick was to find a polymer that could withstand the attack of cyanoacrylate solvents such as acetone and nitroethane. The solution made the choice of cyanoacrylate adhesive much less important.
As this note is meant to be informative as opposed to an advertisement for product, I would be happy to discuss our solution to the 'Super-Glue' problem in more detail off-line.
Regards, Scott D. Holt BUEHLER LTD. 41 Waukegan Rd. Lake Bluff, IL 60044 USA (847)295-4546 Fax (847)295-7942 http://www.buehlerltd.com
We are looking for SEM slow scan image digitizer and TEM image digitizer - for really old microscopes: Philips SEM 503 and TEM 300. We would appreciate any comments, including pricing information.
Vladimir Dusevich Temple University dusevich-at-astro.ocis.temple.edu
The laser is exciting fluorescence in the sample. This is what is being viewed just as we view blue light when exciting with UV using fluorescence stains such as DAPI. We just went through establishing a laser saftey program on our campus. The saftey dept. determined that if all equipment using lasers for visual inspection of samples had saftey features built in such that the user could not be exposed to the laser. The notable exception was during service.
Tommy Sewall
} } } "Frieda Christie" {f.christie-at-rbge.org.uk} 04/30/97 04:10pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Forwarded message:
I use ethanol and sonicate for awhile. Sometimes the stubs need to be rubbed on a paper towel even after this treatment, but the adhesive is fairly easily removed in this way. But I agree, it would be nice to have something that would miraculously make the adhesive instantly dissolve! Regards, Melanie Behrens
If I am off-base with this response, I apologize. I have used nothing but carbon tape ever since it became readily available through regular EM suppliers but I always dreaded having to clean it off my sample holders because it was so sticky and tore so easily. Being a simple minded person, I just took a 3/8" wooden dowel, sharpened one end(like a chisel) and used it to roll the tape up into a wad that I could remove with my fingers. All the adhesive seems to comes off with the tape, but if any should be left behind it should easily come off with a mild solvent. (I always polish my holders, so I'm not sure about residual adhesive.) I also cut a piece of latex tubing to fit on the end of the rod to cushion my hand. This works for the carbon tape, I'm not sure about the other types of adhesive tapes. For what it's worth.
Bill -- ============================================================= Bill Chissoe III Electron Microscopist,University of Oklahoma E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
There are are few 3rd party organization that service Tracor/Noran Equipment.
We use Evex Analytical to service our equipment. They are affordable and provide prompt service. The Evex Headquarters is in Princeton, NJ and have field engineer located throught the country. They can be reached at 609-252-9192.
With reference to Holt's response to Henriks' post: We prep over 1500 specimens per year and have used Ross superglue, a cyanoacrylate, for years with no problem until they recently changed their formulation. The old Ross bonded very well with glass. We have been evaluating replacements and have no recommendation at this time (Read: everything we tried so far s%#*s!).
Scott: If you have a replacement that works, share it on line. I'm sure you can tell us something and not make it sound commercial!
Please contact Scion Corporation or go through their web site. http://www.scioncorp.com/
On Thu, 1 May 1997 kna101-at-utdallas.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi all, } } Yes, one more request for info on the best camera to use with our } microscope-computer set up. I'm sorry for not keeping all the info that } has been out on this subject a few months back, but I thought there was } no way the money would show up for one any time soon. Wrong. } We have an Olympus BH microscope and a Power Mac computer. We } are interested in using the NIH image software with this set up, but we } need a CCD camera with at least good resolution and any software/hardware } to link the camera and computer. Notes on personal experience with these } systems would be appreciated. I will need to know how much the camera and } interface will cost too. } Once again, sorry for not paying closer attention the first time. } } Karen }
Tseng-Ming Chou (Alex) Dept. of Materials Science and Engineering Stevens Institute of Technology Castle Point on Hudson, Hoboken, NJ 07030 e-mail: tchou-at-attila.stevens-tech.edu tchou-at-menger.eecs.stevens-tech.edu The Microstructure Group of Stevens
Hello, I need your assistance in acquiring several cell cross sections prepared by ultra-microtomy. It is my hope that some of our BIO-TEM friends have specimens on grid that are no longer of interest. We are not particular as to the type of cell but for reference would like to know what it is & the basic prep. technique. Anything taht has been=20 viewed in a TEM would be fine. Contrast enhancement technique are not required. Carbonless grids might be preferable. Our EM group consist of Material Scientist, Nano Particle Chemist & Physical Electronics Scientist, thus we lack the equipment & skills to do this our selves so we=92re bumming.=20 The application is 109nm (VUV) holography. We will gladly take care of expenses incurred on our behalf.
Thank you in advance for any anecdotal experiences with the various visible and fluorescent image capture and archiving systems available. I am looking for a combination of camera/video board/storage media/software for organizing images that will allow long term indexable storage of high resolution images. I have tried to look through the archives of the listserver for the past two months, but didn't find anything about this (but did find out a lot about Bremstrahlung).
Sincerely,
G. Steven Bova Departments of Urology and Pathology Johns Hopkins University
I need information about the basic principles and applications of the different microscopy techniques ( LM, SEM and TEM ) applied to the characterization of pure metals (like copper).
I need answers to simple questions like: What kind of information can I get with each technique? What makes one better than another and for what purposes? How to decide which one to choose for characterizing a pure metal like copper?
I would appreciate very much any help or reference that you could give me. Thanks.
Addendum to this message sent a few hours ago: I need to use a PC platform- Thanks to Michael Shaffer for pointing out this omission.-GSB
Dear All,
Thank you in advance for any anecdotal experiences with the various visible and fluorescent image capture and archiving systems available. I am looking for a combination of camera/video board/storage media/software for organizing images that will allow long term indexable storage of high resolution images. I have tried to look through the archives of the listserver for the past two months, but didn't find anything about this (but did find out a lot about Bremstrahlung).
Sincerely,
G. Steven Bova Departments of Urology and Pathology Johns Hopkins University
I've always just scraped off the old specimen and sonicated the stubs in acetone or ethanol. If all of the crud isn't removed, it also doesn't usually matter, since it is covered up by any new tape/silver or carbon paint and the specimen.
For analytical WDX or EDX, then, yes, removing the tape, adhesive, and a few layers of aluminum is useful.
Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
I am sorry, but the stock number I gave in my previous comments about plastic pages sold by 20th Century Plastics Co to hold Polaroid prints is for those made of vinyl, which are not recommended for archival storage. The corresponding pages made of polypropylene, which are recommended for archival storage, have stock number EZV450A-00. Each page has four pockets suitable for holding 4.25" x 5.25" prints, giving a capacity of 8 prints (back to back) per page. The holes for the binder rings are in a tab that runs along the side of the page, completely outside the pockets, thereby eliminating any need for having to punch holes in the prints. I have no commercial interest, just trying to set the record straight.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Advanced Imaging Concepts, Inc. provides just such systems, we produce the databasing & archiving software "Image Central" and supply that as just software, sell it with a frame grabber board or supply complete turnkey systems for archiving, or full blown image networks, or enable existing networks. We handle a large array of cameras, printers, grabbers, and image analsysis software to complete your digital workstation please contact me at AIC I will be glad to discuss your requirements with you and set up a demonstration if desired.
Scott E. Berman Sales Manager Advanced Imaging Concepts, Inc. Princeton, NJ phone(609) 921-3629 x26 fax(609) 924-3010 email Scott E57-at-aol.com
We use Rubber Cement Thinner to remove adhesive. I don't know what's in it but it is readily available and more effective than single solvents.
Jacob Bastacky, M.D. Room 116 Donner Laboratory Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
Negafile has polaroid 4x5 pages 4 per sheet. I'm not sure if they have gone to one size now or if they still have the two 4x5 sizes. You used to have to specify an "L" in the part number when ordering, but the number on the sheet itself, 4504, was the same for both. Check with them. I do not have their phone number, but their address is P.O. Box 78, Furlong, PA 18925, USA
- -Scott Walck
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POST-DOCTORAL RESEARCH FELLOW Princeton University
A post doctoral research fellow position is available in the Princeton Materials Institute and the Princeton Center for Complex Materials at Princeton University. The appointment is for one year initially with the possibility of renewal. It involves the use of a Philips CM-200 FEG transmission electron microscope equipped with a Gatan Image Filtering System in studying carbon nanotubes, and other nanostructured materials of interest. Candidates should have a Ph.D. in the physical sciences, and with strong EM background (HREM, Electron diffraction, and EELS). Experience in image analysis and UNIX workstations, and a strong math background are highly desirable.
Applicants should send a detailed curriculum vitae, copies of 2 selected publications, and names and addresses of three referees by June 15, 1997 to:
Nan Yao Princeton Materials Institute Princeton University Bowen Hall, 70 Prospect Avenue Princeton, NJ 08540
Princeton University is an equal opportunity employer.
=============================================================== Nan Yao Princeton Materials Institute Princeton University Bowen Hall, 70 Prospect Ave. Princeton, New Jersey 08540-5211
Hello, I need your assistance in acquiring several cell cross sections prepared by ultra-microtomy. It is my hope that some of our BIO-TEM friends have good cell sections on grid that are no longer of interest. We are not particular as to the type of cell but for reference would like to know what it is & the basic prep. technique.=20 Anything that has been viewed in a TEM would be fine. Contrast enhancement technique are not required. Carbonless grids might be preferable. Our EM group consist of Material Scientist, Nano Particle Chemist & Physical Electronics Scientist, thus we lack the equipment & skills to do this our selves so we=92re bumming.=20 The application is 109nm (VUV) holography. We will gladly take care of expenses incurred on our behalf.
Just a few more comments in an attempt to answer questions that have been raised about the characteristics of different diffusion pump fluids and their use in electron beam systems (matters which are discussed in some detail in Sect. 5.4 of my book, "Vacuum Methods in Electron Microscopy").
We ran into the problem of identifying Lion-S oil a number of years ago, and finally decided that it is most probably the synthetic fluid di(2-ethyl-hexyl) sebacate, and not a hydrocarbon oil. This same compound is sold as a pump fluid by several other companies under various trade names, most of which contain the letter S, such as Octoil-S (CVC) Diffoil-S (Lesker), etc. It has a vapour pressure at 20=B0C of about 3 x 10-6 Pa, and a boiling point at the normal boiler pressure of diffusion pumps (about 100 Pa) of about 205=B0C. It has better resistance to thermal and oxidative degradation than hydrocarbon oils, but will still break down if exposed to air at pressures much above 100 Pa while at operating temperature. We had trouble obtaining Lion-S oil when we serviced the DP on one of our SEMs and so used Octoil-S in its place, and had no problems as a result.
DC-704 is a synthetic silicone compound (tetraphenyl-tetramethyl-trisiloxane) that has a vapour pressure at 20=B0C o= f about 3 x 10-6 Pa and a boiling temperature at 100 Pa of about 220=B0C. Thu= s the properties of DC-704 are so nearly the same as those of di(2-ethyl-hexyl) sebacate that a pump designed for one fluid will work quite well with the other, without modification of heater wattage. The silicone fluids are very resistant to thermal, oxidative, and radiative degradation, hence their great advantage in a number of industrial processes. As pointed out by others, however, under electron bombardment they break down to form non-conducting siliceous deposits which are very insoluble, and so are not normally used in electron microscopes these days. I remember when RCA used silicone oils in some of their early microscopes. These instruments had platinum apertures, and in order to clean the aertures we had to flame them first (removing the organic component of the contamination) and them soak them in hydrofluoric acid to get rid of the siliceous residue.
The level of oil contamination produced by an diffusion pump depends not only on the type of pump fluid used, but also critically on the design of the system, and of the pump itself (see. Vac Meth p. 190-198). Several manufacturers of diffusion pumps devoted a tremendous amount of effort to developing pump designs that minimize backstreaming, and so systems equipped with their pumps will give minimal trouble in this regard. However, early models of diffusion pumps did not incorporate these critical design improvements, and so if you happen to have a system equipped with one of these pumps you can expect lots of oil in your system. Also, it seems that in the 'early days' some manufacturers of electron microscopes made their own diffusion pumps, and , of course, these often produced high levels of contamination, because they did not incorporate the design features needed to reduce backstreaming. The use of cold caps, water-cooled baffles and liquid nitrogen traps above the diffusion pumps will also reduce contamination rates (Vac Meth p. 190-198). The problem is, the design of the pump and its associated accessories is pretty much set by the manufacturer, and so it is rather difficult for the user of an instrument to do much about its basic characteristics. However, there are several mishaps that can occur during the operation of a diffusion pump (i.e. loss of cooling water, failure of the backing pump, exposure to air at high pressure, loss of electric power) which can lead to severe degradation of the pump oil (e.g. see Fig. 5.15 Vac Meth, p 218) and produce horrendous levels of backstreaming (Vac Meth , p. 214-223), and users should be extremely carful to guard against the occurrence of any such events.
=46inally, as I mentioned before, experience shows that in many cases the major source of oil contamination in systems with oil diffusion pumps is the breakdown of the hydrocarbon oil in the rotary vane backing pump (Vac Meth p. 144)
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
} Date: Thu, 1 May 1997 10:45:13 -0400 (EDT) } From: Vladimir Dusevich {dusevich-at-astro.ocis.temple.edu} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Image digitizers
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Colleagues } } We are looking for SEM slow scan image digitizer and TEM image digitizer - } for really old microscopes: Philips SEM 503 and TEM 300. We would } appreciate any comments, including pricing information. } } Vladimir Dusevich } Temple University } dusevich-at-astro.ocis.temple.edu
A possibility for the SEM is a device called ImageSlave sold in Australia by OED (Fax +61 (0)2 9482 1196) I understand this has been fitted to a number of older SEM's around the world
Ian
(I have no financial interesed in OED or ImageSlave)
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
With the recent traffic concerning uranyl acetate and lead citrate, it seems an appropriate time to ask a question that has puzzled me for some time. It concerns using Uranyl acetate as an en bloc stain. Protocol for en bloc staining vary. Some people use 2% UA in water after osmium postfixation. Others do the UA block stain during the dehydration (ie; alcoholic UA) while yet another group go to the trouble of making up a Maleate buffered UA block stain. Maleate buffered UA block stain seems to be the most commonly recommended one, yet I have not been able to find out why it is preferred to the others. Can anyone suggest why Maleate buffered UA is preferred over both aqueous and alcoholic UA when it is used as an enbloc stain?
Thanks in advance,
Allan Mitchell, Technical Manager South Campus EM Unit Otago School of Medical Sciences Dunedin New Zealand
---------- } From: Steve Beck {becks-at-sunynassau.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Denton Desk II Sputter Coater } Date: Thursday, May 01, 1997 12:09 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Colleagues, } } I am seeking some input and suggestions on a problem that I am having with } our Denton Desk II cold sputter coater. } } About 2 weeks ago, a student came to inform me that the sputter button } wasn't "popping" when depressed as usual - I believe the popping is the } opening of the solenoid valve which admits Argon to the chamber. After } confirming that the problem existed, I noted that the outlet valve from the } regulator had been turned off (I'm not sure if this contributed to the } situation or not). Upon opening the back of the unit, I found a blown 2 Amp } slo-blow fuse. } } After replacing the fuse, the sputter valve has been working just fine, } however, after replacing it, a new problem arose - when the 45mA current } was applied to the gold target (at 50 millitorr), the typical plasma glow } was not evident even though the indicator read 45mA - I did notice some } initial arcing, then nothing! } } After consulting a very helpful sales/service representative (who suggested } that I take the sputterhead apart and clean the teflon spacer/insulator) I } was able to restore normal function - at least it seemed to be fixed for a } few trial runs! } } Now, no matter how many times I take the head apart and clean each piece, } the coater fails after only a few runs or less - it arcs violently and the } solenoid valve automatically shuts down (cycles to off position). } } Has anyone had similar problems with the unit and/or any advise? This unit } was purchased in 1992 and is not used excessively (1 to 2 SEM courses per } year) - I recently replaced only my second gold target in that amount of } time. } } Thanks in advance for your kind assistance. } Dear Steve,
We had the same problem with our Denton Desk II, to the extent that the gold foil target was torn by the arcing. We "solved" our problem by increasing the argon pressure to 70 millitorr during sputtering. This does not appear to have had any adverse effects on the quality of the coating and has eliminated the arcing problem completely.
At HRL we are urgently seeking a circuit board for the Motorised Stage Drive (ISM-MSD40-2) for our Jeol 840 SEM. The required circuit board is the computer interface (RS232) for external control, ie a EDXA control of the SEM stage. Should anyone know the whereabouts of a spare board or can get the schematic of this board we will be glad to here from you.
Warren Straszheim wrote: ================================================ What are some effective solvents for removing adhesive tape, spots, etc. from aluminum SEM mounts. Acetone is marginal as are other ketones. I've tried several alcohols without success. Caustics are out because they eat the aluminum. Pure and mixed acids don't work either. I've even tried WD- 40 and liquid dish soap. ================================================ Even though we have manufactured SEM mounts for others for more than 25 years, we ourselves have always had a program of cleaning and recycling our own mounts used internally. What you end up using depends a lot on just how much elbow grease you are willing to expend at the beginning in terms of removing the material physically. And Andy Blackwood in our lab just told me, some times they "give the mounts a 'lick' on 600 grit on the metallographic wheel, thus removing the whole surface."
But what ever solvents are eventually used, the real question is, just how many "rinsings" are enough in order to reduce the level of organics on the surface to an "acceptable" level. And I have not ever figured out how to make that determination.
Therefore, for critical samples, requiring the highest performance, samples that might not be receiving any further gold or carbon coatings, for example , we expose the mounts for a few minutes to an oxygen plasma in our SPI Plasma Prep II plasma etcher. Call it "etching" or call it "plasma cleaning", but in order to be absolutely sure there are no remaining organic residues, this is the only method literally guaranteed to remove the last remains of any organic residues.
PS: Even brand new SEM mounts, while they look and feel brand new, for high performance work, not other wise using applied conductive coatings, should be given some kind of a cleaning, the best one being this kind of a plasma cleaning treatment.
Information about plasma etchers for this kind of cleaning can be found on our website given below. We think that recycling mounts in this kind of situation makes a lot of good sense, provided the time, labor, and other wise unnecessary exposure to solvents exceed the benefits from the recycling .
Disclaimer: SPI Supplies manufactures the SPI Plasma Prep II plasma etcher and has a vested interest in seeing more uses for this system!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I have had a query from a collegue who wonders how much success people have had running MAC versions of NIH Image with the PC MAC emulator Executor. I would appreaciate any comments and/or contact details of anyone with experience of this combination.
Thanks in Advance
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
Dear Harold and all, I have found that the best way to remove the carbon adhesive discs from Al stubs is to soak the stubs in acetone for at least 1/2 hour, then sonicate. When you rub the wet stub on a paper towel, the disc will slip off. Methanol is my solvent of choice for tape residues, but often works poorly if the residue is old. Most of the rubber cement-type glues respond to chloroform. My $0.02CAN ($0.014US) worth. Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Dear All, I find oil on the EDX detectors in both my SEM's, but if you think of the conditions inside the SEM, it is almost inevitable. The majority of the molecules remaining in the vacuum of an average SEM at 10 -5 torr consists of diffusion pump oil. This oil will gradually condense on all surfaces in the SEM, but on the coolest surface first. In most SEM's this is the EDX snout. One EDX manufacturer has solved the problem by warming up the snout with a small heater. The oil just goes somewhere else, but if it forms a thin film over all surfaces it is not so visible or worrying. The cleaning method recommended by the manufacturer of my EDX's is to gently run clean-grade iso-propanol over the snout, then allow to air-dry. They used to recommend Freon, but that is no longer permitted or available. Hope this helps, Mary
Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
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Dear colegues, I found organelles that look like clathrin coated vesicles in the location where for some reasons coated vesicles should not be. Do somebody know about other organelles that have similar structure? I'll appreciate any information, Thank you in advance, Elia
Elia Beniash, PhD Student, Dept of Stuctural Biology, Weizmann Institute of Science, 76100, Rehovot, Israel
Hi, We are considering the purchase of an processor for TEM samples. We currently process about 10 to 15 samples a week. Would individuals please recommend or discuss with us their experiance using these instruments. Thanks, Robert Cox Shriners Burn Institute Galveston Tx, rcox-at-sbi.utmb.edu or 409-770-6655
I am interested in buying a color camera for phase, brightfield, and fluorescence imaging, which will be our only camera. At this point I don't absolutely need to do any ratioing or math involving intensities. I am interested in low noise but probably don't need exposures longer than a couple seconds (in fact motion of my wet-mounted samples may limit exposure time). I want to focus and make exposure adjustments in near-real time from the computer or video screen, not through a viewfinder. Expense is a consideration, too. With our currrent cheap monochrome video camera (Sony XC-75, TCX frame grabber) we have plenty of illumination but still lots of noise.
Are there any good systems (camera+grabber) that give greater than 8 bits per color after digitization?
The Diagnostic Instruments Spot camera is a 24 bit cooled ccd camera using a single Kodak KAF1400 chip. It uses some type of switchable liquid crystal filters to take 3 sequential images, one each RGB. Has anyone had experience with it, and compared it to the alternatives? I expect that due to the filters it is significantly less sensitive than others, is that true? How does noise compare with images digitized from color video cameras, cooled or not? Are there any other considerations?
For video cameras, what frame grabbers/drivers give histograms of light intensity to aid in optimizing exposure settings?
Thanks for the prompt delivery of the last order. I need to make another now. I need 2 #PA0050 It is a tonor cart. for a Lexmark Optra-R printer( a hateful machine). The price at the last order was $165 but if that has changed it is OK just e-mail me back the price. I need this billed out as soon as you can even if the items are not available. The blanket PO's need to be closed out for the fiscal year here soon. I can wait on delivery. Thanks mucho!!
PO# L06681 cust. # 6646947
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
} } } } } ------- Forwarded Message Follows ------- } } } } } Does anyone know the dangers associated with a technique called Confocal } } } Microscopy or something like that? In our Chemistry Dept. we have a } } } research group that works with a Class 4 LASER. To the best of my } } } understanding, the LASER beam is directed toward a sample that is sitting } } } under a microscope. Some of the beam is reflected and filtered so the } } } power or strength of the beam is reduced before it hits the sample } } } (a biological sample). The researcher then looks through the microscope } } } objective at the sample. } } } } } } Currently, we have a LASER safety program in place. This particular } } } procedure concerns me because this technique allows the researcher to } } } look directly at the LASER beam. This does not sound too good. Also, } } } the microscope objective may magnify the beam and direct the beam } } } toward the researchers eye. } } } } } } Currently, the LASER that is being used is an Class 4 ARGON Ion LASER } } } (Continuous Wave) with a wavelength of 0.488 um (ultraviolet range) and a } } } beam power of 1.5 W. } } } } } } Does anyone know where I can get more information on this subject. I am } } } particularly interested in the safety aspects of this technique. It } } } may be the case that the LASER beam hitting the sample has a low enough beam } } } power to classify it as a Class 1 LASER. } } } } } } Thanks in advance for your help. } } } } } } Laurie Princiotto } } } Laboratory Safety Specialist } } } Indiana University - Dept. of Environmental Health and Safety } } } Phone: (812)-855-6115 } } } Fax: (812)-855-7906 } } } E-mail: lprincio-at-indiana.edu } } } } Laurie, } } This does not sound like a very safe activity to me. I would be very } } concerned until proven safe. } } } } Regarding the lasers used in confocal microscopes...I've worked with the } } Bio Rad commercial system and I believe most use a class II Ar or Ar/Kr } } laser at ~1mW power (457-648nm). The laser image cannot be directly } } observed but is displayed in a computer screen. } } } } Your best info sources would be the manufacturers of confocal microscopes. } } } } Leica - Ph: 415-348-2233 } } Bio-Rad - Ph: 800-4BIORAD } } Molecular Dynamics - http://www.mdyn.com ph:800-333-5703 or 408-773-1222 } } Zeiss 800-233-2343 } } } } good luck } } } } Edward J. Basgall, PhD } } The Pennsylvania State University } } Surface Chemistry Group ejb11-at-psu.edu } } Materials Research Institute Building Ph: 814-865-0493 } } University Park, PA 16802-7003 FAX: 814-863-0618
Laurie,
I have never heard of a laser confocal microscope that involves direct observation. To my knowledge, all of the laser confocal microscopes are scanning devices and the scan-derived image is displayed on a monitor -- the user never looks directly into the microscope.
Since you did not identify the make or model of the confocal scope, it is not clear to me that you have even spoken to the users about your safety concerns. That is generally a good place to start. They may know more about laser safety than you give them credit for. After that, talk to the company that makes the system. After that, send up a posting to the list with enough relevant background information, and I am sure you will receive informative responses.
As far as whether the laser is Class 4 or Class 1, it is not a good idea to look directly at the beam of any laser. Just in case anyone asks you that, in your capacity as a safety officer.
Duane McPherson Dept. of Biology SUNY at Geneseo Geneseo, NY
Advanced International Immunofluorescence Course Gargnano '97 (Italy)
The Advanced International Immunofluorescence Course is a post-doctorate theoretical/practical course, with propedeutical lectures and practical stages on traditional and confocal immunofluorescence microscopy and image and ion analysis. The course will take place in Gargnano (Lake of Garda) from 7 to 10 October 1997. Further information and registration details will be found at the following Web address
http://imiucca.csi.unimi.it/endomi/ACIF.html
Program:
Morning Tuesday, October 7, 1997
9.00-10.00 Registration of partecipants
10.00-10.45 AN INTRODUCTION TO IMMUNOCYTOCHEMISTRY AND IMMUNOFLUORESCENCE (I.Barajon)
10.45-11.15 Coffee Break
11.15-12.00 LIGHT SOURCES AND FLUOROCHROMES (G. Bottiroli)
12.00-12.45 THE USE OF FLUORESCENCE MICROSCOPE (C.Rumio)
Advanced International Immunofluorescence Course Gargnano '97 (Italy)
The Advanced International Immunofluorescence Course is a post-doctorate theoretical/practical course, with propedeutical lectures and practical stages on traditional and confocal immunofluorescence microscopy and image and ion analysis. The course will take place in Gargnano (Lake of Garda) from 7 to 10 October 1997. Further information and registration details will be found at the following Web address
http://imiucca.csi.unimi.it/endomi/ACIF.html
Program:
Morning Tuesday, October 7, 1997
9.00-10.00 Registration of partecipants
10.00-10.45 AN INTRODUCTION TO IMMUNOCYTOCHEMISTRY AND IMMUNOFLUORESCENCE (I.Barajon)
10.45-11.15 Coffee Break
11.15-12.00 LIGHT SOURCES AND FLUOROCHROMES (G. Bottiroli)
12.00-12.45 THE USE OF FLUORESCENCE MICROSCOPE (C.Rumio)
There are many issues here when making a choice on a CCD camera, among which are: 1) color vs BW (price and resolution factors) 2) resolution 3) sensitivity to low intensity signals (fluorescent applications?) 4) and therefore cooled vs not 5) dynamic range (8 - 12bit) 6) software These are not all but I think the major ones and they are all interelated. It depends on the applications the camera will be used and will it need to be pushed to its specs. Photometrics (on their website) and Princeton Instruments (catalogue) have most of the information in easily understandable language to help you make a decision. ( I have no commercial interests in these companies.) If you want to contact me and I cangive you my reasons for our recent decision on a CCD. Hank Adams EML New Mexico State U. 505-6463600
Hi Robert, About a year or so ago we purschased a RMC tissue processor and have used it pretty much on a daily basis. The main reasons for the RMC over the Leica were: 1) price and 2) I liked the reagent tube caps for each tube, and not the rubber ring over all the tubes. Have had a few minor problems with it, and RMC has been very helpful with repairs.
Any more questions, please feel free to contact me.
Ed Calomeni Medical College of Ohio Dept. Pathology Toledo, OH 43699 emlab-at-opus.mco.edu 419-381-3484
Many thanks for the great response to my request for Bio TEM specimens. Several parties have indicated that they will send a few over. These should more than meet our need.
LocTite 460 is a superior super glue for bonding to all materials. It doesn't bubble and the mechanical strength is the best compared to any other LocTite product available. This is according to what they have told me and my customers are very happy with it.
Bug and tar remover at Chief Auto Parts works very well for removing and dissolving wax from stubs if acetone is not satisfactory. Perhaps a new wax is in order also.
Yours Truly,
Gary Liechty Product Application Specialist Allied High Tech Products, Inc. 2376 E. Pacifica Place Rancho Dominguez, Ca. 90220
please subscribe me to the microscopy listserver Dr. Vickie A. Kimler Assistant Professor of Biology and Allied health Mercyhurst College Erie, PA 16546
e mail: vkimler-at-paradise.mercy.edu Vickie A. Kimler, Ph.D. Biology and Allied Health Department Mercyhurst College Erie, PA 16546 1-814-824-2169
Does anyone have or know of an SEM with cathodoluminescence imginging avaliable for service work (hourly or daily)? I do not need a cold stage, but energy filtering is needed. East coast location would be prefered, southest ideal.
Thanks for any leads or ideas, Phil Russell
Phillip E. Russell Analytical Instrumentation Facility Box 7531, Room 318 EGRC North Carolina State University Raleigh, NC 27695-7531
As this mail list's token optical engineer I guess I should put in a few good words. The fact is that there are many different types of laser confocal microscopes, some of which are direct view, some use a detector and a video monitor, and some are fluorescence. Since the original poster did not specify what microscope she was worried about we have to assume that it is a direct view machine. These use spinning disks (Nipkow disks) or equivalent to raster scan.
Two points should be made: first, and least interesting, is that the manufacturer knows what it is doing and safety of looking into the microscope was worked out at its end and concerns could easily be handled by a call to them.
Second, remember that the laser itself is classified based on the output beam. When that beam is modified by the microscope optics it will have less power spread over a much larger area. Thus the brightness of the beam can be reduced many orders of magnitude before it hits the eye.
It is not intrinsically dangerous to look into a laser. The danger depends on the brightness of the beam. A few months ago a poster on the optics news group reported hooking up deflection and intensity modulators to a video signal and watching TV by writing directly onto his retina. Not something that I would be interested in doing myself, but afterwards he could see good enough to at least type his message.
best regards mark
Mark W. Lund, PhD Director } } 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
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
My laboratory has not used 35 mm film for 12 months. We purchased the SONY DKC5000 camera and have it connected by SCSII drive to a MAC computer. Works great, is very easy to handle and even though the photos do not look so hot when first captured, by using Adobe Photoshop 4.0, and the levels adjustment, within 2 minutes the photo is outstanding. We are printing to a Kodak DS 8650PS printer that has high resolution.
If you would like to view a plate containing 19 photos that never saw film go to the following URL using Netscape:
__________________________________________ Rex A. Hess, Ph.D., Associate Professor, Director, Center for Microscopy and Imaging University of Illinois, Veterinary Biosciences, 2001 S. Lincoln, Urbana, IL 61802-6199 217-333-8933; FAX 217-244-1652; email: r-hess-at-uiuc.edu homepage-- http://www.cvm.uiuc.edu/HomePages/rhess/hesspage.html Center for Microscopy & Imaging -- http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html Society of Andrology-- http://godot.urol.uic.edu/~androlog/
I'm real new to wedge thinning but as of this morning, I'm having reasonable luck with Loctite's general purpose instant adhesive. It comes in an applicator that actually looks like it may last as long as the glue supply. It's called a SuperBonder Gluematic Pen. Got it at my favorite toy store, W.W.Grainger (www.grainger.com). About $2.50 list.
No interests (other than as a consumer) in either company
cheers, John heckman-at-pilot.msu.edu } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Folks, I have become very concerned that all our talk on pH and Pbcitrate will cause some people to go to their pH meters with their lead solutions to find out the pH. Please do not do this! It almost certainly is not good for the electrode. You will not get an accurate reading anyway, since the pH is will be so high, and the solution is not buffered. What might happen is that you will assess erroneous information from your pH meter to be correct and become embroiled in a vicious cycle of repeatedly making up lead stain attempting to get a correct pH reading. If you follow Reynolds directions, you will automatically get a correct pH. (That is, if you get a true 1N solution of NaOH either from pellets, or from a commercially titrated product). Next week, I will address this problem one more time, and then we all need to go on to something else. Bye, Hildy
Dear Folks, I have become very concerned that all our talk on pH and Pbcitrate will cause some people to go to their pH meters with their lead solutions to find out the pH. Please do not do this! It almost certainly is not good for the electrode. You will not get an accurate reading anyway, since the pH is will be so high, and the solution is not buffered. What might happen is that you will assess erroneous information from your pH meter to be correct and become embroiled in a vicious cycle of repeatedly making up lead stain attempting to get a correct pH reading. If you follow Reynolds directions, you will automatically get a correct pH. (That is, if you get a true 1N solution of NaOH either from pellets, or from a commercially titrated product). Next week, I will address this problem one more time, and then we all need to go on to something else. Bye, Hildy
Allan Mitchell asks why some protocols call for sodium maleate buffers for enbloc staining. There are several reasons: 1. Phosphate and Cacodylate buffers used just previously to UA enbloc have been known to precipitate the UA 2. Rinsing tissue (washing) with maleate buffer, pH 5.2, prevents precipitation of UA in the tissue, and it prevents the minor (but perhaps important) movement of osmium in or out of the tissue. 3. Mixing maleate buffer pH 6.0 with a 3% UA solution 1:1, brings the pH of the UA solution to approximately pH 5.2. 4. This is a refinement, not a necessity, for the achievement of the goal of superior preservation and beautiful micrographs. I have used both concepts to a large extent (no maleate, or maleates) depending on the results I needed. It is more to do if you use maleates. But, if the decision is made to use maleates, they can be made in huge quantities and kept in a freezer. It is called having fun with EM. All the above appeared in detail in the EMSA journal about 10 years ago. It also came up in a workshop given at EMSA by Janet Boyne previous to the journal article. Perhaps it can be located by those interested. Bye, Hily
We will be purchasing a confocal microscope in the near future. It's purpose will be for biological research (no material sciences) and I was wondering if anybody had any strong feelings (positive or negative) about the type of confocal system that they were using, the service from the seller, etc.
We will be purchasing a confocal microscope in the near future. It's purpose will be for biological research (no material sciences) and I was wondering if anybody had any strong feelings (positive or negative) about the type of confocal system that they were using, the service from the seller, etc.
Allan Mitchell asks why some protocols call for sodium maleate buffers for enbloc staining. There are several reasons: 1. Phosphate and Cacodylate buffers used just previously to UA enbloc have been known to precipitate the UA 2. Rinsing tissue (washing) with maleate buffer, pH 5.2, prevents precipitation of UA in the tissue, and it prevents the minor (but perhaps important) movement of osmium in or out of the tissue. 3. Mixing maleate buffer pH 6.0 with a 3% UA solution 1:1, brings the pH of the UA solution to approximately pH 5.2. 4. This is a refinement, not a necessity, for the achievement of the goal of superior preservation and beautiful micrographs. I have used both concepts to a large extent (no maleate, or maleates) depending on the results I needed. It is more to do if you use maleates. But, if the decision is made to use maleates, they can be made in huge quantities and kept in a freezer. It is called having fun with EM. All the above appeared in detail in the EMSA journal about 10 years ago. It also came up in a workshop given at EMSA by Janet Boyne previous to the journal article. Perhaps it can be located by those interested. Bye, Hily
In my experience, most of the molecules in a conventional SEM chamber under normal operating conditions are attributable to water and air, less so to ethanol and isopropanol followed by diffusion and roughing pump oils. Monitoring by residual gas analyzer (RGA) reveals all but the air can be significantly reduced by maintaining the liquid nitrogen baffle above the diff pump. Never open the gate valve to the chamber without the liquid nitrogen trap filled and routinely let the trap warm up with the gate valve closed. Regular maintenance of the molecular sieve trap on the roughing line is also critical.
I hope the EDS manufacturer refered to responds because I don't believe the heater warms the snout or window such that oil would be evolved but rather that is heats the chip to drive ice off the chip face and into the bowels of the dewar. If it does drive off the oil do you really want that oil all over the inside of your SEM? Seems like the solvent trickle method might be best - I have never done it has anyone else had experience with this tricky procedure. Check with your EDS manufacturer before trying it.
