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I am looking for the best options on the market for hardware and software to create video presentations. I want to be able to create video from series of TIFF images (or whatever format one uses) and edit these videos, add text and effects of different kinds...
I would appreciate relevant comments from you about: - Hardware for this that goes with a PC - Software for this that runs under Windows NT 4.0
I am simply interested to hear what you may use for this and comments about it.
Thanks, Martin
----------------------------------------------- Martin K=F6hler Department of Molecular Medicine Rolf Luft Center for Diabetes Research L6B:1 Karolinska Hospital S-171 76 STOCKHOLM SWEDEN phone 46-8-51775732 46-8-51775727 fax 46-8-51773658 E-mail mk-at-enk.ks.se ---------------------------------------------
Under ideal (and reasonably attainable conditions), what is the detection limit (in grams) for EDX and WDX? Thanks.
#################################################################### 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 ####################################################################
} ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ----------------------------- } -----------------------------------------. } } Under ideal (and reasonably attainable conditions), what is the } detection } limit (in grams) for EDX and WDX? Thanks. } } ...
The question you ask is not specific enough ... to many factors to be considered with regard to which elements you are interested in, and (e.g.) ... how sensitive your specimen is to long count times and high beam currents ... ... ask again ...
cheerios, shAf -- ~~~~~~~~~~~~~~~~~~~~~~~ cogito, ergo zZOooOM ~~~~~~~~~~~~~~~~~~~~ Michael Shaffer - Geological Sciences - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I need a solution to electro polishing a Cu, Ni, Al alloy (~ 80:14:4 wt%)for EBSP work. A CrO3 + HClO4 has been suggested, however does anyone know of any other suitable solutions.
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.
Postdoctoral Studentship in Surface Science and Engineering
Applications are invited for Postdoctoral Research Studentships in the following subjects of study:
- micro and nanotribology - study of laser processed materials using transmission electron microscopy.
The Studentships are tenable for one year, renewable for a second year and include a maintenance grant of about US$20,000 per year. The Studentship will be supported by a research contract. Applicants should have a Ph.D. degree in an appropriate subject, or expect to obtain it before receiving the grant. Experience in transmission electron microscopy of metallic materials or ceramics is essential. Formal applications including a CV, names of two referees and specification of areas of interest and past experience should be sent to:
Prof. R. Vilar Departamento de Engenharia de Materiais Instituto Superior Ticnico Av. Rovisco Pais, 1096 Lisboa Codex, Portugal Tel. no. 351-1-8418121, fax no. 351-1-8418121 Email address: pcrvilar-at-alfa.ist.utl.pt.
New additions have been posted at http://members.aol.com/ImagProcTK/updates.htm for both Mac and Windows users. The Measure/Size&Shape routine previously available for Mac is now provided for Windows. Routines to draw grids of lines (horizontal or vertical, or radial with either uniform or sine weighting) are useful for stereological counting or to AND with binary images for measurement (both platforms). Coming next month: Measure/Select for Windows, Color space conversion routines, and more...
} } } Hello all, } I need to look at the surface morphology of some yeast } cells and as I'm a physics sort of person I need a little help. } } I going to be using an ESEM, but what is the best way to } prepare yeast cells for this sort of examination? } } A step by step answer or reference would be great. }
Don't prepare the samples! Mount them on a peltier stage stub, cool them, get the chamber water vapour stabilised and view them as they are. For anything more detailed please mail me direct.
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 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
Greetings, OK, this sounds a bit off topic, but honestly, I encountered this problem in a procotol for extracting polymerized butyl-methylmethacrylate resin from sections. The protocol, from the early 1950's, simply calls for "amyl alcohol" and "amyl acetate". Nowadays, there is a very popular solvent called "ISOamyl alcohol" (or more formally, 3methyl-1butanol), and I think this might have been what was being refered to in the fifties as simply amyl alcohol. Contrarywise, there is a solvent that is still called in the catalogs "amyl acetate" (more formally, pentacetate), that I guess is what was meant (despite the fact that a compound exists that is now called "ISOamyl acetate". I would like to try this protocol, and I would rather not all of the different amyl alcohols (n-, t-, dlsec- etc). Can anyone with a long memory or a good chemistry background help here? Many thanks! Tobias Baskin
Don't know if this will work for your alloy, but I have used a simple electrolyte that you can use at room temperature for jet polishing various copper alloys for TEM. Might work for your purposes.
25% Phosphoric Acid 25% Ethylene Glycol 50% Distilled Water
Good Luck
Dan Edwards
------------------------------------------------------- Dan Edwards Structural Materials Research Section Battelle Pacific Northwest National Laboratory P.O. Box 999, MSIN P8-15 Richland, WA 99352
} -----Original Message----- } From: Keith Moulding [SMTP:mcmouldk-at-uxmail.ust.hk] } Sent: Tuesday, September 02, 1997 12:06 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Electro polishings of Cu,Ni,Al alloy } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Hello, } } I need a solution to electro polishing a Cu, Ni, Al alloy (~ 80:14:4 } wt%)for } EBSP work. A CrO3 + HClO4 has been suggested, however does anyone } know of } any other suitable solutions. } } 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. } } FAX: (852) 2358 2451 } TEL: (852) 2358 8724 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
There is a Fe-Mo-Ni ternary phase diagram in the Journal of Phase Equilibria 15 (6), 622-626. The article includes references to other papers. } } Dear Sirs: } I am working on the sputtering NiFe/Mo magnetic multilayers by } using HREM. The interface between the NiFe and Mo layers may be a kind } of NiFeMo alloy, but I can't make decision about the idea. So I hope to } know the NiFeMo phase diagram and the solution degree between NiFe and } Mo. } } I am looking for the help from you and please tell me the } references where I can find the NiFeMo phase diagram. } } Yours Sincerely } } Gao Yihua
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
AMT has a "fast track" opportunity for a national field service engineer. The position requires understanding of electron microscopes, digital technology and software. This individual will be responsible for installation and support of AMT's imaging and motion control products for electron microscopy.
Inquiries and questions should be addressed to:
Jim Mancuso Advanced Microscopy Techniques Corp. 3 Electronics Avenue Danvers MA 01923
Acording to my material safety data sheets: Amyl alcohol is 1-pentanol, CAS# 71-41-0, although the name is also used for a mixture. Isoamyl alcohol is 3methyl-1butanol, CAS #123-51-3. Amyl acetate is pentacetate or 1-pentyl acetate or N-pentyl acetate or N-amyl acetate. CAS# 628-63-7. Isoamyl acetate is 3methyl-1butyl acetate or isopentyl acetate, CAS# 123-92-2. } } Greetings, } OK, this sounds a bit off topic, but honestly, I encountered this } problem in a procotol for extracting polymerized butyl-methylmethacrylate } resin from sections. The protocol, from the early 1950's, simply calls for } "amyl alcohol" and "amyl acetate". Nowadays, there is a very popular } solvent called "ISOamyl alcohol" (or more formally, 3methyl-1butanol), and } I think this might have been what was being refered to in the fifties as } simply amyl alcohol. Contrarywise, there is a solvent that is still called } in the catalogs "amyl acetate" (more formally, pentacetate), that I guess } is what was meant (despite the fact that a compound exists that is now } called "ISOamyl acetate". } I would like to try this protocol, and I would rather not all of } the different amyl alcohols (n-, t-, dlsec- etc). Can anyone with a long } memory or a good chemistry background help here? } Many thanks! } Tobias Baskin } } } _ ____ ^ __ ____ Tobias I. Baskin } / \ / / \ / \ \ University ofMissouri } / | / / \ \ \ BiologicalSciences } /___/ /__ /___ \ \ \__ 109 Tucker Hall } / / / \ \ \ Columbia, MO 65211-7400 USA } / / / \ \ \ voice: 573-882-0173 } / /____ / \ \__/ \____ fax: 573-882-0123
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
Traditionally, AMYL ALCOHOL is a mixture of 3-methyl-1-butanol and 2-methyl-1-butanol, derived from the distillation of fermented STARCH (Latin: amylum). This is almost certainly what was meant in the 1950s and even much later. These days, you are likely to find this listed as iso-amyl or iso-pentyl alcohol (or acetate). When catalogues list the n-isomer they generally specify n-amyl or n-pentyl.
For any traditional protocol, the iso-amyl (or pentyl) is what would have been on the shelf at the time.
Useful source: "Chemistry of Organic Compounds" by Noller - the 2nd edition (1957) is particularly good for historical information.
(Incidentally, the higher alcohols are not formed by side-reactions on the glucose from the starch, but are derived from amino-acids derived from the yeast or present in the potatoes, or whatever).
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I need some advice and wonder if there is any one who can help me. We have a Hitachi H-600 tem, the machine is quite old ie. 16 years. A collegue from a neighbouring centre's, Hitachi h-600 is giving problems. They would like to borrow our high voltage board and test it on their machine to make sure that the board is not broken. I am in a dilemma as I don't know if what ever caused thei.r problem will damage our board. I feel I would like to help them out but not if there is a chance that I might be making problems for myself. I would really appreciate any advice as I have not been running our EM lab for very long (2 years), and am still not 100% confident with the machine.(All our senior staff left and we have been unable to replace them, so I don't really have anyone to ask who has a lot of experience with the mechanical running of the Hitachi)
Thanks Helen Ilsley Diagnostic EM Lab UCT Medical School Groote Schuur Hospital Cape Town South Africa e-mail : hilsley-at-chempath.uct.ac.za
The question of detection limits in X-ray spectroscopic analysis is a complex one which generally does not have a simple answer. This matter is discussed in some detail by Heinrich in his book 'Electron Beam X-ray Micro Analysis', Van Nostrand-Reinhold, 1981 (ISBN 0-442-23286-1) p. 193 & p. 216, with the conclusion that the term 'm inimum detectability limit' probably should not be used.
The matter is also discussed by Goldstein, et. al. in their book 'Scanning Electron Microscopy and X-ray Microanalysis', Plenum Prewss,2nd Ed., 1992, (ISBN 0-306-44175-6). In Table 9.17, p. 501, they give calculated values comparing MDLs for several different elements for both the EDS and WDS detection systems. If you don't have this book (you should, however, by all means obtain a copy if you are doing any work in SEM of X-ray microanalysis) I can fax you a copy of this discussion.
Best regards,
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
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Looking for a substitute for methanol for use in electropolishing of metal samples for TEM observation. Additionally it should have a flash point near or above 100 deg. F.
Hi John and Michael et al: I recall that one of your Presidents (Kennedy?) was looking for a one handed science adviser; the ones he had always prevaricated "on the one hand and on the other hand".
Michael is quite right, detection limits vary greatly depending on endless factors. However, it is useful to have some figures as guidelines: Considering say the 10 elements following sodium. Detection of these is about best. For these in EDX the limit for quantitative analysis is about 1%. Detection limit is about 0.1%. Increasing counts and counting times beyond the customary 100 seconds at perhaps 2000cps will scarcely improve either limit.
WDX is near quantitative to its detection limit and that is at least two orders of magnitude greater than is EDX.
I'll enter correspondence only when it concerns errors in excess of five orders of magnitude. 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, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} John J. Bozzola wrote: } } } } Under ideal (and reasonably attainable conditions), what is the } } detection } } limit (in grams) for EDX and WDX? Thanks. } } The question you ask is not specific enough ... to many factors to be } considered with regard to which elements you are interested in, and } (e.g.) ... how sensitive your specimen is to long count times and high } beam currents ... ... ask again ... } cheerios, shAf } -- } ~~~~~~~~~~~~~~~~~~~~~~~ cogito, ergo zZOooOM ~~~~~~~~~~~~~~~~~~~~ } Michael Shaffer - Geological Sciences - University of Oregon } mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Dear Teresa - I waited a couple of days but there was no reply on the listserver. Here is mine: I do not know about your regulations, maybe you must collect the stuff and then dispose of it "properly". But too many regulations are not wise - eg. the dangerous goods shipping laws which appear designed to make things more expensive but not safer.
Uranium occurs in trace amounts throughout the environment, including seawater and especially in granite. U does not concentrate in the food chain like P, K, Pb, Hg or some organic compounds. For very small quantities, as are used in EM labs, prompt disposal with some water into the sewage system appears perfectly reasonable to me. The mistake is to store the stuff and to accumulate larger quantities.
---------- } From: Flores, Teresa {tflore-at-lsumc.edu} } To: Microscopy Newsgroup {Microscopy-at-sparc5.microscopy.com} } Subject: } Date: Saturday, 30 August 1997 4:33
} } What do routine labs do with uranyl acetate waste. Ours has been picked up } by in house safety department and is being stored in drums as there isn't a } company that will pick up to discard. Is anyone else having this problem. } It is a .025mg uranyl acetate to 100 ml of 50% ethanol washed out with } copious amounts of sterile distilled water. Any imput will be appreciated. } Many thanxs } }
Anyone out there with experience exporting a Tencor P-11 Profilometer 3D data set to an external analysis program (e.g. IGOR or Nanoscope AFM)? We are looking for suggestions on successful transfers.