Scott Wight
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
Allan Mitchell asks why some protocols call for sodium maleate buffers for enbloc staining. There are several reasons: 1. Phosphate and Cacodylate buffers used just previously to UA enbloc have been known to precipitate the UA 2. Rinsing tissue (washing) with maleate buffer, pH 5.2, prevents precipitation of UA in the tissue, and it prevents the minor (but perhaps important) movement of osmium in or out of the tissue. 3. Mixing maleate buffer pH 6.0 with a 3% UA solution 1:1, brings the pH of the UA solution to approximately pH 5.2. 4. This is a refinement, not a necessity, for the achievement of the goal of superior preservation and beautiful micrographs. I have used both concepts to a large extent (no maleate, or maleates) depending on the results I needed. It is more to do if you use maleates. But, if the decision is made to use maleates, they can be made in huge quantities and kept in a freezer. It is called having fun with EM. All the above appeared in detail in the EMSA journal about 10 years ago. It also came up in a workshop given at EMSA by Janet Boyne previous to the journal article. Perhaps it can be located by those interested. Bye, Hily
Mark, Plastic Pages-4 pocket for 3 ring binder are available From IMBROS in Moonach, Tasmania Tel 03 6273 1300 If you wish we can supply them from the USA. Ladd CAT# 11-81250
Preece, T. 1978. Toluidine blue: The staining method of Shoremaker..... FBPP News 1:40-40.
Can anyone tell me what "FBPP" stand for? (note: is apparently is not "Federation of British Plant Pathology")
TIA
Robert R. Wise Plant Physiologist and Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
I am interested in procuring a cryostage that is in good working condition for our ZEISS 902 Energy Filtering TEM. If anybody who has one and wants to part with it can contact me directly. Thanks!
M.V.P.
************************************************************************* M.V. Parthasarathy Professor of Plant Biology & Director, Cornell Integrated Microscopy Center (CIMC) Section of Plant Biology 228 Plant Science Building Cornell University, Ithaca, NY 14853 Telephone: Plant Biology Office (607) 255-1734 Fax: " (607) 255-5407 Telephone: CIMC Office (607) 253-3803 Fax: " (6)7) 253-3803 E-mail: mvp2-at-cornell.edu ***********************************************************************
I am interested in procuring a cryostage that is in good working condition for our ZEISS 902 Energy Filtering TEM. If anybody who has one and wants to part with it can contact me directly. Thanks!
M.V.P.
************************************************************************* M.V. Parthasarathy Professor of Plant Biology & Director, Cornell Integrated Microscopy Center (CIMC) Section of Plant Biology 228 Plant Science Building Cornell University, Ithaca, NY 14853 Telephone: Plant Biology Office (607) 255-1734 Fax: " (607) 255-5407 Telephone: CIMC Office (607) 253-3803 Fax: " (6)7) 253-3803 E-mail: mvp2-at-cornell.edu ***********************************************************************
The thread on solvent cleaning of stubs has turned into a solid rope, perhaps we can twist it into a 4 inch hawser. I like Bill Chissoe III's variation: Brute force and not those nasty solvents. A medium-fine flat file, lying on a bit of paper towel served me well to clean the residues from the top of stubs. A suede wire brush is effective for occasional cleaning of the file. I believe that method its less messy and often faster than those solvents.
Economics, however, also come into such considerations. Every now and then somebody rediscovers that by going back in the literature they can find the methods to refilament, make thin film apertures, various standards or even LaB6 and they could make them for the lab. One in a thousand may have good reasons to do so.
Generally, cleaning up stubs is still worthwhile. A stub costs A$0.32 or about US$0.25. Amazingly some labs even manufacture new stubs without a production lathe "in house". An employer has to count "on cost" and the cost of the facilities. Commercially it makes sense to "recover" triple the hourly pay for any task before break-even point is reached. A person on $15/h needs to clean up 180 stubs/h to break even so far as the institution is concerned. Clearly, Professors should work faster or delegate hum drum chores.
Some thirty years ago the most commonly used Cu grids cost A$7.00/100. They now cost A$10 and although our currency has depreciated, it is obvious that grids are much cheaper now in relative terms. I recycled thousands of grids up to about 25 years ago. Today this would make no sense whatever - unless you are working in a cheap labour country.
Of course, we must not forget institutional economics which work to different realities: "We only have so much (too little) money for maintenance; we're paying that person anyway. Productivity? Who cares, we're only trying to balance our budget."
Stop reading your email now and do something, CLEAN THOSE STUBS! Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
} } If I am off-base with this response, I apologize. I have used nothing } but carbon tape ever since it became readily available through regular } EM suppliers but I always dreaded having to clean it off my sample } holders because it was so sticky and tore so easily. Being a simple } minded person, I just took a 3/8" wooden dowel, sharpened one end(like a } chisel) and used it to roll the tape up into a wad that I could remove } with my fingers. All the adhesive seems to comes off with the tape, but } if any should be left behind it should easily come off with a mild } solvent. (I always polish my holders, so I'm not sure about residual } adhesive.) I also cut a piece of latex tubing to fit on the end of the } rod to cushion my hand. This works for the carbon tape, I'm not sure } about the other types of adhesive tapes. For what it's worth. } } Bill } -- } ============================================================= } Bill Chissoe III } Electron Microscopist,University of Oklahoma } E-mail: wchiss-at-ou.edu Ph. (405)325-4391 } =============================================================
The Microbeam Analysis Sciety is pleased to announce the introduction of a new mailing list (listserver) dedicated to electron microprobe analysis. This mailing list is sponsored by MAS and is managed by two MAS Directors, John Mansfield and Greg Meeker. It is being formed in response to several requests from members of MAS. The list is open to anyone interested in microprobe analysis. To subscribe you may send mail to the mailing list at:
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John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 Email: jfmjfm-at-umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
I want to be able to grab S-VHS video in my camputer for sending through the web. I have a video camera mounted on my trinocular Nikon and save as S-VHS on VCR'S. I have heard of an add-on called SNAPPY. Has anyone used this and does it give good images? Can anyone supply names of video cards?
Mike Dingley -- Check out the P.M.C.A. Homepage and Mike's Freshwater Algae Homepage at http://www.pnc.com.au/~dingley
Is anyone aware of a solvent which dissolves polymerized LR White? I have two pieces of aluminum, separated by a small gap, which infiltrated with LR White prior to UV polymerization. Now they are hopelessly stuck together. I've tried methylene chloride with no success, and am now searching for other ideas. I can work in the hood, and considering the importance and value of these parts, I am willing to take what ever precautions a particular solvent might require. Acids which might attack aluminum are unfortunately not an option. Any ideas?
Many thanks,
Doug Keene Shriners Hospital for Children Portland, Oregon ---------------------- Doug Keene DRK-at-shcc.org
N,N,dimethylformamide might be worth a whirl, I think I used to use it years ago to dissolve cured epoxy resins for subsequent chemical analysis. I don't know about LR White, but DMF dissolves lots of things.
cheers
Ritchie
} Is anyone aware of a solvent which dissolves polymerized LR } White? I have two pieces of aluminum, separated by a small } gap, which infiltrated with LR White prior to UV } polymerization. Now they are hopelessly stuck } together. I've tried methylene chloride with no success, } and am now searching for other ideas. I can work in the } hood, and considering the importance and value of these } parts, I am willing to take what ever precautions a } particular solvent might require. Acids which might attack } aluminum are unfortunately not an option. Any ideas? }
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
I would be grateful for information on daily/weekly workloads expected = of technical staff in diagnostic TEM labs (pathology). For example: = number of specimens received and processed (by hand or automatic = processor), sections cut, stained and viewed, photographics (including = printing), other duties included, etc. Also, I am interested in = determining what is considered an efficient diagnostic TEM pathology = service in terms of numbers of specimens processed per year against = total staffing levels (all staff) and specimen turn-around times. If you do not want your information to be seen on the MSA listserver, = mail can be sent direct to: John.Stirling-at-flinders.edu.au
Thanks in advance! John W Stirling Department of Anatomical Pathology Flinders Medical Centre Bedford Park SA 5042 Australia=20
I think that a cup of boiling water may help you to separate the samples. The epoxy might not be dissolved in the hot water, but it would become soft enough to let you tear the aluminum off.
With the best wishes,
Charlie Kong EM Unit, UNSW, Australia
On Mon, 5 May 1997, Doug Keene wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Is anyone aware of a solvent which dissolves polymerized LR } White? I have two pieces of aluminum, separated by a small } gap, which infiltrated with LR White prior to UV } polymerization. Now they are hopelessly stuck } together. I've tried methylene chloride with no success, } and am now searching for other ideas. I can work in the } hood, and considering the importance and value of these } parts, I am willing to take what ever precautions a } particular solvent might require. Acids which might attack } aluminum are unfortunately not an option. Any ideas? } } Many thanks, } } Doug Keene } Shriners Hospital for Children } Portland, Oregon } ---------------------- } Doug Keene } DRK-at-shcc.org } }
Edward J. Basgall wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } } ------- Forwarded Message Follows ------- } } } Does anyone know the dangers associated with a technique called Confocal } } Microscopy or something like that? In our Chemistry Dept. we have a } } research group that works with a Class 4 LASER. To the best of my } } understanding, the LASER beam is directed toward a sample that is sitting } } under a microscope. Some of the beam is reflected and filtered so the } } power or strength of the beam is reduced before it hits the sample } } (a biological sample). The researcher then looks through the microscope } } objective at the sample. } } } } Currently, we have a LASER safety program in place. This particular } } procedure concerns me because this technique allows the researcher to } } look directly at the LASER beam. This does not sound too good. Also, } } the microscope objective may magnify the beam and direct the beam } } toward the researchers eye. } } } } Currently, the LASER that is being used is an Class 4 ARGON Ion LASER } } (Continuous Wave) with a wavelength of 0.488 um (ultraviolet range) and a } } beam power of 1.5 W. } } } } Does anyone know where I can get more information on this subject. I am } } particularly interested in the safety aspects of this technique. It } } may be the case that the LASER beam hitting the sample has a low enough beam } } power to classify it as a Class 1 LASER. } } } } Thanks in advance for your help. } } } } Laurie Princiotto } } Laboratory Safety Specialist } } Indiana University - Dept. of Environmental Health and Safety } } Phone: (812)-855-6115 } } Fax: (812)-855-7906 } } E-mail: lprincio-at-indiana.edu } } Laurie, } This does not sound like a very safe activity to me. I would be very } concerned until proven safe. } } Regarding the lasers used in confocal microscopes...I've worked with the } Bio Rad commercial system and I believe most use a class II Ar or Ar/Kr } laser at ~1mW power (457-648nm). The laser image cannot be directly } observed but is displayed in a computer screen. } } You're best info sources would be the manufacturers of confocal microscopes. } } Leica - Ph: 415-348-2233 } Bio-Rad - Ph: 800-4BIORAD } Molecular Dynamics - http://www.mdyn.com ph:800-333-5703 or 408-773-1222 } Zeiss 800-233-2343 } } good luck } } Edward J. Basgall, PhD } The Pennsylvania State University } Surface Chemistry Group ejb11-at-psu.edu } Materials Research Institute Building Ph: 814-865-0493 } University Park, PA 16802-7003 FAX: 814-863-0618 Dear Ed et. al.,
As past technical marketing manager for Sarastro (now Molecular Dynamics), I can assure you that when these microscopes are designed, they must meet extremely stringent safety standards. One of two options is used: either existing optics are chosen which eliminate any possibility of laser light reaching your eyes (ex: when we used a Nikon microscope as the basis for the Sarastro system, we used an "F-head" in which the full binocular body rotated out of position when the laser was directed to the sample) or optics are designed with fool-proof safety mechanisms. One of the previous respondents had the imaging story partially correct: In confocal microscopy, the laser scans the sample very much like a TV raster scan. However, the light goes to a detector (either a photomultiplier tube or a video camera). From that point, the signal passes to a computer for processing and imaging. There are some "direct view" confocal systems, notably, those produced by Technical Instruments (San Francisco), but these are not laser systems . Instead, they use the same sort of "white light" illuminator you would normally find on a regular light microscope. Hope this helps.
} Jim: } I'd be very interested to know what method you would recommend for } cleaning Be windows on EDS detectors (and probably so also would a lot of } others on the Microscopy Listserver). } } We have done it several times in the past by using a dropper to run a } bit of petroleum ether over the face of the window. You have, of course, } to be very careful not to puncture the window with the dropper, but then } the whole operation is one of high risk to begin with. Pet ether is } basically a low-boiling hydrocarbobn fraction that is a pretty good solvent } for other hydrocarbons, but which does not attack epoxies and which also } evaporates quite readily and cleanly. We have even been able to do this } without removing the detector from the SEM by using a long dropper and } collecting th Pet ether on a tissue as it runs off the detector. I am not } sure how well this might work with silicone oils, because of their limited } solubilities; however, one old-time industrial solvent for them was } kerosene, which is again basically a hydrocarbon material. } } Wilbur C. Bigelow, Prof. Emeritus } Materials Sci. & Engr., University of Michigan } Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; } Fx:313-763-4788; Ph:313-764-3321 *********************************** Wil: Your method was fairly similar to ours. You and I would not use silicone in an EM, but if kerosene was used, a second solvent would be needed to remove the kerosene residues. Because I worked for many years in a rather isolated lab (1500km to the nearest other electron beam instrument) factory maintenance was never an option and for a time we had three EDS. Link suggested that reagent grade methanol was a most suitable solvent. Presumably this was recommended because it did not affect the Be window's glue and it is a good solvent of vacuum oils and fluids. Happily we too could leave the detector in place and I collected the used solvent in a Petrie dish and also with tissue. The significant difference in our methods was the application. Because the window is recessed and our detector had some tilt, it seemed that little solvent would reach the window if the methanol was just dripped from the top.
Use polyethylene Pasteur type pipette with a fairly large bulb. Pull the tip into a fine capillary after brief (5-10 seconds) heating in a low flame. (Tip: move and rotate pipette while heating; pull vertically, this keeps the tip straight while the plastic sets) Cut the tip so that about 100mm of capillary tip and a small bore, perhaps 0.2mm is available. FILL the bulb with methanol; releasing air once or twice by up-turning and squeezing the bulb. Filling is slow but no trouble. Freehand application is possible with care and a steady hand, but most people would be happier mounting the pipette horizontally in a low "retort" stand, at the Be window's level. Slide the stand until the pipette tip is within 20 to 40mm of the window. Hold pipette with one hand (hand should rest, never work mid-air) near the tip and gently squeeze the bulb. The aim is to "hit" with little force the upper rim of the Be window - where is supported. Less force on the window and also slightly better coverage is achieved with the methanol stream directed well away from the perpendicular.
A side benefit of well tilted detectors is that cleaning is much easier: Simply use a suitable vial half filled with solvent and slide snout into this. Slightly swirl and cleaning is done. Regards Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
Hello all; Is anyone out there doing in situ hybridization using the DIG kit from Boehringer-Mannheim? We have been trying to use the kit, and have been having trouble with a couple of things....Has anyone out there ever gotten the blocking agent to dissolve at 0.5% (every time I try, I get what looks like poached egg white, regardless of how gently or vigorously I heat it!) I can't remember who sent the suggestion to someone else to try cold water fish skin gelatin for blocking, but bless you!! It seems to be of some help. Is it possible to "overdetect" with this system? I keep thinking I've overdone it, but when I get the slides dehydrated and Permounted, things aren't very dark (not like what I've seen in the literature) In the DIG applications manual from B-M, I notice that if you're doing blots, the colour is different depending on the membrane used (brownish on nylon, blueish on nitrocellulose). What types of colours are people getting on what types of tissues (I'm looking at soybeans, by the way). Sorry if this sounds incoherent...it is, after all, Monday morning.
Thanks in advance shea
Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
At Advanced Imaging Concepts we make software for archiving and databasing digital images and also handle a wide variety of frame grabbers and systems. The Snappy you mentioned is a parallel port device that only handles composite video, not Y/C. Please contact me and I will be glad to discuss a number of options with you.
Scott E. Berman Advanced Imaging Concepts, Inc. Princeton, NJ Phone(609) 921-3629 Fax(609) 924-3010 email Scott E57-at-aol.com
There is a discussion on this archived at "Tips & Tricks" Go to the web address at the end of this message and click on "Tips & Tricks. In the "TEM" section you will find a link to "Dissolving Methacrylates" which has what you need. Good luck.
At 10:59 PM 5/5/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 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 felt that this would be a good place to ask this question and hopefully I will get lots of good advice:
I want to know if anyone knows where and/or how I can puchase 2.5% calcium gluconate gel.
I have purchased hydroflouric acid from Aldrich Chemical Co. and in their MSDS sheet they recomend this gel as a neutralizing agent if you should happen to get any HF on your skin. I am quite aware of how extremely dangerous HF is and I do not want to use it for any of my etching experiments until I have all my safety precautions taken care of.
I called Aldrich and they do not sell the calcium gluconate and they were of absolutely no help in letting me know where I could find it and they did not give me any readily available alternatives.
Does anybody have any clues or suggestions as to where I can get this calcium gluconate gel or for that matter, do you have any helpful suggestions about dealing with this extremely nasty chemical. If I had my choice I would not use it at all, but I must.
Thank you for your help.
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
I've had some expierience with several confocal systems. Our lab currently has an InSignt and Ultima form Meridian Instruments. The Ultima is more specialized for cytometry, does a wonderful job, but requires a full-time supervisor. The InSight is brand new and looks promising. It has not been used enough to make any judgments.
I personally use an older Zeiss LSM (model 20, I believe) and a Biorad MRC600. Both provide good general purpose use. I would suspect the upgrades of these machines would be very useful. (The limits on these systems for me is that they do not allow ratiometric imaging--that is the older models)
All of this is IMO, alone and I have no financial interest in any of these companies.
Good luck.
Tommy C. Sewall Image Analysis Laboratory College of Veterinary Medicine Texas A&M University
} I want to know if anyone knows where and/or how I can puchase 2.5% calcium } gluconate gel. } The three readily available chemical catalogs--Alfa Aesar, Aldrich and Sigma--all have gluconates, and Aldrich and Sigma have Ca-salts (you could, of course, add CaOH to the acid if need be). I guess that adding the right amount of H2O will give the 2.5% gel.
} I have purchased hydroflouric acid from Aldrich Chemical Co. and in their } MSDS sheet they recomend this gel as a neutralizing agent if you should } happen to get any HF on your skin. I am quite aware of how extremely } dangerous HF is and I do not want to use it for any of my etching } experiments until I have all my safety precautions taken care of. } } I called Aldrich and they do not sell the calcium gluconate and they were } of absolutely no help in letting me know where I could find it and they } did not give me any readily available alternatives.
Obviously, their catalog and sales reps are not on the same page! } } Does anybody have any clues or suggestions as to where I can get this } calcium gluconate gel or for that matter, do you have any helpful } suggestions about dealing with this extremely nasty chemical. If I had my } choice I would not use it at all, but I must. } Use it in a hood, wear a lab coat and polyethylene gloves, and use goggles or a face shield. H2F2 is an inhalation hazard, so make sure your hood has adequate air flow. H2F2 is nasty, but not as nasty as some of the chemicals in use in organic chem labs. Good luck. Yours, Bill Tivol
OK, so Tilex was on sale this weekend, and I bought some. Never mind that when I once used it in my shower years ago I was mightily offended by the smell. Let's put together a list of all the wonderful uses this group has come up with! I'm sure the Clorox Company would be amazed. This sounds like the duct tape of the solvent world. Contains diethylene glycol butyl ether.
*Duct tape is like the Force; it has a dark side and a light side and holds the Universe together.*
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* ****************************************************************************
You may also want to check with manufacturers of such equipment. We maintain a database of sample preparation methods and can often give references fr specific preparation needs. We also publish a bibliography of technical papers dealing with sample preparation which we are pleased to send you at no charge. You can then select papers of interest and we will send you reprints at no charge.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy. Message text written by Juan Marti } Hello EVERYBODY,
Does anyone out there knows if there is a specific list about Surface preparation methods for LM (Grinding, Polishing, etching...etc)?
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Materials technician positions are available immediately to qualified persons as SEM and TEM operators in the Process Characterization Analysis Laboratory at AMD's Austin manufacturing facilities. The positions require demonstrated experience in the preparation of samples associated with the microelectronics industry as well as basic understanding of materials involved in submicron CMOS technology. Good organizational skills are a must. Good interpersonal and communication skills in a multidisciplinary environment are expected. Additional skills in the operation of state of the art TEM or SEM's would be helpful. These are career path positions and allow contribution individually or in teams in a cooperative environment. An Associate degree in electron microscopy, materials science or a physical science is needed. Directly related experience will be considered in lieu of a degree. Send resumes to David Valdez, AMD, 5204 E. Ben White Blvd., M/S 556. Austin, TX - 78741; FAX (512) 602-5108. AMD is an equal opportunity employer.
Mr-Received: by mta RANDD; Relayed; Mon, 05 May 1997 14:25:12 -0500 Mr-Received: by mta MCM$RAND; Relayed; Mon, 05 May 1997 14:25:13 -0500 Mr-Received: by mta RANDB; Relayed; Mon, 05 May 1997 14:25:20 -0500 Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
About three years ago, I spent several months trying to use the Boehringer digoxygenin system on lung tissue sections. The blocking agent was a huge pain to get into solution. I had several conversations with the technical reps about this - and got conflicting recommendations on how to get it in solution. One of the methods was to heat the solution in a microwave processor - the result was a carmelized goopy mess. I was also told to heat water to 68C on a hot plate and add the blocking solution powder while stirring vigorously; the key, I was told, was to avoid a rolling boil. That worked fairly well - until I autoclaved it. When I pulled my containers out of the autoclave, I found something that resembled soupy cottage cheese. As I recall, the next time I made the solution, I used the hot plate method and ignored Boehringer's recommendation to autoclave it. I eventually switched to P-33 and autoradiography, as the mRNA I was looking for was not abundant, so I never became an expert at using the digoxygenin method.
About the color reaction: if you are using BCIP/INT as the chromogen for alkaline phosphatase (as I remember, that's what Boehringer gives you), the reaction product is soluble in organic solvents. You may be losing it if you are mounting your slides in Permount. There are aqueous mounting media that will work better for this chromogen.
Good luck!
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064
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Dear MSA Newsgroup:
I felt that this would be a good place to ask this question and hopefully I will get lots of good advice:
I want to know if anyone knows where and/or how I can puchase 2.5% calcium gluconate gel.
I have purchased hydroflouric acid from Aldrich Chemical Co. and in their MSDS sheet they recomend this gel as a neutralizing agent if you should happen to get any HF on your skin. I am quite aware of how extremely dangerous HF is and I do not want to use it for any of my etching experiments until I have all my safety precautions taken care of.
I called Aldrich and they do not sell the calcium gluconate and they were of absolutely no help in letting me know where I could find it and they did not give me any readily available alternatives.
Does anybody have any clues or suggestions as to where I can get this calcium gluconate gel or for that matter, do you have any helpful suggestions about dealing with this extremely nasty chemical. If I had my choice I would not use it at all, but I must.
Thank you for your help.
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 Microscopy ListServer -- Sponsor: The Microscopy Society of America
Materials technician positions are available immediately to qualified persons as SEM and TEM operators in the Process Characterization Analysis Laboratory at AMD's Austin manufacturing facilities. The positions require demonstrated experience in the preparation of samples associated with the microelectronics industry as well as basic understanding of materials involved in submicron CMOS technology. Good organizational skills are a must. Good interpersonal and communication skills in a multidisciplinary environment are expected. Additional skills in the operation of state of the art TEM or SEM's would be helpful. These are career path positions and allow contribution individually or in teams in a cooperative environment. An Associate degree in electron microscopy, materials science or a physical science is needed. Directly related experience will be considered in lieu of a degree. Send resumes to David Valdez, AMD, 5204 E. Ben White Blvd., M/S 556. Austin, TX - 78741; FAX (512) 602-5108. AMD is an equal opportunity employer.
FESEM Engineer FAB 12 Yield Dept. Materials Lab Intel Corporation
The successful candidate will be a materials analyst in the FAB-12 Materials Lab. Responsibilities will include interacting with process, reliability and materials/LYA lab engineers in resolving materials issues in manufacturing and improving process yields by performing required analysis and providing the team with data interpretation and recommendations. Initial responsibilities will be focused on supporting the Scanning Electron Microscopy (SEM) lab. The candidate will be required to provide direction/supervision for 2-3 technicians in sustaining SEM support. Must have BS or MS in Materials Science, Chemical Engineering or equivalent with work experience or relevant coursework in materials analysis related to the semiconductor industry.
Skills:
Excellent communication/interpersonal skills required. The successful candidate should be able to prioritize multiple tasks to effectively meet customer needs in a changing environment. Must have hands on experience with Scanning Electron Microscopy (SEM).
Intel is an equal opportunity employer. Resumes accepted by fax, email, or snail mail. Interested candidates should contact:
Satish Naidu Manager-F12 Materials Lab Intel Corporation M/S OC2-218 4500 S. Dobson Rd Chandler, AZ 85248-4906 satish_k_naidu-at-ccm.ch.intel.com Fax(602)715-8363
I clean mine by gently dribbling Freon over the window, a gentle stream from a plastic dropper bottle (the sort that has a nozzle as its top and that you squeeze), directing the stream to the metal above the window. I feel a bit bad about using Freon, but it's such a great solvent for grease and oil, and so non-aggressive towards epoxies and things that I justify my continued use of the stocks which I inherited by the absense of any disposal facilities in NZ. When it runs out I think I'll use pet ether.
cheers
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
I do not not if this might help you. I am using HF on a regular basis. Here in Australia I can get the Calcium Gluconate Gel from ORION - Laboratories P/L - Briggs Street - Welshpool - Western Australia.
Hans Brinkies Senior Lecturer SWINBURNE, University of Technology School of Engineering and Science HAWTHORN - Vic. - 3122 - Australia
This reply is mostly a soapbox digression from the microscopy theme of the listserver so DELETE or "click past" now if you're busy. I would suggest sending any replies to me personally rather than cluttering up the list.
To Ritchie and others worried about CFC (freon) use,
Dab away merrily with your freon soaked swab. And do so without guilt. Freon is one of humanity's greatest creations. Inert, non toxic, non-flammible and the planet's most cost effective refrigerant, this compound and other CFCs have gotten a bum wrap. The new CFC-free replacement compounds are corrosive, toxic, expensive and only 75 to 80% as efficient. At least one toxic spill/bird kill has already occurred. My 70's vintage Neslab Cryocool can crank -100 C after 25 years. When it breaks, I can only get the new CFC-free model can achieve no more than -80 C and will not last as long. Absolutely no disrespect to Neslab intended or deserved.
Stratospheric ozone depletion measurements were not taken before the 1970's. "Ozone Depletion" may turn out to be natural ozone variation. We won't know for centuries. O3 to O2 CAN occur via chlorine and fluorine catalytic pathways. That man-made CFCs are the catalysts can never be proved since chlorine and fluorine are abundant natural elements. Maximum reported CFC concentrations in the stratosphere are in the area of 3 parts per trillion (Nasa?). Seems kind of sparse if true. Can anyone tell me if that kind of detection limit is reliable for any instrument?
Maximum increased "plus hundred year" UV fluxes related to projected stratospheric ozone thinning, CFC caused or otherwise, are comparable, in skin cancer health risk to moving a few hundred miles South or a thousand feet up in elevation. Pale northerners taking winter vacations in southern latitudes will suffer radical UV induced cancer risk due to insufficient melanin buildup, regardless of ozone layer variations. (Don't forget to smear the PABA 16 between your toes, the new high risk site for benign skin cancer.) In my understanding malignant melanoma is not directly associated with UV exposure.
The Montreal Protocol back in 89-90 created an agreement among the industrialized nations that CFCs would be banned from manufacture in 1995. E.I. duPont was the principal supplier of CFCs, and yet duPont was an important financial contributor to the physical execution of the meetings for the Montreal Protocol. 1995 was also the year that duPont lost their patent on CFCs. Seem suspicious? I WOULD SAY SO, when you consider that they also hold the new patents on the legal CFC replacements. And... please forgive me or correct me if I am wrong about this, but I have heard in conversation with industrial suppliers that in 1995 duPont fired up a large new CFC manufacturing facility in India to supply refrigerants for the anticipated third world appliance boom. Once again, special interests make the planet go-round.
I have no direct financial stake in CFC issues. I just hate losing another great material and suffering useless regulatory expenses as a result of hasty science and ill conceived politics.
I welcome all elaborations and criticisms on the above. Ozone and asbestos paranoia are my pet peeves. I would only be happier to learn that our sacrifices are not in vain.
We have a new Polaron CPD and are having difficulty maintaining the carbon dioxide level when flushing. A check for leaks, revealed a small leak at the pressure gauge, which new seals fixed. However the level problem still persists. The temperature is kept around 12 degrees celisus. Could this be the inlet valve? Has anyone else had this problem and what did you do to fix it. TIA
Mary Toogood Atlantic Food & Horticulture Research Centre Agriculture and Agri-Food Canada Kentville N.S. B4N 1J5 ToogoodM-at-em.agr.ca
Thanks for all your good solutions (yuk, yuk) for removing tapes/spots from Al mounts. You have given me just the ammunition I need to justify a $50K (US) supercritical CO2 organic extraction system.
Seriously, I used a commercial paint/label remover, Goof-off and a little elbow grease. It worked fine. I then immersed the stubs in an ultrasonic bath of warm (40C) DI water and 0.5% by volume Micro for about five minutes followed by a DI rinse and air dry. I haven't looked at them in the SEM yet, but at optical mags up to 40x, they appear clean even when viewed using a UV lamp.
Thanks again. (Note to vendors: I still buy new ones.)
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
How does one go about getting added (and deleted) from your mailing list?
MG
Dr. Michael G. Gorczyca, Ph.D. Department of Biology Morrill Science Center University of Massachusetts Amherst, MA 01003 tel: (413) 545-1783 or -4627 fax: (413) 545-3243 e-mail: mgor-at-bio.umass.edu
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Hello, I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS spectra has a large noise peak, typically 10 times the dominant real peak. I have been talking with Oxford about this a great deal and they are unsure of the cause. I now ask you for input.
I checked the electrical isolation between the dewar and the column and found there is none (1 Ohm) if the cables are connected and infinite if they are disconnected. To Oxford's suprise they found the same on their system. Is this normal? This means that noise in the SEM is in the EDS!? How should I be grounding this system. We do have a 1 cm diameter copper wire that is a dedicated ground running up to these to systems. Any general tips on electrical issues for EDS would be appreciated.
thanks
Brad Storey Materials Scientist Argonne National Lab - West P.O. Box 2528 Idaho Falls, ID 83403 Ph. 208-533-7685 Fax 208-533-7683 e-mail brad.storey-at-anl.gov
Message-Id: {199705061459.KAA22195-at-umic.sunysb.edu} Comments: Authenticated sender is {greg-at-mail.umic.sunysb.edu}
Mary, Maintaining the level of CO2 is a balancing act. It must be watched!! As the CO2 drains it freezes at the valve blocking the opening. This can slow draining. Then it will blow free and the rate of draining speeds up. The flushing should be done several times for a few minutes each time rather than continually. Hope this helps.
} } We have a new Polaron CPD and are having difficulty } maintaining the carbon dioxide level when flushing. A } check for leaks, revealed a small leak at the pressure } gauge, which new seals fixed. However the level problem } still persists. The temperature is kept around 12 degrees } celisus. Could this be the inlet valve? Has anyone else } had this problem and what did you do to fix it. TIA } } } } Mary Toogood } Atlantic Food & Horticulture Research Centre } Agriculture and Agri-Food Canada } Kentville N.S. B4N 1J5 } ToogoodM-at-em.agr.ca } Gregory Rudomen Greg-at-UMIC.SUNYSB.EDU 516-444-3126 University Microscopy Imaging Center S.U.N.Y. Stony Brook
Dear all, A week ago, I sent the following message to the list.
} Dear all, } I am aware of a number of programs that can produce calculated diffraction } } patterns given the unit cell parameters and the location of all atoms in } the } unit cell. This poses difficulties for some materials with more } complex } structures, however, where site occupancies are not 100% and } where the 'unit } cell' is somewhat of an average structure (this occurs } for some oxides). This } can be overcome if a good crystal structure } refinement has been done from XRD } where average atomic positions and site } occupancies have been determined. Many } structures have not been studied } in this detail, however, and the only } information that is available is } the unit cell parameters and the space group. } Is there any software that } can plot electron diffraction patterns from this } limited information. } } I hope there is someone who can help me with this.
I have received a number of helpful suggestions. These include using Desktop Microscopist (for Macintosh) from Virtual Labs. We have this program here (version 1.05) but I've had problems running it on my computer. Some other prgrams were mentioned that we don't have and haven't been tried. Phillipe-Andr=E9 Buffat at EPFL in Switzerland recommended the use of EMS online via the WWW at http://cimewww.epfl.ch/ and this has proved very useful and I will probably use it at some point.
Thanks to all who responded
Yours sincerely
Ian MacLaren
P.S. A copy of the responses can be received by emailing me.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Dear all, A week ago, I sent the following message to the list.
} Dear all, } I am aware of a number of programs that can produce calculated diffraction } } patterns given the unit cell parameters and the location of all atoms in } the } unit cell. This poses difficulties for some materials with more } complex } structures, however, where site occupancies are not 100% and } where the 'unit } cell' is somewhat of an average structure (this occurs } for some oxides). This } can be overcome if a good crystal structure } refinement has been done from XRD } where average atomic positions and site } occupancies have been determined. Many } structures have not been studied } in this detail, however, and the only } information that is available is } the unit cell parameters and the space group. } Is there any software that } can plot electron diffraction patterns from this } limited information. } } I hope there is someone who can help me with this.
I have received a number of helpful suggestions. These include using Desktop Microscopist (for Macintosh) from Virtual Labs. We have this program here (version 1.05) but I've had problems running it on my computer. Some other prgrams were mentioned that we don't have and haven't been tried. Phillipe-Andr=E9 Buffat at EPFL in Switzerland recommended the use of EMS online via the WWW at http://cimewww.epfl.ch/ and this has proved very useful and I will probably use it at some point.
Thanks to all who responded
Yours sincerely
Ian MacLaren
P.S. A copy of the responses can be received by emailing me.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
When you say "noise peak", are you referring to the strobe at 0 keV? Ours is generally comparable to the major peaks in our spectra for an Isis 300 system on a Hitachi 2460N SEM. But we typically run several thousand counts per second for most of our samples. If we turn our beam off, we can still get our strobe peak. Therefore, I wonder if the issue is one of count rate. What is your typical rate. I am not familiar with a DSM960a so I don't know what your normal conditions are. I hope this helps some.
At 08:47 AM 5/6/97 -0700, you wrote: } Hello, } I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS spectra } has a large noise peak, typically 10 times the dominant real peak. I } have been talking with Oxford about this a great deal and they are } unsure of the cause. I now ask you for input. } } I checked the electrical isolation between the dewar and the column and } found there is none (1 Ohm) if the cables are connected and infinite if } they are disconnected. To Oxford's suprise they found the same on their } system. Is this normal? This means that noise in the SEM is in the } EDS!? How should I be grounding this system. We do have a 1 cm } diameter copper wire that is a dedicated ground running up to these to } systems. Any general tips on electrical issues for EDS would be } appreciated. } } thanks } } Brad Storey } Materials Scientist } Argonne National Lab - West } P.O. Box 2528 } Idaho Falls, ID 83403 } Ph. 208-533-7685 } Fax 208-533-7683 } e-mail brad.storey-at-anl.gov ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
} I have a Reichert Ultracut E that needs service. Does anyone know } who does repairs on these things? I'm on the west coast, is there someone } local?
Paul Swerdlick, of Leica Inc.'s service group, is based in California. He recently worked on our two Ultracut E's. Contact me privately for his phone number, if you need it.
} Keith Porter died last Friday. For those of you who aren't familiar with } this name, Porter truly deserved the title 'pioneer'. While he was at the } Rockefeller University, he helped develop the use of electron microscopy } for biological materials. Thus, together with other giants like George } Palade, he discovered cellular organelles that we now take for granted, } like the endoplasmic reticulum. He also helped invent the microtome } with which ultrathin sections could be cut. For years it was called the } Porter-Blum ultramicrotome. After the introduction of glutaraldehyde as } a fixative, he helped discover microtubules and thus played a key role in } igniting interest in the cytoskeleton. While his work primarily } centered on animal cells, Porter also investigated a variety of other } organisms, including plants. Together with Myron Ledbetter, he discovered } cortical microtubules in plant cells and therefore helped establish the } microtubule-cellulose microfibril paradigm. } Porter literally helped start the modern era of cell biology. He } moved from Rockefeller to Harvard and attracted a generation of new } scientists to the emerging field including Peter Hepler, Ben Bouck, Dick } McIntosh, and Lew Tilney, all of whom do or have worked on plants and } algae. Our own David Porter was an alumnus of the Keith Porter lab. Later, } Keith switched to the University of Colorado to found the Department of } Molecular, Cellular and Developmental Biology, one of the first if not the } first department of its kind. He thus started another trend: we now have } many departments of molecular and/or cellular biology all around the } world. } Porter was also president (and I believe a founder) of the } American Society for Cell Biology, one of the most active scientific } societies in the world, and he helped start the Journal of Cell Biology. } He also received the National Medal of Science. Sadly, when the Nobel } Prize in Medicine and Physiology was handed out years ago to George } Palade, Albert Claude and Chistian DeDuve for pioneering studies on cell } ultrastructure, Porter was snubbed. It was a great injustice. } Keith Porter was also known for his wit and sarcasm. Having } witnessed his sparkle from meeting halls and to elevators, I can tell you } that he was one of a kind. } In his last years, Keith was an emeritus professor at the } University of Pennsylvania. I am told that he was close to Kenneth } Thimann, a legend in plant physiology (identified indole acetic acid } as auxin) who also spent his last days at Penn and who, coincidentally, } also recently passed away. } Keith Porter's obituary appears in today's New York Times } (www.nytimes.com).
This message was sent to everyone in my Department today. Thought I should share it with y'all.
Best regards, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
One of my students, a talented young boy finishing his third year of studying Materials Engineering is looking for a job for the summer. He is ready to do anything for two months at a university or research institute abroad, speaks English quite well, and is experienced in SEM as well as other analytical techniques. He will be satisfied if he can pay his accommodation and living expenses while working since he just wants to gain experience and face the challenge. Please answer me directly and not through the List.
Thanks,
Kris
************************************************************************* Dr. Kristof KOVACS Associate Professor University of Veszprem Department of Silicate and Materials Engineering, Central Laboratory P.O.Box 158, Veszprem, HUNGARY H-8201 Phone: +36-(88)-421-684 *************************************************************************
Has anyone used those knife cleaners that swirl water back and forth? Does anyone have a better idea? Even trying to keep the knife clean I seem to get pieces of resin on the back side that is dificult to get off with a jet streem of water from a bottle or syringe. Thanks in advance.
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How does one go about getting added (and deleted) from your mailing list?
MG
In case you didn't receive a copy of the message, the answer is found above.
Brad Storey wrote: } } Hello, } I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS spectra } has a large noise peak, typically 10 times the dominant real peak. I
} have been talking with Oxford about this a great deal and they are } unsure of the cause. I now ask you for input. } } I checked the electrical isolation between the dewar and the column and } found there is none (1 Ohm) if the cables are connected and infinite if } they are disconnected. To Oxford's suprise they found the same on their } system. Is this normal? This means that noise in the SEM is in the } EDS!? How should I be grounding this system. We do have a 1 cm } diameter copper wire that is a dedicated ground running up to these to } systems. Any general tips on electrical issues for EDS would be } appreciated. } } thanks } } Brad Storey } Materials Scientist } Argonne National Lab - West } P.O. Box 2528 } Idaho Falls, ID 83403 } Ph. 208-533-7685 } Fax 208-533-7683 } e-mail brad.storey-at-anl.gov }
Dear Brad, I think the noise can occur for four reasons:
1. Microphone effect from small crystals of ice, which accumulated at the bottom of dewar. They can be seen at the bottom in a moment when the nitrogen in dewar is almost over. I got rid of them with the help of some litres of boiling water, which filled in instead of liquid nitrogen. For realization of this procedure it is necessary to take off the detector from microscope, to pour out nitrogen, to fill boiled water and shake up approx. 10 seconds. Then it is necessary to dry up the dewar with the help of compressed air or other similar gas. All operations should be done quickly, that the sensor plate and FET transistor inside the detector had no time to heat up.
2. Microscope and EDS are connected to different phases of 3-phases power mains. Check they are connected to the same phase. If microscope uses all three phases, EDS and low voltage electronics of the microscope should be connected to the same phase.
3. Grounding. The detector and column should be electrically isolated from each other in order to be grounded only in one point. Your situation is OK. In general the good grounding is complex technical problem. Sometimes it is better (in sense of noise) to work with neutral connection instead of grounding (but all metal subjects in a room should be connected equally), sometimes without it at whole (but it is dangerous for health!). It is very difficulty to advise something about grounding from far away.
4. Dust on high-voltage circuits. Sometimes a dust accumulates on high-voltage circuits, then microdischarges of high voltage appear. These microdischarges cause occurrence of noise on a spectrum. Therefore these HV circuits need sometimes to be cleaned. They are located inside the rack with electronics and in preamplifier, which hangs on dewar. Caution! It is necessary to wait about half-hour after turning off the instruments until the charge flows down.
Attention! All above works should be carried out by the qualified personnel.
} Hello, } I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS spectra } has a large noise peak, typically 10 times the dominant real peak. I } have been talking with Oxford about this a great deal and they are } unsure of the cause. I now ask you for input.
Ground loops can be a source of major noise peaks such as this, but I have seen them caused both by faults in the preamp, or incorrectly adjusted preamps (some EDS preamps have trim pots for adjustment - don't touch unless you have detailed instructions for set up)
} I checked the electrical isolation between the dewar and the column and } found there is none (1 Ohm) if the cables are connected and infinite if } they are disconnected. To Oxford's suprise they found the same on their } system. Is this normal?
I would be worried about this, because it is exactly as it should be - find somebody else at Oxford to talk to:)
} This means that noise in the SEM is in the } EDS!? How should I be grounding this system. We do have a 1 cm } diameter copper wire that is a dedicated ground running up to these to } systems.
The EDS detector should be grounded through its control electronics, the power supply of which then goes back to the same source as the SEM this is where you dedicated ground should connect in. Make sure the grounding scheme is 'singly connected', that is any point in the syste can only get to ground by one route - you may have to spend a lot of time disconnecting and one-by-one reconnecting various accessories.
} Any general tips on electrical issues for EDS would be appreciated. } } thanks } } Brad Storey } Materials Scientist } Argonne National Lab - West
I'd first suggest taking the detector off the SEM and powering it all up on the bench. Its nice to have a radioactive source to allow you to test the EDS on the bench. If the noise peak disappears, it's got to be something to do with the SEM, although if as you say the EDS detector is correctly isolated from the SEM, I'd expect the peak to stay there.
You don't actually specify where you see the noise peak - I'm assuming it is right at the low energy cut off of the spectrum. I'm not familiar with the latest systems but certainly with earlier units, there is always a noise peak at the low energy end - it is usually not apparent because it is electronically cut off at the preamp - one of the trim pots I mentioned earlier. I've also seen systems where the output from the noise peak seems to swamp the preamp in some way, so that as you adjust the position of the low end cut off, and remove the noise peak, the count rate in the real peaks goes up.
Brad Storey wrote: } } Hello, } I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS spectra } has a large noise peak, typically 10 times the dominant real peak. I
} have been talking with Oxford about this a great deal and they are } unsure of the cause. I now ask you for input. } } I checked the electrical isolation between the dewar and the column and } found there is none (1 Ohm) if the cables are connected and infinite if } they are disconnected. To Oxford's suprise they found the same on their } system. Is this normal? This means that noise in the SEM is in the } EDS!? How should I be grounding this system. We do have a 1 cm } diameter copper wire that is a dedicated ground running up to these to } systems. Any general tips on electrical issues for EDS would be } appreciated. } } thanks } } Brad Storey } Materials Scientist } Argonne National Lab - West } P.O. Box 2528 } Idaho Falls, ID 83403 } Ph. 208-533-7685 } Fax 208-533-7683 } e-mail brad.storey-at-anl.gov }
Dear Brad, I think the noise can occur for four reasons:
1. Microphone effect from small crystals of ice, which accumulated at the bottom of dewar. They can be seen at the bottom in a moment when the nitrogen in dewar is almost over. I got rid of them with the help of some litres of boiling water, which filled in instead of liquid nitrogen. For realization of this procedure it is necessary to take off the detector from microscope, to pour out nitrogen, to fill boiled water and shake up approx. 10 seconds. Then it is necessary to dry up the dewar with the help of compressed air or other similar gas. All operations should be done quickly, that the sensor plate and FET transistor inside the detector had no time to heat up.
2. Microscope and EDS are connected to different phases of 3-phases power mains. Check they are connected to the same phase. If microscope uses all three phases, EDS and low voltage electronics of the microscope should be connected to the same phase.
3. Grounding. The detector and column should be electrically isolated from each other in order to be grounded only in one point. Your situation is OK. In general the good grounding is complex technical problem. Sometimes it is better (in sense of noise) to work with neutral connection instead of grounding (but all metal subjects in a room should be connected equally), sometimes without it at whole (but it is dangerous for health!). It is very difficulty to advise something about grounding from far away.
4. Dust on high-voltage circuits. Sometimes a dust accumulates on high-voltage circuits, then microdischarges of high voltage appear. These microdischarges cause occurrence of noise on a spectrum. Therefore these HV circuits need sometimes to be cleaned. They are located inside the rack with electronics and in preamplifier, which hangs on dewar. Caution! It is necessary to wait about half-hour after turning off the instruments until the charge flows down.
Attention! All above works should be carried out by the qualified personnel.
} } I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS spectra } } has a large noise peak, typically 10 times the dominant real peak. I } } have been talking with Oxford about this a great deal and they are } } unsure of the cause. I now ask you for input. } } } } I checked the electrical isolation between the dewar and the column and } } found there is none (1 Ohm) if the cables are connected and infinite if } } they are disconnected. To Oxford's suprise they found the same on their } } system. Is this normal? This means that noise in the SEM is in the } } EDS!? How should I be grounding this system. We do have a 1 cm } } diameter copper wire that is a dedicated ground running up to these to } } systems. Any general tips on electrical issues for EDS would be } } appreciated. } }
The dewar must be electrically isolated from the column or you will get a ground loop between the EDS electronics and the microscope grounds. The ground path must be via the pre-amp cabling, so this is correct. I'm amazed that the Oxford people that you are talking to did not know this, it's one of the things that is checked when you install a detector. For general grounding rules, think meca, all grounds must have one path to the main ground. If you have two paths, then that's a ground loop.
Sounds like your pulse processor's discriminators are not adjusted correctly. You need to monitor the input or fast channel cps to adjust the discriminators. The adjustment is easy, much better to tune than Noran or Kevex pulse processors. Check your manual for how to adjust the pulse processor (if you don't have one, get Oxford to FedEx you one, you should have it). You will either have a 2010, 2020, 2040, XP2 (computer adjusted), or XP3 (same as XP2 but standalone and manual adj). The new digital (DXP50) is most likely computer adjusted too).
The zero strobe goes off at about 500-600 cps. The zero strobe rate will decrease as the count rate is increased. It should be at zero eV in energy. With the beam off, it's all you should see (or rather 1/2 of the peak since the peak should be at zero eV).
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
The Electron Microscopy Laboratory of New Mexico State University announces an opening for an Electron Microscopy Specialist. Qualifications: a B.S./M.S. degree with at least 4 years of experience in electron microscopy. The ideal candidate should have advanced experience in SEM/EDX. Proven experience in digital image analysis is a plus. The candidate should also be comfortable with the following techniques: metal evaporation, sputter coating, critical point drying, support film production, low temperature embedding, and film processing and printing. The successful candidate must be able to work well with people both as a collaborator and as a teacher. Duties include: record keeping, fixation, embedding, ultra-thin sectioning, staining, operation and routine maintenance of transmission and scanning electron microscopes and other laboratory equipment. Salary: $28,000 to $30,972. Benefits include: group medical and hospital insurance, group life insurance, state educational retirement, workmen's comp, sick leave, and unemployment compensation. Deadline for Applications: screening of applications will begin May 15 and will continue until a candidate is chosen. Applications should include a resume, letter of interest and 3 letters of recommedation. Send applications to: Arts and Science Research Center Box 30001/Dept. RC Las Cruces, NM 88003-8001 Tel. (505) 646-2611 Fax (505) 646-6095
I omitted the name of the person who wrote the article on Keith Porter. Dr. Barry Palevitz wrote the email that I forwarded to the list yesterday. I apologize for that omission.
Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Does anyone know of a commercial source of antibodies which could be used to identify golgi activity. My specimens ( P. carinii ) have been fixed with 4% paraformaldehyde - 0.1 % glutaraldehyde and are embedded in LR white resin. You can send replies directly to me at: mgoheen-at-indyvax.iupui.edu.
Thanks
Mike Goheen Dept of Pathology Indiana University School of Medicine
} ... } } I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS } spectra } } has a large noise peak, typically 10 times the dominant real peak. } ...
Noise can arise from nitrogen boiling in the dewar ... which can be an indication the vacuum in the dewar has degraded. Ice boining around in the dewar can also cause noise ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
To everyone on this listserv, I have reposted the position for EM Specialist just to get those who might have missed it while on vacation or were glued to their tem binoculars. I am sorry if I have confused anyone.
I recently came across a procedure that Hildy Crowley gave me at a M.S.A. meeting some time ago in which the lead stain is kept in a 60 ml syringe that is fitted with 2 Acrodisc filters...and kept refrigerated. What I don't know is how often do I need to change out those filters. It would seem to me that a precipitate would form that would eventually cause artifact in the stain, showing up like "black boulders" under the TEM. Has anybody used this system and how long does the stain stay "good"?
Thanks in advance,
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
A colleague of mine mentioned seeing an article recently (1-4 weeks ago) in The Chronicle of Higher Education regarding economy measures and electron microscopy units in universities/colleges. Does anyone have the reference so I can check it out in the library? I can imagine what it says .......
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
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Sharron, I use the same system. I have a syringe needle on the filter which I change daily, my naive thinking being that any precipitates will go with the needle and when the syringe is empty I start afresh with a 20ml syringe and 0.2=B5m nylon Acrodisc filter. I use the same system for uranyl acetate (saturated in 50% methanol), this time with a 0.45=B5m Acro LC3S filter. I've been using this system for years and it's never let me down giving clean crisp staining. Ian.
I am collecting information on the currently valid standards applied in metallographic examinations in different countries and corresponding to the following standards of ASTM, ISO nad JIS:
In april 20-24 1998 in Amsterdam, the Netherlnads is made "7th European Congress For Strereology" title "trends and challenges in stereology"
adres local organizing committee:
Congress Secretariat VU Conference Service De Boelelaan 1105 1081 HV Amsterdam The Netherlands tel: +31 (0)20 4445790 fax: +31 (0)20 4445825 e-mail:vu_conference-at-dienst.vu.nl
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Structural and Physical Research Laboratory
We have embedded bacteria in LR White successfully but now are trying to embed a fungus, Ustilago hordei, so that we can do immunogold work with it. We had embedded it for years in various epoxies and had few problems, but with the LR White we have large holes not only in the cells but outside, not random but so that it appears that the cells are loose in the acrylic instead of embedded in it. Does anyone have experience preparing similar materials for immuno work? We don't have the equipment for frozen sections. Joyce Craig Chicago State University jcraig-at-csu.edu
I am working on a potato storage project and trying to conquer embedding of amyloplast starch in tuber samples. I've been using Spurr's resin, after 2% glutaraldehyde fixation in sodium cacodylate buffer and osmium tetroxide post fixation. I use propylene oxide after ethanol dehydration series to ensure dry samples, but still most of the starch granules and plastids fall out during sectioning and post staining.
Has anybody else worked with starch or potato tuber with success? I'm looking for any protocol hints that will better preserve starch and amyloplasts. Has anybody tried microwave fixation procedures on starch samples?
Thanks for any help you can give. I'll send 5# of the fantastic Minnesota russets to anyone who provides information that leads to success (but you'll have to wait until October to get them!).
Darryl Krueger Mn. Ag. Experiment Station EM Lab Univeristy of Minnesota St. Paul, MN 55108 (612)625-8249 darrylk-at-puccini.crl.umn.edu
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Subject: Time:1:14 PM OFFICE MEMO POSTDOCTORAL POSITION - ELECTRON... Date:5/8/97
POSTDOCTORAL POSITION - ELECTRON MICROSCOPY OF ELECTRONIC OXIDES
A post-doctoral position is available for electron microscopy studies of electronic oxide materials. The materials of interest include high-temperature superconductors, diffusion membranes, and colossal magneto-resistive compounds.
A Ph.D. in Materials Science/Physics or related area is required. Candidates with a strong background in interfaces, crystallography, and the effects of crystal defects on properties, especially in oxides, are preferred as are those with experience in one or more of the materials systems described above. Extensive hands-on experience in both analytical electron microscopy and HREM is required as is a demonstrated proficiency in TEM specimen preparation of bulk and thin film samples.
Additional skills in x-ray diffraction, in-situ experimentation in either EM or XRD, or measurement of electrical and/or ionic transport properties are also highly desirable.
Interested candidates should send a resume, publication list, and the names of three references to:
Dean Miller Materials Science Division Argonne National Laboratory Argonne, IL 60439
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The Goodyear Tire & Rubber Company, a world leader in tire development and manufacturing, has an excellent opportunity for a professional skilled in Microscopy in its Analytical Services Department, Corporate Research Division, located in Akron, Ohio. This position involves the analysis and characterization of tire components, polymers and other materials used in the rubber industry.
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An Equal Opportunity Employer, M/F/D/V Applicants must be lawfully authorized to work in the U.S.
------------------------------------------------------------------------- John A. Wise jawise-at-goodyear.com The Goodyear Tire & Rubber Co. Research Analytical Services - D415B Voice 216.796.8716 142 Goodyear Blvd Fax 216.796.3304 Akron, OH 44305 U.S.A. -------------------------------------------------------------------------
We've down potato tuber by extending 4 to 5X the amount of time spent in each of the dehydration/ infiltration steps. Also, you might give a try using LR White. In the end we find there're still some problems as you mentioned but greatly reduced.
} Darryl wrote: } I am working on a potato storage project and trying to } conquer embedding of amyloplast starch in tuber samples. } I've been using Spurr's resin, after 2% glutaraldehyde } fixation in sodium cacodylate buffer and osmium tetroxide } post fixation. I use propylene oxide after ethanol } dehydration series to ensure dry samples, but still most } of the starch granules and plastids fall out during } sectioning and post staining. } } Has anybody else worked with starch or potato tuber with } success? I'm looking for any protocol hints that will } better preserve starch and amyloplasts. Has anybody tried } microwave fixation procedures on starch samples?
} Darryl, You might try a graded series of P.O. and Spurr and extend your infiltration times. I have used this on bone and it works fine. Or try Epon 812. I know it is viscous but it worked on yeast where Spurr did not. Best of luck!
Gregory Rudomen University Microscopy Imaging Center S.U.N.Y. Stony Brook Greg-at-UMIC.SUNYSB.EDU 516-444-3126
Those of you in the colleges and universities throughout the world are likely struggling with the task of grading final exams about this time of the year, while most of the rest of us will be rejoicing about the fact that we no longer have to take them, and so I thought all of you might enjoy this answer to a history exam question that was printed in the Winter 97 issue of The Hexagon, published by the Chemistry Honors Society, Alpha Chi Sigma. They didn't give the wording of the question exactly, but here's the answer:
"Renaissance was an age in which more individuals felt the value of their human being. Martin Luther was nailed to the church door at Wittenbert for selling papal indulgences. He died a horrible death, being excommunicated by a bull. It was the painter Donatello's interest in the female nude that made him the father of the Renaissance. It was an age of great inventions and discoveries. Gutenberg invented the Bible. Sir Walter Raleigh is a historical figure because he invented cigarettes. Another important invention was the circulation of blood. Sir Francis Drake circumcised the world with a 100-foot clipper."
How'd you like to have to assign a grade to that?
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
I have a query for this renowned group of microscopists. In attempting to account for time spent in the lab in response to" why isn't he sitting in front of the microscope more often" from a remotely situated administrator/bean counter, I thought I'd calculate a general (or range) ratio for the number of hours spent on the microscope (TEM) to the number of hours spent processing, sectioning (and semi-thinning? to find the block with the appropriate region), staining (conventional), development of negs/ printing (or digitally printing) cataloguing/marking the negs and prints, sending prints off and possibly writing reports, descriptions then phone calls, etc.on biological specimens (plant and animal tissue). I realize that plant material usually entails longer processing times and beam time (as it usually is 95% empty), but an average would be ok for plant and animal. If you do the final printing and plate prep for publications, add that in also. If you have just a couple of minutes to quickly compute the ratio of beam time to the pre/post time spent to completion I would appreciate your estimate . TIA, Hank Adams EML NMSU
Dear all, Thanks to all who replied to my recent queries to the list regarding maleate buffered-uranyl actetae enbloc stain and negative scanners.
My colleagues and I appreciate everyones suggestions and comments,
Yours,
----------------------------------------------------------------------- Richard Lander Senior Technician South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
It seems to be generally known that perfluorinated-polyether pump oils and vacuum greases (e.g., Krytox, Fomblin) can cause micro-discharges on high-voltage insulators and thus lead to high-voltage instabilities. (This despite the fact that their beam-degradation mechanisms produce volatile fragments which should be pumped out of the system.)
Though I've gotten the above information from several authoritative sources (such as Wil Bigelow's book) I've never heard an explanation. Specifically: (1) What is the reason why these compounds create more of a high-voltage problem than normal hydrocarbon compounds?; and (2) What is the contamination mechanism? (Direct transfer of the compounds themselves? Deposit of molecular fragments? )
Microscopists: Perhaps you would like to flame that "free money" moron. I have send him a choice message. Apparently he is using our server "ultra.net" and I have asked the service provider to use a little influence too. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
---------- } From: friend-at-noreply.com.ultra.net.au } To: friend-at-noreply.com.ultra.net.au } Subject: FREE MONEY! } Date: Friday, 9 May 1997 3:00 } } Dear Friend, } } Yes, you read it right! FREE MONEY!!!!! Up to $600 per day!! How can that be } possible, you ask? A new money making concept generates income for YOU, } automatically. snip . . . . .
You could try fixing with 2 - 3% potassium permanganate (aqueous) which gives excellent fixation and staining of amyloplasts - at least it did for root tips and endosperm of wheat. However my notes say not to embedd in Spurr's. We used araldite but I should imagine Epon would be OK.
my understanding of LR white is that, like all acrylics (I'm sure someone will correct me if I'm wrong), it does not cross polymerise well. This is one of it's advantages for immuno work because it should cause less damage to antigens than the epoxide resins. However it also tends to mean that it is less effective in embedding more difficult specimens and is less stable in the beam. I would suggest that, if you do not already do so, you should use coated grids to support the less stable sections and it may solve some of your problems.
I apologise if you know all this but it is a problem I've had for years - basically I use LR White for routine embedding of bacteria and cell pellets, and immuno work but I still use Spurr's for virtually everything else because it is reliable and versatile, despite the hazards.
Malcolm Haswell University of Sunderland UK ----------
Darryl Try the following it works in our lab.
Fix in 3% Glutaraldehyde in 0.05M sodium cacodylate buffer. (8 hours or overnight in fridge.)
2% osmium in 0.05M sodium cacodylate buffer - 2 hours maximum 3 x 30 minute wash in 0.05M sodium cacodylate buffer Block stain 2% aqueous uranyl acetate- 45 minutes 2 x 10 minutes wash in double distilled water Dehydration in ethanol series 10 minutes each. 2 x 30 minutes in propylene oxide
Infiltration - use epon/araldite resins To make up. 1 PART EPON 812 1 PART ARALDITE CY212 3 PARTS DDSA ( DODECENLY SUCCINIC ANHYDRIDE) Stir well with a glass stirrer.
25% resin* - 75% propylene oxide - 2 hours 50% resin* - 50% propylene oxide - 2hours 75% resin* - 25% propylene oxide -overnight with caps off in a fume cupboard 100% resins* -24 hours (2 changes in between)
POLYMERIZATION 100% RESIN IN ALUMINIUM DISHES OR EMBEDDING MOULDS IN A OVEN AT 70 DEGREES CENTIGRADE FOR 48 HOURS.
Very important : *Remember to add 1 drop of (DMP-30) 2,4,6 tri (dimethyl aminomethyl) phenol, per 1 ml epon each time. ie. every step for infiltration (25% -100%) If you miss out DMP-30 you might have to discard your specimen and start all over again.
All our fixation and infiltration steps are done in a glass pill vials in a fume cupboard.
I have also tried spurrs resins and found epon to be the best.
You can extend your infiltration time at 100% resins. Best of luck
Vijay H. Bandu Centre for Electron Microscopy Private Bag X01 Scottsville 3209 Kwa Zulu Natal South Arfica E-Mail bandu-at-emu.ac.za
Hi Darryl; My experience with potatoes is strictly at the light microscopy level, embedding in glycol methacrylate. While I realise that GMA isn't suitable for EM, there are, I believe, acrylics that are, such as LR White. My samples were fixed in 4% glutaraldehyde in phosphate buffer, dehydrated through a solvent series (not one I think you'd like for EM, but an ethanol series should do fine) and infiltrated in methacrylate for a week or so in the fridge. Starch granule preservation is quite nice, and I have even been able to observe the "growth rings" in various preparations. I do realize that phosphate buffer is not compatible with EM preps... but I doubt if the buffer would be your problem.
Good luck shea
Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
I am Resia Pretorius from South Africa and am currently working on a PhD )the phylogeny of internal structures of the superfamily Scarabaeoidea ((insects))
I would like to use a SEM to photograph one of the internal chitinous structures (metendosternites) to which flight and leg muscles are attached, and do calculations of different lengths for morphometric analysis.
These muscles are attached firmly to the metendosternites and I would like to remove them, but so far without luck. I have tried Sodium perborate (different concentrations) they use this chemical to clean muscles from snake skulls, but if only loosens the muscles slightly, I still need to pull and tuck at the muscles to remove them.
I have also tried tripsin a 0.5% without it working. Are there any suggestions from anyone, I read in an 1890 article that one can use nitric acid for the macercation of muscles but that was obviously before the SEM was invented. Can one use an acid or would it harm the actual structure of the metendosternites.
I realy need help!! my e-mail address is:
resia-at-mcd4330.medunsa.ac.za
Thanks, Resia Pretorius Lecturer (Biology) Medunsa South Africa
I am doubtful that its a true infiltration problem. Poor fixation emulates infiltration problems. I assume that you are fixing in the cold and that will not do for most yeasts and plants with "massive" cell walls. Plants which store well generally are in that category.
Up to about 25 years ago most of such tissues were fixed in 1-4% KMnO4, without any buffers. Its a lousy fixatives and only preserves membranes, but try it for say 30 minutes at about 20 degrees. I expect that your sectioning and physical retention of starch and plastids will be much better.
To get better fixation I suggest to forget the GA, it does not penetrate those plant cell walls. Use the Osmium at at least 20 degrees and try 30, 60, 120 minutes.
Botanists working on storage tissues have for years struggled with fixation problems, mostly because they copied the biomedical "in ice" doctrine. Autolysis is much more rapid in animal cells than it is in plants. Keeping the temperature low dramatically slows enzyme reaction in animal cells and hence slows autolysis.
See it as a race: Low temperature slows fixation somewhat, but in animal cells much slower autolysis is the winner. In those "difficult" plant tissues cold fixation is disproportionately slower because of the massive cell walls. Additionally, plant enzyme's activities varies much less in the considered temperature range than would animal enzyme's activities. Accordingly, higher (20 - 35 degree) temperatures, rather than shorter fixation times are preferable for those "difficult" tissues.
I am sorry that I could not accept the offer of Minnesota russets, if offered. Australian Quarantine would have an instant baby, without a pregnancy what's more. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au ---------- } I am working on a potato storage project and trying to conquer embedding of } amyloplast starch in tuber samples. I've been using Spurr's resin, after 2% } glutaraldehyde fixation in sodium cacodylate buffer and osmium tetroxide post } fixation. I use propylene oxide after ethanol dehydration series to ensure dry } samples, but still most of the starch granules and plastids fall out during } sectioning and post staining. } } Has anybody else worked with starch or potato tuber with success? I'm looking } for any protocol hints that will better preserve starch and amyloplasts. Has } anybody tried microwave fixation procedures on starch samples? } } Thanks for any help you can give. I'll send 5# of the fantastic Minnesota
} russets to anyone who provides information that leads to success (but you'll } have to wait until October to get them!). } } Darryl Krueger } Mn. Ag. Experiment Station } EM Lab } Univeristy of Minnesota } St. Paul, MN 55108 } (612)625-8249 } darrylk-at-puccini.crl.umn.edu }
The wall of the fungus and may prevent good embedding. I have also had Aspergillus (esp. the spores) that would pull out if faced with a razor blade. We routinely sectioned these by doing all the block facing with a diamond knife (use an old one) and starting fully 50 microns above the sample. The sections were collected on formvar.
Need used SEM with W or LaB6 gun, prefer stepper motor stage.
................................................................. Dave Pierce pierce-at-magic.geol.ucsb.edu WB6LRN Geological Sciences (805) 893-2466 (voice/message) University of California (805) 893-2314 (fax) Santa Barbara, CA 93106
I often have trouble observing fine powder on my SEM. Powder that is less than 1 micron in size. I'll first either pack it lightly on carbon tape or sprinkle some over silver paint, then coat. When the electron beam hits it, I can see the powder being blown off and I get charging. What are some good techniques for observing light in weight and small powder on the SEM?
Mark E. Darus (216) 266-2895 General Electric Co. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I often have trouble observing fine powder on my SEM. Powder that is } less than 1 micron in size. I'll first either pack it lightly on carbon tape } or sprinkle some over silver paint, then coat. } When the electron beam hits it, I can see the powder being blown off and } I get charging. ...
The particle undersides are not apparently being coated ... but the problem is each particle sensing its neighbor and being repelled by it. I think if the particles were dispersed with increased distance, distributed individually on the carbon tape, you'd see "less effect" from charging ... Hope this helps ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
} } I often have trouble observing fine powder on my SEM. Powder that is } less than 1 micron in size. I'll first either pack it lightly on carbon tape } or sprinkle some over silver paint, then coat. } When the electron beam hits it, I can see the powder being blown off and } I get charging. What are some good techniques for observing light in weight } and small powder on the SEM? } } Thanks } Mark Darus Dear Mark,
For our automated particle classification analysis by size, morphology and composition we disperse small amount of powder in appropriate liquid (water, oil, etc.) in ultrasonic bath and filter suspended particles onto 0.2 micron filter. Filter is transferred on a holder, coated, etc. Regards,
Igor C. Ivanov, SIMS,Auger,XPS,TEM,SEM Senior Scientist Analytical Services RJ LeeGroup, Inc. Contract Research 530 McCormick Str. (510)-567-0480 phone San Leandro, CA 94577 (510)-567-0488 FAX
I suggest you do the other way around: put a small amount of your powder on filter paper, then apply gently the SEM sample holder with carbon tape. Thus in theory only the particles in contact with carbon tape will adhere. In order to get rid of any other particle you may also want to blow dry nitrogen though I do not believe it is necessary, and if you don't, at least you'll be sure you won't get any contamination from N2. You must get a kind of monolayer of particles.
Then coat.
On Fri, 9 May 1997, Mark E. Darus wrote:
} I often have trouble observing fine powder on my SEM. Powder that is } less than 1 micron in size. I'll first either pack it lightly on carbon tape } or sprinkle some over silver paint, then coat. } When the electron beam hits it, I can see the powder being blown off and } I get charging. What are some good techniques for observing light in weight } and small powder on the SEM?
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Mark:
I'm not certain how applicable this is, but SPI Supplies offers a product called Tacky Dot Slides which, as I understand it, have some very small and precisely spaced "tacky dots" which will grab individual particles. You can probably get the information from their web site which I believe is at http://www.2spi.com. If I'm wrong about the web site, you can access it through my "related Links" section from my web site.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy.
Message text written by "Mark E. Darus (216) 266-2895 General Electric Co." } I often have trouble observing fine powder on my SEM. Powder that is less than 1 micron in size. I'll first either pack it lightly on carbon tape or sprinkle some over silver paint, then coat. When the electron beam hits it, I can see the powder being blown off and I get charging. What are some good techniques for observing light in weight and small powder on the SEM?
Dear Folks, I have relished our interchanges regarding the situation with Pbcitrate and NaOH. Here is a short summary and some ideas. 1. Good citrate staining is the product of many contributions. If there are problems with it however, the simplest and most likely cause is the incorrect production of the stain. Therefore - 2. If you are happy with your stain, if it is reliable, and you never worry about it working, and you do not have to make it up often, leave it alone no matter how you make it. 3. If you are anxious about it, sometimes it stains, sometimes it is very good, other times marginal, and sometimes it may even precipitate, do the following simple things: Use commercially prepared 1 N NaOH instead of pellets, and shake and invert your volumetric for 25 minutes. Then store immediately in plastic syringes. 4. If you frequently come to a dead stop, frequently have sections ruined or unpublishable, it is time to get out the Reynolds paper and follow instructions carefully, but do not rinse your sections after staining in anything but plain dwater. 5. If the above does not improve the situation, there may be a major problem with your embedding process. If so, please call me at 303-871-3026, and I will be glad to see if I can help.
In summary it has always seemed to me that it is imperative that all basic operations of an EM lab such as processing, embedding, cutting, staining, scoping, photomic, must, must, must run flawlessly, simply, and reproducibly without worries, so that we can save our emotional and intellectual energies for our experimental conditions. Therefore I believe in making up quantities (500ml) of Pbcit with 1.0N NaOH (do not pH it) storing it in syringes in the refrigerator according to the method of Reynolds, and then using it for at least six months. I have been asked what the pH should be. I do not know. It is not important (and is probably counterproductive as it leads to using the pH meter which should not be done). I know absolutely that small differences in pH will change the staining properties of Pbcit. (Pellets are hard to weigh, they absorb water, etc, sometimes they work and sometimes they are off - it is just not worth fussing with them.) If someone has stubborn problems, please call me, and I will see if I can be of help. Bye now, Hildy
We do a lot of TEM of keratinocyte cell cultures using this procedure:
EMBEDDING PROCEDURE /cell culture plates
PBS rinsex3 (discard leftover tissue media down sink with bleach)
1/2 KARNOVSKYS 3hrs to overnight
0.1M NaCaco 15minx2
1%OsO4 1hr
dH2O 15minx2
1%UA 11/2 hrs
35%ETOH 15minx2
70%ETOH 15minx2
95%ETOH 15minx2
100%ETOH 15minx2
100% ETOH 30minx2
3:1 ETOH:EPON 6-8hrs
2:1 ETOH:EPON 12-16hrs (overnight w/ caps off)
1:1 ETOH:EPON 6-8hrs (w/ caps off)
100%EPON 6-8hrs
bake for 24-48 hours in 600 C oven
DO NOT USE PROPYLENE OXIDE!!! IT DISSOLVES THE PLASTIC TISSUE CULTURE PLATES
Bob Underwood Morphology Core Univ of Washington
On 8 May 1997, Manoj Misra wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Wonder if someone has a protocol for preparing keratinocyte } (skin cell) culture for TEM. } } Thanks, } } Manoj Misra }
I did a time study 10 years ago while working as an EM tech in a hospital pathology lab with a Siemens TEM. The pathologist attempted the scanning and photography at the microscope. Results: 16% tissue preparation 30% cutting and staining 25% photo processing 12% equipment maintenance 9% lab management 5% continuing education and professional activities 2% training, demonstration, public relations Final result: Closure of the lab and loss of my position, thanks to the bean counters.