Michael T. Dineen The Dow Chemical Company 1897 Building Midland MI 48667 (517/636-4008 4 517/638-6443 + mtdineen-at-dow.com
Hello, I need to replace the 12 inch bell jar on a Denton 502. Denton wants $627 for a new one. Does any one know a source of used bell jars, etc. for this equipment? Thanks Wallace Ambrose
Jim, U in the USA must be disposed of in a way other than "down the sewer". It may occur in trace amounts in "nature" but the concentrations we work with are much higher than that. Besides the legal problems there are the moral responsibilities to not unnecessarily endanger others with the chemicals we use. True, uranium is one of the milder toxins we handle....if handled properly, but it still is a beta emitter and can do extensive tissue damage if allowed to come into contact with any soft tissues. Flushing this waste down the sewer does not guarantee that this will not happen to some unsuspecting individual. That is why there are regulations in place for the proper handling and disposal of this chemical. I also empathize with Teresa's dilema because we are in the same position....radiation waste handlers won't take it because it is a naturally occurring isotope and hazardous waste handlers won't take it because it is radioactive....a perfect catch 22. If anyone can come up with a practical solution to this dilemma that satisfies the current clean water act regulations we would be more than happy to listen.
Jim Darley wrote: } Dear Teresa - } I waited a couple of days but there was no reply on the listserver. Here is } mine: } I do not know about your regulations, maybe you must collect the stuff and } then dispose of it "properly". But too many regulations are not wise - eg. } the dangerous goods shipping laws which appear designed to make things more } expensive but not safer. } } Uranium occurs in trace amounts throughout the environment, including } seawater and especially in granite. U does not concentrate in the food } chain like P, K, Pb, Hg or some organic compounds. For very small } quantities, as are used in EM labs, prompt disposal with some water into } the sewage system appears perfectly reasonable to me. } The mistake is to store the stuff and to accumulate larger quantities. } } ---------- } } From: Flores, Teresa {tflore-at-lsumc.edu} } } To: Microscopy Newsgroup {Microscopy-at-sparc5.microscopy.com} } } Subject: } } Date: Saturday, 30 August 1997 4:33 } } } } } What do routine labs do with uranyl acetate waste. Ours has been picked } up } } by in house safety department and is being stored in drums as there isn't } a } } company that will pick up to discard. Is anyone else having this problem. } } It is a .025mg uranyl acetate to 100 ml of 50% ethanol washed out with } } copious amounts of sterile distilled water. Any imput will be } appreciated. } } Many thanxs } } } }
I agree, EDX detection limit somewhere just less than 1%...but
I work with polymer fibers and by ashing I can get great analysis of additives at 100 ppm in the polymer. This doesn't change the EDX detection limit per se, but there is more than one way to skin a cat.
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Hi John and Michael et al: I recall that one of your Presidents (Kennedy?) was looking for a one handed science adviser; the ones he had always prevaricated "on the one hand and on the other hand".
Michael is quite right, detection limits vary greatly depending on endless factors. However, it is useful to have some figures as guidelines: Considering say the 10 elements following sodium. Detection of these is about best. For these in EDX the limit for quantitative analysis is about 1%. Detection limit is about 0.1%. Increasing counts and counting times beyond the customary 100 seconds at perhaps 2000cps will scarcely improve either limit.
WDX is near quantitative to its detection limit and that is at least two orders of magnitude greater than is EDX.
I'll enter correspondence only when it concerns errors in excess of five orders of magnitude. 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, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} John J. Bozzola wrote: } } } } Under ideal (and reasonably attainable conditions), what is the } } detection } } limit (in grams) for EDX and WDX? Thanks. } } The question you ask is not specific enough ... to many factors to be } considered with regard to which elements you are interested in, and } (e.g.) ... how sensitive your specimen is to long count times and high } beam currents ... ... ask again ... } cheerios, shAf } -- } ~~~~~~~~~~~~~~~~~~~~~~~ cogito, ergo zZOooOM ~~~~~~~~~~~~~~~~~~~~ } Michael Shaffer - Geological Sciences - University of Oregon } mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I am looking for a set of circuit diagrams for a Zeiss EM-10A transmission electron microscope. If anyone is willing to part with an old set, or will allow me to copy a set, please email me at:
One of our customers has asked for some advice. He's picking up 1 inch diameter, .040" thick aluminum discs and exposing them to a chemical process which requires the least amount of the disc to be obstructed. He now uses triceps to hold the discs but they lose their memory after a mumber of uses. Can anyone suggest a better way to handle the discs?
Thanks, John Arnott Ladd Research ladres-at-worldnet.att.net
Teresa, I was hoping that someone experianced in haz. waste disposal methods would comment on your question. In my experiance, the problem with UAC waste disposal is if the radioactive material is in methanol. I have no trouble having the waste picked up if the UAC is in water. I have been told by Environmental health and Safety personnel that no disposal option exists for the UAC in methanol. You have brought up a very important topic and hopefully comments from other labs and especially from individuals experianced in waste disposal may follow.
Robert Cox Shriner Hospital Galveston Tx. -------- -----------------------------------------------------------------------.
Dear Teresa - I waited a couple of days but there was no reply on the listserver. Here is mine: I do not know about your regulations, maybe you must collect the stuff and then dispose of it "properly". But too many regulations are not wise - eg. the dangerous goods shipping laws which appear designed to make things more expensive but not safer.
Uranium occurs in trace amounts throughout the environment, including seawater and especially in granite. U does not concentrate in the food chain like P, K, Pb, Hg or some organic compounds. For very small quantities, as are used in EM labs, prompt disposal with some water into the sewage system appears perfectly reasonable to me. The mistake is to store the stuff and to accumulate larger quantities.
---------- } From: Flores, Teresa {tflore-at-lsumc.edu} } To: Microscopy Newsgroup {Microscopy-at-sparc5.microscopy.com} } Subject: } Date: Saturday, 30 August 1997 4:33
} } What do routine labs do with uranyl acetate waste. Ours has been picked up } by in house safety department and is being stored in drums as there isn't a } company that will pick up to discard. Is anyone else having this problem. } It is a .025mg uranyl acetate to 100 ml of 50% ethanol washed out with } copious amounts of sterile distilled water. Any imput will be appreciated. } Many thanxs } }
The problem of Ur disposal will continue to exist until someone develops a plan on how to dispose of the material AND makes a decision that is legally binding. One way to start the ball rolling is to formulate some possible ways of dealing with the material and then to forward the package to appropriate legislators.
Here is what I propose: take some epoxy resin and coat the inside of a polypropylene beaker with the epoxy by building up layers and polymerizing the layers. If one uses outdated, quite viscous epoxy monomers, one can achieve several mm of thickness in a couple days. Now, use the epoxy coated vessel as a waste container into which you pour your uranium containing liquids. Keep the vessel in a warm oven or fume hood (if volatiles are involved) to speed up evaporation. Allow the uranium liquids to evaporate to dryness (taking care to avoid generating dust) and apply another layer of epoxy over the uranium salts. What one gets is a layered system of epoxy/uranium/epoxy/uranium etc. Keep this up until the polypropylene vessel is completely filled with epoxy. The enrobed uranium can then be disposed in a toxic waste burial site where it might be further enrobed.
If users scale back on the use of uranium using tiny amounts and minimize rinse volumes, some of the problem will already be solved.
Key ingredients: scale down use, encapsulate dried salts in epoxy.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Are you replacing the bell jar because the rim is chipped ?. If so have you thought of having the rim cut down to remove the damaged area ? We had that done several times by a glass blower at a fraction of the cost of a new bell jar.
Jordi Marti ----------------------- Hello, I need to replace the 12 inch bell jar on a Denton 502. Denton wants $627 for a new one. Does any one know a source of used bell jars, etc. for this equipment? Thanks Wallace Ambrose
How about a trivet, like metalsmiths use for enamelling? (Check the metals program at your nearest college's art dept.) The tips can be modified by a little judicious filing to minimize the areas of contact between the disc and the legs of the trivet.
Phil
} To All, } } One of our customers has asked for some advice. He's picking up 1 inch } diameter, .040" thick aluminum discs and exposing them to a chemical } process which requires the least amount of the disc to be obstructed. He } now uses triceps to hold the discs but they lose their memory after a } mumber of uses. } Can anyone suggest a better way to handle the discs? } } Thanks, } John Arnott } Ladd Research } ladres-at-worldnet.att.net
I have no financial interest in VWR, etc.... } } } 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}
I have no financial interest in VWR, etc.... } } } 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}
Uranium in the USA must be disposed of in a way other than "down the sewer". It may occur in trace amounts in "nature" but the concentrations we work with are much higher than that. Besides the legal problems there are the moral responsibilities to not unnecessarily endanger others with the chemicals we use. True, uranium is one of the milder toxins we handle....if handled properly, but it still is a beta emitter and can do extensive tissue damage if allowed to come into contact with any soft tissues. Flushing this waste down the sewer does not guarantee that this will not happen to some unsuspecting individual. That is why there are regulations in place for the proper handling and disposal of this chemical. I also empathize with Teresa's dilema because we are in the same position....radiation waste handlers won't take it because it is a naturally occurring isotope and hazardous waste handlers won't take it because it is radioactive....a perfect catch 22. If anyone can come up with a practical solution to this dilemma that satisfies the current clean water act regulations we would be more than happy to listen.
Jim Darley wrote: } Dear Teresa - } I waited a couple of days but there was no reply on the listserver. Here is } mine: } I do not know about your regulations, maybe you must collect the stuff and } then dispose of it "properly". But too many regulations are not wise - eg. } the dangerous goods shipping laws which appear designed to make things more } expensive but not safer. } } Uranium occurs in trace amounts throughout the environment, including } seawater and especially in granite. U does not concentrate in the food } chain like P, K, Pb, Hg or some organic compounds. For very small } quantities, as are used in EM labs, prompt disposal with some water into } the sewage system appears perfectly reasonable to me. } The mistake is to store the stuff and to accumulate larger quantities. } } ---------- } } From: Flores, Teresa {tflore-at-lsumc.edu} } } To: Microscopy Newsgroup {Microscopy-at-sparc5.microscopy.com} } } Subject: } } Date: Saturday, 30 August 1997 4:33 } } } } } What do routine labs do with uranyl acetate waste. Ours has been picked } up } } by in house safety department and is being stored in drums as there isn't } a } } company that will pick up to discard. Is anyone else having this problem. } } It is a .025mg uranyl acetate to 100 ml of 50% ethanol washed out with } } copious amounts of sterile distilled water. Any imput will be } appreciated. } } Many thanxs } } } }
The Duniway Stockroom Corp., 1305 Space Park Way, Mountain View, CA 94043, Tel. 800-446-8811, e.mail: info-at-duniway.com, deals in new, used, and rebuilt vacuum equipment. You might copntact them.
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, 04 Sep 1997 14:11:03 +0200 } From: Catherine Goffaux {catherine.goffaux-at-wkb.be} } To: njoschko-at-datagram.be, claudejp-at-esvax.dnet.dupont.com, orion-at-infoboard.be, } deschuyt-at-sbbio.be, bdd.translations-at-skynet.be, labio-at-telecom-plus.sn, } sandra.rens-at-vulcan.be } Subject: (Fwd) (Fwd) Virus Warning (fwd) -RETRANSMIS } } Received: from mserv.rug.ac.be by wkb.be (SMI-8.6/SMI-SVR4) } id NAA01015; Sun, 31 Aug 1997 13:12:47 +0200 } Received: from eduserv2.rug.ac.be by mserv.rug.ac.be with SMTP id AA14051 } (5.67b/IDA-1.5 for {inge.vanderhaegen-at-wkb.be} ); Sun, 31 Aug 1997 13:21:01 +0200 } Received: from localhost by eduserv2.rug.ac.be (SMI-8.6/SMI-SVR4) } id NAA03840; Sun, 31 Aug 1997 13:20:59 +0200 } Message-Id: {Pine.SOL.3.94.970831131452.3715A-100000-at-eduserv2.rug.ac.be} } Date: Sun, 31 Aug 1997 13:20:59 +0200 } From: Kristof Van der Haegen {Kristof.VanderHaegen-at-rug.ac.be} } To: Karl.Theeten-at-rug.ac.be } Cc: Peter.Forret-at-keyware.be,Bartel.Van.Der.Haegen-at-mobile.belgacom.be, } Benedikt.Ameloot-at-rug.ac.be, Gunther.Heene-at-rug.ac.be, } Luc.VanSintJan-at-rug.ac.be, Samuel.VanBelle-at-rug.ac.be, } inge.vanderhaegen-at-wkb.be } Subject: (Fwd) (Fwd) Virus Warning (fwd) } Mime-Version: 1.0 } Content-Type: text/plain } Content-Disposition: inline } } PLEASE read the following warning. } WARNING!!!!!! If you receive an e-mail titled "JOIN THE CREW" } DO NOT open it! } It will erase EVERYTHING on your hard drive! Send this letter out to } as many people you can.......this is a new virus and not many people } know about it! } } This information was received this morning from IBM, please share it } with anyone that might access the Internet. } } Also, } } If anyone receives mail entitled; PENPAL GREETINGS! please delete it } WITHOUT reading it!! This is a warning for all Internet users - } there is a dangerous virus propagating across the Internet through an } e-mail message entitled "PENPAL GREETINGS!". } } DO NOT DOWNLOAD ANY MESSAGE ENTITLED "PENPAL GREETINGS"!! } } This message appears to be a friendly letter asking you if you are } interested in a penpal, but by the time you read this letter, it is } too late. The trojan horse" virus will have already infected the boot } sector } of your hard drive, destroying all of the data present. It is a } self-replicating virus, and once the message is read, it will } AUTOMATICALLY forward itself to anyone who's e-mail address is present } in YOUR mailbox! } } This virus will DESTROY your hard drive, and holds the potential to } DESTROY the hard drive of anyone whose mail is in your in box, and } who's } mail is in their in box and so on. If this virus keeps getting } passed, it has the potential to do a great deal of DAMAGE to computer } networks worldwide!!!! } } Please, delete the message entitled "PENPAL GREETINGS!" as soon as you } see it! And pass this message along to all of your friends, relatives } and the other readers of the newsgroups and mailing lists which you } are on so that they are not hurt by this dangerous virus!!!! } } Please pass this along to everyone you know so this can be stopped. } PASS THIS ON TO YOUR FRIENDS!!! WARNING !!! } There is a new virus going around in the last couple of days!!! } DO NOT open or even look at any mail that you get that says: "Returned } or Unable to Deliver" This virus will attach itself to your computer } components and render them useless. Immediately delete any mail items } that says this. AOL has said this is a very dangerous virus, and there } is NO remedy for it at this time, Please Be Careful, And forward to } all your on-line friends A.S.A.P. } } Forward this A.S.A.P. to every single person you know!!!!!!!!! } *************************************************** } } } }
Best regards,
Paul Vanderlinden. Sales Manager.