Hi! Are there any specific stains that react only with nucleic acids that can be used for negative staining in transmission electron microscopy of plasmid DNA without additional shadowing with tungsten/platinum/palladium etc? Thanks, Yamini Dalal
On Fri, 9 May 1997, Mark E. Darus (216) 266-2895 General Electric Co. wrote:
} -----------------------------------------------------------------------. } I often have trouble observing fine powder on my SEM. Powder that is } less than 1 micron in size. I'll first either pack it lightly on carbon tape } or sprinkle some over silver paint, then coat. } When the electron beam hits it, I can see the powder being blown off and } I get charging. What are some good techniques for observing light in weight } and small powder on the SEM? }
I think it is a matter of conductivity on those unbond or loose particles. Normally I sprinkle the powder over sticky-tab on stub and use a Duster to blow those unbond particle away. After sputter coating you will get a beautiful image of particles without charging or being blown off. This is my two cents tip and wish you luck.
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Another variation: Drop the powder onto the carbon taped stub, then rap the side of the stub to make the powder "slide" around. Finally, dump/jar/blow the loose powder off the tape surface. This tends to also give a "mono-layer" of powder on the stub with fewer charging problems. Just for comparison I also usually press the powder down in one area with a small spatula blade before removing the excess (sort of a hedge against any weird settling phenomena occurring.) The pressed area virtually always charges much worse than the rest of the stub surface.
----------------------------------------------------- Bede Willenbring Research Chemist
H.B. Fuller Company 1200 Willow Lake Blvd Vadnais Heights MN 55110-5132
} } } Yves Maniette {yves-at-giga.sct.ub.es} 5/9/97 13:15 } } } I suggest you do the other way around: put a small amount of your powder on filter paper, then apply gently the SEM sample holder with carbon tape...
} ... I often have trouble observing fine powder on my SEM. Powder that is } less than 1 micron in size. I'll first either pack it lightly on carbon tape } or sprinkle some over silver paint, then coat....
} ... } Microscopists: Perhaps you would like to flame that "free money" moron. I } have send him a choice message. Apparently he is using our server } "ultra.net" and I have asked the service provider to use a little influence } too. } Jim Darley }
The domain given in the body of the "free money" message, neo-quest.com, is registered to the lowlife Sanford Wallace, of the detested cyberpromo.com, who makes a living spamming with false From: addresses. From a WHOIS search:
Neo-Quest NEO-QUEST-DOM 3900 Moorpark Ave.-Suite 27 San Jose, CA 95117 USA
Dear Mark, Whenever I observe powder in the SEM, I first determine the average size. For small powders like yours, the best method is to suspend it in a solvent such as alcohol, disperse by sonicating, then put a drop on a smooth surface, such as a glass coverslip or glassy carbon. Wait until it dries, then coat as usual. The powder is then thinly dispersed. Most of your problem is because the powder is piled up too high. I always say that if you can see it, its too thick. You wrote:
} I often have trouble observing fine powder on my SEM. Powder that is } less than 1 micron in size. I'll first either pack it lightly on carbon tape } or sprinkle some over silver paint, then coat. } When the electron beam hits it, I can see the powder being blown off and } I get charging. What are some good techniques for observing light in weight } and small powder on the SEM? } } Thanks } Mark Darus }
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Dear Mark, when I worked on Auger microscope, I needed frequently to investigate of a non-conducting powder without any coating. Best it resulted if the powder is rolled up in In foil. Please work with In in gloves. Regards, Victor
Victor Sidorenko ANTRON Co. Ltd., scientific service Moscow, Russia antron-at-space.ru
Thank you to all of you who have answer to my question. It seems that no such a list exist on the web, anyway I got some useful information that I post below:
Response 1
If you are referring to metals, then there are several good books published by ASM (American Society for (Metals/Materials). There is the ASM Handbook, several guides to metallography as well as George Vander Voorts (?) guide to metallographic preparation and etchants.
Good Luck.
Alan Stone {as-at-mcs.com} ASTON
Response 2
There are a number of good reference books on sampe preparation for LM. I have listed a few of them below.
1) "Metallography & Microstructures" American Society for Materials, Volume 9
2) "Metallographic Polishing by Mechanical Methods" L.E. Samuals, American Society for Materials
3) "Metallographic Specimen Preparation" James L. McCall & William M. Mueller, Plenum Press ISBN 0-306-30791-x
4) "Metallography of Advanced Meterials" Henry J. Cialone et al, American Society for Materials.
You may also want to check with manufacturers of such equipment. We maintain a database of sample preparation methods and can often give references fr specific preparation needs. We also publish a bibliography of technical papers dealing with sample preparation which we are pleased to send you at no charge. You can then select papers of interest and we will send you reprints at no charge.
I hope this helps!
Best regards-
David Henriks (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.
Response 3
Look on the List web server . This server is located at: http://www.liszt.com/.
Henrik
-- Henrik Kaker SEM-EDS Laboratory, Metal Ravne d.o.o., Slovenia, mailto:Henrik.Kaker-at-guest.arnes.si
We've been doing this on Tingids for Dr Gerry Cassis here at the Australian Museum. He's in a team which includes experts on Crustacea, Arachnids, Trilobites, etc. who are looking at the evolutionary origins of the arthropod groups.
The final protocol worked out by Sue Lindsay was;
Digest the muscle tissues in either KOH or NaOH (3 pellets in 3 or 4 mls water in watchglass at 70 degrees C). Leaving it in this solution after the tissue has digested will also start to clear the exoskeleton.
Partially clear the exoskeleton for examination under transmitted light microscopy by either leaving it in the caustic solution for much longer or by BREIFLY boiling the solution. I'm told that Lactic Acid is also useful for clearing exoskeletons. Light microscopy was useful for detecting overlapping of sternites, relative thickness of sternites, difference between sternite and arthrodial membrane, etc.
Clean in ultrasonic bath for extended time (say 5 minutes) to get all the digested ooze out - obviously with some escape path such as the holes where the head &/or abdomen were.
Dry the specimen. We got our best results with Critical Point Drying, though this was because the tingids were a bit flimsy. Your scarabs might me more robust, in which case HMDS (HexaMethyl DiSilazine) might work well, or even just air drying from a volatile solvent (Acetone or Ethanol?).
Dissect the specimen. We use eye surgery scissors, very small blade length which worked on even the smallish specimens. Cut along the dorsal and ventral midlines for two mirror-image samples; in our case, one for SEM and the other for light microscopy.
Comments: We found the order to be important, since cutting the specimen first and then digesting the tissue lead to the skeleton just rolling up on drying. But this probably depends entirely on your particular specimen. If given the choice I'd rather cut first and digest second, making digestion quicker and allowing you to help it along by picking away at the muscle with forceps. The ultrasound cleaning would also be easier. As always in EM, you've just got to find what works with your particular beasts.
Geoff Avern Manager Microscopy Labs Australian Museum Sydney, Australia
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Hi,
I am Resia Pretorius from South Africa and am currently working on a PhD )the phylogeny of internal structures of the superfamily Scarabaeoidea ((insects))
I would like to use a SEM to photograph one of the internal chitinous structures (metendosternites) to which flight and leg muscles are attached, and do calculations of different lengths for morphometric analysis.
These muscles are attached firmly to the metendosternites and I would like to remove them, but so far without luck. I have tried Sodium perborate (different concentrations) they use this chemical to clean muscles from snake skulls, but if only loosens the muscles slightly, I still need to pull and tuck at the muscles to remove them.
I have also tried tripsin a 0.5% without it working. Are there any suggestions from anyone, I read in an 1890 article that one can use nitric acid for the macercation of muscles but that was obviously before the SEM was invented. Can one use an acid or would it harm the actual structure of the metendosternites.
I realy need help!! my e-mail address is:
resia-at-mcd4330.medunsa.ac.za
Thanks, Resia Pretorius Lecturer (Biology) Medunsa South Africa
Message-Id: {1.5.4.32.19970512034746.00693dbc-at-pop3.unsw.edu.au} X-Sender: s7001031-at-pop3.unsw.edu.au X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Tempfix is good for powders. Its an adhesive which melts around 80 deg. We melt some onto a stub, then warm it until the adhesive is just liquid, then press it into the powder. Let it cool, then blow with an air duster. If it blows off it wasn't stuck down. Most EM supplier stock it.
Hello dear subscribers I am a new comer to this listserver. I am interested to take training in Electron Microscopy. I have already taken preliminary training in EM in KKUH, and KSU. I am looking addresses of Institutions/Univresities/Organizations where I can undertake further training in Electron Microscopy . Pls send any information at my following email address: F40C028-at-KSU.EDU.SA I appreciate and be solicited for providing informations. Thanking you in anticipation. Abdulrahman N.S.Al-Ohali
I try to immunostain A431 cells with the preembedding method. Staining works well with 1nm gold and IGSS staining (IntenSE) on the LM level. On the EM level I can not detect any staining. Samples are fixed in glutaraldehyd and osmium after IGSS staining and conventionally dehydrated and embedded in Epon. It seems that during the fixation and embedding procedure the staining is washed out. Is there anyone who has made the same experience?
Dear Microscopists, I am working on the dermis of echinoderms on which I plan to perform a high-pressure freezing followed by a freeze-substitution in two steps: firstly in a solution containing glutaraldehyde and tannic acid and secondly in osmium tetroxide. I am wondering if I would better use tannic acid (i.e. a mixture of different gallotannins) or 3,4,5-Trihydroxy-benzoic-acid (C7H6O5), the active component of the so called tannic acid. Has anybody experience in the subject? I thank you for your help,
Uranyl acetate will stain DNA, but that is a positive stain and is not highly specific for nucleic acids. UA and several other heavy metal salts can be used as negative stains, but in that case you do not want the DNA to stain specifically; this is usually done when you want to visualize proteins associated with DNA (eg Griffith, J. Science 187: 1202 and 201: 525). DNA replicated in the presence of BrdU or biotin-labeled nucleotides can be specifically stained with anti-BrdU and gold-labeled secondary antibodies or avidin.
Also, DNA can be spread on a surface, picked up on a film, positively stained with UA, and embedded in a thin layer of the detergent photoflo.
-Dennis Goode
} Reply-to: lsdalal-at-scifac.indstate.edu Organization: ISU To: Microscopy-at-Sparc5.Microscopy.Com Subject: EM and DNA
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Hi! Are there any specific stains that react only with nucleic acids that can be used for negative staining in transmission electron microscopy of plasmid DNA without additional shadowing with tungsten/platinum/palladium etc? Thanks, Yamini Dalal Dr. M. Dennis Goode Phone (301) 405-6917 Department of Zoology Fax (301) 314-9358 University of Maryland e-mail goode-at-zool.umd.edu College Park MD 20742 ************************************************************* "If the Lord Almighty had consulted me before embarking upon the creation, I should have recommended something simpler." -Alphonso X of Castile, 15th Century
Thank you to all who responded to my query regarding slides imprinted with grid squares. Several people requested information be forwarded on, so the following is a quick summary of responses.
Field Finder Slides - a standard slide imprinted with grid squares intended to be used along with the (separate) specimen slide McCrone 'England Field Finder' 800-622-8122 Fisher 'Field Finder' 800-766-7000
Several coverslips were recommended, made by: Eppendorf 'cellocate' Bellco 800-257-7043 (USA) 800-445-7051 (Canada)
Thank you to IMR and McCann Imaging for their thoughtful and helpful advice as well as Charles Garber (SPI) and Stacie Kirsch (EMS) who pointed me to websites at: cccbi.chester.pa.us/spi/new/ptfesld.html (SPI) emsdiasum.com (EMS)
Thanks to all of you we now have the slides we needed. My regards to all of you who responded.
************************* * Marilyn Wadsworth * * mwadswor-at-zoo.uvm.edu * *************************
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PHILADELPHIA SOCIETY FOR MICROSCOPY MAY 1997 MEETING NOTICE Wednesday, May 28, 1997
MICROSCOPY IN THE MUSEUM at the Philadelphia Museum of Art
The preservation of the collections for future generations is the first obligation of the Philadelphia Museum of Art as a public institution. The Museum's Conservation Department is entrusted with this responsibility. The Museum's staff of 17 conservators employ a wide range of analytical techniques for assessing the condition of objects in the collection as well as monitoring the success of treatments that are carried out. Many of these techniques are microscopy-based.
At this meeting, members of the conservation staff will give an overview of microscopy and other analytical techniques employed at the Philadelphia Museum of Art, which include PLM, visible light and UV microscopy, Micro-FTIR and SEM-EDS. Recent work ranging from the analysis of Czanne's paintings to William Penn atop City Hall will be discussed.
Following the presentations, there will be a "Behind the Scenes" tour of the Conservation and Analytical Labs. The Museum's galleries will be open for viewing as well, until 8:45 PM.
The Speakers: Marigene H. Butler Head of Conservation Andrew Lins Senior Conservator of Decorative Arts and Sculpture Beth Price Conservation Chemist Peter Eastman Conservation Information Specialist
*****PLEASE NOTE CHANGES FOR THIS SPECIAL EVENT*****
DATE: Wednesday, May 28, 1997
PLACE: Philadelphia Museum of Art Benjamin Franklin Parkway at Eakins Oval Philadelphia, PA
PARKING: Parking around the Museum will be difficult because of special Wednesday night events. Free parking may be available around the building before 5:30 PM. Additional parking can also be found along Pennsylvania Avenue (street level to the north), inside Eakins Oval (below the steps on the east side) or behind the Museum in the Azalea Garden (west side). There is parking and access for persons with disabilities along the south side of the Museum.
TIME: 5:15 - 6:30 PM Check-in. The check-in table will be located inside the West door (facing the Schuylkill River) during this time. You must check in to receive your Museum admission badge and pay for your meal. A Museum map of the galleries and the outdoor dinner tent will be provided.
5:30 - 7:00 PM Dinner will be served in the outdoor tent on the East Terrace. Members $12 Student members $6 Non-members $15
Menu: Penne pasta with roasted eggplant, red and yellow peppers and ricotta cheese Chicken provencal with olives, red and yellow peppers, zucchini and tomatoes Green beans, Roasted potatoes Sliced seasonal fruit Chocolate truffle cake Coffee, decaf or tea
6:55 PM Gather at check-in table before going to Conservation Lab
7:00 - 8:00 PM Presentations, in the Conservation Lab
8:00 - 8:30 PM "Behind the Scenes" tour through conservation laboratories
NEWS FLASH: Reservations can now be sent by e-mail by writing to PSM-RESERVATIONS-at-INAME.COM. Be sure to include your name and affiliation in the body of the message. PSM prefers that you use this convenient method to RSVP. If you don't have access to e-mail, phone reservations will be taken by Ms. Pat Overend at the University of Pennsylvania, 215/898-8337.
Deadline for reservations will be WEDNESDAY, May 21. At the Museum's request, we must insist that you adhere to this deadline. If you have any questions regarding the meeting please feel free to contact Rollin Lakis of the Executive Council T 215/898-8718. Cancellations must be received no later than 5:00 PM, May 23, 1997.
Bonus: The first 25 people to make reservations will receive complimentary tickets to the Rodin and Michelangelo exhibit, which will be open until 8:45 PM.
You are not alone. In fact this was talked about some time ago (last year?); check the archives for that discussion. I have had no luck with conventional embedding and IGSS. It seems that the osmium changes the Ag-Au which then washes out during UAc treatment. I've tried cold short osmium and alcoholic UAc, but it still happens. So I gold tone all my tissue before embedding. Adds contrast and the tissue looks "grainy" at high mag, but otherwise it's fine and the Ag-Au remains in place. The alternative (for me) is to use osmium only, no Uac, and increase the staining time with UAc on the grids. I find for my tissue that the contrast is still not good enough, but it may work on yours. If you want the gold toning method, Email me and I'll send it. Good luck.
Diana van Driel Dept Ophthalmology Sydney University C09 AUSTRALIA 2006
Dear All, I am currently using antibodies to localise fungi in semi-thin resin sections of Tomato. When using a gold conjugate system I have no problems, but I am also using WGA and another antibody both linked to Rhodamine, and there is a huge amount of autofluorescence from the plant and the fungus when observed under the green light. There is a similar problem with the filter blocks we have for FITC. I tried treating the sections with KOH solution and this worked really well for the WGA labelled sections reducing the problem, but obviously that is not an option with the antibody. Does anyone have any suggestions? Thanks,
Mark Munro. The Soil Biology Unit SAC e-mail m.munro-at-ab.sac.ac.uk.
Dear subscribers! I have several questions for the list. 1. Why everybody uses for section constrasting uranyl acetate which is less soluble than uranyl nitrate or uranyl sulfate? 2. Is fluorescence of green fluorescent protein (GFP) affected by low concentration of glutaraldehyde? 3. Is fluorescence of GFP affected after embedding into epoxy or acrylate resins? 4. Have epoxy and acrylate resins autofluoresence?
Sincerely yours, Alexander Mironov
Unit of Morphology Consorzio Mario Negri Sud S.Maria Imbaro (Chieti), 66030, Italy Fax: +39 872 578 240
Dear All, I am currently using antibodies to localise fungi in semi-thin resin sections of Tomato. When using a gold conjugate system I have no problems, but I am also using WGA and another antibody both linked to Rhodamine, and there is a huge amount of autofluorescence from the plant and the fungus when observed under the green light. There is a similar problem with the filter blocks we have for FITC. I tried treating the sections with KOH solution and this worked really well for the WGA labelled sections reducing the problem, but obviously that is not an option with the antibody. Does anyone have any suggestions? Thanks,
Mark Munro. The Soil Biology Unit SAC e-mail m.munro-at-ab.sac.ac.uk.
Dear All, I am currently using antibodies to localise fungi in semi-thin resin sections of Tomato. When using a gold conjugate system I have no problems, but I am also using WGA and another antibody both linked to Rhodamine, and there is a huge amount of autofluorescence from the plant and the fungus when observed under the green light. There is a similar problem with the filter blocks we have for FITC. I tried treating the sections with KOH solution and this worked really well for the WGA labelled sections reducing the problem, but obviously that is not an option with the antibody. Does anyone have any suggestions? Thanks,
Mark Munro. The Soil Biology Unit SAC e-mail m.munro-at-ab.sac.ac.uk.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America The important thing with Tannic acid is the original source. For E.M. the best is Turkish, available from Mallinckrodt and not the Chinese from Sigma. Ian.
Dear friends: Department of AME of Un. of AZ has received SEM S-650(Hitachi) as a gift. Howereve, there is't manual or documentation and we are not able run it. We would greatly appreciate your help in supplying us such manual or any information. Thanks. Yury Shipilov Research Assosiste ph.(520) 626-4470
Dear subscribers! I have several questions for the list. 1. Why everybody uses for section constrasting uranyl acetate which is less soluble than uranyl nitrate or uranyl sulfate? 2. Is fluorescence of green fluorescent protein (GFP) affected by low concentration of glutaraldehyde? 3. Is fluorescence of GFP affected after embedding into epoxy or acrylate resins? 4. Have epoxy and acrylate resins autofluoresence?
Sincerely yours, Alexander Mironov
Unit of Morphology Consorzio Mario Negri Sud S.Maria Imbaro (Chieti), 66030, Italy Fax: +39 872 578 240
I need to buy 2 new elemental standards blocks for use in our SEMs. Required dimensions are 1" diameter by { 1" thick. These must NOT be carbon coated. Any recomendations or products not to consider?
AccuMed International, Inc., has an immediate opening for a TECHNICAL = MANAGER to oversee the development, scientific and clinical testing, and = production of computerized, microscope-based IMAGING workstations for = clinical and research applications. The successful candidate will = shepherd new instruments from R&D to finished product. The position = requires a minimum of a Bachelor's degree and a strong = scientific/technical background, with at least 3-5 years experience in = developing and utilizing imaging workstations and applications for light = microscopy. Specific areas of responsibility will include:
- managing the development of new hardware and software products - establishing and maintaining timelines for delivery of prototype = instruments and finished products, including software-based applications - coordinating activities in various departments including R&D, = engineering, software development and manufacturing - assisting with applications to regulatory agencies
AccuMed International, headquartered in Chicago, is a global laboratory = diagnostic products company and an emerging leader in the field of = automated cytopathology. EOE.
Please respond via mail, FAX or email (NOT telephone) to:
Marc M. Friedman, Ph.D. Vice President, Scientifc and Technical Affairs AccuMed International, Inc. 900 N. Franklin, Suite 401 Chicago, IL 60610 Tel: (312) 642-9200 FAX: (312) 642-8684 email: marc.friedman-at-accumed.com
I am using immunostaining protocols to localize several different intracellular, some intranuclear proteins. I was hoping to benefit from both the ease and high sensitivity of the avidin-biotin system and the advantages of recording fluorescent images with a confocal microscope.
The tissue is frozen sections of paraformaldehyde fixed mouse retina. Following a blocking step, I used 2 different antibodies, one a mouse monoclonal, the other, a rabbit polyclonal. After overnight incubation with the primary antibodies, both at R.T. and 4 deg., I applied biotinylated mouse/rabbit IgG, followed by fluorescein conjugated streptavidin. To my greatest diasppointment, all I saw was a high uniformly fluorescent background throughout the entire retina for both antibodies.
Am I leaving out a critical step?
Any suggestions would be highly appreciated.
Judy Trogadis Eye Research Institute of Canada and University of Toronto ph: 416-603-5088 fax: 416-603-5126 email: judy-at-playfair.utoronto.ca
Do any of the universities, companies, hospitals or research institutions have any policies regarding the use of electron microscopy by pregant employees? Is there any literature available on the topic?
Is the advice not to use EM at all during the 9 months, or maybe only during critical periods of development?
In searching the archives I only found one reference on this topic.
Any help would appreciated.
Patrick Diehl Materials Science Department University of North Texas
Dear Patrick, I have sheparded three pregnancies (users, not me) on my 200 kV TEM and I found it a good way to get the instument monitored and thoroughly tested. The last one wore a accumulation monitor at all times and found a curious phenomenon: she accumulated more radiation when sitting at her desk, next to the cinder-block wall in the next room, than when she was working at the TEM at 200 kV. These instruments, when assembled and used properly and monitored in case of leaks after maintenance, leak less x-rays than a TV. I believe the regulations regarding radiation exposure are considered valid for pregnant women.
You wrote:
} Do any of the universities, companies, hospitals or research institutions } have any policies regarding the use of electron microscopy by pregant } employees? Is there any literature available on the topic? } } Is the advice not to use EM at all during the 9 months, or maybe only } during critical periods of development? } } In searching the archives I only found one reference on this topic. } } Any help would appreciated. } } Patrick Diehl } Materials Science Department } University of North Texas } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Dear Patrick, I have sheparded three pregnancies (users, not me) on my 200 kV TEM and I found it a good way to get the instument monitored and thoroughly tested. The last one wore a accumulation monitor at all times and found a curious phenomenon: she accumulated more radiation when sitting at her desk, next to the cinder-block wall in the next room, than when she was working at the TEM at 200 kV. These instruments, when assembled and used properly and monitored in case of leaks after maintenance, leak less x-rays than a TV. I believe the regulations regarding radiation exposure are considered valid for pregnant women.
You wrote:
} Do any of the universities, companies, hospitals or research institutions } have any policies regarding the use of electron microscopy by pregant } employees? Is there any literature available on the topic? } } Is the advice not to use EM at all during the 9 months, or maybe only } during critical periods of development? } } In searching the archives I only found one reference on this topic. } } Any help would appreciated. } } Patrick Diehl } Materials Science Department } University of North Texas } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I am resending this message to the group becuase of an "undleiverable" message from the orginator's mail system.
HJC
} ---------- } From: Crossman, Harold } Sent: Tuesday, May 13, 1997 3:26PM } To: 'timet-htl' } Subject: RE: Elemental Standards } } We use standards from: } Geller Microanalytical Labs } 426e Boston St. (Rt. 1), Topsfield, MA 01983-1212 } 508 887-7000, fax 508 887-6671, sales-at-gellermicro.com } Website: http:/www.gellermicro.com } } Geller has a varity of elements and will custom fabricate standards if they } can. } You might also want to consider their magnification standard as well. We use } it for both optical and electron scopes. You may be surprised at the } differences between what the scopes reads/images, and what is really there! } } I have no financial interest in promoting Geller. My company has been a } satisfied customer for years. } } } ------------------------------------------------ } Opinions or statements expressed herein, rational or otherwise, do not } necessarily reflect those of my employer. } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com} }
Dear Patrick, I have got two healthy daughters (the youngest one is 10 weeks). During the whole period of both my pregnancies I have worked with electron microscopy (TEM, SEM, low vacuum SEM and environmental SEM). Risoe National Laboratory is a former Atomic Energy Commission Research Establishment, so we still have health physicists around. They were consulted as soon as I realised that I was pregnant. The health physicist measured the radiation level at all our TEMs and SEMs. Only the background level of approx. 0.06 microSv/h was found. During the my pregnancies I wore two badges that checked the radiation continuosly. Nothing above background level was detected.
Charlotte C. Appel Materials Research Department Risoe National Laboratory DK-4000 Roskilde DENMARK radiation
It sounds like the problem may be using a monoclonal mouse antibody on mouse tissue. When you come in with your secondary against mouse IgG it will find a whole bunch of it in the tissue. If you don't have any choice but to use a monoclonal on the mouse tissue you can come in with a 10x excess of anti-mouse fab fragments to block all the mouse IgG, then cone in with your Primary.
Robert Underwood Morphology Core U of Washington
On Tue, 13 May 1997, Judy Trogadis wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Fellow immunostainers, } } I am using immunostaining protocols to localize several different } intracellular, some intranuclear proteins. I was hoping to benefit from } both the ease and high sensitivity of the avidin-biotin system and the } advantages of recording fluorescent images with a confocal microscope. } } The tissue is frozen sections of paraformaldehyde fixed mouse retina. } Following a blocking step, I used 2 different antibodies, one a mouse } monoclonal, the other, a rabbit polyclonal. After overnight incubation } with the primary antibodies, both at R.T. and 4 deg., I applied biotinylated } mouse/rabbit IgG, followed by fluorescein conjugated streptavidin. To my } greatest diasppointment, all I saw was a high uniformly fluorescent } background throughout the entire retina for both antibodies. } } Am I leaving out a critical step? } } Any suggestions would be highly appreciated. } } Judy Trogadis } Eye Research Institute of Canada and } University of Toronto } ph: 416-603-5088 } fax: 416-603-5126 } email: judy-at-playfair.utoronto.ca }
Radiation exposure should be limited to a fetus, particularly during the first 3 months. The Nuclear Reg. Comm. limits this exposure to {500 mRem for the 9 months. This SHOULD be a huge value compared to any exposure from any EM unless it is a HVTEM that is improperly setup (you must stay out of the chamber when running {g} ). A SEM running 30 kV. cannot produce any x-rays over 30 keV - Most are well under. These relatively soft x-rays are (typically) completely shielded by the column and chamber walls....Consult your manufacutrer for detailed exposure rates. I think you will find that a cross country air flight (or 9 months in Denver vs. a sea level city) will expose you to more radiation than will 9 months in front of the EM.
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Do any of the universities, companies, hospitals or research institutions have any policies regarding the use of electron microscopy by pregant employees? Is there any literature available on the topic?
Is the advice not to use EM at all during the 9 months, or maybe only during critical periods of development?
In searching the archives I only found one reference on this topic.
Any help would appreciated.
Patrick Diehl Materials Science Department University of North Texas
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Dear Rui:
I assume you are looking for Dr. Charles Garber of Structure Probe/SPI Supplies. I have copies below the signature block from one of his messages.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI Structure Probe, Inc. FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy. Message text written by Rui Costa }
I am looking for Dr Charles Graber (or Graber). Sorry I do not if the last name is correct.
After a few hours using the TEM (Philips CM30) with the cold trap filled with LN2, it appears that some vapors in the microscope column get trapped, as it should be.
Now the problem is: have you ever noticed any problem AFTER removing the LN2 dewar, thus heating the trap, provoking a degassing in the column. We have noticed that after several hours working with the cold trap, the IGP gas pressure level increases a lot after removing the dewar (it happens that HT switches off during the night because of that IGP pressure increasing above the "safety level"), and some users here tell me that it should not be that way.
Question: did you ever notice such a problem, and if yes, how long time would you work without heating the trap in order not to get any dramatic increase of IGP pressure after removal of the dewar.
} After a few hours using the TEM (Philips CM30) with the cold trap filled } with LN2, it appears that some vapors in the microscope column get } trapped, as it should be. } } Now the problem is: have you ever noticed any problem AFTER removing the } LN2 dewar, thus heating the trap, provoking a degassing in the column. We } have noticed that after several hours working with the cold trap, the IGP } gas pressure level increases a lot after removing the dewar (it happens } that HT switches off during the night because of that IGP pressure } increasing above the "safety level"), and some users here tell me } that it should not be that way. } } Question: did you ever notice such a problem, and if yes, how long time } would you work without heating the trap in order not to get any dramatic } increase of IGP pressure after removal of the dewar.
Yes, we notice this on both our Philips CM-12 and CM-20. Since our lab does a lot of cryoTEM, it would be especially bad if we did not deal with the problem. At the end of each day we turn off the high tension and then use the Philips CRYO function to turn off the ion getter pump and open V6 so that the diffusion pump pumps the column. We then replace the cold-trap Dewar vessel with a vessel filled with warm water and permit the diffusion pump to pump away all the contaminants (it takes about 30 min.) We then switch off the CRYO function, which turns the ion getter pump back on. After about another 15-30 min when the vacuum has recovered, we switch on the HT (on conditioning) and leave the microscope for the night. We get very little downtime by following this procedure. I should point out that the power in Rochester is very reliable, so we have only had one unplanned power outage in the last five years. Under these conditions, the microscope seems to perform best when the high tension is left on continuously. -- Best Regards, John Minter
Eastman Kodak Company Phone: (716) 722-3407 Analytical Technology Division FAX: (716) 477-3029 Room 2112 Bldg 49 Kodak Park Site email: minter-at-kodak.com Rochester, NY 14562-3712 calendar: via PROFS
} Do any of the universities, companies, hospitals or research institutions } have any policies regarding the use of electron microscopy by pregant } employees? Is there any literature available on the topic? } } Is the advice not to use EM at all during the 9 months, or maybe only } during critical periods of development? } } In searching the archives I only found one reference on this topic. } } Any help would appreciated.
Dear Patrick,
Everyone who uses radiation-producing equipment should be monitored. We have ion-chamber dosimeters for occasional users and film badges for people who are around the instrument every day. The limit for exposure for pregnant women is 500 millirem over the course of the pregnancy, but this amount should not happen in a short period. There is an unofficial limit of 50 millirem per month. These numbers come from the experts in the NY State Department of Health. For exposures to x-rays, rad and rem are equi- valent, and 1 rad (Radiation Absorbed Dose) is 100 ergs per gram. If you have no capability for monitoring users, the next best thing is to use a detector, such as a Geiger counter or an ion chamber, to deter- mine whether the radiation near the microscope is different from background. If there is no difference between the radiation measured before the scope is turned on and when it is cranked up fully, then you can be reasonably sure there is no danger (assuming you have a detector which is sensitive to the radiation your scope produces); however, this probably won't satisfy any legal requirements. Yours, Bill Tivol
There is no danger from contemporary microscopes which are well maintained and monitored. The problem exists with chemicals. Cacodylate buffer if used in a laboratory eventually contaminates bottles, hoods, etc., with fine arsenic dust. Then there is the lead stain and the UA. Anyone pregnant should not be weighing out any chemical containing heavy metals. Extreme care needs to be taken with other noxious things such as glut, paraform, xylene, propylene, etc. During my time in another lab, there occurred a chemical spill - the holding tank backed up into the lab. A mixture of the above seeped up through the floor drains. My coworker attempted to contain the spill with a properly fitted mask. At the time she was 12 days pregnant. At seven months she gave birth to a badly deformed baby which lived only for an hour. After exhaustive investigation, it was decided that the uterine fluid had been contaminated with heavy metals. The enormous quantity of fluid gave her the appearance at seven months of a 10 months preganancy, at least. Anyone who is not protected by birth control and might be pregnant at any time should not be weighing out questionable material, and should take extreme care with being gloved, etc., at all times. So long, Hildy
In message {Pine.LNX.3.93.970514184137.22551B-100000-at-giga.sct.ub.es} Yves Maniette writes: } } After a few hours using the TEM (Philips CM30) with the cold trap filled } with LN2, it appears that some vapors in the microscope column get } trapped, as it should be. } } Now the problem is: have you ever noticed any problem AFTER removing the } LN2 dewar, thus heating the trap, provoking a degassing in the column. We } have noticed that after several hours working with the cold trap, the IGP } gas pressure level increases a lot after removing the dewar (it happens } that HT switches off during the night because of that IGP pressure } increasing above the "safety level"), and some users here tell me } that it should not be that way. } } Question: did you ever notice such a problem, and if yes, how long time } would you work without heating the trap in order not to get any dramatic } increase of IGP pressure after removal of the dewar. } } Yves MANIETTE } Universitat de Barcelona
Yves, Yes I see this problem once and awhile with our CM-12. Seems to occur if we've had samples in the column that are volatile and thus the cold trap collects more junk, which can be released suddenly into the column when the cold trap warms up.
Two thoughts: 1. Instead of removing the dewar, leave it on and allow the liquid nitrogen to evaporate with the dewar installed. I think the warm-up rate would be reduced, compared to removing the dewar and exposing the copper braid to room air, such that the IGP and other pumps may be able to handle the gas load without crashing the vacuum system.
2. But if you still get crashes, turn off the IGP until the diffusion pump has time to recover the system. I had my service engineer install a simple toggle switch on the back side of the column cover to allow me to shut off the IGP, as my instrument did not come configured with any software switch to do this.
Typically, in my lab, a crash would occur late afternoon after my last TEM user has left and the cold trap warms up. Then I shut off the IGP for overnight and turn it on the next morning.
Hope this helps.
Gib
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Mn Microscopy Society Home page: http://charfacnu.cie.umn.edu/MnMicSoc.html
Microscopy & Imaging Consortium Home page: http://biosci.umn.edu/MIC/consortium.html
DEPARTMENT OF PHYSICS, UNIVERSITY OF ILLINOIS AT CHICAGO
Two postdoctoral positions are 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 detecto= r (Z-contrast), Gatan Imaging Filter, and Noran EDS; a VG HB501A =46ield-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 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, Fischione 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. The two research positions are as follows:
(I) Atomic Resolution Imaging and Analysis Techniques
With the demise of VG as a supplier of state-of-the-art analytical electron microscopes, it is essential that atomic resolution imaging (Z-contrast) and analysis techniques (EELS and EDS) are developed on other instruments. The JEOL 2010F has the potential to exceed both the imaging and analytical performance of the VG instruments, with a resolution at 200kV of 1.4=C5. Through an NSF award, UIC has recently purchased this microscope with the primary aim of developing the next generation of high resolution imaging/analytical microscope. Coupled with the purchase of this microscope is a postdoctoral position to perform the initial instrument characterization and coordinate its application to research programs at UIC and at neighboring institutions (Argonne National Laboratory, Northwestern University and Illinois Institute of Technology). This position therefore offers the potential to develop a wide range of research programs, in addition to a unique opportunity to address fundamental issues concerning incoherent vs coherent imaging and microanalysis. The postdoctoral position will also have primary responsibility for the VG HB501A STEM and JEOL 3010 TEM. At the end of the 2-year funding by NSF for this position, UIC will be seeking a permanent electron microscope facility manager.
(II) Grain Boundaries in Ceramics
As part of a DOE funded program, a postdoctoral position is available for the investigation of the structure-property relationships at grain boundaries in ceramics. The aim of this program is to correlate the experimental results from atomic resolution imaging and microanalysis to produce structural models which can be compared with theoretical simulations. Of particular interest is the use of EELS to characterize the 3-dimensional structure of the grain boundary and quantify the number of vacancies and dopant atoms present in the structure. This information can only be obtained through the use of EELS. This program necessarily includes the use of multiple scattering analysis of the EELS fine-structure, the use of bond-valence sum analysis to produce preliminary models and the application of the CASTEP simulation codes for detailed analysis of the boundary structure. Prior experience in these techniques is not essential, only a willingness to learn how to use the commercially available software packages.
Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound 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 and publication list to Professor N. 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.
*************************************************************** Nigel D. Browning PhD, Assistant Professor, Interface Physics Group, Department of Physics (M/C 273), University of Illinois at Chicago, 845 West Taylor Street, Chicago. Il 60607-7059
} The last one wore a accumulation monitor at all times and found a curious } phenomenon: she accumulated more radiation when sitting at her desk, next to } the cinder-block wall in the next room, than when she was working at the TEM } at 200 kV.
Dear Mary, This is not so curious. The potassium-40, naturally occurring in concrete is a well-known source of radiation. If the user had her desk in Denver, there would have been an additional component from the greater cosmic ray flux at high altitude. Yours, Bill Tivol
You may consider to put in the specimen and let it pump for about 15 min. before filling the cold trap. } All, } } After a few hours using the TEM (Philips CM30) with the cold trap filled } with LN2, it appears that some vapors in the microscope column get } trapped, as it should be. } } Now the problem is: have you ever noticed any problem AFTER removing the } LN2 dewar, thus heating the trap, provoking a degassing in the column. We } have noticed that after several hours working with the cold trap, the IGP } gas pressure level increases a lot after removing the dewar (it happens } that HT switches off during the night because of that IGP pressure } increasing above the "safety level"), and some users here tell me } that it should not be that way. } } Question: did you ever notice such a problem, and if yes, how long time } would you work without heating the trap in order not to get any dramatic } increase of IGP pressure after removal of the dewar. } } Yves MANIETTE } Universitat de Barcelona
Ke Han Center for Materials Science Los Alamos National Laborotory Los Alamos, New Mexico NM87545, USA Tel: 505-6650771 Fax: 505-6652992
We see exactly the same phenomenon on two separate TEM's (Hitachi H500 and H7100). After an initial improvement in vacuum due to application on the LN2 on the specimen cold trap (but oddly, not the DP traps), we get good performance for 2-3 hr. Then, after the LN2 level goes below the transfer probe or braid, the vacuum deteriorates to the point that the HT kicks off. Very predictable: so we simply keep adding LN2 after 2 hr until the HT is turned off for the evening. Now, if one does not add the LN2 to the specimen trap to begin with, the phenomenon never happens. So, I conclude that the loss of HT must be due to the sudden release of a lot of contamination (water vapor, organics from the specimen). We do not see this with the DP traps because the vapors must be removed by the DPs. Wil Bigelow probably knows what's going on here, so I will wait for his comments.
} After a few hours using the TEM (Philips CM30) with the cold trap filled } with LN2, it appears that some vapors in the microscope column get } trapped, as it should be. } } Now the problem is: have you ever noticed any problem AFTER removing the } LN2 dewar, thus heating the trap, provoking a degassing in the column. We } have noticed that after several hours working with the cold trap, the IGP } gas pressure level increases a lot after removing the dewar (it happens } that HT switches off during the night because of that IGP pressure } increasing above the "safety level"), and some users here tell me } that it should not be that way. } } Question: did you ever notice such a problem, and if yes, how long time } would you work without heating the trap in order not to get any dramatic } increase of IGP pressure after removal of the dewar. } } Yves MANIETTE } Universitat de Barcelona
John Bozzola 71 Concordia Drive Carbondale, IL 62901 Internet address: JBozzola-at-aol.com
------------------------------------- I need information on linking the video signal of a SEM (1V p-p) composite video single to a LECO imagin system that can accept a variety of formats (PAL, NTSC) etc. Any ideas or experience with this? Thanks Craig
Hi all I have a question for all the "digital microscopysts: I need to mount a ccd system to my OLYMPUS BX 50 microscope. I want to do optical sectioning and fluorescence imaging. Some people tell my that the more convenient is a digital ccd camera B&W, whit a resolution of 758 x 512 or 1024 x 1024. What is your opinion of these points?. thanks in advance =================================================== Fernando D. Balducci Laboratorio de Microscopia Electronica Facultad de Ingenieria - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077 ===================================================
In reply to Yves Maniette query on cold trap degassing:
Philips told me to keep the HT of my CM20 on all the time. Unfortunately, when the cold trap warms up at the end of the day, off goes the HT! Of course, the more samples going in and out the microscope during the day, the further the IGP(ion getter pump) reading goes up as one would expect. I normally take off the cold trap dewar when I have finished for the day since I wish to reduce the nitrogen bill - I put the nitrogen in the ion mill dewar.
I also would be interested to know the best way to tackle the problem. So far all I have got is shrugs!
I also think that there should be a toggle switch to take the ion getter off, removing and replacing the back panel is an unnecessary pain. Mike
Michael J Witcomb PhD Electron Microscope Unit University of the Witwatersrand Private Bag 3 WITS 2050 South Africa
} In reply to Yves Maniette query on cold trap degassing: .../... } } I also think that there should be a toggle switch to take the ion } getter off, removing and replacing the back panel is an unnecessary } pain. } Mike
You can ask your local maintenance engineer to place a switch on the microscope SIDE panel near the IGP pump. This switch shoult be connected to the small socket down the IGP pump. Any Philips Engineer also knows that trick and will install it for you if you ask him. The only point is NOT to place the switch on the back panel, because that will be a hindrance when you really want to remove that panel...
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I have noticed such a problem with a CM30 at a previous worksite. It was first evident after installation of a new filament which had an asymmetrical emission pattern within a couple of days of operation. I hooked a dual channel chart recorder to X10, the ion pump current monitor test point, and to the output of a nude ion gage installed near the column vacuumn system. After leaving the cold trap unattended at 5PM I found on monitoring the chart recorder that the cold trap would regenerate during the night, early hours of morning, with concommitent increase of ion pump current and ion guage voltage. Depending on the number of sample exchanges made during the day the column backing pipe pressure would go as high as 1E-5 Torr according to to the ion guage.
By my experience you should an have at least an hour or so of use after removing the LN2 before the cold trap begins to warm up. Unfortunately, the CM30 has a filament preheat voltage that remains after the filament knob is turned all the way down and is not interrupted until the IGP interlock circuit cuts off the high voltage. If you experience high tension trips frequently you may have a vacuum leak and the filament may die prematurely if it is LaB6.
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All,
After a few hours using the TEM (Philips CM30) with the cold trap filled with LN2, it appears that some vapors in the microscope column get trapped, as it should be.
Now the problem is: have you ever noticed any problem AFTER removing the LN2 dewar, thus heating the trap, provoking a degassing in the column. We have noticed that after several hours working with the cold trap, the IGP gas pressure level increases a lot after removing the dewar (it happens that HT switches off during the night because of that IGP pressure increasing above the "safety level"), and some users here tell me that it should not be that way.
Question: did you ever notice such a problem, and if yes, how long time would you work without heating the trap in order not to get any dramatic increase of IGP pressure after removal of the dewar.
In order to answer this accurately, need to know a bit more about the SEM video than it's amplitude. Is it NTSC? Slow Scan? The 1 volt p-p video is a standard "line" level.... Woody
------------------------------------- I need information on linking the video signal of a SEM (1V p-p) composite video single to a LECO imagin system that can accept a variety of formats (PAL, NTSC) etc. Any ideas or experience with this? Thanks Craig
On May 14, 6:51pm, Yves Maniette wrote: } Subject: TEM Cold trap degassing } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } All, } } After a few hours using the TEM (Philips CM30) with the cold trap filled } with LN2, it appears that some vapors in the microscope column get } trapped, as it should be. } } Now the problem is: have you ever noticed any problem AFTER removing the } LN2 dewar, thus heating the trap, provoking a degassing in the column. We } have noticed that after several hours working with the cold trap, the IGP } gas pressure level increases a lot after removing the dewar (it happens } that HT switches off during the night because of that IGP pressure } increasing above the "safety level"), and some users here tell me } that it should not be that way. } } Question: did you ever notice such a problem, and if yes, how long time } would you work without heating the trap in order not to get any dramatic } increase of IGP pressure after removal of the dewar. } } Yves MANIETTE } Universitat de Barcelona } -- End of excerpt from Yves Maniette
It is same with our CM300FEG and CM120, especially after a few hours cryo work or change sample several times. But I think this is normal. As far as the dewar is filled with LN2, you can work as long as you want. With our CM300FEG, there is an extra valve V7 which should be manually closed after finishing the work. Soon after you remove the dewar or run out the LN2, the IGP1 will increase quite a lot (} 19, which is the safe value for open the V7), and then come down below 19 after a while. If V7 is closed, there should be no harm to the TEM. If you have the cryo option in the vacumm page, you can also turn off the IGP1 for a while (I usually turn it off for half hour after remove the dewar), and let ODP to pump the column, which is more efficient to remove the water vapor from the column. But this works only when your P3 is very good, definitely before changing film.
Yifan Cheng
--
************************************************************************** * Dr. Yifan Cheng * Phone: +1-904-644-4104 * * Institute of Molecular Biophysics * Fax: +1-904-561-1406 * * Florida State University * Email: ycheng-at-sb.fsu.edu * * Tallahassee, FL 32306-3015, U.S.A. * http://www.sb.fsu.edu/~ycheng * *------------------------------------------------------------------------* * Home address: * * * 313 Pennell Circle #4 * Phone: +1-904-575-3620 * * Alumni Village * * * Tallahassee, FL32310, U.S.A. * * **************************************************************************
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Pregnancy in EM 5/15/97 7:54 AM
I was wondering what most EM labs do for precautionary measures when someone in the lab is pregnant. I have a colleague who is pregnant now, and she does not do resin embedding. She does cut cured blocks,stain them with uranyl acetate and lead citrate, and look at them on the TEM. Is this normal procedure in other labs? Should she wear a mask when staining? Linda Chicoine Center for Cell Imaging Yale University New Haven, CT
Greetings, I wonder anyone knows of a source for Technovit resin in the USA? This resin is made by Hareus (spelling?) in Europe. Is there perhaps a similar if not identical resin available here? Also, does anyone know if Technovit is glycol methacrylate? Thanks in advance for any leads, Tobias Baskin
On the Philips microscopes there is a socket in the lower part of the IGP. if this socket is disconnected then IGP will be disconnected, and also the vacuum valves will open in such a way that the ODP (low chamber) will be more openly connected with the column and gun. This allows to help "cleaning" the IGP gun.
But the socket is hidden behin the rear panel so that you have to remove it in order to remove the socket, something not very practical. The hint is to connect the socket to a switch that you place at any convenient place, and the side panel is a convenient one.
On Thu, 15 May 1997, Ann-Fook Yang wrote:
} I follow the thread with interest. } What is the switch, you talked about, for? } When do you use it? } Just curious. } } Ann Fook } } } } } } Yves Maniette {yves-at-giga.sct.ub.es} 05/15/97 06:18am } } } } ------------------------------------------------------------------------ } } You can ask your local maintenance engineer to place a switch on the } microscope SIDE panel near the IGP pump. This switch shoult be } connected } to the small socket down the IGP pump. Any Philips Engineer also knows } that trick and will install it for you if you ask him. The only point is } NOT to place the switch on the back panel, because that will be a } hindrance when you really want to remove that panel... } } Yves MANIETTE } Universitat de Barcelona } }
Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Lluis Sole i Sabaris, 1-3 E-08028 BARCELONA ESPANYA
I am interested in purchasing a set of ZEISS Plan Apo objectives for my Photomicroscope II.
These must be 160mm tube length and be in mint condition. I need the following: 4X, 10X, 20X, 40X, 63X and 100X (the 63 and 100X are O.I.). Will consider other Zeiss types but prefer Plan Apo.
Will also consider purchasing a complete vintage (1975/1990) Zeiss scope.
Thanks for any leads that may result in a favorable purchase.
Cordially, Gil Groehn ULTRAMED, INC. - RESEARCH DIV.
-- ============================================================= ULTRAMED, INC. Research Div. ad408-at-detroit.freenet.org Grosse Pointe Farms, MICH 48236 USA PHONE: 313-884-1139 ===============================================================
Electron Microscopy Technician - EM technician needed by state-of-the-art laboratory studying the organization of proteins and nucleic acids in cell nuclei. The individual should be highly competent in thin sectioning with a diamond knife, fixation and embedding methods, darkroom work, immunofluorescence microscopy and/or fluorescence in situ hybridization. Experience in cell culture and immunogold labeling is highly desirable. Please send resume and two letters of reference to: Dr. David L. Spector, Cold Spring Harbor Laboratory, P.O. Box 100, Cold Spring Harbor, New York 11724.
} Appello Urgentissimo - VERY URGENT APPEAL } } Urge aiuto per una bambina italiana di sei anni Francesca De Lunas } FRANCESCA DE LUNAS IS A 6 YRS OLD DAUGHTER WHO URGENTLY NEEDS HELP. } } ricoverata presso il reparto rianimazione dell'ospedale Brotzu di } Cagliari } (Sardegna-Italia) Dott. Pettinao. } SHE'S NOW UNDER THERAPY BY DR. PETTINAO AT THE "REPARTO RIANIMAZIONE" } [REANIMATION DEPT.], } OSPEDALE BROTZU, CAGLIARI (SARDINIA, ITALY) } } La bambina presenta una sindrome caratterizzata da: } SHE SHOWS A SYNDROME WHICH PECULIARITIES ARE: } } - ipercoagulazione del sangue; } BLOOD HYPERCOAGULATION } } - trombi che si formano in circa 15 secondi di colore rosso con nucleo } bianco } di lunghezza di circa 2-3 cm. } RED THROMBI WITH WHITE NUCLEUS, ABOUT 2-3 CM LONG, WHICH FORM IN 15 } SECS. } } Alla bambina e' stata gia' amputata la gamba sinistra e, finora non e' } stato possibile effettuare diagnosi di alcun tipo. } THE DAUGHTER HAS ALREADY BEEN AMPUTATED THE LEFT LEG. IT HAS NOT BEEN } POSSIBLE } TO MAKE ANY DIAGNOSIS UNTIL NOW. } } Chiunque sia in grado di ipotizzare una diagnosi o di riconoscerla } si metta urgentemente in contatto con le seguenti e-mail: } ANYBODY ABLE TO MAKE A DIAGNOSIS OR RECOGNIZE THIS DISEASE PLEASE GET } IN } TOUCH } URGENTLY BY E-MAIL TO THE FOLLOWING ADDRESSES: } } lukrezia-at-mbox.vol.it } } stefania-at-alanet.it } } schintu-at-pan.bio.uniroma1.it } } psico-at-mbox.vol.it } ********************************* } THANK YOU VERY MUCH FOR YOUR COOPERATION ____________________________________________________________________________ ____
Dr. Cristiano Rumio Istitute of Human Anatomy Via Mangiagalli 31 20133 Milan Italy Tel. -39-2-2663683 Fax. -39-2-2364082 E-mail: crylsm-at-imiucca.csi.unimi.it URL: http://imiucca.csi.unimi.it/~endomi/confocal.html Immunofluorescence Course http://imiucca.csi.unimi.it/endomi/ACIF.html ____________________________________________________________________________ ____
Appello Urgentissimo - VERY URGENT APPEAL } } Urge aiuto per una bambina italiana di sei anni Francesca De Lunas } FRANCESCA DE LUNAS IS A 6 YRS OLD DAUGHTER WHO URGENTLY NEEDS HELP. } } ricoverata presso il reparto rianimazione dell'ospedale Brotzu di } Cagliari } (Sardegna-Italia) Dott. Pettinao. } SHE'S NOW UNDER THERAPY BY DR. PETTINAO AT THE "REPARTO RIANIMAZIONE" } [REANIMATION DEPT.], } OSPEDALE BROTZU, CAGLIARI (SARDINIA, ITALY) } } La bambina presenta una sindrome caratterizzata da: } SHE SHOWS A SYNDROME WHICH PECULIARITIES ARE: } } - ipercoagulazione del sangue; } BLOOD HYPERCOAGULATION } } - trombi che si formano in circa 15 secondi di colore rosso con nucleo } bianco } di lunghezza di circa 2-3 cm. } RED THROMBI WITH WHITE NUCLEUS, ABOUT 2-3 CM LONG, WHICH FORM IN 15 } SECS. } } Alla bambina e' stata gia' amputata la gamba sinistra e, finora non e' } stato possibile effettuare diagnosi di alcun tipo. } THE DAUGHTER HAS ALREADY BEEN AMPUTATED THE LEFT LEG. IT HAS NOT BEEN } POSSIBLE } TO MAKE ANY DIAGNOSIS UNTIL NOW. } } Chiunque sia in grado di ipotizzare una diagnosi o di riconoscerla } si metta urgentemente in contatto con le seguenti e-mail: } ANYBODY ABLE TO MAKE A DIAGNOSIS OR RECOGNIZE THIS DISEASE PLEASE GET } IN } TOUCH } URGENTLY BY E-MAIL TO THE FOLLOWING ADDRESSES: } } lukrezia-at-mbox.vol.it } } stefania-at-alanet.it } } schintu-at-pan.bio.uniroma1.it } } psico-at-mbox.vol.it } ********************************* } THANK YOU VERY MUCH FOR YOUR COOPERATION } send to your mailing list
Hello: Anybody who has the experience in using liquid helium stage for either TEM or SEM, or has the information on where I can find this type of stage, please send me an email at wtao-at-mtu.edu. Thanks.
A colleague is interested in purchasing a used, inexpensive light microscope. Nothing fancy: standard LM with 10x, 40x and 100x objectives and reasonable resolution. To be used for examination of biological and/or clinical materials (smears, sections, etc). Private individuals or vendors should contact:
Contact: Dr. Faiqa Hassan St. Louis, Mo. Phone: 314-522-0083 Fax: 314-322-0083
At 10:22 AM 5/15/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a small business which deals mostly with electron optics but when ever I have needed help with glass optics, I contact Mel Sobel Microscopes. Their address is 29 Louis Street, Hicksville, New York 11801 Phone: 516/935-7794 FAX 516/935-6131. These people seem to have parts for any optical microscope ever made and their prices are very reasonable.
Good luck.
I am not associated with Mel Sobel Microscopes. } } } } } I am interested in purchasing a set of ZEISS } Plan Apo objectives for my Photomicroscope II. } } These must be 160mm tube length and be in mint } condition. I need the following: 4X, 10X, 20X, } 40X, 63X and 100X (the 63 and 100X are O.I.). } Will consider other Zeiss types but prefer Plan Apo. } } Will also consider purchasing a complete vintage } (1975/1990) Zeiss scope. } } Thanks for any leads that may result in a favorable } purchase. } } Cordially, } Gil Groehn } ULTRAMED, INC. - RESEARCH DIV. } } -- } ============================================================= } ULTRAMED, INC. Research Div. ad408-at-detroit.freenet.org } Grosse Pointe Farms, MICH 48236 USA PHONE: 313-884-1139 } =============================================================== } } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Alexander Greene Scientific Instrumentation Services, Inc. Number 499, Post Office Box 19400 Austin, Texas 78760 Phone: 512/282-5507 FAX 512/280-0702
REASONABLY PRICED ELECTRON MICROSCOPE REPAIR ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Microscopy and Microanalysis '97 is less than three months away. The meeting this year will be held August 10-14 in Cleveland, Ohio. The deadline for early registrations is July 15. If you are plan to attend, please consider making plans as soon as possible. Cleveland has recently become a popular place to visit and hotel rooms WILL be at a premium if you don't act soon. A popular attraction, the Cleveland Indians, will be playing at home that week. The baseball games and other attractions such as the Rock and Roll Hall of Fame have already lead to a huge demand for hotel space for that week.
Information on the Meeting proceedings can be found on the world wide web at http://www.bright.net/~strecker/msno/mm97.html. The forms you will need for early meeting registration and hotel registration are available in pdf format for downloading here. You will also find information on hotels, events and local attractions available to attendees of the meeting at this site. If you require additional information or are unable to access the WWW, you can contact the MSA business office by e-mail at BusinessOffice-at-MSA.Microscopy.com or by phone at (800) 538-3672.
Once again, please register early so you dont miss out on Microscopy and Microanalysis 97. I hope to see you in Cleveland this year.
A colleague studying trace metals in San Francisco Bay has noticed more osmium in the sediments than expected.
He knows EM-types use osmium and quizzed me regarding possible sources for the Os in the bay. He wondered how many EM labs are around the bay and if they could be sources of Os in the waste stream. I said probably not, most EM-types are careful about disposing of all toxic/hazardous waste in an approved manner. He was still curious so I offered to pose a few questions to the group on his behalf.
If you have any suggestions or information to add to his research please send them my way. Here are some of his specific questions:
1. How many EM labs that use OsO4 are there around the SF Bay area?
2. About how much OsO4 do they use in a year?
3. How do the labs dispose of waste OsO4 and does that include every last bit, even that from rinsing used containers?
4. If the Os he sees is not from EM labs, then where could it be coming from? Does anyone know of industrial or other applications that use a lot of Os?
Stay tuned for further details as he develops his theories.
Thanks,
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
A colleague has asked me to post this request. He is seeking information on short courses which may be available in confocal laser scanning microscopy (general principles, applications, etc. in the biological and biomedical sciences). The course(s) should preferably be offered in the continental U.S. The first choice would be in the Rocky Mountain or Desert Southwest areas.
I've perused the (published) lists of the Lehigh and Woods Hole courses, and nothing jumps out at me as addressing specifically confocal laser microscopy. Perhaps I've missed something??
Does anyone have a recommendation as to who my colleague should contact? If so, please e-mail me directly. I will pass the info along.
BTW, it might also be worth posting your reply to the Microscopy Listserver. If there is such a course coming up this year there may be others who would be interested.
it's glycol methacrylate and is the same stuff as Historesin under a different name.
At 08:42 AM 5/15/97 -0600, Tobias Baskin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Delilah Wood United States Department of Agriculture Western Regional Research Center 800 Buchanan Street Albany, CA 94710
I would like to make contact with those of you who use beryllium grids fairly often or occasionally in your work, for the purpose of discussing some particular issues with those grids. I am concerned, really, about the toxicity of that material. Are there any viable alternatives? Or is the hazard, to operator or environment, in fact not so worrisome that anyone really need be concerned about looking for something else? (Opinions?) I'm also interested to learn for what work it is generally felt that one should even be considering a beryllium grid, since they are pretty costly to boot!
Cheers,
Cam
____________________________________________________________________________ P.O. Box 1932 Main Station T: 1 403 913 3850 Cameron Sorlie, President Edmonton, AB T5J 2P3 Canada F: 1 403 433 9376 General Microdevices, Inc. ----------------------------------------------- generalmicro-at-incentre.net ______________________________________________________ Microfabrication technology applications
} I would like to make contact with those of you who use beryllium grids } fairly often or occasionally in your work, for the purpose of discussing } some particular issues with those grids. I am concerned, really, about the } toxicity of that material. Are there any viable alternatives? Or is the } hazard, to operator or environment, in fact not so worrisome that anyone } really need be concerned about looking for something else? (Opinions?) I'm } also interested to learn for what work it is generally felt that one should } even be considering a beryllium grid, since they are pretty costly to boot! } } } Cheers, } } Cam
Beryllium grids can be very useful when doing X-ray microanalysis - especially if you are looking for Cu, or anything with a peak that might be swamped by Cu. Also potentially useful if you are looking for light elements whose lines might be swamped by the Cu L line.
There are alternatives - Al grids resolve most of the above problems. Carbon coated nylon and carbon composite grids are also available from the usual suppliers - Agar, SPI, etc. However, for really critical analysis, I'd say Be grids are to be preferred.
I don't believe there is any serious hazard associated with these grids. Be is a problem if it get into the lungs (or trapped under the skin), where it leads to medical conditions similar to those caused by asbestos. So don't use abrasives on it, and I guess you want to avoid exposure to any strong acids that might attack the oxide surface (and don't eat them:)
a new tipe of SPM tip artifact is described and put on the WWWW:
http://newton.phy.bme.hu/pub/corfu93/index.html
may be it is interesting for some of you!
Best regards: Peter
*************************************************************************** * dr. Nagy, Peter (physicist) * * MTA (Hungarian Academy of Sciences) * * KFKI (Central Research Institute for Physics * * ATKI (Research Institute for Materials Sciences) * * STM (Scanning Tunneling Microscopy Group) * * H-1525 Budapest-114, P.O.Box.49 Tel.: +36-1-3-959-220/19-68 * * Budapest, Konkoly T.M. u. 29-33 Fax.: +36-1-3-959-284 * ***************************************************************************
I saw a fiber name recently that is unfamiliar to me and thought someone might help me out. It's "Marquesa Lana", no doubt a trade or brand name? Supposedly very durable as a chair fabric? Appreciate any leads.
} Hello: } Anybody who has the experience in using liquid helium stage for either } TEM or SEM, or has the information on where I can find this type of } stage, please send me an email at wtao-at-mtu.edu. Thanks. } } Weimin Tao
Oxford Instruments supply both SEM and TEM liquid He stages. Gatan Inc supplies a liquid He TEM stage.
When I worked for Gatan, I used their TEM He stage a few times. The biggest hassle was actually arranging the He supply and cooling the stage down. Once you'd done that, I didn't see any real problems using it, although I never spent a lot of time working with it.
Hello all; We have developed some problems with heat polymerization of our GMA for light microscopy, and would like to do some testing of UV polymerization. One of my co-workers remembers that in another lab she worked in, distance from the light source was an important factor. I believe Wayne England, who used to be at Carleton University, worked out a lot of bugs on this system... some input from him, and anyone else on the list would be most welcome. If you feel the information is not of interest to the general list, please contact us directly:
Lu-Ann Bowman bowmanla-at-em.agr.ca or Shea Miller millers-at-em.agr.ca
Thanks in advance shea
p.s. Thanks a bunch for my SOS about in situ hybridization using DIG- we got some great suggestions/tips, and will be posting a summary to the list next week.
Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
I am trying to obtain some price estimates of using PIXE (proton-induced X-ray emission) microanalysis. What does it cost per hour to use such a microprobe? Is it a lot costlier than using conventional microprobes?
Thanks a lot for your help -
Karin E. Limburg Dept. of Systems Ecology University of Stockholm, Sweden
I just did a quick web search using Alta Vista and found:
Marquesa lana is indeed a wear-resistant chair fabric (olefin) but I couldn't find a manufacturer's name.
Marquesa lana is also the name of a horse scratched in the 8th race at Sam Houston Race Park on March 30th. Perhaps she was rubbed too much and wore out?
Harry Crossman
} ---------- } From: David_R_Stadden-at-armstrong.com[SMTP:David_R_Stadden-at-armstrong.com] } Sent: Friday, May 16, 1997 8:12AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Fiber ID? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I really appreciate very much all your answers, ideas and suggestions about mechanical and diffusion pumps fluids. We have asked quotations to different companies for differents brands (all of them taken from from your messages) and finally we have decided, according to the prices and facilities + duties to be paid to be imported in Argentina, to buy Fisherbrand #19 and Santovac 5. Thanks to your time and typing effort we have gotten more money than what we had expected. Thanks very much. Greetings from the very south of America.
Silvia Montoro Centro Regional de Investigacion y Desarrollo Santa Fe - Argentina
Position for an electron microscopy technician is availible at Emory University neurology department (Atlanta, Georgia). Minimum qualification required includes B.S in a related field and 1 year experience in an anatomical research lab. The technician will be involved in studies on Fragile X syndrome and Huntingtons disease. Routine work includes morphological and immunocytochemical sample processing for light and electron microscopy, ultramicrotoming, examining sections on electron microscope, and digitizing results on computer. The starting date of this job will be on July 1. 97. Interested individuals please send CV to the following address:
Hong Yi Rm 6223 Woodruff Memorial Building Department of Neurology Emory University 1639 Pierce Dr. Atlanta, GA 30322
Hi Kaye; I have experienced this problem as well, and haven't figured out where it comes from. Initially I thought it was age, (old sections) but have also had problems with freshly sectioned material, (including freshly embedded/polymerized). Something that seems to help... I soak the slide with attached sections in a coplin jar with acetone for 10 minutes or so (I have gone up to 30 or 40 minutes), rinse with water, and then apply my stain while the sections are still wet. This may be a problem for some of the stuff you want to look at. Generally the cracks seem to disappear, or are at least minimized, and it no longer interferes with the microstructure of whatever I'm looking at. Hope this helps shea Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
Hello! Does anyone have a used Ion Tech ion mill lying around that needs a good home? I have a personal project that requires one. Please reply directly - thanks!
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Page 6773 of the tenth edition of the Merck Index states the uses of osmium tetroxide as "an oxidizing agent " , "catalyzes chlorate, peroxide, periodate and other oxidations". Osmium is used "as an alloy with iridium for pen points and fine machine bearings; as catylyst in the synthesis of ammonia ; as catylyst in hydrogenation of organic compounds ". I suggest that could explain the presence of Os in SF bay from industrial uses. Another explanation is the incredible sensitivity of the detection equipment. How much is more that expected? Kate Connolly
Karin I have no experience of using the technique myself but I met a man who did. If I recall correctly the cost was higher because of the need of a proton source and the specialist design of the associated equipment. There was a group at Oxford 5 years ago (I assume they are still there) and you could try contacting: F. Watt, Scanning Proton Microprobe Unit, Nuclear Physics Laboratory, Department of Physics, Keble Road, Oxford OX1 3RH, UK. I'm sorry that I don't have a phone, fax or e-mail address but I'm sure that there will probably be a Web site. Malcolm Haswell University of Sunderland UK ----------
llo microscopists,
I am trying to obtain some price estimates of using PIXE (proton-induced X-ray emission) microanalysis. What does it cost per hour to use such a microprobe? Is it a lot costlier than using conventional microprobes?
Thanks a lot for your help -
Karin E. Limburg Dept. of Systems Ecology University of Stockholm, Sweden
From Microscopy-request-at-sparc5.microscopy.com Fri May 16 11:07:51 1997 Received: from srvr20.engin.umich.edu (root-at-srvr20.engin.umich.edu [141.212.2.26]) by srvr5.engin.umich.edu (8.8.5/8.8.5) with ESMTP id LAA05680; Fri, 16 May 1997 11:07:50 -0400 (EDT) Received: from judgmentday.rs.itd.umich.edu (0-at-judgmentday.rs.itd.umich.edu [141.211.83.37]) by srvr20.engin.umich.edu (8.8.5/8.8.5) with ESMTP id LAA17361; Fri, 16 May 1997 11:07:49 -0400 (EDT) Received: by judgmentday.rs.itd.umich.edu (8.8.5/2.2) with X.500 id LAA13249; Fri, 16 May 1997 11:07:49 -0400 (EDT) Received: from Sparc5.Microscopy.Com by judgmentday.rs.itd.umich.edu (8.8.5/2.2) with SMTP id LAA13110; Fri, 16 May 1997 11:07:29 -0400 (EDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id JAA04028 for dist-Microscopy; Fri, 16 May 1997 09:42:03 -0500 Received: from imsd-exchange.nrc.ca (imsd-exchange.nrc.ca [132.246.160.7]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id JAA04023 for {microscopy-at-MSA.microscopy.com} ; Fri, 16 May 1997 09:42:01 -0500 Received: by imsd-exchange.nrc.ca with SMTP (Microsoft Exchange Server Internet Mail Connector Version 4.0.994.63) id {01BC61E6.2205D070-at-imsd-exchange.nrc.ca} ; Fri, 16 May 1997 10:44:30 -0400 Message-ID: {c=CA%a=_%p=NRC%l=NRC/IMSD/001293E4-at-imsd-exchange.nrc.ca} From: "McCaffrey, John" {John.McCaffrey-at-nrc.ca} To: worldwide BB for microscopist {microscopy-at-sparc5.microscopy.com} Subject: Used Ion Tech ion mills Date: Fri, 16 May 1997 09:48:00 -0400 X-Mailer: Microsoft Exchange Server Internet Mail Connector Version 4.0.994.63 MIME-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Content-Transfer-Encoding: 7bit Errors-to: Microscopy-request-at-sparc5.microscopy.com Status: RO
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Hello! Does anyone have a used Ion Tech ion mill lying around that needs a good home? I have a personal project that requires one. Please reply directly - thanks!
From Microscopy-request-at-sparc5.microscopy.com Fri May 16 11:07:51 1997 Received: from srvr20.engin.umich.edu (root-at-srvr20.engin.umich.edu [141.212.2.26]) by srvr5.engin.umich.edu (8.8.5/8.8.5) with ESMTP id LAA05680; Fri, 16 May 1997 11:07:50 -0400 (EDT) Received: from judgmentday.rs.itd.umich.edu (0-at-judgmentday.rs.itd.umich.edu [141.211.83.37]) by srvr20.engin.umich.edu (8.8.5/8.8.5) with ESMTP id LAA17361; Fri, 16 May 1997 11:07:49 -0400 (EDT) Received: by judgmentday.rs.itd.umich.edu (8.8.5/2.2) with X.500 id LAA13249; Fri, 16 May 1997 11:07:49 -0400 (EDT) Received: from Sparc5.Microscopy.Com by judgmentday.rs.itd.umich.edu (8.8.5/2.2) with SMTP id LAA13110; Fri, 16 May 1997 11:07:29 -0400 (EDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id JAA04028 for dist-Microscopy; Fri, 16 May 1997 09:42:03 -0500 Received: from imsd-exchange.nrc.ca (imsd-exchange.nrc.ca [132.246.160.7]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id JAA04023 for {microscopy-at-MSA.microscopy.com} ; Fri, 16 May 1997 09:42:01 -0500 Received: by imsd-exchange.nrc.ca with SMTP (Microsoft Exchange Server Internet Mail Connector Version 4.0.994.63) id {01BC61E6.2205D070-at-imsd-exchange.nrc.ca} ; Fri, 16 May 1997 10:44:30 -0400 Message-ID: {c=CA%a=_%p=NRC%l=NRC/IMSD/001293E4-at-imsd-exchange.nrc.ca} From: "McCaffrey, John" {John.McCaffrey-at-nrc.ca} To: worldwide BB for microscopist {microscopy-at-sparc5.microscopy.com} Subject: Used Ion Tech ion mills Date: Fri, 16 May 1997 09:48:00 -0400 X-Mailer: Microsoft Exchange Server Internet Mail Connector Version 4.0.994.63 MIME-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Content-Transfer-Encoding: 7bit Errors-to: Microscopy-request-at-sparc5.microscopy.com Status: RO
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Hello! Does anyone have a used Ion Tech ion mill lying around that needs a good home? I have a personal project that requires one. Please reply directly - thanks!
I gave the wrong fax number in the earlier version of this message. Sorry.
A colleague is interested in purchasing a used, inexpensive light microscope. Nothing fancy: standard LM with 10x, 40x and 100x objectives and reasonable resolution. To be used for examination of biological and/or clinical materials (smears, sections, etc). Private individuals or vendors should contact:
Contact: Dr. Faiqa Hassan St. Louis, Mo. Phone: 314-522-0083 Fax: 314-522-1179
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
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I saw a fiber name recently that is unfamiliar to me and thought someone might help me out. It's "Marquesa Lana", no doubt a trade or brand name? Supposedly very durable as a chair fabric? Appreciate any leads.
} I am trying to obtain some price estimates of using PIXE } (proton-induced X-ray emission) microanalysis. What does it cost per } hour to use such a microprobe? Is it a lot costlier than using } conventional microprobes? } Hi Karin:
Try to contact the PIXE group (Prof. Malmqvist et al.) in Lund. You might get a free access. They are helpful. The address is: Nuclear Physics Fax: +46 46 222 4709 E-mail: nuclear-at-nuclear.lu.se
Regards,
B.E.