======================================================================= See our web site: http://www.microscopy-uk.org.uk
Jim Darley's posting is hardly furthering good science.
1: Detection limits (John's original question). Modern EDX systems are easily capable of quantitative analysis in the sub-1wt% range. See, for example http://prism.mit.edu/facltis/stem/stmexam.htm (the second illustration on that page) where I was getting analyses for Cr in steel of the order of 0.6+/-0.1 wt%. I don't know about you, but I consider this quantitative. This, mind you, was in a measurement where I was attempting to optimize spatial resolution rather than sensitivity, and the acquisition time was 60sec. per data point. This example, of course, was from a STEM. There are many more examples in the literature.
Using beam gating techniques such as those developed by Charlie Lyman and colleagues, EDX data can readily be acquired at 20,000-40,000 counts per real second, if spatial resolution is sacrificed. Combine this with an acquisition time of 1,000 seconds (by no means unrealistic for an important measurement) and the detection limit is in the rage of, or better than, 0.01wt%.
Of course, in the SEM, which may have been the point of John's original question, the situation is not the same, the beam voltage is lower (resulting in poorer peak/bremmstrahlung ratios) and the emission of x-rays from a solid sample is different from that in a thin foil, but these are some of the variables that Michael quite rightly pointed out must be considered.
2: Comparison between EDX and WDX.
This is like comparing apples and oranges, because the instruments designed with them are generally intended for different purposes.
WDX has a much better peak resolution than WDX, which results in better measured P/B ratios (and hence improved statistics), as well as much better capability in resolving nearby x-ray lines. Also, because the x-ray counting and wavelength analysis are different functions in the crystal spectrometer, available countrates have traditionally been higher in WDX than EDX (although modern EDX detectors are an order of magnitude faster than they were fifteen years ago). Against this must be set the fact that WDX is inherently a serial technique (although multiple spectrometers help here), while EDX is a parallel technique (compare the advantages of PEELS over SEELS - although this is not a totally fair comparison). Anyway, the advantage of WDX (ignoring the capability of resolving peak overlaps) is improved statistical precision. Is this useful?
Well, maybe.
By far the most significant parameter which affects electron-induced x-ray emission spectra is sample geometry. Two extreme cases are where the sample is a thin foil (as in the STEM) where, to a first approximation, the x-ray spectrum recorded by the detector is the same as that emitted, and to a second order, it is possible to derive a reasonable thickness correction for cases where the error is small. Alternatively, when the sample has dimensions large compared with the volume irradiated by the electron beam, and has an accurately known geometry compared to the incident beam and the detector (such as a polished flat sample in the microprobe), correction programs such as ZAF can do a reasonable job of extracting a quantitative analysis.
What about where the sample geometry is unknown (for example, a rough surface such as might be examined in the SEM)? In that case the uncertainty in the analysis is far, far worse than any uncertainty caused by the poor statistics of the EDX spectrum, so there is no point or advantage whatever in using WDX to try to improve things, because it won't. This is why typically an SEM has an EDX detector - it is cheaper and gives just as good an analysis (except for the somewhat poorer detection limit). In the microprobe, great care is taken in polishing and mounting the sample, so the advantage of the WDX detector can be realised.
Where does this leave us? The advantage of a WDX detector over an EDX detector *ON THE SAME SAMPLE* is limited - perhaps an order of magnitude in detection limit, and, on a flat, polished sample, also perhaps approaching an order of magnitude in precision. The WDX detector can also resolve many cases where peaks would overlap in EDX. On general rough SEM samples, the only advantages of WDX are a small improvement in detection limits and the ability to resolve overlaps (which could be important if a trace element peak overlaps a major peak in the EXD spectrum.
The President's science advisors were right - it all depends. I seem to remember that one of Jim Darley's Prime Ministers (Gough Whitlam?) had to be dismissed because Parliament would not pass his budget - but does that have anything to do with Science?
Tony Garratt-Reed
} Hi John and Michael et al: } I recall that one of your Presidents (Kennedy?) was looking for a one } handed science adviser; the ones he had always prevaricated "on the one } hand and on the other hand". } } Michael is quite right, detection limits vary greatly depending on endless } factors. However, it is useful to have some figures as guidelines: } Considering say the 10 elements following sodium. Detection of these is } about best. } For these in EDX the limit for quantitative analysis is about 1%. Detection } limit is about 0.1%. Increasing counts and counting times beyond the } customary 100 seconds at perhaps 2000cps will scarcely improve either } limit. } } WDX is near quantitative to its detection limit and that is at least two } orders of magnitude greater than is EDX. } } I'll enter correspondence only when it concerns errors in excess of five } orders of magnitude. } 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, 400+ Links, MSDS } ************************ http://www.proscitech.com.au } } } John J. Bozzola wrote: } } } } } } Under ideal (and reasonably attainable conditions), what is the } } } detection } } } limit (in grams) for EDX and WDX? Thanks. } } } } The question you ask is not specific enough ... to many factors to be } } considered with regard to which elements you are interested in, and } } (e.g.) ... how sensitive your specimen is to long count times and high } } beam currents ... ... ask again ... } } cheerios, shAf } } -- }
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
We received an JEOL 6400 SEM without an instruction manual. If anyone can help us out with a copy of a manual, we'd greatly appreciate it!
************************************************************************* Lucille A. Giannuzzi, Ph.D.
Assistant Professor Dept. of Mechanical, Materials, and Aerospace Eng.
Director, Cirent/UCF Materials Characterization Facility President, Florida Microscopy Society (a local affiliate of MSA)
University of Central Florida phone (407) 823-5770 PO Box 162450 fax (407) 823-0208 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu Orlando, FL 32816-2450 USA *************************************************************************
Before sending chills of panic up and down the spines of all of us computer freaks by posting information regarding an alleged virus one might check the latest information compiled by the Department of Energy at their web site dedicated to computer security.
The URL is http://ciac.llnl.gov/
The "viruses" listed are hoaxes according to CIAC.
Bob Craig OSRAM SYLVANIA Products Inc. Beverly, MA 01915
Paul Vanderlinden's posting included warnings about some computer viruses. While computer viruses can create many problems for the person or organization where an infection occurs, computer virus HOAXES can create their own problems as well.
According to the DOE, the PENPAL GREETINGS virus is a hoax, and possibly the other ones mentioned in Paul Vanderlinden's posting are also. Information about viruses can be found at the U.S. Department of Energy Computer Incident Advisory Capability (DOE-CIAC) web site which is at http://ciac.llnl.gov/ and the hoaxes page at http://ciac.llnl.gov/ciac/CIACHoaxes.html
Anyone who receives warnings about computer viruses should follow the DOE's recommended action...
"Users are requested to please not spread unconfirmed warnings about viruses and Trojans. If you receive an unvalidated warning, don't pass it to all your friends, pass it to your computer security manager to validate first. Validated warnings from the incident response teams and antivirus vendors have valid return addresses and are usually PGP signed with the organization's key." (from the DOE-CIAC web page on Internet Hoaxes)
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Try a vacuum forceps. They are available from all microscopy supply = houses and places like Edmund scientific etc. It is a pen like implement = which one puts various size needles on depending on the size one wants to = pick up. Suction is created by a very small motor. No microscopy lab = should be without one.
Judy M.
Judy Murphy, PhD Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207
Phone: 209/954-5284 FAX: 209/954-5600 e-mail: murphy-at-sjdccd.cc.ca.us program web page: http://www.sjdccd.cc.ca.us/ElectMicro/sjdc.html
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To All,
One of our customers has asked for some advice. He's picking up 1 inch diameter, .040" thick aluminum discs and exposing them to a chemical process which requires the least amount of the disc to be obstructed. He now uses triceps to hold the discs but they lose their memory after a mumber of uses. Can anyone suggest a better way to handle the discs?
Thanks, John Arnott Ladd Research ladres-at-worldnet.att.net
------------------ RFC822 Header Follows ------------------ Received: by sjdccd.cc.ca.us with ADMIN;4 Sep 1997 14:14:35 -0800 Received: from Sparc5.Microscopy.Com (206.69.208.10) by = ms.sjdccd.cc.ca.us with SMTP (Eudora Internet Mail Server 1.2); Thu, 4 Sep 1997 14:13:51 = -0800 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) id LAA02799 for dist-Microscopy; Thu, 4 Sep 1997 11:09:10 = -0500 Received: from mtigwc03.worldnet.att.net (mtigwc03.worldnet.att.net = [204.127.131.34]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id = LAA02796 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 4 Sep 1997 11:09:09 = -0500 Received: from server ([207.116.37.66]) by mtigwc03.worldnet.att.net (post.office MTA v2.0 0613 ) with SMTP id AAA640 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 4 Sep 1997 16:16:22 +0000 Message-ID: {340EDE5A.6A22-at-worldnet.att.net} "Paul VANDERLINDEN" {orion-at-euronet.be} X-Mailer: Mail*Link SMTP/QM 3.0.0
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} (Fwd) (Fwd) Virus Warning (fwd) -RETRANSMIS
See http://ciac.llnl.gov/ciac/CIACHoaxes.html for information on the PenPal hoax.
-Mike --------------------------------------
} Date: Thu, 04 Sep 1997 14:11:03 +0200 } From: Catherine Goffaux {catherine.goffaux-at-wkb.be} } To: njoschko-at-datagram.be, claudejp-at-esvax.dnet.dupont.com, orion-at-infoboard.be, } deschuyt-at-sbbio.be, bdd.translations-at-skynet.be, labio-at-telecom-plus.sn, } sandra.rens-at-vulcan.be } Subject: (Fwd) (Fwd) Virus Warning (fwd) -RETRANSMIS } } Received: from mserv.rug.ac.be by wkb.be (SMI-8.6/SMI-SVR4) } id NAA01015; Sun, 31 Aug 1997 13:12:47 +0200 } Received: from eduserv2.rug.ac.be by mserv.rug.ac.be with SMTP id AA14051 } (5.67b/IDA-1.5 for {inge.vanderhaegen-at-wkb.be} ); Sun, 31 Aug 1997 13:21:01 +0200 } Received: from localhost by eduserv2.rug.ac.be (SMI-8.6/SMI-SVR4) } id NAA03840; Sun, 31 Aug 1997 13:20:59 +0200 } Message-Id: {Pine.SOL.3.94.970831131452.3715A-100000-at-eduserv2.rug.ac.be} } Date: Sun, 31 Aug 1997 13:20:59 +0200 } From: Kristof Van der Haegen {Kristof.VanderHaegen-at-rug.ac.be} } To: Karl.Theeten-at-rug.ac.be } Cc: Peter.Forret-at-keyware.be,Bartel.Van.Der.Haegen-at-mobile.belgacom.be, } Benedikt.Ameloot-at-rug.ac.be, Gunther.Heene-at-rug.ac.be, } Luc.VanSintJan-at-rug.ac.be, Samuel.VanBelle-at-rug.ac.be, } inge.vanderhaegen-at-wkb.be } Subject: (Fwd) (Fwd) Virus Warning (fwd) } Mime-Version: 1.0 } Content-Type: text/plain } Content-Disposition: inline } } PLEASE read the following warning. } WARNING!!!!!! If you receive an e-mail titled "JOIN THE CREW" } DO NOT open it! } It will erase EVERYTHING on your hard drive! Send this letter out to } as many people you can.......this is a new virus and not many people } know about it! } } This information was received this morning from IBM, please share it } with anyone that might access the Internet. } } Also, } } If anyone receives mail entitled; PENPAL GREETINGS! please delete it } WITHOUT reading it!! This is a warning for all Internet users - } there is a dangerous virus propagating across the Internet through an } e-mail message entitled "PENPAL GREETINGS!". } } DO NOT DOWNLOAD ANY MESSAGE ENTITLED "PENPAL GREETINGS"!! } } This message appears to be a friendly letter asking you if you are } interested in a penpal, but by the time you read this letter, it is } too late. The trojan horse" virus will have already infected the boot } sector } of your hard drive, destroying all of the data present. It is a } self-replicating virus, and once the message is read, it will } AUTOMATICALLY forward itself to anyone who's e-mail address is present } in YOUR mailbox! } } This virus will DESTROY your hard drive, and holds the potential to } DESTROY the hard drive of anyone whose mail is in your in box, and } who's } mail is in their in box and so on. If this virus keeps getting } passed, it has the potential to do a great deal of DAMAGE to computer } networks worldwide!!!! } } Please, delete the message entitled "PENPAL GREETINGS!" as soon as you } see it! And pass this message along to all of your friends, relatives } and the other readers of the newsgroups and mailing lists which you } are on so that they are not hurt by this dangerous virus!!!! } } Please pass this along to everyone you know so this can be stopped. } PASS THIS ON TO YOUR FRIENDS!!! WARNING !!! } There is a new virus going around in the last couple of days!!! } DO NOT open or even look at any mail that you get that says: "Returned } or Unable to Deliver" This virus will attach itself to your computer } components and render them useless. Immediately delete any mail items } that says this. AOL has said this is a very dangerous virus, and there } is NO remedy for it at this time, Please Be Careful, And forward to } all your on-line friends A.S.A.P. } } Forward this A.S.A.P. to every single person you know!!!!!!!!! } *************************************************** } } } }
Best regards,
Paul Vanderlinden. Sales Manager.