Benyam Estifanos Dept. of Mineralogy & Petrology University of Lund Soelvegatan 13 S-223 62 Lund, Sweden INTERNET: estif-at-gemini.ldc.lu.se / Benyam.Estifanos-at-Geol.lu.se Tel.:+46 46 2224597; Fax:+46 46 121477
YUHUI XU requested Info on DNA imaging; below is a few articles Ive found helpful....
SALLY BURNS e-mail: burnssal-at-pilot.msu.edu
REVIEW ARTICLES
Davis, R., Simon, M., and Davidson N. 1971 Electon Microscope Heteroduplex methods for Mapping regions of Base sequence homolgy in Nucliec acids Methods in Enzymology #21 419-428 (QP601.C733)
Burkhardt and Lurz Electron microscopy: CHP 6 in the book: Adv Molec Genetics 1984 (QH 430.A38 1984)
Spiess, E., and Lurz R. 1988 EM analysis of nucleic acids and nucleic acid protein complexes. Methods in microbiology 20: 293-323. ........................................................ ------------------------Other relevant articles
Backert, S., Lurz,R., and Borner T. 1996 E.M investigation of mitochondrail DNA from Chenopodium album (L.) Curr Genet 29:427-436
Backert, S. Dorfel, P., Lurz, R., and Borner, T. 1996 Rolling circle...... Mol and Cellular Biol. vol 16 no 11 p6285-6294.
DAVIS, RW AND HYMAN,RW. 1971 A study in evolution : ..... J. MOL BIOL 62,287-301.
Kolodner, RD and Tewari,K. 1974 Denaturation Mapping Studies on the Circular Chloroplast deoxyribonucleic Acid from Pea Leaves J Biological Chem 250313 4888-4895
Kolodner, RD and Tewari, K. 1975. Presence of Displacement loops..... J. Biol Chm vol 250, No22 8840-8847.
****LANG,D., and MITANI, M. 1970 ******** Simplified Quantitiative Electon Microsopy of Biopolymers Biopolymers Vol 9 373-379.
Wessel Muller, H. and Hoffman Berling (1990) ELECTRON MICROSOPY OF DNA-HELICASE I COMPLEXES IN THE ACT OF STRAND SEPERATION Eur J Biochm 189,277-285 ......... (uses SSB and formamide.)
I have just become the principal operator of a Zeiss DSM-960, and would like to hear any comments or insights from anyone having experience with Zeiss SEM's, especially pertaining to the use of LaB6 sources. I have experience on other vendors instruments, but the design of cathode assembly is different than what I am familiar with. Specifically, there is no height adjustment, other than a shim. The instrument manuals provide little information and simply state the LaB6 is not operator replacable and is factory serviced as an assembly. The lab has been sending the entire assembly to FEI for rebuilding. Is this really necessary? Does anyone replace their own sources on a Zeiss?. I can only find reference to a Zeiss base for the DENKA M1 and M7 in any of the supplier catalogs I have, although I haven't talked to any suppliers yet.
Thanks Jim Mabon _____________________________________________________ James C. Mabon Center for Microanalysis of Materials Frederick Seitz Materials Research Laboratory 104 South Goodwin Avenue Urbana, Illinois 61801 (217)333-4265 *Fax(217)244-2278 email: mabon-at-mrl7.mrl.uiuc.edu {that's the letter l} _____________________________________________________
With regard to the question from Cam on the matter of Be grids: ------------------------------------------------------ I am concerned, really, about the toxicity of that material. Are there any viable alternatives? Or is the hazard, to operator or environment, in fact not so worrisome that anyone really need be concerned about looking for something else? (Opinions?) I'm also interested to learn for what work it is generally felt that one should even be considering a beryllium grid, since they are pretty costly to boot! ------------------------------------------------------
and the reply from Larry Stoter: ------------------------------------- There are alternatives - Al grids resolve most of the above problems. Carbon coated nylon and carbon composite grids are also available from the usual suppliers - Agar, SPI, etc. However, for really critical analysis, I'd say Be grids are to be preferred.
I don't believe there is any serious hazard associated with these grids. Be is a problem if it get into the lungs (or trapped under the skin), where it leads to medical conditions similar to those caused by asbestos. So don't use abrasives on it, and I guess you want to avoid exposure to any strong acids that might attack the oxide surface (and don't eat them:) =================================================
There are "alternatives", but most really are not as acceptable as Be. The Al grids give a strong x-ray line that can swamp the system much like Cu and also, coatings (e.g. "Formvar", etc.) don't seem to want to stick as well as on Be. Oxide coatings on the Al tend to reduce conductivity even further. The carbon composite grid, while conductive, gives high Bremsstrahlung background radiation reducing experimental sensitivites. The Nylon grids have to be carbon coated to reduce charging, but generally it is never eliminated completely and charging does continue to be at least somewhat of a problem. Also, the Nylon grids (at least the ones we have tried) are further plagued by a low level additive of TiO2 which gives additionally, low level instensities due to Ti. And there is still the problem with the high Bremsstrahlung background radiation. Probably diamond grids come the closest to being an acceptable substitute, and the Bremsstrahlung background is not significant, except that they are even more expensive than Be. But for those who operate in institutional environments where Be is not permitted under any circumstances, the diamond grids would be the alternative of choice, at least technically, even if it would fail economically.
Now a few other comments about Be grids:
a) what is toxic is the oxide, not the metal. There is widespread appreciation that BeO is highly toxic. However, what is not toxic is the metal itself. So if what is being done, in the handling of the Be grids does not result in the formation of an oxide scale or film, something that could in theory, "come off", then so far as we know, there is no problem. I am unaware or any explanation of how the oxide could form while being irradiated in the column and I am unaware of anyone who has ever shown that the oxide has indeed formed on any of the Be grids.
I myself feel much more comfortable offering for sale the Be grids, for example, than our Be planchettes. While it is true that (at least our) planchettes are accompanied with all kinds of safety instructions, the fact is, I have this nagging concern that a Be planchette once carbon coated, for example, tends to take on the appearance like any other planchette or mount made of aluminum, and very clearly, as evidenced by the recent postings on the topic of the cleaning of SEM mounts, there is quite a bit of grinding and recycling of Al mounts. Hence if one or more Be planchettes ended up accidentally being mixed in with the Al mounts, and received these same "grinding and polishing" treatments, then there very well could end up being a BeO dust problem, from the grinder and polisher for sure when they became dried out.
Now, from my own personal contacts in the Be industry (remember, such contacts might not give out objective information!), I have been told that even in the case of the drying out of a polishing table, the levels of dust released would be at such low levels that it would be an extraordinary thing for someone to suffer any of the classic symptoms of berylliosis. However, they do point out that some small percentage of us are in fact highly reactive to such low levels, and the problem is that no one knows in advance who is going to be a reactor and who would not react.
Quite possibly, at least some of the concern about the grids flows from this very real concern related specifically to the planchettes but which would not be applicable to the grids since grids are not "ground and polished".
b) With regard to the "high cost", there is nothing that can be done about that, however, from all reports we have ever heard, as well as our own experience in our own laboratory, Be grids in a small beaker of acetone in an ultrasonic shaker for some period of time, tends to result in a fairly quick cleaning up for reuse.
Also, plasma cleaning as a final step, is a possibility as well, however, in this instance I am a bit less certain. Some years ago, we did an oxygen plasma exposure of several minutes to some Be grids and found no change. This was not a rigorous "looking" for oxide formation, but to the eye, and also, at stereo zoom LM level there was no apparent change. In other laboratories this seems to be a standard way of cleaning for the reuse of Be grids. So while there might be "sticker shock" when Be grids are first purchased, the ability to reuse them over and over again, tends to mitigate that high cost and furthermore, the enhancement to data quality (as opposed to the alternatives) is quite significant to say the least.
c) All Be grids were not created equal, so the results that are obtained from one supplier may or may not be the same as from all other suppliers. I am referring to both the quality of the etching of the Be foil and also, the purity of the Be starting foil material. So far as I know, the Be grids from Agar, as well as those from SPI, are made from the highest purity Be foil starting material available for this kind of use.
In any case, we are frequently asked about these alternatives and there is no one place, at least that I am aware of, where one could find the "answers".
Disclaimer: SPI Supplies is a supplier of Be grids, diamond grids, as well as the other grids mentioned so it would be in our own best interests to see more people doing this kind of work.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Can anyone refer a source for laser printer enhancement software, similar to LaserPix, which is compatible with WINDOWS NT 32 bit architecture? Now using HP Laserjet 4.
Thanks for the help,
Bob Hessler Hessler Technical Services Stamford, CT PHONE/FAX: (203) 358-0266 E Mail: hestec-at-ix.netcom.com
Sorry to bring up this subject again but we have just managed to get the = funds to purchase a negative scanner. We need to scan 35mm, 120 and TEM = (6.5x9cm) negatives so we need a versatile system which will run on a = PC.
From what I have been able to get so far, dedicated negative scanners = are better than flatbeds with transparency adaptors, so we've decided to = restrict ourselves to these. However if you know of any flatbeds which = match the performance of the dedicated ones please respond.
There are 3 which I have managed to find available in Australia within = our price range. These are:
Nikon LS 4500 AF - had some problems but I've been assured that these = have been fixed
Kodak RFS 3570 - don't know much about these
Polaroid SprintScan 45 - don't know much about these either.
So, if you have had any experience (good or bad) running any of the = above on a PC, or if you know of any other suitable scanners please tell = me.
Many thanks in advance.
Colin Veitch
Colin.Veitch-at-dwt.csiro.au CSIRO Division of Wool Technology PO Box 21 Belmont Vic 3216 Australia Tel: +61 (0) 3 5246 4000 Fax: +61 (0) 3 5246 4057
At 08:42 AM 5/15/97 -0600, Tobias Baskin asked: } I wonder anyone knows of a source for Technovit resin in the USA? } This resin is made by Hareus (spelling?) in Europe. Is there perhaps a } similar if not identical resin available here? Also, does anyone know if } Technovit is glycol methacrylate?
We (Energy Beam Sciences) took over distribution of the Technovit resins from Delaware Diamond Knives about two months ago. They are manufactured by Heraeus Kulzer in Germany. There are three resins:
Kulzer has given us some very nice technical/applications brochures on these products; please contact me back-channel if you would like one.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Dear colleagues, I would like to add two works to the "DNA biblio" list:
Preparation and length measurements of the total deoxyribonucleic acid content of T2 bacteriophages. 1962 (classical article) Kleinschmidt-AK; Lang-D; Jacherts-D; Zahn-RK Biochim-Biophys-Acta 1989; 1000: 41-48
Preparation and length measurements of the total DNA content of T2 bacteriophages: a commentary on 'Darstellung und Langenmessungen des gesamten Desoxyribonucleinsaure-Inhaltes von T2-Bakteriophagen' [published erratum appears in Biochim Biophys Acta 1989 Dec 8; 993 (2-3):309] Kleinschmidt-AK Biochim-Biophys-Acta. 1989; 1000: 35-40
A lot about beginning of EM of DNA can be found in these articles.
Best regards O. Benada +---------------------------------------------------------------+ Dr. Oldrich Benada Acad. Sci. CR Phone: +420-2-4752399 Institute of Microbiology Fax: +420-2-4715743 Videnska 1083 E-mail: benada-at-biomed.cas.cz 142 20 Prague 4 - Krc Czech Republic +---------------------------------------------------------------+
Dear colleagues, I would like to add two articles to the "DNA biblio" list:
Preparation and length measurements of the total deoxyribonucleic acid content of T2 bacteriophages. 1962 (classical article) Kleinschmidt-AK; Lang-D; Jacherts-D; Zahn-RK Biochim-Biophys-Acta 1989; 1000: 41-48
Preparation and length measurements of the total DNA content of T2 bacteriophages: a commentary on 'Darstellung und Langenmessungen des gesamten Desoxyribonucleinsaure-Inhaltes von T2-Bakteriophagen' [published erratum appears in Biochim Biophys Acta 1989 Dec 8; 993 (2-3):309] Kleinschmidt-AK Biochim-Biophys-Acta. 1989; 1000: 35-40
A lot about beginning of EM of DNA can be found in these articles.
Best regards O. Benada +---------------------------------------------------------------+ Dr. Oldrich Benada Acad. Sci. CR Phone: +420-2-4752399 Institute of Microbiology Fax: +420-2-4715743 Videnska 1083 E-mail: benada-at-biomed.cas.cz 142 20 Prague 4 - Krc Czech Republic +---------------------------------------------------------------+
If anyone can help Matt out, please reply to him directly
} From: Abrcrombee-at-aol.com } Date: Sat, 17 May 1997 21:25:53 -0400 (EDT) } To: sbarlow-at-sunstroke.sdsu.edu } Subject: Hello } } I found your email address while searching for information on Trichonympa. } My biology teacher assigned my class semester reports, and mine is on that. } Would you possibly have any information that you could send me about } Trichonympa? I would appreciate it if you would send something.....anything } at all, for I have found barely anything on this organism. Thanks...Matt } McCluskey }
Dr. Steven Barlow EM Facility/Biology Department 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/EM_Facility
I do not purport to be an expert on Be but we do handle a fair amount of it around here for other purposes than TEM grids. I believe your response may promote too cavalier an attitude towards handling Be as opposed to BeO. Our ES&H people do not make a distinction between the two regarding handling and toxicity. The latest MSDSs I have for the two indicate roughly the same hazards (NFPA ratings of 2,0,0 for BeO and 2,1,1 for Be). The toxicological sections of the MSDS also indicate roughly the same hazard, around 20mg/kg producing tumors in rats. While I agree that on the scale of all hazardous things in the world, Be is only a moderate hazard for most people (the ~95% who are not allergic), I would be more cautious as regards recycling Be grids. I think they {italic} should be {/italic} recycled to reduce the volume of Be waste generated, but people should pay particular attention to any operation that has the potential for producing respirable dust. Our people who produce sputtered Be films must wear full face respirators every time they open the vacuum chamber even though the material is deposited as a film on the inside of the chamber and not very likely to produce dust.=20 As an aside we have noted that sputtered Be films are fairly reactive, almost always producing substiochiometric oxide, which may not necessarily be applicable to the TEM grid case, but is some indication of its reactivity.
We have a Reichart Ultracut E with the cryo attachment FC 4. The flange on the liquid nitrogen vessel which is used to attach the corresponding flange on the refill dewar head has developed a (nitrogen) leak. The leak developed in the joint between the flange and the dewar . We actually located the point where it leaks . We tried adding some epoxy to that area, but it did not solve the problem.This joint seems to be an epoxy . The manufacturer wants to replace the whole dewar at considerable cost. Has anyone encountered a similar problem . I would appreciate any comments and possible (alternate) solutions.
In a number of German microtechnique texts, including D. Gerlach's "Botanische Mikrotechnik", I have seen references to Pfeiffer's Mixture as being ideally suited to fixing of algae, particularly the filamentous forms. However, I'm confused about one of the constituents of this mixture, given in German as "roher Holzessig". I thought this might be the same as tannic acid, but I'm not sure. I'm also not sure of the concentration and the meaning of the term "roher" (raw) in the description.
Can anyone help clarify the formula Pfeiffer's Mixture? I have not found any references to Pfeiffer's Mixture in any English or American texts on botanical microtechnique - are there fixing mixtures which are as good or better for filamentous algae?
I have several thousand confocal images (TIFF single files, 8bit grey, 24 bit RGB, z-series etc.) and I was wondering whether there is some relatively cheap software/shareware around that I could use to catalogue the images. I want to be able to save some information with the images and thumnail sketches would be useful.
Alternatively, if someone knows how I can use Microsoft Access as an image database without having to embed the full resolution picture in each record I would be grateful.
Any ideas? contact Mark Auty Mark Auty DPC Moorepark Fermoy Co. Cork Ireland
As this is perhaps the very first enquiry on PIXE microanalysis, let me try to give a few details.
Firstly, you were given the mail address to Lund nuclear microprobe group. The full address is: Dept. of Nuclear Physics, Lund Institute of Technology, Box 118, SE-221 00 Lund, Sweden In addition to the e-mail which was given to you, the contact with Jan Pallon (one of the senior members of this group) is: Pixejan-at-outis.lucas.lu.se or: Jan.Pallon-at-pixe.lth.se
To answer directly your question on running costs. At our nuclear microprobe, which is situated in South Africa, at National Accelerator Centre in Faure, close to Cape Town, the full cost is ca. 100 USD per hour. This is however the maximum price, for industrial applications with full confidentiality of results, if required. We are the so-called national facility and users from local universities are encouraged to apply for the beam time, and in many cases this comes free of charge. Our preferred collaboration is under projects in many research fields (geology, materials science, botany and life sciences). This means longer collaboration, sharing students if possible, full access to data. We welcome any international collaboration, if suitable for this type of equipment.
You can contact me at:
PRZYBYLOWICZ-at-nac.ac.za Address: Dr. W.J. Przybylowicz National Accelerator Centre Van de Graaff Group P O Box 72 Faure 7131 South Africa Fax: +27-21-8433543
Web site (does not give all details; some information, e.g. publications, is not fully updated) is: http://www.nac.ac.za xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz National Accelerator Centre PO Box 72 Faure 7131 South Africa E-mail: PRZYBYLOWICZ-at-nac.ac.za Fax: +27-21-8433543 Phone: +27-21-8433820 xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
I run a morphology core where a TEM is used every day. Last year when I was pregnant, I called Health Physics who monitored the scope and me for a period of time. There was absolutely no leakage fromm the column. Although every scope is different, ours is brand new, you can always have your environmental people come out and check for leaks. I believe them to be safe - my son had all of his fingers and toes.
Cheri Owen
On Tue, 13 May 1997, PATRICK DIEHL wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Do any of the universities, companies, hospitals or research institutions } have any policies regarding the use of electron microscopy by pregant } employees? Is there any literature available on the topic? } } Is the advice not to use EM at all during the 9 months, or maybe only } during critical periods of development? } } In searching the archives I only found one reference on this topic. } } Any help would appreciated. } } Patrick Diehl } Materials Science Department } University of North Texas }
If you have a single or double tilt heating holder (for Philips 400 series microscopes)that is in working order and you want to part with it, contact me privately so we can discuss arrangements.
John C. Wheatley Lab Manager Center for High Resolution Electron Microscopy BOX 871704 Tempe, AZ 85287-1704 Phone: (602) 965-3831 FAX: (602) 965-9004 John.Wheatley-at-ASU.Edu
Thank you to all who have responded to my e-mail. If anyone wants copies of the replies, please e-mail me privately. I am unsubribing from the microscopy list.
I am anxiously seeking microscopy video footage for a project which deal with
blood pressure, the contraction of blood vessels and arteries and catherters imagery. Specifically, I need: -blood traveling through arteries (not capillaries) -footage from inside arteries -video from catharter: somethind like a 3/4" view which illustrates pressure which closes and opens valves (looks like open and closing circle) - perhaps this is from an endoscopy?
Please contact me at mhallmedia-at-aol.com with any information you may forward. Many thanks, Heather
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I'm in need of some advice on delineating grain boundaries in a plated copper film. I've tried several ASTM wet etches and a few different CF4 plasma etches, but am not satisfied with the results. Grain boundaries are somewhat defined but not clearly enough to confidently measure their parameters. Does anyone have an etch they are satisfied with for this application? I'm using an FESEM for imaging and have had very good results delineating various alloys, but this copper film of roughly 7 microns thick has me puzzled. X-ray diffraction shows the film is crystalline. Any help is greatly appreciated.
********************************************************** Jake Schaper Product Analysis Lab Advanced Digital Consumer Division Motorola, Inc. 1300 N. Alma School Rd. Chandler, Arizona 85224 Mail Drop CH240 Phone 602-814-4756 **********************************************************
Hi: Our school have received in donation recently a sphectrophotometer BECKMAN 24, that has arrived whitout the technicals handbooks. If some people can send us a copy of all , please contact me thanks in advance =================================================== Fernando D. Balducci Laboratorio de Microscopia Electronica Facultad de Ingenieria - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077 ===================================================
Our department has 2 academic positions currently advertised of which it is intended that one will be filled by a person with an active interest in cell biology and in particular the development of microscopy related research techniques in cell biology (ie Confocal Microscopy, Electron Microscopy, Image Processing and Anaylsis). A copy of the advertisement follows;
LECTURERS IN ANATOMY AND STRUCTURAL BIOLOGY
Two lectureships in the Department of Anatomy and Structural Biology will be available from September 1997. These are confirmable positions. Appropriately qualified applicants could be considered for appointment at Senior Lecturer level.
Applicants will be expected to hold a PhD or equivalent qualification, be committed to research, have proven research ability, and have an on-going and active research program. Their interests should fit within one of the areas of research excellence within the Department: neuroscience, cell biology, reproductive biology, developmental biology and functional anatomy.
The successful applicants will also play an active role in teaching. The Department has teaching commitments to medical, dental, physiotherapy, medical laboratory science, pharmacy, physical education and science students. The appointees will teach in two or more of the following areas: cell biology, histology, gross/functional anatomy, reproductive biology, and/or neuroscience, at both the undergraduate and postgraduate level.
The Department will be looking to make one appointment in the area of cell biology and one in functional anatomy (with responsibility for physiotherapy teaching).
Further information about the position and the Department can be obtained from Professsor DG Jones, Department of Anatomy and Structural Biology (Tel: 64 3 479 7364; Fax: 64 3 479 7254; email: gareth.jones-at-stonebow.otago.ac.nz) and/or from the Department's homepage: http://gandalf.otago.ac.nz:800/Anatomy/home-page.html.
Salary of Lecturer (Non-medical): $NZ42,750 - $53,250 Salary of Senior Lecturer (Non-medical): $NZ56,250 - $66,250
Reference number A97/32. Closing Date 4 July 1997.
METHOD OF APPLICATION
Further details regarding this position, the University, and the application procedure, are available from the Deputy Director, Personnel Services, University of Otago, PO Box 56, Dunedin, New Zealand (facsimile 64-3-474 1607 or e-mail laurie.hibbert-at-stonebow.otago.ac.nz).
Applicants should send two copies of their curriculum vitae together with the names, addresses and fax numbers of three referees, to the Deputy Director of Personnel Services by the specified closing date, quoting the appropriate reference number.
If an applicant is short listed for interview, whanau support will be welcome.
Equal opportunity in employment is University policy.
E tautoko ana Te Whare Wananga o Otago i te kaupapa whakaorite whiwhinga mahi.
One of our students is doing a project involving the production of artificial caries on teeth, in vitro. She would like to use Quinoline as one of the stains to help delineate the structure of the lesion but we cannot find any information on what concentration to use. Articles which mention Quinoline do not give any information as to what concentration was used.
To all UK TEM users We have a Philips 301 TEM here at the dept. of Materials Engineering & Materials Design, Nottingham University, that is surplus to our requirements. This instrument has been out of use for several years and we need the space it is taking up for a new ESEM-FEG. The TEM will need some repairs to get it operational again and may be most suitable for spare parts. We urgently need to move this microscope out of the department and are prepared to give it away, if you are prepared to pay removal costs.
All interested parties please contact Ms.Nicola Bock, Dept.of ME&MD,University of Nottingham. Tel: (0115)9513759 Email: emznjb-at-emn1.nott.ac.uk
I use PC-Image Release Beta 1 under Windows NT 4.0. When I change the clipboard size from the default setting 400k in Options/Preferences NIH-image terminates. What might be the reason? Any ideas?
We are interested in purchasing a portable multi-media projector to present microstructures, either in form of digital images or in form of .avi (or similar) movie clips from in-situ SEM or TEM analysis. Is the expense to go from Polysilicon to DLP technology worthwhile? How portable are they really, i.e. how easy it is to set up the display at any location? How much of the details of an image is lost, if any?
Thanks for the help Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
Hi guys, Does anyone know of any shareware that will allow me to convert quicktime movies into individual frames. Thanks in advance,
--Ciprian Have fun and keep the sun on your back and a smile on your face. __________________________________________________________ Ciprian A. Almonte University of Pittsburgh Center for Biologic Imaging Pittsburgh, PA 15261
Visit my web site at http://www.pitt.edu/~calmonte Laboratory's website: http://sbic6.sbic.pitt.edu __________________________________________________________
You could try GifBuilder - this is used for making animated GIF89 image files, but it can read in QuickTime movies and export single frames as GIF files (which can then be converted to other formats with Photoshope, etc...). You can find info and download the program at
http://member.aol.com/royalef/toolbox.htm
This has links to other software as well. Hope it's useful,
Rick Powell
****************************************************************** * NANOPROBES, Incorporated | Tel: (516) 444-8815 * * 25 East Loop Road, Suite 124 | Fax: (516) 444-8816 * * Stony Brook, NY 11790-3350, USA | nano-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
Our department has 2 academic positions currently advertised of which it is intended that one will be filled by a person with an active interest in cell biology and in particular the development of microscopy related research techniques in cell biology (ie Confocal Microscopy, Electron Microscopy, Image Processing and Anaylsis). A copy of the advertisement was posted yesterday;
LECTURERS IN ANATOMY AND STRUCTURAL BIOLOGY UNIVERSITY OF OTAGO DUNEDIN NEW ZEALAND
Sorry, the institution at which these positions were available was not posted yesterday.
I need help fighting the bean counters! We are currently working our way thru a new cost recovery evaluation. Based on the results off this evaluation I am being asked to raise my rates for Basic TEM and SEM services to a level which seems outrageously high to me. I would like to find out what other labs are charging for scope time, as well as basic sample preperation. If you don't want to post this information to the listserv you could E-mail me directly at fnksd1-at-aurora.alaska.edu Thank you! Kim
I just borrowed an Apollo system for a local microscopy meeting. (Sorry, I don't know the model number or where they bought it.) We used it to interactivelly display a Photoshop 4.0 demonstration (1024x780). It has a remote mouse joystick with a laser pointer in it. The mouse operated from about 25-30ft by pointing it at the screen and bouncing it into the unit. We operated it off a laptop. We also used it to play output from a VCR unit onto the screen with sound. (BTW, I highly recommend MSA's LAS speaker, Conly Rieder to local societies-He comes with a great understandable video presentation with sound.) The unit weighed about 25 pounds and was easily transported by its handle. ( I carried it a long way across a parking lot and also carried it on a bus without any trouble or pausing for breath.
This thing was just fantastic. I understand that the price tag is about $13,000 though.
Let me know if you need details, I'll try to get them from our computer guys.
- -Scott Walck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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I just borrowed an Apollo system for a local microscopy meeting. (Sorry, I don't know the model number or where they bought it.) We used it to interactivelly display a Photoshop 4.0 demonstration (1024x780). It has a remote mouse joystick with a laser pointer in it. The mouse operated from about 25-30ft by pointing it at the screen and bouncing it into the unit. We operated it off a laptop. We also used it to play output from a VCR unit onto the screen with sound. (BTW, I highly recommend MSA's LAS speaker, Conly Rieder to local societies-He comes with a great understandable video presentation with sound.) The unit weighed about 25 pounds and was easily transported by its handle. ( I carried it a long way across a parking lot and also carried it on a bus without any trouble or pausing for breath.
This thing was just fantastic. I understand that the price tag is about $13,000 though.
Let me know if you need details, I'll try to get them from our computer guys.
- -Scott Walck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I just borrowed an Apollo system for a local microscopy meeting. (Sorry, I don't know the model number or where they bought it.) We used it to interactivelly display a Photoshop 4.0 demonstration (1024x780). It has a remote mouse joystick with a laser pointer in it. The mouse operated from about 25-30ft by pointing it at the screen and bouncing it into the unit. We operated it off a laptop. We also used it to play output from a VCR unit onto the screen with sound. (BTW, I highly recommend MSA's LAS speaker, Conly Rieder to local societies-He comes with a great understandable video presentation with sound.) The unit weighed about 25 pounds and was easily transported by its handle. ( I carried it a long way across a parking lot and also carried it on a bus without any trouble or pausing for breath.
This thing was just fantastic. I understand that the price tag is about $13,000 though.
Let me know if you need details, I'll try to get them from our computer guys.
- -Scott Walck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I just borrowed an Apollo system for a local microscopy meeting. (Sorry, I don't know the model number or where they bought it.) We used it to interactivelly display a Photoshop 4.0 demonstration (1024x780). It has a remote mouse joystick with a laser pointer in it. The mouse operated from about 25-30ft by pointing it at the screen and bouncing it into the unit. We operated it off a laptop. We also used it to play output from a VCR unit onto the screen with sound. (BTW, I highly recommend MSA's LAS speaker, Conly Rieder to local societies-He comes with a great understandable video presentation with sound.) The unit weighed about 25 pounds and was easily transported by its handle. ( I carried it a long way across a parking lot and also carried it on a bus without any trouble or pausing for breath.
This thing was just fantastic. I understand that the price tag is about $13,000 though.
Let me know if you need details, I'll try to get them from our computer guys.
- -Scott Walck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I just borrowed an Apollo system for a local microscopy meeting. (Sorry, I don't know the model number or where they bought it.) We used it to interactivelly display a Photoshop 4.0 demonstration (1024x780). It has a remote mouse joystick with a laser pointer in it. The mouse operated from about 25-30ft by pointing it at the screen and bouncing it into the unit. We operated it off a laptop. We also used it to play output from a VCR unit onto the screen with sound. (BTW, I highly recommend MSA's LAS speaker, Conly Rieder to local societies-He comes with a great understandable video presentation with sound.) The unit weighed about 25 pounds and was easily transported by its handle. ( I carried it a long way across a parking lot and also carried it on a bus without any trouble or pausing for breath.
This thing was just fantastic. I understand that the price tag is about $13,000 though.
Let me know if you need details, I'll try to get them from our computer guys.
- -Scott Walck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I just borrowed an Apollo system for a local microscopy meeting. (Sorry, I don't know the model number or where they bought it.) We used it to interactivelly display a Photoshop 4.0 demonstration (1024x780). It has a remote mouse joystick with a laser pointer in it. The mouse operated from about 25-30ft by pointing it at the screen and bouncing it into the unit. We operated it off a laptop. We also used it to play output from a VCR unit onto the screen with sound. (BTW, I highly recommend MSA's LAS speaker, Conly Rieder to local societies-He comes with a great understandable video presentation with sound.) The unit weighed about 25 pounds and was easily transported by its handle. ( I carried it a long way across a parking lot and also carried it on a bus without any trouble or pausing for breath.
This thing was just fantastic. I understand that the price tag is about $13,000 though.
Let me know if you need details, I'll try to get them from our computer guys.
- -Scott Walck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I hope, that someone on the list can help me to find suitable PC-software (DOS or Windows) or other tools for the following problem: I have to reconstruct the three-dimensional structure of different organs of microarthropods based on serial sections of resin embedded specimen. Up to now I go the time consuming and not exact way of drawing complete transverse, saggital and horizontal series and then to "reconstruct" the 3D-structure more or less "by hand". Any idea is welcome. Heike Dr. Heike Buecking University of Bremen UFT Physiological Plant Anatomy Leobener Str. D 28359 Bremen Germany TEL: +49-421-218-2954 or TEL: +49-421-218-7283 FAX: +49-421-218-3737 e-mail: heibueck-at-uft.uni-bremen.de
} } I use PC-Image Release Beta 1 under Windows NT 4.0. } When I change the clipboard size from the default setting 400k in } Options/Preferences NIH-image terminates. } What might be the reason? Any ideas? } } Reinhard } } I had the same problem with the WIN95 version. I overcame it by manually editing the .ini file where buffer size= XXXX Hope this helps
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171
Polysilicon offers a couple of benefits. The colors are truer, and the units are brighter (higher lumens/$). The DLP technology offers smoother images(colors/tones and edges) without the 'screen door effect' that is inherent with LCD technology(not a defect, just a function of the technology that enables you to see each and every pixel on the screen). This smoothness is more apparent in video with movement than a digital signal.
The units are portable in that they have handles and/or cases(generally with wheels) available. As far as setup goes, they are plug and play, just as if you are plugging a monitor. It is necessary to sync them with a digital signal since computers have different signals. This is most easily done with a checkerboard(black and white) wallpaper pattern.
The image that is projected is the same image/signal that you would see on your monitor. In some cases, depending on the specs of your monitor, it may actually be better. Most projectors have controls to adjust the picture, similar to monitors.
Another opportunity to consider is resolution. The demand for VGA(640 x 480) has fallen off and the current demand is highest for SVGA(800 x 600). There is some demand for XGA(1020 x 768) and SXGA(1280x 1040). Depending on the output of your computers currently and for the foreseeable future, you need to look at this as well. Generally, projectors are downwards compatible in resolution. There is a new technology, 'intelligent' or 'smart' compression that offers display of XGA signals on a unit that is technically SVGA. To keep it simple, this is essentially 'loss-less' compression of the XGA signal. I am aware of it only being available on DLP models.
As I suggest with any imaging system or device, have a dealer(s) bring in the two technologies and see for yourself. Specifications are great to compare but 'a picture is worth...'
John D. Warren Area Sales Manager Digital Products Polaroid Corporation "See What Develops" 4525 Leonard Parkway Richmond, Virginia 23221-1809 804 254 1011 804 254 1013 Fax warrenj1-at-polaroid.com
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We are interested in purchasing a portable multi-media projector to present microstructures, either in form of digital images or in form of .avi (or similar) movie clips from in-situ SEM or TEM analysis. Is the expense to go from Polysilicon to DLP technology worthwhile? How portable are they really, i.e. how easy it is to set up the display at any location? How much of the details of an image is lost, if any?
Thanks for the help Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
Try checking Alcheny Mindworks (Yahoo will turn up the address). If I Recall correctly, they offer something along that line as inexpensive shareware..
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Hi guys, Does anyone know of any shareware that will allow me to convert quicktime movies into individual frames. Thanks in advance,
--Ciprian Have fun and keep the sun on your back and a smile on your face. __________________________________________________________ Ciprian A. Almonte University of Pittsburgh Center for Biologic Imaging Pittsburgh, PA 15261
Visit my web site at http://www.pitt.edu/~calmonte Laboratory's website: http://sbic6.sbic.pitt.edu __________________________________________________________
AutoQuant Imaging Inc. is a world leader in the research and development of deconvolution software products for 3D visualization in biology and ophthalmology. AutoQuant is a growth company with excellent opportunities for future advancement, and is planning a major expansion. We have the following 2 openings at our Troy NY laboratory.
Description: The candidate is expected to have completed his/her graduate degree in Electrical Engineering, Biomedical Engineering, or a closely related field from an accredited university. A background is preferred in biomedical imaging, although strong candidates with other qualifications listed below will be considered as well. All levels of experience will be considered. Responsibilities will be a diversity of three areas having equal emphasis: (1) Basic research, (2) product development, and (3) customer support. The candidate must have strong writing skills and strong interpersonal skills. Industrial experience will be considered a plus. A technical background or desire to work in any of the following areas will be considered a plus: graphical software development using C/C++, MFC and Visual C++, signal processing, image processing, mathematical modeling, optimization methods, detection-and-estimation methods. Although not essential, a background in optics, 3D microscopy, confocal microscopy, ophthalmic imaging, or radiological imaging (MR, PET, CT) will be considered a plus as well. The ideal candidate will be a self-starter, self-motivated, highly responsible, organized, capable of being self-managed and will require very little supervision. He/she will be diverse and capable of handling a multitude of projects, tasks and responsibilities at once.
The 2 openings are as follows: (1) R&D Scientist: An individual having a Ph.D. or equivalent degree and matching the above description. This individual will be responsible for leading new research initiatives, creating new ideas and for applying for external R&D contracts. A strong publication record or record of presentation at professional conferences is expected. (2) R&D Engineer: A creative and resourceful individual having a Masters degree and matching the above description. A strong publication record or industrial experience will be a plus, although not absolutely required.
AutoQuant is located adjacent to the Rensselaer Polytechnic Institute campus and collaborates in research with the Rensselaer Polytechnic Institute, the Wadsworth Center for Laboratories and Research and the Albany Medical College. The R&D Scientist will be expected to develop similar collaborations with these or other collaborating institutions. The opportunity exists for the R&D Engineer to pursue a Ph.D. degree on a part time basis.
The City of Troy is part of the historic tri-city Capital District of Upstate New York. The area features numerous recreational opportunities for the family, including good dining, the arts, music, hiking, wilderness, water sports and winter skiing nearby. This area is in a valley lying among the Adirondack, Catskill and Berkshire Mountains. Boston and New York City are 2-1/2 hour drives. Rochester, New York and Montreal are 4 hour drives. The area is noted for its many outstanding school districts.
Salary and benefits are competitive and commensurate with training and experience. AutoQuant is an equal opportunity employer, and will not discriminate based on race, color or national origin.
Please mail or fax your resume, a brief description of interests and the contact information for at least 3 references. If you respond by E-mail, please be sure to follow the E-mail with a regular mailing or fax. Please avoid encoded E-mail attachments. Respond to:
Human Resources AutoQuant Imaging, Inc. 1223 Peoples Ave. fax: 518-276-6380 Troy NY 12180-3536 E-mail: holmes-at-aqi3.aqi.com
It sounds like the Neurolucida software from MicroBrightField would handle your problem very well. You can get more information from them at info-at-microbrightfield.com or from their web site at www.microbrightfield.com/microb. You can also correspond with me; I know the system's details intimately.
On Thu, 22 May 1997, Heike Buecking wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I hope, that someone on the list can help me to find suitable PC-software } (DOS or Windows) or other tools for the following problem: } I have to reconstruct the three-dimensional structure of different } organs of microarthropods based on serial sections of resin embedded } specimen. Up to now I go the time consuming and not exact way of drawing } complete transverse, saggital and horizontal series and then to } "reconstruct" the 3D-structure more or less "by hand". Any idea is } welcome. } Heike } Dr. Heike Buecking } University of Bremen } UFT } Physiological Plant Anatomy } Leobener Str. } D 28359 Bremen } Germany } TEL: +49-421-218-2954 or } TEL: +49-421-218-7283 } FAX: +49-421-218-3737 } e-mail: heibueck-at-uft.uni-bremen.de }
Edmund Glaser, D. Eng. Dept. Physiol. Univ. Md. School. Med. Baltimore, MD 21201 USA Ph: (410) 706-5041 Fax: (410) 706-8341
Dear Colleagues, I have another question regarding to LR White: I am doing some immunostaining on semithin sections embedded in LR White. Although I know it is not absolutely necessary to remove LR White before the staining, I wonder whether there is an effective way to dissolve the cured resin but do not damage the tissue's fine structure. I faintly recall someone mentioned 70% alcohol can do the job. Is this correct? Is there other way which can be more effective? Thank you in advance for the help. Best regards, Yuhui Xu
I have been trying to locate a source for purchasing Lacomit Varnish and the Solvent and I haven't met with too much success. The one company I found does not have it in stock and has absolutely no ideal when they could get it. They said anywhere from now to maybe even next year. I have been waiting since last February and I think that is now long enough. Do any of you know where I can get it and is it in stock. I really don't want to place any more orders unless I know when I can get it. Thanks so much.
Peggy 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
Heike Buecking wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I hope, that someone on the list can help me to find suitable PC-software } (DOS or Windows) or other tools for the following problem: } I have to reconstruct the three-dimensional structure of different } organs of microarthropods based on serial sections of resin embedded } specimen. Up to now I go the time consuming and not exact way of drawing } complete transverse, saggital and horizontal series and then to } "reconstruct" the 3D-structure more or less "by hand". Any idea is } welcome. } Heike } Dr. Heike Buecking } University of Bremen } UFT } Physiological Plant Anatomy } Leobener Str. } D 28359 Bremen } Germany } TEL: +49-421-218-2954 or } TEL: +49-421-218-7283 } FAX: +49-421-218-3737 } e-mail: heibueck-at-uft.uni-bremen.de
We use the VoxBlast software to do just what you are asking for. It works well for us. You can get more information from the WWW site http://www.vaytek.com
Held alongside the CYTO 97 meeting, CRYO 97 promises to be a comprehensive update on the state of the art in Low Temperature Microscopy and Analysis. Come to York in July and enjoy a taste of both meetings and the large joint Trade Exhibition.
(this programme is easier read - I hope - if you maximise your email reader's window!)
CRYO 97 - Low Temperature Microscopy & Analysis - Programme
Sunday 6 July 12:00 - 14:00 REGISTRATION P.Monaghan Inst of Cancer Research, Sutton Introduction
SESSION A: SPECIMEN PREPARATION E Kellenberger Univ of Lausanne Retro-and perspectives of cryotechniques applied to sections: beliefs, dreams, misconceptions expectations, surprises and disappointments! M Miller To be advised. M Thijssen Wageningen Agricultural Univ Comparison of embedding fluids for high pressure freezing of Petunia ovules folled by freeze substitution. J Lepault CNRS, Gif surYvette Electron cryo-microscopy of biological tissues. D Studer Univ of Berne High pressure freezing of thick specimens.
Monday 7 July SESSION B: CRYO-MICROSCOPY H Saibil Birkbeck College, London To be advised. B Gowen EMBL, Heidelberg Visualising virus/vesicle fusion by time-resolved cryo-TEM. H Delacroix Univ P et M Curie, Gif sur Yvette Crystallographic analysis of freeze-fracture electron micrographs: application to the structure determination of cubic lipid systems. R Henderson MRC Lab of Mol. Biol., Cambridge Atomic resolution structure determination of biological macromolecules by electron microscopy. B Papahadjopoulos- Univ of the Pacific, USA Electron microscopic investigations of the fine-structure of lipid-coated DNA fibrils formed during Sternberg cationic liposomes/DNA interaction. N Atkin Univ of York Determination of salt-induced gellan polymer conformation by ultra-rapid freezing and low angle rotary shadowing. M van Heel Imperial College, London To be advised. D Studer Univ of Lausanne Electron beam induced changes in frozen hydrated specimens. S Butcher MRC Inst of Virology, Glasgow To be advised.
Tuesday 8 July SESSION C: CRYO-SEM W Jongebloed Univ of Groningen Cryo-preparation versus tannic acid/arginine/osmium tetroxide non-coating preparation observed with field emission SEM. J Nijsse Wageningen Agricultural Univ Cryo-SEM observations on frozen hydrated lettuce seeds during inhibition. K Robinson British Antarctic Survey Combination - To be confirmed. SESSION D: ELEMENTAL ANALYSIS G Roomans Univ of Uppsala Cryotechniques for preparation of tissue for X-ray microanalysis. P Echlin Univ of Cambridge Low temperature, low voltage microanalysis of diffusible elements in bulk frozen hydrated bio-organic samples. B Wolf Univ Freiburg Drug targeting and metabolic investigations on tumour cells with TEM-EELS analysis on cryo-prepared material. H Elder Univ of Glasgow Parameters for optimally freeze-drying cryosections of muscle for ultrastructural and microanalytical studies. A Pogorelov State Univ at Pushchino X-ray microanalysis of potassim deficit in cardiomyocyte induced by ischemia. I Karydis Univ of Cambridge Elemental changes presage apoptosis in human monocytes challenged with oxidised low density lipoprotein. SESSION E: APPLICATIONS L Edelmann Univ des Saarlandes Electron microscopy of freeze-dried biological material. I Lambrichts Limburgs Univ Centrum Cryomicroscopy of human periodontal ligament cells. N Hajibaheri ICRF, London High pressure frozen/freeze-substituted yeast and mammalian cells/tissue for immunocytochemistry. S Brookes Inst of Animal Health, Pirbright Cryo-electron microscopy of African swine fever virus: a novel pathway of virus morphogenesis as particles are wrapped by the Endoplasmic Reticulum. R Hermann ETH Zurich Structural and immunological studies of the nematode Heterorhabditis sp. and its host bacterium Photorhabdus luminescens, employing enhanced sampling techniques, followed by high pressure freezing, freeze substitution and plastic embedding.
POSTER PRESENTATIONS PC01 P Bennett Structural changes in samples cryo-fixed by contact with cold metal block. PC02 H Ekwall Cryo-electron microscopy of frozen boar semen PC03 B Martin Controlled freeze-drying for the preservation of fungal extracellular matrices PC04 E Shimoni A new micro biopsy system for high pressure freezing. PC05 P Wild Cryo-immobilisation and freeze-substitution of cell monolayers. PC06 C Winters A portable freezing and freeze-substitution assembly: anhydrous cryo-preparation in the field of a metal-binding freshwater moss for X-ray microanalysis. PC07 T Ishikawa Structural change of troponin induced by calcium ions are revealed by three dimensional cryo-electron microscopy. PC08 C Moores The interaction between utrophin N-terminus and F-actin: structural determination by cryo-electron microscopy. PC09 S Stoylova Structural of native PSII complex. A cryo-electron microscopy and crystallography study. PC10 G Daniel Use of HR-cryo-FE-SEM for studies on microbial degradation of wood and wood fibre structure. PC11 A Minnocci LTSEM and EDX X-ray microanalysis of wheat protoplast: a possible model for environmental stress investigations. PC12 A Minnocci LTSEM stomatal analysis in olive plants exposed to ozone. PC13 R Petacchi LTSEM sensilla studies on Dicyphus (D) erran (Wolff) (Heteroptera: Miridae). PC14 K Robinson An application of LT-SEM: how do hitch-hiking mites attach to coastal sand-hoppers? PC15 K Robinson Application of low temperature scanning electron microscopy: how eggs of a tardigrade respond to humidity changes. PC16 A Johnson Cryoprocessing for microanalysis of magnesium in the heart. PC17 A Morgan Hyperbaric freezing of invertebrate tissues: morphology and X-ray mapping of ion-transporting and metal-sequestering epithelia PC18 A Scully Microanalysis of freeze-substituted dentine materials. PC19 L Tetley The use of cryotechniques and energy filtering TEM for improved 3D cellular reconstruction: novel structure revealed in coccidian parasite infective stages. PC20 A Warley Freeze-drying and the preparation of cryosections for electron probe X-ray microanalysis. PC21 E Zellmann Cryo in-column EFTEM for free choice of detectors. PC22 D Carter Slam-frozen bone mineral. PC23 K Hovananyan Cryo-ultramicrotomy for electron microscopy study relation entamoeba with bacteria and liposomes. PC24 K Jennings Characteristics of beta amyloid peptide: a combined TEM, AFM and SEM study employing ambient and cryo-preparative techniques. PC25 C Milanesi New tips for freeze-substituting and immunogold labelling pollen grains from different species. PC26 N Perusinghe Ultrastructural immunocytochemistry of gap junctions in lactating mouse mammary gland. PC27 E L Punnonen Immunolabelling of 46 kDa mannose 6-phosphate receptor on cryo-sections using antibodies against the luminal domain and the cytoplamic tail. PC28 L Tetley Characterisation of polymeric glycol chitosan - a new drug delivery system - using cryo electron microscopy
Dr Laurence Tetley IBLS EM Centre Joseph Black Building University of Glasgow Glasgow G12 8QQ
We have a customer that is having difficulties with reembedding and would like some help. The following is what our customer wants to do: They want to reembed epon sections cut at 1-5 microns so that they can be resectioned at 60 nm. What is the best way to do this? The most effective with the least time and the fewest steps. Is thermanox the best material on which to collect these sections? Is there a mounting medium, or some other technique that will seal the sections to the Thermanox, but not create a thick layer on top of the section? Will the section sealed to the Thermanox actually cut at 60nm or will it just come off the Thermanox at some point? If the section on the Thermanox is inverted onto the top of a beem capsule fulled with unpolymerized epon and baked, how can the Thermanox be removed so that the section remains on the epon side? Will this procedure allow the section to remain flat and close to the surface during baking? Is there a reference that describes a useful protocol for this technique? I look forward to all responses.
} I have been trying to locate a source for purchasing Lacomit } Varnish and the Solvent and I haven't met with too much success. The one } company I found does not have it in stock and has absolutely no ideal when } they could get it. They said anywhere from now to maybe even next year. I } have been waiting since last February and I think that is now long enough. } Do any of you know where I can get it and is it in stock. I really don't } want to place any more orders unless I know when I can get it. Thanks so } much. } } } } Peggy Bisher
I would have expected most of the EM consumables suppliers to have - try SPI or Agar.
Dear all, We have had someone recently contact us regarding the use of a photographic-fixer rejuvenating service run by a company called Continu-Fix. I believe it is Canadian based. Essentially what is being offered is, we send our used film and paper fixes to this outfit, they will run it through some sort of refining process and then we buy back the fix as 'good as new' fixer. This then saves heaps of used fixer going down the drain and out into the environment, and saves us buying in new fix in 'un-environmentally friendly' bottles.
What we would like to hear from people regarding this is;
Is there any EM/LM/photo labs out there using this process or something similar? Do you believe that it is cost effective? Does the fix still have adequate fixing properties for archival quality? (ie; are we going to find that 6 months/6 years down the track, people will be coming back to us with black/brown prints).
We would appreciate any comments from anyone who has used this process,
Thanks in advance,
Rich.
----------------------------------------------------------------------- Richard Lander Senior Technician South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
American Lab/News has provided an opportunity for us to review any new equipment/software/etc. which will be exhibited at the upcoming Microscopy & Microanalysis meeting. If you did not receive our fax, please contact our offices *immediately* and/or send words (50-75) and images (35 mm slides or prints) ASAP. We go to press next week.
Look forward to hearing from you. Barbara Foster, editor "Focus on Microscopy" c/o MME 53 Eton Street Springfield, MA 01108 (413)746-6931 fax: (413)746-9311 email: mme-at-map.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear colleagues,
A working group of the Dutch Society for Microscopy is collecting information on accreditation of microscopical work.
We would like to have information on:
1 Organisations that set formats for accreditation and certification in microscopy 2 certified standards for microscopy callibration 3 laboratories in microscopy that work under a accreditation regime yet 4 other interested parties
Thanks in advance,
Marcel Paques (chairman working group) Dutch Society for Microscopy
A working group of the Dutch Society for Microscopy is collecting information on accreditation of microscopical work.
We would like to have information on:
1 Organisations that set formats for accreditation and certification in microscopy 2 certified standards for microscopy callibration 3 laboratories in microscopy that work under a accreditation regime yet 4 other interested parties
Thanks in advance,
Marcel Paques (chairman working group) Dutch Society for Microscopy
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } It sounds like the Neurolucida software from MicroBrightField would } handle your problem very well. You can get more information from them } at info-at-microbrightfield.com or from their web site at } www.microbrightfield.com/microb. You can also correspond with me; I know } the system's details intimately. } } On Thu, 22 May 1997, Heike Buecking wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } I hope, that someone on the list can help me to find suitable PC-software } } (DOS or Windows) or other tools for the following problem: } } I have to reconstruct the three-dimensional structure of different } } organs of microarthropods based on serial sections of resin embedded } } specimen. Up to now I go the time consuming and not exact way of drawing } } complete transverse, saggital and horizontal series and then to } } "reconstruct" the 3D-structure more or less "by hand". Any idea is } } welcome. } } Heike } } Dr. Heike Buecking } } University of Bremen } } UFT } } Physiological Plant Anatomy } } Leobener Str. } } D 28359 Bremen } } Germany } } TEL: +49-421-218-2954 or } } TEL: +49-421-218-7283 } } FAX: +49-421-218-3737 } } e-mail: heibueck-at-uft.uni-bremen.de } } } } Edmund Glaser, D. Eng. } Dept. Physiol. } Univ. Md. School. Med. } Baltimore, MD 21201 USA } Ph: (410) 706-5041 } Fax: (410) 706-8341 } }
We have a customer that is having difficulties with reembedding and would like some help. The following is what our customer wants to do: They want to reembed epon sections cut at 1-5 microns so that they can be resectioned at 60 nm. What is the best way to do this? The most effective with the least time and the fewest steps. Is thermanox the best material on which to collect these sections? Is there a mounting medium, or some other technique that will seal the sections to the Thermanox, but not create a thick layer on top of the section? Will the section sealed to the Thermanox actually cut at 60nm or will it just come off the Thermanox at some point? If the section on the Thermanox is inverted onto the top of a beem capsule fulled with unpolymerized epon and baked, how can the Thermanox be removed so that the section remains on the epon side? Will this procedure allow the section to remain flat and close to the surface during baking? Is there a reference that describes a useful protocol for this technique? I look forward to all responses. Stacie Kirsch EMS
Hello all reembedders,
I have been reembedding 1-5 micron epon sections for years in the following way:
I section the tissue at 1-5 microns(the thicker the better) as if using for LM. That is I pick up the sections and place them on a drop of water on a glass slide. I heat the section as usual on a hot plate to afix it to the glass. After the slide cools off, I carefully scrape the section off the slide with a razor blade. I immediately pick the section up with tweezers and superglue the section onto a blank epon block. The blank block must be smooth, flat, and clean of course. I let the superglue dry well, and then the tissue is ready to be sectioned again. I have successfully then gotten ultra thin sections in the usual way.
Others in our lab have done the above, except they use epon itself rather than superglue to afix the 1-5 micron section to the blank block. I don't like this method because it takes longer ( you have to put the block combo back in the oven to polymerize ). This method is also good for those more patient than I.
Hope this works well for you. Hope I didn't leave anything out of the technique.
Sections can be collected on untreated glass microscope slides, dried on a hotplate, stained with toluidine blue if desired, then covered with an inverted BEEM capsule (the small ones work best) filled with unpolymerized resin. Place them in a 90C oven for 60 - 90 minutes, then peel the blocks off while the blocks and slides are still hot. If the slides cool off, put them back into the oven for 5 - 10 minutes and try to peel the blocks off again. The blocks may need a little more time in the oven to fully polymerize after they've been removed from the slides.
The tricks are to use plain microscope slides (no poly-L-lysine, silanes, or acid cleaning) and to remove the blocks while the plastic is still slightly soft.
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064
We have routinely used parlodion coated grids for our EM thin sections. 2.5% parlodion in amyl acetate dropped on to distilled water, then lowered on to the grids and dried. However, lately we are having problems with uneven thickness. We have tried new amyl acetate and different percentages of parlodion but still no luck on solving the uneven thickness.
Any ideas?! Or protocols that work for you?
Thanks in advance for any help.
Bob Underwood Morphology Core U of Washington Seattle, USA
If I am not mistaken, a lab on the floor above us has the Polaroid Sprintscan (it is definitely a Polaroid, whether it is the same model Sprintscan I don;t know). The resolution is fine. It is easy to use. It does not advance the film automatically. It is slow at 2700 DPI, but otherwise decent speed.
It only take 35 mm or smaller, so it does not meet your specified needs. It can be used, if you cut a custom size metal insert or buy one from a company that has been advertising heavily (but I forgot the name anyway) to scan microscope slide.
-------------------------------------------- email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 --------------------------------------------
The following message which was sent by Analytical Imaging Facility {aif-at-telico.bioc.aecom.yu.edu} had incomplete information concerning Polaroid scanners, so I thought I'd respond even though I have no financial interest in Polaroid. The SprintScan 45 is a multi-format film scanner which scans film as large as 4x5 inch to 35 mm format. The SprintScan 35 is a scanner for 35 mm format films only. See Polaroid's web page at http://www.polaroid.com/products/scanners/index.html. } } If I am not mistaken, a lab on the floor above us has the Polaroid } Sprintscan (it is definitely a Polaroid, whether it is the same model } Sprintscan I don;t know). The resolution is fine. It is easy to use. } It does not advance the film automatically. It is slow at 2700 DPI, but } otherwise decent speed. } } It only take 35 mm or smaller, so it does not meet your specified needs. } It can be used, if you cut a custom size metal insert or buy one from } a company that has been advertising heavily (but I forgot the name } anyway) to scan microscope slide. } } -------------------------------------------- } email sent from an account of the Analytical Imaging Facility } The Albert Einstein College of Medicine of Yeshiva University } 1300 Morris Park Ave. Bronx, NY 10461 } (718) 430-2890 FAX: (718) 430-8996 } --------------------------------------------
Russell E. Cook Scientific Associate 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
} We have routinely used parlodion coated grids for our EM thin sections. } 2.5% parlodion in amyl acetate dropped on to distilled water, then lowered } on to the grids and dried. However, lately we are having problems with } uneven thickness. We have tried new amyl acetate and different } percentages of parlodion but still no luck on solving the uneven } thickness.
Several causes of uneven thickness in Parlodion (Collodion, cellulose nitrate, etc) are:
1. too rapid evaporation rate of the solvent carrying the parlodion on the water surface (possibly due to the water being too warm, or the room too warm, or air currents - as from a fume hood - blowing over the water surface),
2. incomplete dissolution of the parlodion in the amyl acetate (possibly due to not waiting long enough to dissolve, or slightly moist parlodion flakes/chips),
3. the parlodion solution and the water should be the same temperature (20-24 deg C).
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Marcel Paques wrote: ========================================== } A working group of the Dutch Society for Microscopy is collecting } information on accreditation of microscopical work. } } We would like to have information on: } } 1 Organisations that set formats for accreditation and certification
} in microscopy } 2 certified standards for microscopy callibration } 3 laboratories in microscopy that work under a accreditation regime yet } 4 other interested parties ================================================ In the USA, the American Association for Laboratory Accreditation (A2LA) has been accrediting laboratories in "microscopy" under the "chemical" discipline. The accreditation in microscopy started in 1980 and today there are some number of laboratories accredited by A2LA and also, the accreditation today is being done to the standard of ISO Guide 25.
For SEM calibration, there is the NIST Certified Reference Material. But for TEM it is more complicated, and what accredited laboratories end up using seems to be determined in part by the preferences of the one doing the assessing. The two main approaches are either a) the use of a diffraction grating from which one figures out the precise number of lines from the optical properties and b) dimensional samples, such as calibrated spheres that are reported traceable to NIST.
Are there laboratories actually operating under this accreditation regimen? Absolutely! Are there very many? Not too many. An up-to-date list can be obtained through A2LA:
American Association for Laboratory Accreditation 656 Quince Orchard Rd. #620 Gaithersburg, MD 20878-1409 301-670-1377, FAX 301-869-1495 http://www.a2la.org/ E-mail: You can contact them through their website above
Internested parties? Other than A2LA above, ACIL would certainly fall into that category. You can reach ACIL at {http://www.acil.org/} , which is the professional and trade association of independent testing, analytical and engineering laboratories in the USA.
One might ask whether this program "works". Yes, it does work and it works very well. And I speak from first hand experience since the laboratory part of our business has been accredited by A2LA since the inception of their program in 1980.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I am collecting information on the currently valid standards applied in metallographic examinations in different countries and corresponding to the following standards of ASTM;
A247 - 67 Test Method for Evaluating the Microstructure of Graphite in Iron Casting
E3 - 95 Preparation of Metallographic Specimens
E7 - 96 Metallography
E45 - 95a Determining the Inclusion Content of Steel
E1245 - 95 Determining the Inclusion or Second-Phase Constituent Content of Metals by Automatic Image Analysis
E1268 - 94 Assessing the Degree of Banding or Orientation of Microstructures
E1382 - 91 Determining Average Grain Size Using Semiautomatic and Automatic Image Analysis
E1558 - 93 Electrolytic Polishing of Metallographic Specimens
ISO 9042 - 88 Steels: manual point counting method for statistically estimating the volume fraction of a constituent with a point grid.
JIS G 0555 - 77 Microscopic Testing Method for the Non-metallic Inclusions in Steel
If such information is available kindly send me the data arranged in the following sequence; No. of standards according to ASTM, ISO, JIS - number of the corresponding national standard, its title (English and national), date of publication and the date of last revising.
If you think that in the list given above someother relevant standards have been omitted, kindly let me know their numbers and titles.
Thanking you in advance for your kind co-operation and assistance, I remain
yours sincerely
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Structural and Physical Research Laboratory
Recently there was discussion of Aclar film and its myriad uses, including reembedding thick sections. I recently bought some from ProSciTech (haven't used it yet) and it was said to be available from Electron Microscopy Sciences. From memory, the Aclar is cut into slide size pieces and used instead of an ordinary slide. The section is stained as usual, then inverted over an embedding mould with resin and cured. The Aclar can be sectioned, is light and electron lucent and is chemically inert. The ProSciTech web page has some info and references about Aclar: http://www.proscitech.com.au/l35.htm#l105. Diana
Diana van Driel Dept Ophthalmology Sydney University C09 AUSTRALIA 2006
Here are some references which describe the methods for re-sectioning semi-thin (or thick) resin sections. Our procedure included : (1) prepare 4um thick sections with a histo diamond knife and mount on microscope slides and stain with toluidine blue. (2) stick re-faced resin blocks on the mounted sections with a drop of epon-araldite or crazy glue. (3) leave in oven for 2 days at 60C (4) "pop off" the blocks after leaving on dry ice for 45 minutes.
References : Di Sant'Agnese & De Mesy-Jansen (1984). Ultrastruct. Pathol 6: 247-253
Milroy (1985). J. Electron Microsc. Tech. 2: 399-400
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We have a customer that is having difficulties with reembedding and would } like some help. The following is what our customer wants to do: } They want to reembed epon sections cut at 1-5 microns so that they can be } resectioned at 60 nm. } What is the best way to do this? The most effective with the least time and } the fewest steps. } Is thermanox the best material on which to collect these sections? } Is there a mounting medium, or some other technique that will seal the } sections to the Thermanox, but not create a thick layer on top of the } section? } Will the section sealed to the Thermanox actually cut at 60nm or will it just } come off the Thermanox at some point? } If the section on the Thermanox is inverted onto the top of a beem capsule } fulled with unpolymerized epon and baked, how can the Thermanox be removed so } that the section remains on the epon side? Will this procedure allow the } section to remain flat and close to the surface during baking? } Is there a reference that describes a useful protocol for this technique? } I look forward to all responses. } } Stacie Kirsch } EMS }
We are interested in adding a 2k x 2k image digitiser to a non digital SEM. If you know of such a system, could you advise us of the supplier, and, preferably, the price? 1k x1k systems are not appropriate.
thanks,
Mel Dickson Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
We have a JEOL 200CX which is currently running on Freon. Are there any points or pitfalls which we should know about before we undertake a conversion.
Thanks in advance.
Regards,
Keith. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Dear Mel, I have the Quartz PCI system, which captures images up to 4096 X 4096, depending upon the output of the SEM. They can hook up to any old SEM and uses a Pentium computer for display. It costs about $15,000CAN. In Canada the supplier is NSC, the Hitachi microscope supplier. You can contact the Quartz company at adbrown-at-qrtz.com (Andrew Brown) to ask about Australian suppliers. The lastest version does lots of neat things. You wrote: } We are interested in adding a 2k x 2k image digitiser to a non digital SEM. } If you know of such a system, could you advise us of the supplier, and, } preferably, the price? 1k x1k systems are not appropriate. } } thanks, } } Mel Dickson
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I believe that ELI, a company here in Belgium, has what you are looking for. They have a very good product for your application and they are reliable. I believe that they sell worldwide and you can contact them for more info via E-mail at: orion-at-infoboard.be
The Sales Mgr is Paul Vanderlinden.
Disclaimer: I am not affiliated nor have any interest with that company, I am just a satisfied customer of one of their products.
Good luck and best 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. ****************************************************
There is a very good, reliable, well designed DISS system from German company Point Electronic. System consists from one specially designed PCB installed in standard PC with enough configuration. Can scan with adjustable resolutions from 64x64 till 4096x4096 pixels. It is mains-locked and stops on each scan point integrating signal. PCB has 8 video inputs and 6 digital TTL inputs for xray maps etc. allowing simultanous aquisition. Digitizing goes with 12 bit a/d converter. The newest software is very clever design and runs under windows as TWAIN driver - so for image manipulation one uses any PhotoStyler, Shop, etc software and selects for Aquire-command the SEM driver - it works excellent. Price range fro system is below 30.000 DM but includes PC, instalation (interfacing to SEM) etc. - in case of Australia will be probably different. There are few hundreds of such systems allready running around Europe. - contact is Dr W.Joachimi Point ElectronicGmbH Koethener Str 34, 06118 Halle/Saale, Germany phone: (+49 345) 202 3996 fx: (+49 345) 4235254
kind regards Krzysztof M. Herman (we are polish distributor of Point Electronic) LabSoft S.c. 05-500 Piaseczno, 21 Kosciuszki Str. Poland tel/fx: (48 22) 7502024, 7502028, 7570671 fax only: (48 22) 483787, Email: labsoft-at-ikp.atm.com.pl
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My method is very similar to that used by Jane. When you have found the section of choice cut the slide into small pieces and lay the piece, with the section, on top of a Beem capsule slightly overfilled with resin then polymezire. Remove the block from the Beem capsule, briefly dip the glass into liquid nitrogen which will then fall away leaving block with section attached. Ian.
Wanting to control the presence of esterified pectins in plant cells I've tried the hydroxyalmine/ferric chloride reaction as described by Reeve (1959). I suppose this reaction is routinely used by many of us but.... I have a precipitation problem when I add to the fresh hydroxylamine reagent - i.e. a solution of equal volumes of sodium hydroxide (14g in 100ml EtOH 60%) and hydroxylamine hydrochloride (14g in 100ml EtOH 60%) - concentrated HCl in EtOH in order to acidify the reaction mixture. The precipitate I obtain is of cristalline structure and rather sticky... If you met this problem to when following exactly Reeve's protocole, if you were lucky in solving this problem or if you were not, etc... Please help ! Thanks Pascal
************************************ Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 Fax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ************************************
In a message dated 97-05-26 23:39:41 EDT, M.Dickson-at-unsw.edu.au (Melvyn Dickson) writes:
} We are interested in adding a 2k x 2k image digitiser to a non digital SEM. } If you know of such a system, could you advise us of the supplier, and, } preferably, the price? 1k x1k systems are not appropriate. } }
You might want to give EOS and/or Evex Analytical (609-252-9192) a call. I know Evex has put together a harware & software package including Image Pro. The hardware is 4k by 4k, 12 bit, active imaging system. I believe the PCI system is passive
The price is $9,000 to $20,0000 depending on system configuration.
} There is a very good, reliable, well designed DISS system from German } company Point Electronic.
In the interests of not boring the rest of the subscribers to tears, I thought we vendor types were going to refrain from posting spec sheets to the whole list in response to requests for information.
Let's agree to send this kind of post just to the person making the request, please, and the list will be more enjoyable and useful for all of us...
Hello from Polaroid -- I am Brooks Corl, Senior Applications Manager.
You are clearly describing the Polaroid "SPRINTSCAN 35" scanner, which scans 35mm transparencies or negatives. There is also another model, "SPRINTSCAN 45", that scans up to 4x5 transparencies or negatives. That was the model, I think, that the original question concerned.
Please check our web site (www.polaroid.com) for more information, or feel free to get back directly to me via cc-mail (corlb-at-polaroid.com) if you have questions I can answer or research for you about these scanners. Thanks!
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If I am not mistaken, a lab on the floor above us has the Polaroid Sprintscan (it is definitely a Polaroid, whether it is the same model Sprintscan I don;t know). The resolution is fine. It is easy to use. It does not advance the film automatically. It is slow at 2700 DPI, but otherwise decent speed.
It only take 35 mm or smaller, so it does not meet your specified needs. It can be used, if you cut a custom size metal insert or buy one from a company that has been advertising heavily (but I forgot the name anyway) to scan microscope slide.
-------------------------------------------- email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 --------------------------------------------
This may be too obvious, but did you make sure the surface of the water that you are dropping the parlodion onto is as level as possible? Uneven thickness occurs if the surface is concave or convex. If the uneven thickness is random or patchy, you probably have a contamination problem.
Karen Pawlowski
On Fri, 23 May 1997, Robert Underwood wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello fellow microscopists, } } We have routinely used parlodion coated grids for our EM thin sections. } 2.5% parlodion in amyl acetate dropped on to distilled water, then lowered } on to the grids and dried. However, lately we are having problems with } uneven thickness. We have tried new amyl acetate and different } percentages of parlodion but still no luck on solving the uneven } thickness. } } Any ideas?! Or protocols that work for you? } } Thanks in advance for any help. } } Bob Underwood } Morphology Core } U of Washington } Seattle, USA } } }
Gatan offers DigiScan, a system that converts your analog SEM to Digital. It will digitize up to 8k x 8k and will do more than that, it will drive the scan coils of your SEM. It is versatile and user friendly. The PCI system is a passive capture system and will not give control of the instrument. We have recently installed DigiScan on our 20 year old AmRay 1000A and it drives like a new microscope!
Gatan has a website which provides information and a demo copy of their micrograph image processing software, Digital Micrograph.
Disclaimer: We working with Gatan on developing remote microscopy but have no financial interest in the product or company.
Jacob
Jacob Bastacky, M.D. Room 116 Donner Laboratory Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
Is there an imaging system or a program specifically made for bone morphometry? We are using a regular multipurpose one presently , but it takes long time and certain measurements we still have to do by hand. Thanking you in advance,
WE are sending, via federal express for am delivery, which should arrive at your offices in Springfield by Wednesday May 28, 1997, a photograph and descriptive copy for our Tacky Dot (tm) slides.
Please include this information in your review of new products to be published in the Microscopy & Microanalysis issue of American Lab/News.
THANKS for The Opportunity to expose our products to your audience!!!
I'm trying to locate David J. Barber, recently at the Physics Dept., Hong Kong Univ of Sci and Tech. I tried his e-mail there and it wasn't delivered. He isn't listed on their www site.
If any one knows where he is please send me a note off line.
Medjet is a public compamy for Refractive Surgery Instruments. Need to hire a person with histology skills (light microscopy, scanning microscopy), computer skills and experience in animal experiments; full-time job. Our phone is (908) 7383990; our fax is (980) 7383984
First, thanks to all of you who responded with suggestions on how to fix the leak in my nitrogen dewar. Most of the responses suggested I use a silicone sealant or an epoxy compound.
I do have a follow-up question though. Any suggestions on what's the best way to remove the flange ?. I think that in order to properly fix the leak I should first remove the flange from the dewar (rather than just applying an epoxy or sealant on the surface). The way I understand it, the bottom part of the flange fits into the dewar and an epoxy is used to seal it in place. In my case the leak seems to be right in this epoxy seal. Is there a proper way to remove this flange without damaging the rest of the flange or the dewar ? I don't think that brute force will do it, but what about heat ? If I heat up the neck of the dewar to burn off the epoxy, will I be damaging insulation in the dewar or in the flange ?
German overseas newscast reported the death of Manfred von Ardenne today. During the '30s he was involved in cathode ray work and later lived in East Germany and did significant cancer research. Not mentioned was his conceptual development of the SEM in the late 1930s. He outlined the underlying principles of SEM operation: an electron probe scanning a small region of the specimen, the emitted electrons are captured, amplified and time-sequentially displayed on a cathode ray tube. Magnification is the ratio between the area scanned and displayed. His was the fantastic notion of a microscope without a magnifying lens. It is interesting to contemplate that the now more complex TEM was built and developed from the mid '30s and was a high performer in the early '60s. It was at that time when the Cambridge group, under Oatley, was able to build the first SEM. A lot of technology had to mature before SEM was possible and the Cambridge group deserves great credit. But the development of several unique concepts of a then quite futuristic instrument is to von Ardenne's enduring credit. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
Dear Mr Mott and all vendors/distributors who trace the user's discussions OK about this.... no problem for future ....but why to read boring things and cry? ... just click delete message like by SUBSCRIBE / UNSUBSCRIBE ones!! the real mess would be to give NON REQUESTED ADVERTISING which we not do. regards Krzysztof M. Herman LabSoft S.c. 05-500 Piaseczno, 21 Kosciuszki Str. tel/fx: (48 22) 7502024, 7502028, 7570671 fax only: (48 22) 483787, Email: labsoft-at-ikp.atm.com.pl
I'd like to chime in on another development of Manfred von Ardenne.
Back in my foundry days, I ran a marvelously engineered piece of machinery; an 800kW electron beam furnace. The heart of this machine was array of independently-controlled beams coming from four von Ardenne-produced guns. The furnace was five stories tall and could melt and purify a ton of tantalum an hour. Absolutely amazing technology.
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
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My wife has finished her class and should be getting her high school certification soon, provided that the rules don't change again. Up to this point, I have compiled the answers we've received from the Microscopy server. This was by far the richest source of responses. I would be happy to provide a copy of the responses from the server to anyone who is interested.