======================================================================= See our web site: http://www.microscopy-uk.org.uk
Course Announcement: "Optimizing Light Microscopy" When/Where: (a) New York City, November 3,1997 (b) Springfield, MA November 5, 1997 (c) Boston, MA November 7,1997 What: a lively, fast-paced slide lecture and demonstraton for anyone how uses or plans to use a light microscope: students, teachers, medical technologists, clinicians, pathologists, and lab managers. Beginning to more experienced practictioners welcome.
For details... (a) read below (b) send for brochure (c) visit the Microscopy/Microscopy Education booth at MSA - #502
Program: 1. A quick tour around the microscope - getting to know the bits & pieces 2. Koehler illumination & you: 4 critical steps for aligning you and your microscope to reduce headaches, fatigue, and errors 3. Care and cleaning 4. Useful principles for understanding and optimizing imaging 5. Putting the basics to work: a. Troubleshooting b. Understanding Phase and Hoffman Modulation Contrast 6. The Video connection: cameras, computers, and your microscope 7. Bringing out the best: quick, easy, and often free techniques for improving contrast 8. Advanced contrast techniques: Fluorescence and DIC 9. Becoming a better consumer: matching your microscope to your application 10. Questions, Answers, and Information Exchange (Note: Instructor may vary class content slightly to meet the needs of participants)
Free with your tuition: "Optimizing Light Microscopy for Biological and Clinical Laboratories" (Kendall-Hunt, 1997). Nearly 200 pages of helpful hints, quick experiments, and new iedas for getting the best from your microscope. Hot off the presses!
CEU's: 0.6 CEUs, 6 P.A.C.E CEU's Microscopy/Microscopy Education adheres to the guidelines established by the IACET.
Pricing: $150 (includes tuition, breaks, course materials, and copy of book) *****Save $25 if paid by 10/17/97.***** Send three from the same facility and save $50 on tuition for the third person.
Refund policy: Full refund for cancellations made by 10/17/97. After that date, 50% refund or full credit for future class. Substitutions accepted.
Questions: call Barbara Foster or Dr. Ken Piel at MME: (413)746-6931
Registration: Download the form below and fax to (413)746-9311 or call (413)746-6931 and ask for Ken.
Check course you will be attending: ___ New York City, November 3 (#971103) ___ Springfield, November 5 (#971105)* ___ Boston, November 7 (#971107)
Method of Payment: ____ Check enclosed for $ _______________ ____ Visa ____ Mastercard Name on credit card: __________________________________ Credit card number: ___________________________________ Expiration date: ______________________________________ ***If billing address is different from one shown above, please show billing address below: _______________________________________________________ _______________________________________________________ _______________________________________________________
The PENPAL GREETINGS! Hoax shown below appears to be an attempt to kill an e-mail chain letter by claiming that it is a self starting Trojan that destroys your hard drive and then sends copies of itself to everyone whose address in in your mailbox. Reading an e-mail message does not run it nor does it run any attachments, so this Trojan must be self starting. Aside from the fact that a program cannot start itself, the Trojan would also have to know about every different kind of e-mail program to be able to forward copies of itself to other people. This warning is totally a hoax.
FYI!
Subject: Virus Alert Importance: High If anyone receives mail entitled: PENPAL GREETINGS! please delete it WITHOUT reading it. Below is a little explanation of the message, and what it would do to your PC if you were to read the message. If you have any questions or concerns please contact SAF-IA Info Office on 697-5059.
This is a warning for all internet users - there is a dangerous virus propogating across the internet through an e-mail message entitled "PENPAL GREETINGS!". DO NOT DOWNLOAD ANY MESSAGE ENTITLED "PENPAL GREETINGS!" This message appears to be a friendly letter asking you if you are interestedin a penpal, but by the time you read this letter, it is too late. The "trojan horse" virus will have already infected the boot sector of your hard drive, destroying all of the data present. It is a self-replicating virus, and once the message is read, it will AUTOMATICALLY forward itself to anyone who's e-mail address is present in YOUR mailbox! This virus will DESTROY your hard drive, and holds the potential to DESTROY the hard drive of anyone whose mail is in your inbox, and who's mail is in their inbox, and so on. If this virus remains unchecked, it has the potential to do a great deal of DAMAGE to computer networks worldwide!!!! Please, delete the message entitled "PENPAL GREETINGS!" as soon as you see it! And pass this message along to all of your friends and relatives, and the other readers of the newsgroups and mailing lists which you are on, so that they are not hurt by this dangerous virus!!!!
Join the Crew
Circulating the Internet is an email message entitled "Join the Crew". For a virus to spread, it must be executed. Reading a mail message does not execute the mail message. Trojans and viruses have been found as executable attachments to mail messages, but they must be extracted and executed to do any harm.
CIAC still affirms that reading E-mail, using typical mail agents, can not activate malicious code delivered in or with the message.
IMPORTANT - VIRUS Alert!!!
Take note !
Someone got an email, titled as JOIN THE CREW. It has erased his hard drive. Do not open up any mail that has this title. It will erase your whole hard drive. This is a new email virus and not a lot of people know about it, just let everyone know, so they won't be a victim.
Please e-mail this to everyone you know!!! Remember the title : JOIN THE CREW
Variants of this email message are circulating the Internet. If you receive an email message entitled "Join the Crew" and it has an attachment, CIAC recommends that you delete the message and the attachment. If you receive just the message, delete the message. Please DO NOT circulate unvalidated virus alerts.
************************************************************************** Mike O'Keefe
Course Announcement: "Optimizing Light Microscopy" When/Where: "Optimizing Light Microscopy" Hosted by Providence College Dept. of Biology (Providence, RI) October 3, 1997 What: a lively, fast-paced slide lecture and demonstraton for anyone who uses or plans to use a light microscope: students, teachers, medical technologists, clinicians, pathologists, and lab managers. Beginning to more experienced practitioners welcome.
For details... (a) read below (b) send for brochure Program: 1. A quick tour around the microscope - getting to know the bits & pieces 2. Koehler illumination & you: 4 critical steps for aligning you and your microscope to reduce headaches, fatigue, and errors 3. Care and cleaning 4. Useful principles for understanding and optimizing imaging 5. Putting the basics to work: a. Troubleshooting b. Understanding Phase and Hoffman Modulation Contrast 6. The Video connection: cameras, computers, and your microscope 7. Bringing out the best: quick, easy, and often free techniques for improving contrast 8. Advanced contrast techniques: Fluorescence and DIC 9. Becoming a better consumer: matching your microscope to your application 10. Questions, Answers, and Information Exchange (Note: Instructor may vary class content slightly to meet the needs of participants. Instructor for this course: Barbara Foster of Microscopy/Microscopy Education)
Free with your tuition: "Optimizing Light Microscopy for Biological and Clinical Laboratories" (Kendall-Hunt, 1997). Nearly 200 pages of helpful hints, quick experiments, and new ideas for getting the best from your microscope. Hot off the presses!
CEU's: 0.6 CEUs, 6 P.A.C.E CEU's Microscopy/Microscopy Education adheres to the guidelines established by the IACET.
Pricing: $150 (includes tuition, breaks, course materials, and copy of book) *****Save $25 if paid by 9/22/97 ******* Send three from the same facility and save $50 on tuition for the third person.
Refund policy: Full refund for cancellations made by 10/17/97. After that date, 50% refund or full credit for future class. Substitutions accepted.
Questions: call Barbara Foster or Dr. Ken Piel at MME: (413)746-6931
Registration: Download the form below and fax to (413)746-9311 or call (413)746-6931 and ask for Ken.
Check course you will be attending: ___ Optimizing Light Microscopy (#971003)
Method of Payment: ____ Check enclosed for $ _______________ ____ Visa ____ Mastercard Name on credit card: __________________________________ Credit card number: ___________________________________ Expiration date: ______________________________________ ***If billing address is different from one shown above, please show billing address below: _______________________________________________________ _______________________________________________________ _______________________________________________________
This is a MIME-encapsulated message If you read this, you may want to switch to a better mailer --__==========00000000158236==cellbio.duke.edu==__ Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 8bit
************** Attention all microscopists who might wish to spend a weekend on the coast of North Carolina at THE most beautiful time of the year!!! **********
YOU ARE CORDIALLY INVITED TO PARTICIPATE IN ONE OF THE MORE FUN MEETINGS YOU ARE LIKELY TO ATTEND DURING THE NORMAL COURSE OF EVENTS!
I have attached a textfile for easy download, and also include the complete text as part of this message for those of you who prefer it this way!
Hope to see you in October!
THE NORTH CAROLINA SOCIETY FOR MICROSCOPY AND MICROBEAM ANALYSIS
presents the
SIXTEENTH ANNUAL SYMPOSIUM ON ADVANCES IN MICROSCOPY
"Correlative Microscopy in Biological and Physical Sciences" Blockade Runner Resort, Wrightsville Beach, North Carolina October 17-19, 1997
**************** FOR MORE INFORMATION PLEASE CONTACT PETER INGRAM OR ANN LEFURGEY AT: p.ingram-at-cellbio.duke.edu or a.lefurgey-at-cellbio.duke.edu *************** or READ ON!
Symposium Description The Sixteenth Annual Symposium, sponsored by the North Carolina Society for Microscopy and Microbeam Analysis (NCSMMA), has been planned with a theme of "Correlative Microscopy in Biological and Physical Sciences." Continuing with the tradition of the symposium, the guest lecturers are composed of both nationally and internationally distinguished scientists. The meeting has several purposes, not the least of which is to draw attention of the scientific community to emerging developments in the practical and basic research aspects of exciting new fields, and to bring people together from diverse disciplines to discuss how innovative techniques will be relevant to the future direction of microscopy and microprobe analysis. In particular, this year, special emphasis will be placed on how correlations between the many forms of microscopy are having significant impact in the biological and physical sciences. The symposium also offers an opportunity for interested participants to submit abstracts of related studies for poster display. Three special workshops/tutorials (SEE BELOW) will be offered at no additional charge to participants in the Symposium: (a) Introduction to Scanning Probe Microscopy, ( b) Confocal Microscopy, and (c) Immuno-labeling. These are practical, introductory workshops/tutorials and no previous experience or knowledge is necessary. The annual business meeting of NCSMMA will he held on Saturday, October 18th just prior to lunch.
NCSMMA Student Prizes Student prizes will be awarded at this annual meeting. A total of five awards will be granted to five different students in the following categories: Biological sciences - one abstract (platform presentation) and two posters; Physical sciences - one abstract (platform presentation) and one poster
September 29th is the submission deadline for abstracts of platform and poster presentations. The five winning students will each receive a $100.00 check as well as complimentary registration fees. All abstracts will be pre-judged by NCSMMA members, with platform presentations by the winners, and the posters will be judged at the time of the meeting. Candidates must be members of NCSMMA and be full-time students to be eligible for the competition, and must submit an abstract by the deadline to be considered for either platform or poster awards.
Registration Fees, Hotel rates The $90 ($100 on site) per person and $50 for students ($60 on site) registration fee includes: symposium attendance and materials, Saturday lunch, breaks, and Friday and Saturday evening meals. Additional Friday evening tickets are available for Adults - $20; Children 10 years of age and under - $10. Additional Saturday evening tickets are available for Adults - $20; Children 10 years of age and under - $10. There is a $15 fee for all cancellations. Blockade Runner Resort Hotel special rates start at $69/night including full breakfast.
For questions or further information, please telephone Betty Gooch, Duke University Medical Center: (919) 684-3534 or email: b.gooch-at-cellbio.duke.edu.
PROGRAM
Friday, 17 October, 1997 4:00 - 6:00 pm REGISTRATION and refreshments - Blockade Runner at Wrightsville Beach 6:30 - 8:30 pm Evening Buffet - pool side at the Blockade Runner, Wrightsville Beach Courtesy of JEOL (USA) Inc. and GATAN, Inc. Beverages courtesy of AMRAY Inc. (Complimentary with Registration; Adult and Children Guests $20/$10).
Saturday, 18 October 1997
8:30 - 11:30 am Special Workshops/Tutorials on: (a) Introduction to Scanning Probe Microscopy (hands on participation) (b) Confocal Microscopy (hands on participation) (c) Immunolabeling with Colloidal Gold 9:00 - 11:00 am Coffee, juice, and cookies - Blockade Runner 10:00 - 11:30 am Poster Session, Exhibitors' Displays - Blockade Runner 11:30 - 12:00 Noon NCSMMA Annual Business Meeting 12:00 noon- l:00 pm Lunch - Casual buffet - Poolside
1:00 - 1:15 pm - Welcome -
1:15 - 1:45 pm Correlative Microscopy in Biological Problem Solving Ralph Albrecht 2:00 - 2:30 pm Applications of Scanning Probe Microscopy Chuck Mooney 2:45 - 3:15 pm COFFEE BREAK 3:15 - 3:45 pm Diagnostic Microscopy in the Pharmaceutical Industry Ruth Lightfoot 4:00 - 4:30 pm Electron Backscatter Diffraction in the SEM Joe Michael 4:45 - 5:15 pm Project MICRO: Its Realization and Implementation Caroline Schooley
5:30 - 6:30 pm Poster Session, Exhibitors' Displays 6:30 - 7:00 pm Cocktails, refreshments 7:00 pm Evening Buffet- Al Fresco at the Blockade Runner, Wrightsville Beach Supported in part by Oxford Instruments, Leo and Zeiss
Sunday, 19 October 1997
8:20 - 9:00 am Student Awards and Presentations 9:00 - 9:30 am Correlative Microscopy in Materials Science Mike Kersker 9:45 - 10:15 am Cell Growth Studies with Confocal Microscopy Nina Allen 10:30- 11:00 am COFFEE BREAK 11:00 - 11:30 am EFTEM Techniques in Materials Science Jim Bentley 11:45 - 12:15 Noon Medical Applications of Correlative Microscopy David Howell 12:30 pm Finis
Accommodations Special arrangements have been made to provide a wide variety of accommodations. Use the reservation form immediately and send it directly to the hotel of your choice (except Blockade Runner, which must be telephoned directly) or telephone another hotel directly. Make sure to mention that you are attending the Duke Microscopy Symposium so that you will receive the special rates provided for our registrants.
MAKE YOUR RESERVATION NOW!
A one (l) night's deposit is required to hold reservations. All rates quoted are excluding tax.
Wrightsville Beach, NC (Lumina Avenue) * *BLOCKADE RUNNER RESORT. 1-800-545-5494. Ask for Code #5067. Harbor front $69/single; $77 dbl; ocean front, $89 single/$97 dbl. All rooms include breakfast. * Waterway Lodge (located at drawbridge). 1-800-677-3771. $55/ two dbl. beds or queen size. Condo $65/night. Includes queen and sleeper sofa plus full kitchen.. * Shell Island (at Wrightsville Beach). 1-800-689-6765. Double occupancy suites only. $129/night. * Hampton Inn, 1989 Eastwood Road. 1-919-0256-9600. $89/king or two queen beds plus includes continental breakfast with local calls. * Meeting site
Wilmington, NC (Market Street) * Greentree Inn, 1-910-799-6001. $43.95//two dbl. beds w/ continental breakfast. * Holiday Inn of Wilmington, 1-910-799-1440. $75/king or two double beds. * Howard Johnson Plaza, 1-800-833-4721. 89/night, two double beds or one king size bed. * Days Inn, 1-910-799-6300. $58.88//two dbl. beds. or 1 queen
************* Send checks payable to: "Sixteenth Annual Symposium on Advances in Microscopy" Send to: Betty P. Gooch, Symposium Coordinator Analytical Electron Microscopy Facility Box 3709 Duke University Medical Center Tel: (919) 684-3534 Durham, NC 27710 email: bgooch-at-cellbio.duke.edu Fax: (919) 681-8419
PLUS!!!!!
3 SPECIAL FREE WORKSHOPS!
TO BE HELD IN CONJUNCTION WITH THE
SIXTEENTH ANNUAL SYMPOSIUM ON ADVANCES IN MICROSCOPY
Sponsored by the North Carolina Society for Microscopy and Microbeam Analysis at the Blockade Runner Beach Resort Wrightsville Beach, North Carolina October 17-19, 1997
INTRODUCTION TO SCANNING PROBE MICROSCOPY: Application to Materials Science & Biology Chuck Mooney, Park Scientific Instruments, Sunnyvale, Ca.
Basics of STM and AFM
* Contact, non-contact and tapping microscopy * Problems and solutions * Lateral force microscopy * Operating parameters ***** Hands on participation ******
CONFOCAL MICROSCOPY Nina Allen, North Carolina State University, Raleigh, NC
Basic Operating Principles
* Problems, solutions and limitations * Laser sources * Confocal versus conventional fluorescence imaging * Slit and pinhole apertures and deconvolution * Fluorescence throughput * 3-D imaging * Alignment of optics * Judicious use of dyes * Sensitivity and background subtraction
***** Hands on participation ******
IMMUNOLABELING WITH COLLOIDAL GOLD Ralph Albrecht, University of Wisconsin, Madison, WI
* Production of gold colloids * Conjugation of antibody/gold ligand to colloids * Basics of labelling with colloidal gold conjugates * Techniques for silver enhancement of gold colloids * Problems, solutions and limitations
Use of gold conjugates as labels in correlative microscopy
We are interested in buying a used cryo-ultramicrotome. LKB, Reichert, or RMC acceptable. Please contact: Marek Malecki, PI Phone: 6082638481. Fax: 6082654076. Email: malecki-at-macc.wisc.edu
I was recently given the following homework assignment: Actual resolution as determined by measuring the smallest distance between two distinguishable points in a suitable specimen (resolution standard), is usually greater than the linear resolution of the microscope. Describe the factors that contribute to this discrepency. Find and list suitable good references. Can you give me any help in solving this problem? Do you know of any good sources to speak to? Do you know the answer? Thank you for your help, Fred Meisenkothen
I think that there must be somebody who have got experience with EDS for Al-alloys. I hope you can shine some lights on my silly questions.
We have several students working on Al-alloys (e.g. Al-Fe-Ti, Al-Fe-Cr et al) using arc melting. The problem is that the EDS results showed that all the alloys (as-cast or following solution annealing) lost about 20at% Al (for example, {20% for the nominal composition 25%)?! We tried on two machines, An-10000 EDS on JEOL 840 (over 10 years old) and Oxford ISIS EDS on FESEM Hitachi S-4500 (only 1 year old). They showed similar results at work conditions of 20kV, } 1000cps, for 100s on well polished samples.
Of course, there are several posibilities for the large amount of Al loss. Weight ratios have been double checked. Melting under Argon would cause little loss of Al because the melting point of Al is reletively lower than others, and some dark dust did appear in the furnace, but this error should be limited in +/-0.5%. Where did the Al go?
So, I wonder if there is anything wrong with the standardless analysis of EDS. We have tested the EDS with stanless steel sample. The results were well consistant with the nominal compositions. Is that because there are something wrong with the standard data of Al in the computer, or calibration, or work conditions, or something else? Any idea? Please help.
} Could I suggest the original poster try the Safety group. The address is } } LISTSERV-at-UVMVM.UVM.EDU } Write SUB SAFETY in the text area.
Good suggestion. There was a discussion within the last few years on this topic on the safety listserv. The folks at the University of Vermont who run the list also have an excellent website at: {http://siri.org} where they archive messages from the listserv, as well as maintain access to MSDS information and other safety related information. Full instructions for subscribing to the list are also there.
Among mostly serious information it also contains the following spoof of the Good Times virus hoax that is just too good not to repost here, although it of course has nothing whatsomuchever to do with translation. Same could be said about Join the crew, I believe...
READ THIS:
Goodtimes will re-write your hard drive. Not only that, but it will scramble any disks that are even close to your computer. It will recalibrate your refrigerator's coolness setting so all your ice cream goes melty. It will demagnetize the strips on all your credit cards, screw up the tracking on your television and use subspace field harmonics to scratch any CD's you try to play.
It will give your ex-girlfriend your new phone number. It will mix Kool-aid into your fishtank. It will drink all your beer and leave its socks out on the coffee table when there's company coming over. It will put a dead kitten in the back pocket of your good suit pants and hide your car keys when you are late for work.
Goodtimes will make you fall in love with a penguin. It will give you nightmares about circus midgets. It will pour sugar in your gas tank and shave off both your eyebrows while dating your girlfriend behind your back and billing the dinner and hotel room to your Discover card.
It will seduce your grandmother. It does not matter if she is dead, such is the power of Goodtimes, it reaches out beyond the grave to sully those things we hold most dear.
It moves your car randomly around parking lots so you can't find it. It will kick your dog. It will leave libidinous messages on your boss's voice mail in your voice! It is insidious and subtle. It is dangerous and terrifying to behold. It is also a rather interesting shade of mauve.
Goodtimes will give you Dutch Elm disease. It will leave the toilet seat up. It will make a batch of Methanphedime in your bathtub and then leave bacon cooking on the stove while it goes out to chase gradeschoolers with your new snowblower.
Listen to me. Goodtimes does not exist.
It cannot do anything to you. But I can. I am sending this message to everyone in the world. Tell your friends, tell your family. If anyone else sends me another E-mail about this fake Goodtimes Virus, I will turn hating them into a religion. I will do things to them that would make a horsehead in your bed look like Easter Sunday brunch.
Colleagues, I am wondering how one would keep the particles from falling through the open holes since the particles in general are smaller than the size of any holes in a holey film. Also, how do your "Ultra-thin carbon films" differ from the ones I purchase from Agar Scientific in the UK and from SPI in the USA? Are they really "thinner" on the basis of some measurement and if so, what measurement do you use?
Sincerely
.attila(L)
Attila L. Toth ------------------------------------------ MTA MFKI Research Institute for Technical Physics of the Hungarian Academy of Sciences H-1325 Budapest POB 76 tel: (36.1) 169-2100 x 226 fax:(36.1) 169-8037 ------------------------------------------ email: tothal-at-mufi.hu (EUDORA:=CDrj =E9kesen!) ------------------------------------------
Really need a bit more information, but... Have you examined the material for homogeniety? BSE imaging of a polished surface would be a good start. If you have more than one phase / precipitates, apparent composition will be a strong function of where you analyze and the size of the analysis volume relative to the phases/inclusions. Generally I have found that inhomogenous materials will compromise analysis accuracy.
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
Dear All I was experimenting with new mail filtering and inadvertenetly sent mail to a number of addressess that were not supposed to be mailed. Many apologies if you receive that mail either directly or via the list
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 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
} Here is what I propose: take some epoxy resin and coat the inside of a } polypropylene beaker with the epoxy by building up layers and polymerizing } the layers.
} Allow the uranium liquids to evaporate } to dryness (taking care to avoid generating dust) and apply another layer } of epoxy over the uranium salts.
I have to emphasize the point about avoiding dust. Uranium is an alpha-emitter (not a beta-emitter as another poster said, although some of the daughter nuclei emit betas), and the range of these alphas is so short that they will not penetrate the dead layer of the skin. Thus, uran- ium is not dangerous unless it is ingested or inhaled. If, however, uran- ium is inhaled, particles sitting on the lung cells will provide a very large dose to those cells, and are a serious carcinogen. It is much safer for everyone if there is NO chance of producing dust or droplets containing uranium; I'd even be very careful about pouring the liquid into a beaker. Yours, Bill Tivol
Attila L. Toth wrote: ================================================== I am wondering how one would keep the particles from falling through the open holes since the particles in general are smaller than the size of any holes in a holey film. Also, how do your "Ultra-thin carbon films" differ from the ones I purchase from Agar Scientific in the UK and from SPI in the USA? Are they really "thinner" on the basis of some measurement and if so, what measurement do you use? ================================================= You are right, CdS nano- particles, at least the ones we have seen, those that would be thin enough to "see" through, are smaller than the smallest holes we have ever seen anyone able to make in a "lacy" film. So I think that there might have been some misinformation accidently posted about this a while back (see Aug. 11 posting DUNNTEM-at-aol.com working for Ted Pella, Inc .). If I am wrong about this please correct me and set the record straight .
With regard to so-called "ultra-thin carbon films", obviously carbon films can be made almost infinitely thin, but they do have to have enough mass to be self-supporting on the grid mesh being used. That minimum amount of mass then is going to be a function of the mesh size, the lower the mesh size (e. g. larger the hole), the more durable (a.k.a. thicker) must be the carbon film. If thickness of the carbon film is crucial, e.g. you want it completely minimized, I would recommend our finest, which is our 2000 mesh grid. This simple reality is often times missed by people worried about film thickness (which in fact ought to not even be relavent in the case of a lacey or holely film, because after all, you are getting your information through the holes, not through the "lacy" areas). So the whole need to worry about "thickness" per se really ought to be, in the case of lacey films, a non-issue!