Attached is my wife's thank you. Thank you all very much for your help.
Chuck
P.S.
Dear Answer to a Prayer Scientists,
Thanks for taking the time to answer my survey. In all we had 17 responses from scientists in all walks of life. Many of you had some of the same ideas with slightly different twists. For the most part, people felt the history of science was important in education and research. The ideal classroom was a hands on experimental setting. The nature of science responses were varied just as it is in educational circles. Once again, thanks. Debbie Gilbert
------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
Is it possible to do negative staining (2% PTA) on a suspension of virus -- which has been fixed with 1% glut. in cacodylate buffer? In the past I've only used unfixed materials. Thanx.
John Hardy E.M. Lab City of Hope Medical Center jhardy-at-smtplink.coh.org
First I want to thank everybody who responded to my question on suitable software to reconstruct the 3D-structure of organs (based on serial sections). I think it's a good idea to summarize all the tips I have got the last week. The list below is far from beeing complete (sorry to everybody who thinks that I have forgot something important).
NIH-IMAGE This popular freeware programme for image analyses is originally written for the Mac, but now is also available as Win95 program. Download: http://www.zippy.nimh.nih.gov/ (Mac)or http://www.scioncorp.com/ (Win95) Many informations e.g. online manual, macros, example-files and additional download possibilities can be found at: http://rsb.info.nih.gov/nih-image/ As an example animated reconstructions of plant cells (based on semithin sections) can be found on the homepage of Gary Chinga: http://www.nvg.unit.no/~gary Informations about the NIH-Image mailing list can be found at http://www.soils.agri.umn.edu/infoserv/lists/nih-image/
NEUROLUCIDA Informations about Neurolucida are available at http://www.microbrightfield.com/microb Although Neurolucida is primarily designed for the needs of neurobiologists, it has all the facilities which I need for the planed reconstruction based on serial tissue sections (Data entry by camera or digitizing tablet, alignment, tracing of areas of interest, 3D-modelling and volume measuring).
VOXBLAST Voxblast is available for Unix, Mac and Windows-Sytems. Informations about this software, e.g. instructive demo-versions and a quick-reference-quide can be found at http:/www.vaytek.com/ In order to transfer and prepare (enhancement and alignment of the slices) the images for the use with Voxblast, suitable image analyses software, e.g. NIH-image or ImageProPlus from Mediacybernetics (http://www.mediacy.com/) is needed.
T3D (formerly SLICER) Included in the data-management-package NOESYS of Fortner research. Product informations can be found at http://www.fortner.com/
SLICER DICER Informations about this volumetric data visualization software of Visalogic (available for MAC, Win95 and WinNT) could be found at http://www.visualogic.com/
MONTAGE This 3D reconstruction package is a UNIX-programm, but works also on a PC using the free Unix-system LINUX. It's available by anonymous FTP at ftp://retina.anatomy.upenn.edu:/pub/mont.linux.tgz Data entry is simply by a Summagraphics Digitizing Tablet.
A list of links to the distributors of the above mentioned software and many other programs for image analyses can be found at: http://ddsdx.uthscsa.edu/dig/sites.html
This is all I get up to now from the internet and of course I haven't tested all the programs listed above for their suitability.
Sincerely,
Jens Buecking
--------------------------------------------------------------- Dr. Jens Buecking Tel.: +49-(0)421-218 3745 University of Bremen Fax.: +49-(0)421-218 4042 Department of Biology email: jbueck-at-biologie.uni-bremen.de Leobener Str. - NW2 D- 28359 Bremen Germany ---------------------------------------------------------------
Dr. Heike Buecking University of Bremen UFT Physiological Plant Anatomy Leobener Str. D 28359 Bremen Germany TEL: +49-421-218-2954 or TEL: +49-421-218-7283 FAX: +49-421-218-3737 e-mail: heibueck-at-uft.uni-bremen.de
One of the scientists here wishes to use one micron gold particles as carriers for a plant virus in some transmission studies. He has asked me for suggestions re how to bind the virus to the gold. I don't have any references dealing with this size of gold particle, but have several questions: Can the virus be directly bound to these large gold particles? How? Could the specific antibodies be bound to the gold (as with smaller colloidal gold)? Is there perhaps a commercial supplier of one micron gold conjugated to an anti-rabbit antibody? Or something that will bind a virus?
I am currently looking at catepillars by SEM and would like to do TEM. Does anyone know of a specific stain that could be used prior to embedment to make the nerve cells in the taste receptors stand out? Is there a specific stain that could be used after embedment rather than using UA and PbCitrate? Thanks in advance.
Peace through Christ,
Phil Rutledge, Director e-mail: prutle1-at-gl.umbc.edu Center for Microscopy voice: (410) 455-3582 UMBC Dept. of Biology fax: (410) 455-3875 Catonsville, MD 21228 ///// / / / / /////// /////// / / /////// /////// / / / / / / / / / / / / /////
} Is it possible to do negative staining (2% PTA) on a suspension of } virus -- which has been fixed with 1% glut. in cacodylate buffer? In } the past I've only used unfixed materials. Thanx. }
Yes, this can be done with the following caveats:
1. If the virus was fixed while in suspension (not pelletized or in tissue) you should be OK otherwise the virus will be cross-linked to other virus or tissues in an inseparable pastiche that the beam will not penetrate.
2. The glut/cacodylate should be washed out, if possible, to prevent salt crystals from forming during the drying of the grid. An easy way of doing this would be to float the coated grid first on a suspension of the fixed virus in the glut/cacodylate for several minutes followed by floating the grid on 2% PTA for several seconds.
It certainly wouldn't take much to check it out.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
In message {9704288648.AA864839277-at-smtplink.coh.org} "HARDY, JOHN" writes: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Is it possible to do negative staining (2% PTA) on a suspension of } virus -- which has been fixed with 1% glut. in cacodylate buffer? In } the past I've only used unfixed materials. Thanx. } } John Hardy } E.M. Lab } City of Hope Medical Center } jhardy-at-smtplink.coh.org
Give it a try, but before you set up the neg stain, pellet the virus in a microfuge or ultracentrifuge (whatever it takes to get you enough g's), discard fixative supernatant, and resuspend either in distilled water or very weak buffer, ie, about 0.01 to 0.05 molarity. Otherwise the glut and the buffer will generally clog up the prep., coat the virus and mess things up.
Good luck!
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
I have a questions which has been asked of me in the past: Why is it that there is less charging in a Field emission electron microscope as oppossed to a standard LaB6 or W system?
At about 1keV for most insulators, there is a charge balance condition between the number of electrons coming in (incident current) and the number of backscattered and secondary electrons (backscattered and secondary currents) so that I0= Iback + Isec. The absorbed current is zero (that's why it is an insulator). You can operate a FEG-SEM at lower voltages and get more electrons than with either LaB6 or W thermionic sources. The current available in these decreases with decreasing voltage. With a W source, there just isn't enough current density giving a sufficient signal to noise ratio to do anything. It gets better with a LaB6 and there's plenty with a FEG.
The answer to your question is that there isn't less in a FEG. The charging is dependent on the operating voltage of the microscope. If the voltages are the same in the different microscopes, then the above equation holds. In the W and LaB6 machines, you have to crank up the voltages to see anything in the image and you no longer have the charge balance condition. You can increase the voltage that you can work at by tilting the sample to high angles. For example if the insulators have charge balance at 1 kV 0 deg tilt, then the balance condition will be about 3 kV at 70 deg tilt. This condition can be used in an scanning Auger microscope for insulators if a orthoganal detector is used. (JEOL SAM's)
The question that I have about charging in an SEM is this. If you set up the imaging conditions for the sample not to charge at a high scan rate i.e. TV rate, and then drop it to a slow scan (as you do in an analog machine when recording an image), why does the image now charge?
- -
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} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have a questions which has been asked of me in the past: } Why is it that there is less charging in a Field emission electron } microscope as opposed to a standard LaB6 or W system? } } ...
I can think of only two reasons ... (1) FE guns acquire noise-free images at lower beam currents ... or, (2) FE guns can't deliver higher beam currents. However, conventional guns at the same beam current shouldn't charge more ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
How easily can bone be thin sectioned without demineralization? We are interested in viewing the callus area and osteoblasts and want to preserve this area as best as possible. I would appreciate any information or referral to literature. Thanks. Bill Meek, Ph.D. Meekwd-at-okway.okstate.edu
} I am currently looking at catepillars by SEM and would like to do TEM. } Does anyone know of a specific stain that could be used prior to embedment } to make the nerve cells in the taste receptors stand out? Is there a } specific stain that could be used after embedment rather than using UA and } PbCitrate? Thanks in advance. } } Phil Rutledge
Phil,
In brief, I think the answer is "no". When I was looking at amphipod sensory structures, the cells had to be identified by location, structure, and tracing contacts. LaNO3 could be used to detect intercellular spaces and tight junctions (the LaNO3 was supposed to not diffuse past them), but no specific stains for neurons.
Having said that, you might make a go of it by labelling the nerves, or receptor end-organs with horseradish peroxidase and staining by the usual methods. This requires keeping the beasties alive for a few days to let the HRP diffuse. A steady hand, no coffee, and minute dissecting instruments help.
If they're all dead, you might try the Di-I and Di-O fluorescence methods used in tracing mammal and bird nerves. This would give you a LM image of where the taste bud neurons are, and they could then be sought in the TEM. If it works. I had trouble using these on fish, and as I recall, they didn't work well aside from mammals and birds.
This could all be outdated now, and I would be happy to be corrected!
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
[Apologies if this appears twice, my email program hiccuped.) } I am currently looking at catepillars by SEM and would like to do TEM. } Does anyone know of a specific stain that could be used prior to embedment } to make the nerve cells in the taste receptors stand out? Is there a } specific stain that could be used after embedment rather than using UA and } PbCitrate? Thanks in advance. } } Phil Rutledge
Phil,
In brief, I think the answer is "no". When I was looking at amphipod sensory structures, the cells had to be identified by location, structure, and tracing contacts. LaNO3 could be used to detect intercellular spaces and tight junctions (the LaNO3 was supposed to not diffuse past them), but no specific stains for neurons.
Having said that, you might make a go of it by labelling the nerves, or receptor end-organs with horseradish peroxidase and staining by the usual methods. This requires keeping the beasties alive for a few days to let the HRP diffuse. A steady hand, no coffee, and minute dissecting instruments help.
If they're all dead, you might try the Di-I and Di-O fluorescence methods used in tracing mammal and bird nerves. This would give you a LM image of where the taste bud neurons are, and they could then be sought in the TEM. If it works. I had trouble using these on fish, and as I recall, they didn't work well aside from mammals and birds.
This could all be outdated now, and I would be happy to be corrected!
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
Hello again, Requesting help this time on the repair of our Hitachi H-600 film exchange. The films jam by not falling totally into the slot. When the air compressor moves them forward after the exposure, the front left corner of the plate jams on the support and is wedged in place. The entire camera must be opened, I reach inside (in the dark) and unjam the film. The assembly snaps back into place and seems ok for two or more films and then does it again. Our service contract was not renewed by our sage management team, and Hitachi has not yet returned a call on help over the phone, if help this way is available to us at all. Our in house PP&G people want to use the big hammer and lots of lubricant. HELP Linda Fox lfox1-at-wpo.it.luc.edu
} The question that I have about charging in an SEM is this. If you set up the } imaging conditions for the sample not to charge at a high scan rate i.e. TV } rate, and then drop it to a slow scan (as you do in an analog machine when } recording an image), why does the image now charge?
Because in a slower scan mode the beam dwells longer on the specimen permitting more of a static charge to build up; the charge collects with time.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Generally, FESEMs are operated at an accelerating voltage of around 1kV. At this voltage, most none conductive samples will not charge. Similar charging characteristics will arise at higher voltages regardless of electron gun type.
Steve Joens Standard SEM Section Manager Hitachi Scientific Instruments JOENS_S-at-NISSEI.COM
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I have a questions which has been asked of me in the past: Why is it that there is less charging in a Field emission electron microscope as oppossed to a standard LaB6 or W system?
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ATTENTION ALL IMMUNOHISTOCHEMISTS, IMMUNOCYTOCHEMISTS AND OTHER AFFINITY LABELLERS!!! VOTING IS NOW IN PROGRESS FOR A NEW IMMUNOCYTOCHEMISTRY NEWSGROUP!
The official CFV or CALL FOR VOTES for the proposed new newsgroup sci.bio.immunocytochem has recently been posted to news.announce.newgroups and news.groups. If you are keen to see a newsgroup dedicated to discussion of immunocytochemistry and related topics, then please do vote. All you need to qualify as a voter is a valid e-mail address.
To vote for the new group, use your newsreader to access USENET, find {news.announce.newgroups} or {news.groups} and look for the official posting named {CFV:sci.bio.immunocytochem} (posted on 26th May 1997) The VOTING INSTRUCTIONS and the BALLOT can be found at the end of the CFV. You MUST use the official ballot to vote, and it MUST be posted to the official votetaker for it to be counted. Thank-you.
Amanda Wilson, e-mail {awilson-at-aw.u-net.com} Proponent of the proposed newsgroup {sci.bio.immunocytochem}
Well, I'm about to leave Oz and EM to move to Europe and hopefully start a Ph.D. in Archaeology (a long way from Marine Zool. and EM!) but before I go I've got to do an animated sequence from the SEM, in stereo, on a very small spider (a couple of thousand frames). Should look good. The folks here said I should pass on the mounting technique, so here it is.
Tip #1: BSE detection and Carbon I'm shooting with a Robinson BSE detector. It gives a more "real" illumination than SE and has the benefit of not "seeing" carbon. The specimen is normally mounted on top of a minuten (entomological) pin which is clamped in a 12mm "vice-stub" and tilted at 90 degrees in the SEM. On my Cambridge S120, the top of the stage mechanism is now out of view but you still see the bottom plate below. I've covered that with a piece of alfoil which has been painted with thinned-out carbon dag, making it a perfectly black, "studio-shot" background when using BSE.
Tip #2: No unsightly props Further, instead of mounting the specimen on the usual minuten pin, I've fixed it to the end of the lead from a clutch pencil (draughtsman's pencil), so the pin is also virtually invisible under BSE. Doesn't the pencil lead get coated too? I've taken a medium-bore tip from a syringe, held it in an alligator clip, and fed the pencil lead down into it leaving just the specimen exposed for coating.
Tip #3: Accuracy in mounting Mounting very small specimens on a pin-tip is difficult at the best of times. As the animation sequence is to be in stereo, the specimen must be at exactly the right angles to the pin and we don't want to see any excessive blobs of dag. You'll need a small bench vice, micrometer screw guage and double sided tape (DST). Lay your stereomicroscope down on its front with the head reversed so that the eyepieces point up and the light path is (roughly) horizontal. Clamp the side of the screw guage into the side of the vice and place it in front of the 'scope. Secure your specimen to a foam block with crossed minuten pins and secure the block in position to the opposing pole of the screw guage with DST. Fix the mounting pin to the mobile shaft of the screw guage with DST. Watching it all through the 'scope, do a dummy run to ensure that everything lines up. Screw the pin up, apply a meniscus of dag to the end of the pin and screw it down to the specimen. Leave to dry.
Tip #4: Centre of rotation Because this sequence includes rotating the animal 360 degrees, the pin has to be at the centre of rotation of the stage. However, in my 12mm "vice-stub", the vice is offset. Instead, I'm using a pop-rivet. Push the nail out and use the rivet and its collar; the rivet has the same diameter as the shaft on a normal 12mm grooved stub. Put the pin down the bore and dag it into a central position.
Hope you find these tips helpful and if you're in Oz around September, drop into the Australian Museum, Sydney, to see the stereo animation sequence of the spider (unfortunately, I'll be gone by then).
Geoff Avern Manager Microscopy Labs Australian Museum Sydney, Australia
Dear fellow microscopist I have had a suggestion from one of my colleagues about staining of nerve cells. He recommends including tannic acid in the fixative (paraformaldehyde/gluteraldehyde) at a concentration of 0.25% to 2% and then to use phosphotungstic acid to stain sections or use UA as a block stain prior to embedding. I have no idea how well this will work but apparantly tannic acid stains membranes clearly. I hope you have some success
Cheers Nikki Bock Dept. of materials engineering & materials design University of Nottingham Nottingham NG7 2RD Email: emznjb-at-emn1.nott.ac.uk
Can anyone advise on best technique for preparation of Chlorides (polished sections), as these materials are extremely soft and highly rippable resulting in abundant cavities in polished section. Several sample prep techniques have been tried without success.
John Yes it can be done To do this, put a drop of fixed virus suspension on to your coated grid for about 60 seconds. Than very slowly suck up virus suspension with a whatman filter paper.(cut filter paper into triangle, with sharp arrow point like tip.) Leave to dry for about 60 seconds.
Than float grid containing virus onto a drop of 0.05M sodium cacodylate buffer. (2 x 2 minutes wash)
Again wash with double distilled water. (2 x 2 minutes)
Negative stain for 30 seconds with PTA.
Dry and view.
All the best
Vijay H Bandu University of Natal Centre for Electron Microscopy Private Bag X01 Scottsvile 3209 South Africa
Hi Marty I seem to be havivig difficulty with your e-mail address. thought I might reach you here. I am trying to find a source for dowty window seals for our polaron CPD.You suggested All Seals in Santa Ana. Ca. I contacted them, but they require the Dowty part number or their own. Which Idon't have. Do you have that number? Any help would be greatly appreciated. Reply to ToogoodM-at-em.agr.ca
ToogoodM-at-em.agr.ca Agriculture and Agri Food Canada Kentville Research Centre Kentville N.S. B4N 1J5
Message-Id: {3.0.1.32.19970529084940.006d08e8-at-biotech.ufl.edu} X-Sender: sdw-at-biotech.ufl.edu X-Mailer: Windows Eudora Light Version 3.0.1 (32)
I have archived two discussions you may find useful. One is on thinsectioning bone, and the other undecalcified teeth. Go to the url listed at the bottom of this message and click on the "Tips & Tricks " link. Do a search for "bone" and there will be a number of files listed. The two most useful will be "teeth.html" and "thinbone.html". Good luck
At 05:18 PM 5/28/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 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'm currently evaluating replacements for my Kevex EDS detector with a Be window. I use a Kevex 4505P Pulse Processor which dumps the signal down to a 4pi SC-2 multichannel analyzer for spectral analyses and imaging.
My goal is to either change my Be window to a thin film window or replace my current detector with one equipped with a thin film window for light element analyses.
Does anyone know if:
1) The Be window of the Kevex Model 2003 EDS detector is replaceable with a thin film window?
2) Can the Kevex 4505P Pulse Processor handle light element pulse throughput?
3) Are there EDS detectors available with thin film windows which are adaptable to the 4505P Pulse Processor?
Any ideas or references out there would be highly appreciated.
I hope we haven't missed the deadline for the American Lab article. Please find our submission attached. I will Federal Express the corresponding 35mm slides today. Sincerely, -Jennifer Robinson Product Manager, Fluorescence Microscopy
At 07:01 PM 5/28/97 -0400, Scott Walck wrote: } } The question that I have about charging in an SEM is this. If you set up the } imaging conditions for the sample not to charge at a high scan rate i.e. TV } rate, and then drop it to a slow scan (as you do in an analog machine when } recording an image), why does the image now charge?
As John Bozolla said, the beam is dwelling longer at each spot allowing more charge to build up. I think this would be related to the old V=I*R equation. In fast scanning, the peak current (electron dose) before the beam moves away and the sample has a chance to discharge is relatively small so the resultant voltage would be small. At slow scans (say 30-100 times slower) the voltage should be that much higher and would become noticeable. Warren E. Straszheim Co-Director of Ambience for BIBLE-at-VIRGINIA.EDU
He who has knowledge spares his words, and a man of calm understanding is of a calm spirit. - Pr 17:27 Even a fool is counted wise when he holds his peace; when he shuts his lips, he is considered perceptive. - Pr 17:28 It is better to keep silent and be thought a fool than to open your mouth and remove all doubt. - Murphy
You can also add Kheops from Noesis S.A. and Noesis Vision Inc. The product is now being demonstrated commercially, it will be available in June and on display at the Microscopy show in Cleveland in early August.
Regards,
At 08:32 PM 5/28/97 +0200, Heike Buecking wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
---------------------------------------------------------------------------- ------------- Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com http://www.noesisvision.com 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada ---------------------------------------------------------------------------- -------------
Dear Reembedders, To successfully reembed thick sections (Araldite, Epon, LR White) one must understand the process. 1. Interface - The interface between the slide and the sections to be reembedded is of crucial importance. Sections will stick tenaciously to super clean glass slides, but will beautifully depart on command from the correct interface. The best interface is gelatin (no additives when subbing slides - may interfere with the LM stains). The interface cannot present a "soft" mixture of epoxies, perhaps created by improper polymerization due to alcohol, acetone, or propylene oxide left on the slide. 2. Not all epoxy mixtures are suitable for reembedding. Araldite, some epon replacements containing dilutents or plasticicers my remain too elastic, and the face of the section will deform when detaching. 3. Coefficient of thermal expansion - the most successful (ours is 99.9% successful, we do not loose sections) of detachment methods. Some people use cold, we use heat. We prefer heat, it is simple, and the glass becomes fluid, so to speak, preventing cold glass chips from becomeing part of the new block. Briefly, the rates of expansion between block and slide must vary widely. The slide is rapidly cooled or heated, the epon lags behind, and detachment occurs. Detaching hot blocks from hot slides is not as successful. Frequently the sections deform (bad for correlative microscopy). 4. The water tower effect - Capsules which are reversed over sections on slides (capsules are filled with epoxy) must be of a certain size. Skinny, small capsules, result in a lot more seeping epoxy around the edges, than fat short ones (the standard size of embedding capsules). Seeping epoxy causes "collaring" which makes detachment difficult. 5. Correlative microscopy of immunocytochemical materials (DAB) requires flat sections- Sections will move and warp while being remembedded making exact correlations difficult. This is best prevented by using a glass microscope slide (subbed) as a bottom slide on which the sections rests. Thick sections are dried down on this slide, stained and photomicrographed. The capsule is then inverted over the section. The oven temp should not go over 60degC. If sections are free-floating, as in a DAB reaction, they are placed on subbed microscopy slides, carefully wiped clear of excess epoxy, and covered with another slide which has been treated with mold release (Trennmittel from EMS). (I own no stock in EMS). The two slides are than weighted or clamped together, and polymerized. The top slide comes off easily, the sections is flat for photomicrography, and then reembedded with a gelatin capsule full of epon. I have run numerous trials on epon mixtures and found the Luft's medium mixture to be ideal. It is best to allow the epon mixture to rest for 30 min after acceleration, because it inhibits seeping and "collaring" around the section. 6. Original processing of tissue - If the original tissue is poorly infiltrated or suboptimally prepared, it is essentially "over". During the reembedding process, the new epoxy will bind to the old in an unpredictable manner and the section may warp badly, split when pulled off, etc. Many of the above statements have their origins in the writing of Lee and Neville - Handbook of Epoxy Resins, or in the publications of Causton and the people who hold patents for LR White, etc. If anyone wants a step by step method for reembedding of sections from a Vibratome or an ultramicrotome, please request. This is lots of fun, and lots of challenge. Reembedding could be a masochist's pradise! Actually TEM could be a masochist's best bet! Right? Bye, Hildy
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Dear all,
Wanting to control the presence of esterified pectins in plant cells I've tried the hydroxyalmine/ferric chloride reaction as described by Reeve (1959). I suppose this reaction is routinely used by many of us but.... I have a precipitation problem when I add to the fresh hydroxylamine reagent - i.e. a solution of equal volumes of sodium hydroxide (14g in 100ml EtOH 60%) and hydroxylamine hydrochloride (14g in 100ml EtOH 60%) - concentrated HCl in EtOH in order to acidify the reaction mixture. The precipitate I obtain is of cristalline structure and rather sticky... If you met this problem to when following exactly Reeve's protocole, if you were lucky in solving this problem or if you were not, etc... Please help ! Thanks Pascal
************************************ Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 Fax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ************************************
************************************ Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 Fax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ************************************
Sounds like embedding in resin, followed by diamond (or glass) knife sectioning might be worth a try. This technique has been used for decades as a means of preparing soft metals for metallographic examination. Reaction of the chlorides with the standard embedding resins might be an issue. Also, some resins need modest heating (~60 C) for curing.
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada Ottawa, Canada K1A 0G1
______________________________ Reply Separator _________________________________ Subject: SAMPLE PREP. OR MICROSCOPY Author: petra-at-iafrica.com at internet Date: 5/29/97 11:57 AM
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Can anyone advise on best technique for preparation of Chlorides (polished sections), as these materials are extremely soft and highly rippable resulting in abundant cavities in polished section. Several sample prep techniques have been tried without success.
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Technician Certification 5/29/97 12:45 PM
In light of the recent inquiries to accreditation, I would like to let listserver members know there is a Certification Program for biological electron microscopy technicians available through the Microscopy Society of America. It involves a practical and written examination. The practical requires the technician to prepare and embed samples for resin, section and take pictures. The written exam is a comprehensive 100 questions pertaining to fixation, embedding, microscopy equipment and sectioning techniques. If you would like more information you may contact the MSA Business Office at 4 Barlows Landing, Pocassett, MA 02559, telephone 1-800-538-3672, fax 1-508-563-1211 or email at BusinessOffice-at-MSA.Microscopy.Com
Linda Chicoine Center for Cell Imaging Yale University 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 phone 203-785-3646 fax 203-785-7226
I have not prepared chlorides alone, but have prepared (polished) metallic mounts with chloride inclusions. Typical metallographic techniques were employed except decane (high purity kerosene) was used instead of water. The first prep using water disolved the inclusions. Using non-polar lubricants and cleaning agents solved our problem.
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Can anyone advise on best technique for preparation of Chlorides (polished sections), as these materials are extremely soft and highly rippable resulting in abundant cavities in polished section. Several sample prep techniques have been tried without success.
I have not prepared chlorides alone, but have prepared (polished) metallic mounts with chloride inclusions. Typical metallographic techniques were employed except decane (high purity kerosene) was used instead of water. The first prep using water disolved the inclusions. Using non-polar lubricants and cleaning agents solved our problem.
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Can anyone advise on best technique for preparation of Chlorides (polished sections), as these materials are extremely soft and highly rippable resulting in abundant cavities in polished section. Several sample prep techniques have been tried without success.
Hi: Does anyone have any information on the Life Cell Corp which produces the CF-100 slam freezer. I need their phone #. Also has anyone used one or a similar model. Pros and cons would be appreciated.
Thanks, Charlie Murphy cmurphy-at-ggpl.arsusda.gov
There is a regular full-time position available in the Department of Biological Imaging (a Shared Scientific Service) at the Jackson Laboratory in Bar Harbor, ME.
Duties include operation and training of end users on a PC-based image analysis system and the performance of customer-directed analysis of biological research specimens. Routine maintenance and use of a confocal microscopy system will be a major requirement for this position. Also included is the regular maintenance and alignment of upright, inverted and stereo research-level microscopes, with the following optics: brightfield, darkfield, phase contrast, differential inteference contrast and fluorescence. A successful candidate would have a Master's degree (or equivalent experience and qualifications) in biological sciences and a minimum two years experience in confocal microscopy and/or microscopic imaging, experience in both being most desired. Experience in rudimentary computer programming is also preferred. Experience in electron microscopy and/or histology would be an asset. The applicant must be able to work independently in a multi-user facility and deal with people on a one-to-one basis. Individual will be expected to attend seminars and participate in interest groups disseminating information about current microscopic techniques.
Interested applicants may forward resumes to : Joanne Bradt
At 09:18 29/05/1997 -0400, you wrote: } I hope we haven't missed the deadline for the American Lab article. Please } find our submission attached. I will Federal Express the corresponding } 35mm slides today. } Sincerely, } -Jennifer Robinson } Product Manager, Fluorescence Microscopy } } (See attached file: Cell_des.doc) } } Attachment Converted: "C:\EUDORA\Attach\Cell_des.doc" } Even if English is not my mother language, I think this Email is a commercial advertisement. Specially when it's loading on my attach directory some files I must delete by manually acting. We are a lot of commercial company using this mailing list and we must respect the ame the list. Meme si l'anglais n'est pas ma langue maternelle, je pense que cet Email est a vocation commercial. Je n'admet pas de recevoir en attach un fichier que je devrai detruire en recherchant dans mes fichiers. Nous sommes plusieurs societes commerciales a utiliser cette facilite et nous devons respecter le fond de la liste. Best regards. --=====================_864938563==_ Content-Type: application/msword; name="Cell_des.doc"; x-mac-type="42494E41"; x-mac-creator="4D535744" Content-Transfer-Encoding: base64 Content-Disposition: attachment; filename="Cell_des.doc"
I hope your journey back to Germany was uneventful. I am glad we had a chance to discuss some of issues that create challenges for all of us.
One major point that we failed to discuss was the status of UL listing for the DM LB 30 and DM LB 100. This is a Key success factor for the DM LB. These instruments are effectively being kept out of hospital labs and research labs in hospitals because they don't have UL. This a problem in the 3 largest marked in the US. Los Angeles, Ca --- Chicago, IL --- and New York City. What is our progress and when can we expect to see these marks on the instrument? By the way, it would be nice for the LM, LSP and LP as well.
I understand that another vendor is being considered to supply us with an HBO power supply. Let me add that this power supply should also be UL listed. CSA is also acceptable, but the UL is preferred in the States. Of course, just the opposite occurs in Canada.
Jurgen, I've recently been working with AgBr(x)Cl(1-x). What a pig of a material! The best way of making sections is to use a microtome; polishing using diamond impregnated plastic, finishing with syton, works reasonably well if you can avoid embedding any particles in it. Other problems, apart from the softness, is the reactivity (reacts with virtually everything; Au, Ag and Pt are the only 'safe' metals which can make contact with it), and decomposition of the material when hit with light, electron beams (you can watch the composition change using EDX), and ions (so any ion milling is out). Put this together with an incredibly high coefficient of thermal expansion and you have the material scientist's material from hell. Good luck!
Richard Beanland, GMMT Caswell, Towcester, Northants NN12 8EQ UK
} } } Can anyone advise on best technique for preparation of Chlorides (polished } sections), as these materials are extremely soft and highly rippable resulting } in abundant cavities in polished section. Several sample prep techniques have } been tried without success. } } Regards, } Jurgen Paetz } 9 shell Rd. } Bloubergrand 7441 } CAPE TOWN - R.S.AFRICA } } REPLY: Jurgen Paetz } E-Mail: {petra-at-iafrica.com}
Scott D. Walck wrote: } The question that I have about charging in an SEM is this. If you } set up the imaging conditions for the sample not to charge at a high } scan rate i.e. TV rate, and then drop it to a slow scan (as you do } in an analog machine when recording an image), why does the image } now charge?
John J. Bozzola answered: } Because in a slower scan mode the beam dwells longer on the specimen } permitting more of a static charge to build up; the charge collects } with time.
The observations reported in "Observation of voltage contrast at grain boundaries in YSZ" by Charlotte Clausen (now C. C. Appel) and me in Micron and Microscopica Acta vol. 23 (1992) p. 157-158 illustrates the point made by John. We show a sample of YSZ in which you can see the grain boundaries imaged with a dark contrast when the beam current is 6 nA and the SEM is operated at slow scan rate (50 s per frame). The grain boundaries obtain the dark contrast because they conduct the charge away easier than the grain interior and therefore obtain a positive potential with respect to the grain interior.
The contrast disappears if either the beam current is lowered at constant scan rate OR the scan rate is increased at constant beam current.
-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- J. B. Bilde-Soerensen Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
I apologize for posting what was a commercial press release. I was responding to a request for submissions from Barbara Foster and I inadvertantly posting to the entire list instead of only Barbara. Please be assured that it was unintentional and I have the utmost respect for the aim of the list. -Jen R., Scanalytics
I need a lens with a "C"-mount for a Sony Model XC-77 analog video camera. Can anyone recommend a source? A macro lens would probably work the best for my application (image analysis).
Thank you.
*****************************************
Delilah Wood United States Department of Agriculture Western Regional Research Center 800 Buchanan Street Albany, CA 94710
Why is it that when I do double immunostaining with two successive monoclonals I get inconsistant results with 'stains' that are fine by themselves? If I do a polyclonal followed by a monoclonal, no problems. What is the story? Are there ways around this? Thanks in advance!
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Delilah Wood wrote: } } I need a lens with a "C"-mount for a Sony Model XC-77 analog video camera. } Can anyone recommend a source? A macro lens would probably work the best } for my application (image analysis). } } Thank you. } } ***************************************** } } Delilah Wood } United States Department of Agriculture } Western Regional Research Center } 800 Buchanan Street } Albany, CA 94710 } } Tel: 510.559.5653 } Fax: 510.559.5777 } Email: wood-at-pw.usda.govDear Delilah,
We found a wide variety of c-mounts available from Diagnostic Instruments in Sterling Heights, MI. Phone number: (810)731-6000. Their people were also very knowledgeable about different types of applications and which lens was needed for which situation. They also have an intriguing zoom system.
Good luck, Barbara Foster Microscopy/Marketing & Education
Caveat: We are not a vendor or promotor for Diagnostic Instruments; only an interested user.
You can contact LifeCell Corporation at: Tel. (713) 367-5368 Fax (713) 363-3360
I used to work at LifeCell, and helped develop the CF100 and the molecular distillation drying device, so I'm probably not the one to give you an unbiased opinion. I'm sure LifeCell can give you some references to call.
I *will* take this opportunity to say that cryofixation has never been easier or more reproducible than with the CF100. This is because of the microporcessor control, which monitors all of the parameters of cryofixation and permits fixation only after all of the conditions have been satisfied. Also, the microprocessor automatically regenerates the cryofixation surface after it's been used by heating it under a vacuum and then re-cooling it to prepare for another fixation run. The throughput on the CF100 is truly amazing -- as I recall, you can do a cryofixation about once every 2-1/2 to 3 minutes.
Best of luck to you. If I can be of any further assistance, don't hesitate to contact me off-list at RCHIOVETTI-at-aol.com.
The Pathology Department here at the Boston Univ School of Medicine has asked me to post a notice that their Hitachi 11C TEM is freely available. It has been under service contract for most of its life although not during the last five years. Interested folks should respond at busmpath-at-bu.edu
Dr. Steven Barlow EM Facility/Biology Department 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/EM_Facility
Are you sure it is one micron gold particle? It does not sound right .. It is almot three times the size of an average cell or 50x the size of viral particle.. I believe it should be one nano meter.. That I may be able to help you with...
At 02:08 PM 5/28/97 -0400, Sharon Godkin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I did send email to ListServer-at-... for unsubscribe by using the required command several times. But I am still on the list. Could you remove me from the list?
Are you working at the light level or the EM level?
If you are working with immunoEM, it sounds like the problem with your double immunolabeling may be cross-reactivity between the monoclonals and whatever you're using for the subsequent steps. This is frequently the situation with protocols that use, for example, mouse monoclonal primary antibodies, then goat anti-mouse Ig's followed by some method to localize with colloidal gold (maybe biotinylated goat anti-mouse, followed by streptavidin-colloidal gold?).
If you *are* working at the EM level, you might want to consider doing a double-sided labeling. In a nutshell, place the sections on uncoated hexagonal mesh nickel grids. Do one labeling run on one side, going through all the way to distilled water rinsing and drying. Flip the grid over, then perform the second labeling on the other side, using gold of a different size. This "compartmentalizes" each label, and greatly reduces the cross-reactivity.
Hope this helps. Feel free to contact me off-list if you need additional details.
Best regards,
Robert (Bob) Chiovetti, Ph.D. RCHIOVETTI-at-aol.com
} any suggestions where I can buy a water trap for my liquid carbon dioxide } tank attached to my CPD? } At one time Tousimis (spelling?) sold an in-line unit. I saw the set up at the EM facility of University of Wisconsin -at- Madison.
} ______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey fax: (609) 771-2674 Trenton, NJ 08650-4700
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