But addressing the subject of carbon film thickness generally, when one does have the need to "look through" the film, the philosophy of "one thickness fits all" is not one to which we subscribe. One can have the luxury of having that philosophy only if there are in-house facilities available to quality check what has been made, otherwise the customer ends up doing the quality checking. And finding out at the last minute that coated grids are not stable and can not be used is usually not a very pleasant discovery.
So while I can not comment on the methods used by our competitors to measure film thickness, I can tell you how we do the "test" and that is, as we are making them, and we strive for the minimum possible amount, before we have made too many, we walk across the hall and put them into a TEM that is dedicated (after 5:00 pm) for this purpose, that is, to do our own "clinical" test to make sure that the film is sufficiently durable (e.g. thick enough) for that given mesh size being used. Now you might not believe this, but the instrument used for the testing is an RCA EMU 4-B, manufacturerd by RCA in 1969. So we think of this as a worst case test. If the carbon film is stable enough to survive the beam of an RCA EMU-4B, an instrument featuring technology more than thirty years old, surely it ought to be more than acceptable in an instrument of more modern manufacture (assuming minimum column cleanliness). And that kind of test procedure results in a history of almost (note I said "almost") no returns!
Interestingly enough, the films do not have to be "thicker" to be stable in the RCA than in our much newer JEOL TEM. A film with minimum thickness, stable in one, seems to be similarly stable in the other.
More information about our custom coated carbon grids can be found on our website given below.
Disclaimer: SPI has produced custom coated grids for customers for more than twenty years so we have an interest in promoting our carbon coated grids. The posting I characterized as containing "misinformation" came from a competitor, so everything I said should be taken within that context.
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 ############################ ==================================================
Jim Darley wrote: ================================================= But too many regulations are not wise - eg. the dangerous goods shipping laws which appear designed to make things more expensive but not safer. ================================================= We should all be taking the HAZMAT reguations, no matter where we live, seriously. I can not comment about the laws in Australia, but I have a very high regard for the regulations promulgated by the U. S. Dept. of Transportation (DOT) and IATA (for international shipments) as well as the reasons behind them. And I can state, from first hand experience, that if the regulators are presented with sound technical information for a reguation to be changed, they will indeed listen (at least in the US) and sometimes make changes.
If anyone remains unconvinced, about the importance of adherence to all HAZMAT regulations, keep in mind that the ValuJet disaster was caused by someone not taking seriously these same regulations.
While it is correct that one can incur almost unbelievalbly high shipping costs for HAZMATs, this is not always the case. In many instances, such as for the ordering of osmium tetroxide, if one orders "smart", they can do their shipping at costs only nominally more than if the same weight of tweezers, grids, or SEM mounts were being shipped. Now this would not apply for everything (e.g. our SPI Dusters, for example) but it does apply for a surprising number of HAZMATs routinely used in EM labs. We are also in the process of reformulating some of our embedding kits so they too can be shipped at the lower prices.
We have tried to explain how a customer can "order smart" on our website, click on "Hazardous Items(Good News and Bad News)". While savings in shipping costs for domestic US customers are possible, the real beneficiaries are foreign customers now no longer are restricted to the use of air freight (which has associated with it high minimum charges).
And while we are on this subject, just remember, don't ever try to take HAZMATs in checked airline luggage to save some money, it is unconscienable from a moral standpoint and puts at risk everyone on the plane flight.
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 ############################ ==================================================
Jim Darley wrote: ================================================= But too many regulations are not wise - eg. the dangerous goods shipping laws which appear designed to make things more expensive but not safer. ================================================= We should all be taking the HAZMAT reguations, no matter where we live, seriously. I can not comment about the laws in Australia, but I have a very high regard for the regulations promulgated by the U. S. Dept. of Transportation (DOT) and IATA (for international shipments) as well as the reasons behind them. And I can state, from first hand experience, that if the regulators are presented with sound technical information for a reguation to be changed, they will indeed listen (at least in the US) and sometimes make changes.
If anyone remains unconvinced, about the importance of adherence to all HAZMAT regulations, keep in mind that the ValuJet disaster was caused by someone not taking seriously these same regulations.
While it is correct that one can incur almost unbelievalbly high shipping costs for HAZMATs, this is not always the case. In many instances, such as for the ordering of osmium tetroxide, if one orders "smart", they can do their shipping at costs only nominally more than if the same weight of tweezers, grids, or SEM mounts were being shipped. Now this would not apply for everything (e.g. our SPI Dusters, for example) but it does apply for a surprising number of HAZMATs routinely used in EM labs. We are also in the process of reformulating some of our embedding kits so they too can be shipped at the lower prices.
We have tried to explain how a customer can "order smart" on our website, click on "Hazardous Items(Good News and Bad News)". While savings in shipping costs for domestic US customers are possible, the real beneficiaries are foreign customers now no longer are restricted to the use of air freight (which has associated with it high minimum charges).
And while we are on this subject, just remember, don't ever try to take HAZMATs in checked airline luggage to save some money, it is unconscienable from a moral standpoint and puts at risk everyone on the plane flight.
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 ############################ ==================================================
With regard to references the following are quite good for sizing in the optical microscope:
Handbook of Chemical Microscopy by Chamot and Mason
The Particle Atlas, Volume 1 by McCrone and Delly
Particle Size Measurement Vol 1 by Terence Allen
I have some trouble understanding the question, it doesn't seem to address any issue precisely since it does not specify the optics, the resolution standard, the wavelength of light used or the method of measurement. All those factors affect the resolution. Real measurements are also affected by the contrast in the specimen, the calibration standard, and such mundane issues as how well the optics are aligned and adjusted (ie the use of Kohler illumination, etc.) and how young and fit the eyeballs are doing the measurements - even the lighting in the room can affect measurements.
The questioner may be after how diffraction affects particle measurements. The limit of detection of the optical microscope is much less than the resolution limit. I can detect objects much smaller than 0.4 um (the resolution limit of my neofluor 40 times objective with white light) - probably down to 0.1 um, but diffraction effects make them appear larger so that they all look to be about 0.4 um. Allen's book has a nice description of this effect and the other two books have good discussions of the origins of the resolution equation.
Hi- I am forwarding this message for a colleague who asked me this question. I don't have a clue. Any help appreciated. Dave
} The calibration of the cell for pH measurements using fluorescent probes } is done by using ionophores such as nigericin or by ratiometric calibration. } Does any of this method account for the effect of the dielectric constant } of t } the medium on the pKa' of the probe ? If not is the effect of the } dielectric co } nstant of the medium accounted by any other method ? Is it justified to } ignore the effect of the dielectric constant of the medium ? } } Thank you in advance for your help and would be eagerly waiting for the } respo } onses. } Abizer Harianawala }
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu
Dr. Jarett, The following ad has been posted on the Microscopy listserver which exclusively serves the em community. I did not include the email address on purpose to prevent unnecessary postings in your mail box. Hope you have a pleasant weekend. Neelima
ELectron Microscopist:
The University of Pennsylvannia School of Medicine Institutional Electron Microscopy Core is searching for a Co-Director. This Core has the broad support of the Diabetes Center, the Cancer Center, the Department of Pathology and Laboratory Medicine, and School of Medicine. The Core is widely used by medical community, as well as the University as a whole. The candidate should have a Ph.D. in cell and molecular biology and at least 5 years experience with all aspects of Electron Microscopy. The individual should also possess administrative experience and computer skills. The position title will be Senior Research Investigator and will be responsible for helping the Director develop reports for centers, grants, papers etc. Salary will be consistent with experience. Respondents should send their curriculum vitae, bibliography, and 3 references to: Dr. Leonard Jarett, M.D. Chair, Department Of Pathology and Laboratory Medicine Universtiy Of Pennsylvania 6 Gates Building 3400 Spruce Street, Philadelphia, Pa 19104-4283 Regards... : ) ; ) Visit us at http://www.med.upenn.edu/~path/core/EMCMAIN1.HTM
You should always be careful regarding ZAF corrections for mixtures of light and heavy elements. My advice is, if you don´t have a suitable standard to test your ZAF corrections, then don´t trust them. You have not calibrated your ZAF corrections until you test them on a suitable standard. For mixtures of Al-Fe-Cr, use a similar standard, not just Fe-Cr or Fe standards.
Sorry Bill but Uranium emits a whole zoo of radiations, alpha, beta, gamma. A jar of UA placed on a sheet of fast film for a day or two will expose the film through the jar. UA dissolves beautifully in water and the sitting in the lung argument applies more to other forms of U - like in mining. I maintain that poring small quantities (I used to discard 5ml/months of a 2% solution) into the sewage system is the most sensible solution. Just sit down and work out the dilution factor on an annual basis and I expect in most cities the result will be approximately that concentration of U in seawater. 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, 400+ Links, MSDS ************************ http://www.proscitech.com.au
---------- } From: William Tivol {tivol-at-wadsworth.org}
} } Dear John & others, } } } Here is what I propose: take some epoxy resin and coat the inside of a } } polypropylene beaker with the epoxy by building up layers and polymerizing } } the layers. } } } Allow the uranium liquids to evaporate } } to dryness (taking care to avoid generating dust) and apply another layer } } of epoxy over the uranium salts. } } I have to emphasize the point about avoiding dust. Uranium is an } alpha-emitter (not a beta-emitter as another poster said, although some } of the daughter nuclei emit betas), and the range of these alphas is so } short that they will not penetrate the dead layer of the skin. Thus, uran- } ium is not dangerous unless it is ingested or inhaled. If, however, uran- } ium is inhaled, particles sitting on the lung cells will provide a very } large dose to those cells, and are a serious carcinogen. It is much safer } for everyone if there is NO chance of producing dust or droplets containing } uranium; I'd even be very careful about pouring the liquid into a beaker. } Yours, } Bill Tivol }
I would guess that you are trying to push standardless EDS a bit further than it's intended to go. Why don't you use some standards?
Ritchie
} We have several students working on Al-alloys (e.g. Al-Fe-Ti, } Al-Fe-Cr et al) using arc melting. The problem is that the EDS results } showed that all the alloys (as-cast or following solution annealing) lost } about 20at% Al (for example, {20% for the nominal composition 25%)?! } Where did the Al go? } So, I wonder if there is anything wrong with the standardless } analysis of EDS. } Charlie Kong
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
We are continuing to improve the US Histochemistry Society Web page and welcome your suggestions as to how we can be of better service to the community.
Right now, we are running a link to the Immunocytochemistry Discussion Newsgroup. Check out our page for instructions on how to get involved. It is: http://www.hcs.microscopy.com
We are also running a contest for the best Logo for our society. We have ten entries and welcome more. The prize is $200.00. Check out the Logo contest on the above web site and vote for the best and submit your own.
Thanks for your attention!
Gwen Childs
************* Gwen V. Childs, Ph.D. Professor and Vice-Chair Department of Anatomy and Neurosciences University of Texas Medical Branch Galveston TX 77551-1043 gvchilds-at-utmb.edu http://cellbio.utmb.edu/childs/childs.htm (409) 772-1942; FAX 772-3222 Toll Free Pager: 1 888 715-8636
I have an old EDAX 9100 detector with broken Be window and photo diode plus FET. I like to know whether (1) someone could fix it quickly with affordable price or (2) some information on where we could buy the Be window and the diode-FET unit, plus tips to fix the detector. My students and I would appreciate any help from you and we hope we could have our detector back to work so we could continue our experiments.
Thanks so much,
Judy Wu Dept. of Physics Univ. of Kansas Lawrence, KS 66045 (785)864-3240 (phone) (785)864-5262(fax)
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Dear Microscopists:
We would like your help in locating the best candidate for the position of Director for our Applications Laboratory. The following text and attachment describe the opportunity. Would you please post the advertisement and bring it to the attention colleagues?
IMAGING RESEARCH OPPORTUNITY
We are seeking a talented individual to direct our Applications Laboratory, leading research into fluorescence detection, confocal infrared and ultraviolet microscopy, and x-ray microscopy. The incumbent will also participate in the specification and design of novel optical systems developed by the Engineering Department. The Applications Laboratory patents and publishes research findings, supports Microcosm=92s customers=92 in their use of our technology, and communicates the implications of internal and external research to the Marketing and Engineering Departments.
The position requires a Ph.D. in cell biology or a related science, with primary experience in microscopy and imaging, and a strong physical science background. Familiarity with the latest microscopy techniques is mandatory. It is essential that the incumbent has sound knowledge of optics and digital image analysis, both in theory and in practice. Ideally candidates will have experience with several imaging platforms, including isee and dsp/os, NIH-Image, Image Tool, Metamorph & Image-1, as well as the LSM and MPM instruments made by Carl Zeiss.
The compensation package for the right candidate is negotiable. Benefits will include medical and dental insurance, and participation in our profit-sharing 401(k) retirement plan.
Interested persons should send their r=E9sum=E9 to the attention of Dr. Patrick Huddie at the address above. Please direct e-mail to phuddie-at-microcosm.com.
-------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com The Web is:"Vaster than empires and more slow" Andrew Marvell (1621-1678)
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A good place to start is "The Image Processing Handbook" Second Edition. John C. Russ. CRC Press. ISBN 0-8493-2516-1
Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Hi, Thank you who kindly responded to my question on EDS for Al-alloys. Troublesome students always raise extraordinary questions to test the range of knowledge (and/or self-confidence) of their teachers. Sometimes, they even try to meltdown the reputation of modern technique with the spark of their intuition. Now, let's show our royalty to the advanced materials science. The big question mark has not been erased yet, although the simplest answer, which may be the right one and has been selected, is to blame the students who prepared the dummy samples. Who knows whether they swallowed a piece of aluminium down before melting or not? There were some beginners who asked me quite often about the error range and sensitivity of EDS. My answer is that at first it depends upon your sample (Am I a good lawyer?); secondly, in micro-scale few thing is uniform (Do not blame me if the results are unrepeatable); at last, I would say that in ideal testing conditions the error should be less than +/-0.5% in general(I hope so indeed!). They were quite happy to use the machine. Several experts have advised me that we should give up the dependence upon the standardless analysis of EDS, although the others showed strong evidence to prove that it is working well. Faster would never be safer, just like driving a car. This should not imply that the results of "semi-quantitative analysis" can only be trusted in HALF, neither the error may be 50%. How do you feel? Do you mention "semi-" quite often to the users? My former colleague, Bruce, suggested that oily detector window might cause the problem for softer radiation such as Al-Ka. This message struck a spark in my poor memory. I really saw somebody who dared to wipe oil off the thin Be-window of a TN-5400 EDS using a cotton ball with a drop of acetone. That was ten years ago. Let's go to the question: Have you ever cleaned the Be window of your EDS system? If the answer is positive, how often? Or, just wake me up ---- NEVER EVER THINK ABOUT THAT, YOU THE TROUBLE MAKER! I swear I have never been a trouble-maker. I just want to learn something from you. Just talking, no action following. It is too lang. Thank you for your time.
A professorship due to the retirement of Professor Jon Gj=F8nnes at University of Oslo has been announced. The professor is expected to strengthen the reseach activity of the department in materials science/structure physics. The activity is mainly concerned with the application of electron-optical techniques to metallurgical questions, ceramic and semiconductors and with the development of quantitative methods for obtaining structural information (crystal structure, electronic structure and disordered structure).
Here follows the official announcement, including the application procedure. The announcement can also be found in the Journal Nature. The application deadline is October 3 1997. For additional information please contact
Professor Arne Olsen Department of Physics/Centre for Materials Science Gaustadalleen 21 0371 Oslo Norway Tel. (+47) 22 95 87 40 Fax: (+47) 22 95 87 49 e-mail: arne.olsen-at-fys.uio.no
Professor in Physics (Materials science/Structure physics)
The Faculty of Mathematics and Natural Sciences at the University of Oslo invites applications for the position of Professor in the Department of Physics with research interests within the field of materials science/structure physics.
The Department of Physics at the University of Oslo has 83.5 academic staff of which 26 are temporary (17 research associates and 9 adjunct professor positions). Further,there are about 45 funded by external sources. The department has 38 technical staff positions and 9 administrative staff positions.
Teaching in the Department is directed towards the degrees cand. mag. (approx. B.Sc.), siv.ing. (M.Eng), cand.scient. (M.Sc.) and dr.scient. (Ph.D). There are currently 110 students enrolled in Masters programmes and 70 in doctoral programmes.
Research in the department is organised in 8 groups which undertake both experimental and theoretical studies: Biophysics, Electronics, Elementary Particle Physics, Condensed Matter Physics, Nuclear and Energy Physics, Plasma and Space Physics and Structure Physics. In addition there is a Theoretical Physics Group.=20
The vacant professorship is connected to the Structure Physics Group, which also is part of the Faculty's Centre for Materials Science. The Structure Physics Group, localized in the Oslo University Research Park, runs an electron microscopy and metallographic laboratory with two transmission electron microscopes (JEOL 200CX and 2000FX) equipped for analysis (EDS, EELS, TV-system), optical microscopes and image analysis. A new TEM instrument will be installed in 1998. The research group, which numbers 20-25 students and staff, has extensive collaboration with other research groups within the University and in Norwegian industrial and public research laboratories. The group also maintains strong international contacts. The academic staff are engaged in the University's teaching programme for materials science as well as in general physics teaching in the department.=20
Much of the research in the group is linked to the application of electron-optical techniques to metallurgical questions, ceramics and semiconductors and to the development of quantitative methods for obtaining structural information (crystal structure, microstructure, electronic structure and disordered structure).
The professor will be expected to strengthen the research activity in Structure Physics as well as the operation of the group's laboratories. The successful applicant must be able to document scientific expertise in one or more of the group's major activities.=20
The appointee must be able to provide supervision at all levels of teaching. The professor will have responsibility for supervision of masters and doctoral candidates within her/his special field. The language of instruction for undergraduate courses is Norwegian, but English will be accepted for the first three years of appointment. The appointee may also be required to undertake administrative duties as prescribed in the applicable University regulations.
According to current regulations, the evaluation of applicants takes regard of scientific, professional and educational qualifications as well as other activities, which may qualify the applicant. Where several applicants are deemed to have equivalent qualifications after evaluation of the scientific, professional and educational qualifications, a female applicant will be ranked above a male applicant according to the procedure for the appointment of professorial staff.
The application shall specify the candidate's education, previous positions, scientific, professional and educational activities and administrative experience. Curriculum vitae and publications list are therefore to be included.
The application shall furthermore include a short description of the scientific works that the applicant regards as the most significant and on which the evaluation might especially be based. Normally this ought not exceed 10 items.
The application is to be addressed to the Academic Collegium, University of Oslo, and is to be sent with documentation to: Faculty of Mathematics and Natural Sciences, P.b. 1032 Blindern, N-0315 Oslo, Norway by the application date. Within one month of this date, the applicant must have sent to the Faculty's Secretariat:
- 5 complete sets of scientific works, published or unpublished, which one wishes to be considered in the evaluation (normally not exceeding 10)
- 5 copies of the application with documentation (C.V., complete publications list, description of the 10 most important works)
Scientific works which are in preparation on the application date may nonetheless be submitted within three months of the deadline provided the Secretariat is informed when the remaining works are submitted.
After the application date, the University will send applicants instructions for submission of scientific works.
One is otherwise referred to the regulations for appointment of professorial staff approved by the Academic Collegium in accordance with the University Act =A732.
I thank Garratt-Reed for corroberating my previous posting. John Bazzola had asked for a rough guide (he has confirmed that since) on detection limits of EDS versus WDS techniques. Anybody who has any significant experience with microprobe analysis knows that there is no single line correct answer. However, it is imperative for analysts to remember a few general figures so they can advise on appropriate instrumentation and techniques.
John B's initial inquiry deserved a reply and when none was given, I posted mine more than a day later. I believe that my posting is a useful guide for non-specialist analysts. Nothing that GR writes makes nonsence of my posting.
Nobody had asked about other differences between the techniques eg. resolution or simultaneous acquisition. I am pleased that G-R has supplied some information on those topics and on new, very high count-rate acquisition facilities for EDS. G-R is proud of his 0.6% "quantitative analysis" of Cr at an accuracy of +/- 0.1wt%.
I suggested that in EDS the presence of 1% of an element is the approxiamte lower limit for quantitative analysis. G-R has lowered that limit by some 40% - or has he? +/- 0.1% is good when 20% of the element is present, as it represents an accuracy of 0.5%. +/- 0.1% when 0.6% is present is about +/- 16% accuracy; I call that qualitative or at best semi-quantitative.
Another correspondent emailed me and noted that it was President Trueman who had been looking for a one handed adviser. But that was for an economist and not a scientist as I, apparently wrongly, remembered. The correspondent could see my point though; thanks to Brian Demczyk. I think that Trueman had an excellent idea but he should have extended that search to a scientist as well. Certainly microscopists and economists share disciplines which combine art and science. And I should add require 'good judgement'.
Why now was I abused with that opening: "Jim Darley's posting is hardly furthering good science". Am I to believe that good science is advanced by quarelsome nitpicking and that all broadbanding is verboten? I plead not guilty. 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, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} } Subject: Re: detection limits x-ray } } Date: Friday, 5 September 1997 6:21 } } } } Jim Darley's posting is hardly furthering good science. } } } } 1: Detection limits (John's original question). Modern EDX systems are } } easily capable of quantitative analysis in the sub-1wt% range. See, for } } example http://prism.mit.edu/facltis/stem/stmexam.htm (the second } } illustration on that page) where I was getting analyses for Cr in steel } of } } the order of 0.6+/-0.1 wt%. I don't know about you, but I consider this } } quantitative. This, mind you, was in a measurement where I was } attempting } } to optimize spatial resolution rather than sensitivity, and the } acquisition } } time was 60sec. per data point. This example, of course, was from a } STEM. } } There are many more examples in the literature. } } } } Using beam gating techniques such as those developed by Charlie Lyman and } } colleagues, EDX data can readily be acquired at 20,000-40,000 counts per } } real second, if spatial resolution is sacrificed. Combine this with an } } acquisition time of 1,000 seconds (by no means unrealistic for an } important } } measurement) and the detection limit is in the rage of, or better than, } 0.01wt%. } } } } Of course, in the SEM, which may have been the point of John's original } } question, the situation is not the same, the beam voltage is lower } } (resulting in poorer peak/bremmstrahlung ratios) and the emission of } x-rays } } from a solid sample is different from that in a thin foil, but these are } } some of the variables that Michael quite rightly pointed out must be } considered. } } } } 2: Comparison between EDX and WDX. } } } } This is like comparing apples and oranges, because the instruments } designed } } with them are generally intended for different purposes. } } } } WDX has a much better peak resolution than WDX, which results in better } } measured P/B ratios (and hence improved statistics), as well as much } better } } capability in resolving nearby x-ray lines. Also, because the x-ray } } counting and wavelength analysis are different functions in the crystal } } spectrometer, available countrates have traditionally been higher in WDX } } than EDX (although modern EDX detectors are an order of magnitude faster } } than they were fifteen years ago). Against this must be set the fact } that } } WDX is inherently a serial technique (although multiple spectrometers } help } } here), while EDX is a parallel technique (compare the advantages of PEELS } } over SEELS - although this is not a totally fair comparison). Anyway, } the } } advantage of WDX (ignoring the capability of resolving peak overlaps) is } } improved statistical precision. Is this useful? } } } } Well, maybe. } } } } By far the most significant parameter which affects electron-induced } x-ray } } emission spectra is sample geometry. Two extreme cases are where the } sample } } is a thin foil (as in the STEM) where, to a first approximation, the } x-ray } } spectrum recorded by the detector is the same as that emitted, and to a } } second order, it is possible to derive a reasonable thickness correction } for } } cases where the error is small. Alternatively, when the sample has } } dimensions large compared with the volume irradiated by the electron } beam, } } and has an accurately known geometry compared to the incident beam and } the } } detector (such as a polished flat sample in the microprobe), correction } } programs such as ZAF can do a reasonable job of extracting a quantitative } } analysis. } } } } What about where the sample geometry is unknown (for example, a rough } } surface such as might be examined in the SEM)? In that case the } uncertainty } } in the analysis is far, far worse than any uncertainty caused by the poor } } statistics of the EDX spectrum, so there is no point or advantage } whatever } } in using WDX to try to improve things, because it won't. This is why } } typically an SEM has an EDX detector - it is cheaper and gives just as } good } } an analysis (except for the somewhat poorer detection limit). In the } } microprobe, great care is taken in polishing and mounting the sample, so } the } } advantage of the WDX detector can be realised. } } } } Where does this leave us? The advantage of a WDX detector over an EDX } } detector *ON THE SAME SAMPLE* is limited - perhaps an order of magnitude } in } } detection limit, and, on a flat, polished sample, also perhaps } approaching } } an order of magnitude in precision. The WDX detector can also resolve } many } } cases where peaks would overlap in EDX. On general rough SEM samples, } the } } only advantages of WDX are a small improvement in detection limits and } the } } ability to resolve overlaps (which could be important if a trace element } } peak overlaps a major peak in the EXD spectrum. } } } } The President's science advisors were right - it all depends. I seem to } } remember that one of Jim Darley's Prime Ministers (Gough Whitlam?) had to } be } } dismissed because Parliament would not pass his budget - but does that } have } } anything to do with Science? } } } } Tony Garratt-Reed } } } } } } } Hi John and Michael et al: } } } I recall that one of your Presidents (Kennedy?) was looking for a one } } } handed science adviser; the ones he had always prevaricated "on the one } } } hand and on the other hand". } } } } } } Michael is quite right, detection limits vary greatly depending on } endless } } } factors. However, it is useful to have some figures as guidelines: } } } Considering say the 10 elements following sodium. Detection of these is } } } about best. } } } For these in EDX the limit for quantitative analysis is about 1%. } Detection } } } limit is about 0.1%. Increasing counts and counting times beyond the } } } customary 100 seconds at perhaps 2000cps will scarcely improve either } } } limit. } } } } } } WDX is near quantitative to its detection limit and that is at least two } } } orders of magnitude greater than is EDX. } } } } } } I'll enter correspondence only when it concerns errors in excess of five } } } orders of magnitude. } } } 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, 400+ Links, MSDS } } } ************************ http://www.proscitech.com.au } } } } } } } John J. Bozzola wrote: } } } } } } } } } } Under ideal (and reasonably attainable conditions), what is the } } } } } detection } } } } } limit (in grams) for EDX and WDX? Thanks. } } } } } } } } The question you ask is not specific enough ... to many factors to } be } } } } considered with regard to which elements you are interested in, and } } } } (e.g.) ... how sensitive your specimen is to long count times and high } } } } beam currents ... ... ask again ... } } } } cheerios, shAf } } } } -- } } } } } } } } } Anthony J. Garratt-Reed } } MIT Room 13-1027 } } 77 Massachusetts Avenue } } Cambridge, MA 02139-4307 } } United States of America } } } } Ph: 617-253-4622 } } Fax: 617-258-6478 } }
I gave shipping regulations as an example, where regulations frequently do not make much sense. While shipping regulations are not directly a 'microscopy matter' they do affect us all. Clearly, I did not advocate flaunting any regulations; it's foolhardy for an individual and suicidal for a business. But we are free to discuss: Are those rules wise, cost efficient and appropriate. It appears Chuck Garber thinks they are, I beg to differ. I am only concerned with international airfreight and the IATA defined 'dangerous goods' regulations. To make shipping safer, regulations can extend to packaging requirements, maximum quantities shippable, cargo only aircraft requirements for some goods and 'antidote packing'. Generally, packing requirements are reasonable. Weight limits set to exempt 'dangerous goods' are few and generally they are too high. This means tiny quantities of not particularly dangerous goods require very expensive shipping. Antidote packing is basically not used. It makes sense to pack OsO4 in a tin with a little full cream powdered milk. Vapour from a cracked vial would be absorbed and it gives the lab a handy supply of that powder to keep for any laboratory OsO4 problems. We have shipped OsO4 in that manner for many years now. Packing in vermiculate is less effective than newspaper - which would at least absorb some of the vapour. The funniest thing is the paperwork required and this is largely used to justify the expense of DG shipments. Spot checks and appropriate fines for non-compliance would be a better preventative than lots of paper work. To wit: The ValuJet disaster did happen despite those rules. Dangerous goods, certainly on international routes, go mostly by passenger aircraft. Picture the site of a crash - who would scramble through all that useless paper and to what good? The worst is the expense: We work on airfreight from the US at US$10/kg for the gross weight of medium sized shipments. For DG that figure for generally larger shipments is US$25/kg - which frequently is more than the value of the goods. This cost (+paperwork and extra packing) would be the cost the end-user has to bear if it meant substantially improved safety. I can see none. The overall global cost of DG shipments above the cost of normal shipments has to amount to several billion $ annually. It suits IATA and the airlines and apparently Chuck Garber. I submit that we are getting negligible additional safety for thousands of megabucks. Retaining and improving on the rules for packing and marking of boxes but severely limiting the paperwork would be a rather more effective option. 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, 400+ Links, MSDS ************************ http://www.proscitech.com.au
---------- } From: Garber, Charles A. {cgarber-at-2spi.com} } Date: Saturday, 6 September 1997 8:07 } Jim Darley wrote: } ================================================= } But too many regulations are not wise - eg. the dangerous goods shipping } laws which appear designed to make things more expensive but not safer. } ================================================= } We should all be taking the HAZMAT reguations, no matter where we live, } seriously. I can not comment about the laws in Australia, but I have a very } high regard for the regulations promulgated by the U. S. Dept. of } Transportation (DOT) and IATA (for international shipments) as well as the } reasons behind them. And I can state, from first hand experience, that if } the regulators are presented with sound technical information for a } reguation to be changed, they will indeed listen (at least in the US) and } sometimes make changes. } } If anyone remains unconvinced, about the importance of adherence to all } HAZMAT regulations, keep in mind that the ValuJet disaster was caused by } someone not taking seriously these same regulations. } } While it is correct that one can incur almost unbelievalbly high shipping } costs for HAZMATs, this is not always the case. In many instances, such as } for the ordering of osmium tetroxide, if one orders "smart", they can do } their shipping at costs only nominally more than if the same weight of } tweezers, grids, or SEM mounts were being shipped. Now this would not apply } for everything (e.g. our SPI Dusters, for example) but it does apply for a } surprising number of HAZMATs routinely used in EM labs. We are also in } the process of reformulating some of our embedding kits so they too can be } shipped at the lower prices. } } We have tried to explain how a customer can "order smart" on our website, } click on "Hazardous Items(Good News and Bad News)". While savings in } shipping costs for domestic US customers are possible, the real } beneficiaries are foreign customers now no longer are restricted to the use } of air freight (which has associated with it high minimum charges). } } And while we are on this subject, just remember, don't ever try to take } HAZMATs in checked airline luggage to save some money, it is unconscienable } from a moral standpoint and puts at risk everyone on the plane flight. } } 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 } ############################ } ==================================================
You ma want to define and echo this question to the Society of Ultrastructural Pathology's List Server at their web site http://sup.ultrakohl.com.
} } } maria lucia ribeiro caldas {caldasml-at-amcham.com.br} 09/07/97 07:16pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am having technical problems in EM skin biopsies. I would like to have a detailed protocol for processing skin biopsies.
I would never touch the Be window! If oil is present and you need to clean... CAREFULLY drip solvent acrosse the face to rinse the window. Be sure the solvent of choice not only will disolve the oil, but NOT the materials used to construct the Be window. Woody
{snip}
Let's go to the question: Have you ever cleaned the Be window of your EDS system? If the answer is positive, how often? Or, just wake me up ---- NEVER EVER THINK ABOUT THAT, YOU THE TROUBLE MAKER! I swear I have never been a trouble-maker. I just want to learn something from you. Just talking, no action following. It is too lang. Thank you for your time.
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Thank you for your time, D. Reynolds Customer Services.
I enjoyed your discussion on x-ray analysis and the challenge one faces from some students. As the saying goes, "you never really understand something until you teach it." Students will forever keep us on our toes and perhaps even force us to open certain doors that we might otherwise have tip-toed by.
Anyway, you asked does one clean the detector window? Yes, we clean our NORVAR window perhaps once a year or two - depending upon how dirty it has become. Most software has a subroutine for checking the efficience of the window using a standard of some sort (iron, for example). When efficiency drops, then one should carefully clean the window. In our case, we clean the window by removing the detector and slowly allowing 10-15 ml of methanol to flow over the window. The methanol is not directed at the window but onto the metal part of the housing about an inch away from the window. The methanol then gently flows over the window and washes away most of the oil. Check with your manufacturer, however, since different windows will require different treatments and/or solvents. You must be very gentle with the detector since the window and/or electronics will be damaged by mishandling. Definitely talk to a service person before doing this - if you are new to this procedure. We were, but now we feel comfortable cleaning the window.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Morphometry (i.e. measurement of form) in the biomedical field is mostly done on thin sections either at the LM or the EM level. The methods of choice for obtaining morphometric estimates from=20 sections are those of stereology. The "classical" foundations of stereology including assumption-based methods are covered in:
Modern design-based methods, mainly developed during the last 10-15 years, try to focus on the theory of unbiased sampling and on the counting and sizing of particles as well as on the orientation of=20 structures in 3D space. Reviews of modern stereological methods are e.g.:
Gundersen HJG et al.: APMIS 96;379-394 and 857-881 (1988) Cruz-Orive LM, Weibel ER: Am J Physiol 258;L148-L156 (1990) Mayhew TM: Exp Physiol 76;639-665 (1991)
The Journal of Microscopy and Acta Stereologica are the official journals of the International Society for Stereology.
If you have any further questions please contact me directly.
Sincerely,
Matthias Ochs Matthias Ochs, M.D. Dept. of Anatomy Div. of Electron Microscopy University of G=F6ttingen Kreuzbergring 36 D-37075 G=F6ttingen Germany Phone: +49 551 397036 Fax: +49 551 397004
I'm forwarding this message for a colleague in my department. Please resond to me at the address below. TIA.
Owen
} Seeking appropriate polishing procedures for Ge-Sn samples } ---------------------------------------------------------- } } I have eight Ge-Sn samples ( five Ge-10 at% Sn and three Ge-40 at% Sn ) } which have been annealed at 400 C, 500 C, 600 C, 700 C, and 800 C for five } weeks. I need to have good polishing procedures so that good } polished surfaces can be obtained in order to provide a detailed microchemical } analysis on these samples. Sn smearing problems arise when a fine grid, for example, } 0.05 micron alumina, was applied to the samples. This might due to the } fact that we have both soft and hard materials in the samples. } } Any suggestion on how to polish these samples is highly appreciate. Thank } you.
Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
Does anyone over there know the address (fax, phone, whatever...) of the German maker of time programmators called DIEHL. We have been looking for them all around talking with various vendors of electronics material, not to avail. (if someone can tell me the www address of the phone numbers searching tool of Deutsche Telekom or Chamber of Commerce in Germany that could even be a hint).
Jennifer T. Morse wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I was recently given the following homework assignment: Actual } resolution as determined by measuring the smallest distance between two } distinguishable points in a suitable specimen (resolution standard), is } usually greater than the linear resolution of the microscope. Describe } the factors that contribute to this discrepency. Find and list suitable } good references. Can you give me any help in solving this problem? Do } you know of any good sources to speak to? Do you know the answer? Thank } you for your help, Fred Meisenkothen
Jennifer & Fred,
Two of the best, upper level resources on this type of thing are: 1. Advanced Light Microscopy by Maksymilian Pluta (Elsevier - 3 volume set; volume 1 = ISBM 0-444-98939-0) 2. Progress in Microscopy, M. Francon (Row, Peterson & Co./Pergamon Press)
Born & Wolf's book on the Physics of Optics is also a good reference. I have no info on publisher, etc.
Let me know what you find.
Best regards, Barbara Foster Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
RE73-at-aol.com wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I am having trouble finding information concerning the theroy of morphometry } and its benifits to the biological field... any sugjestions.
Suggest the following: 1. John Russ's Handbook (already sent to you via another email) 2. Image analysis in Biology, Donat-P Hader, ed; CRC Press ISBN 0-8493-6033-1 3. Electronic Light Microscopy by David Shotton, ed., Wiley-Liss (ISBN 0-471-56077-4)
If you need just a quick overview of imaging principles, please see: Optimizing Light Microscopy for Biological and Clinical Laboratories, Kendall-Hunt (ISBN 0-7872-3538-5). We have them available, still on show discount from the Microscopy & Microanalysis meeting.
Good luck! Barbara Foster Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Your question is very broad so you will have to narrow the following list to your interests. Enjoy the search!:
Bibliography
Aherne, W.A. and M.S. Dunnill 1982 Morphometry. Edward Arnold (Publishers) Ltd. London,-205.
Bertram, J.F. and R.P. Bolender 1990 Counting cells with stereology: Random versus serial sectioning. J Electron Microsc Techn, 14:32-38.
Bianciardi, G., P. Tanganelli, and G. Weber 1993 Blood cell activation: New perspectives from ultrastructural morphometry. Semin Thromb Hemost, 19:108-114.
Bolender, R.P. 1978 Correlation of morphometry and stereology with biochemical analysis of cell fractions. Int Rev Cytol, 55:247-289.
Bolender, R.P. 1981 Stereology: applications to pharmacology. Annu Rev Pharmacol Toxicol, 21:549-573.
Bolender, R.P. 1982 Stereology and its uses in cell biology. Ann NY Acad Sci, 383:1-16.
Bolender, R.P. 1992a Quantitative morphology for biologists and computer scientists: I. Computer-aided tutorial for biological stereology (version 1.0). Microsc Res Techn, 21:338-346.
Cruz-Orive, L.-M. 1980 On the estimation of particle number. J Microsc, 120:15-27.
Cruz-Orive, L.-M. and E.R. Weibel 1981 Sampling designs for stereology. J Microsc, 122:235-257.
Cruz-Orive, L.-M. 1987 Particle number can be estimated using a disector of unknown thickness: the selector. J Microsc, 145:121-142.
Cruz-Orive, L.-M. and E.R. Weibel 1990 Recent stereological methods for cell biology: a brief survey. Am J Physiol, 258:L148-56.
Dardick, I. and D. Caldwell 1985 Reproducibility of morphometric image analysis. Hum Pathol, 16:1178
Freedman, L.S. 1974 A note on Aherne's method of counting tissue components in relatively thick sections. J Microsc, 100:219-225.
Gundersen, H.J.G. 1977 Notes on the estimation of the numerical density of arbitrary profiles: the edge effect. J Microsc, 111:219-223.
Gundersen, H.J.G., M. Boysen, and A. Reith 1981 Comparison of semiautomatic digitizer-tablet and simple point counting performance in morphometry. Virchows Arch B Cell Pathol, 37:317-325.
Gundersen, H.J.G. 1986 Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William R. Thompson. J Microsc, 143:3-45.
Gundersen, H.J.G. and E.B. Jensen 1987 The efficiency of systematic sampling in stereology and its prediction. J Microsc, 147(3):229-263.
Gundersen, H.J.G. 1988 The nucleator. J Microsc, 151:3-21.
Hammel, I., D. Lagunoff, M. Bauza, and E. Chi 1983 Periodic, multimodal distribution of granule volumes in mast cells. Cell Tissue Res, 228:51-59.
Henderson, W.R. and E.Y. Chi 1985 Ultrastructural characterization and morphometric analysis of human eosinophil degranulation. J Cell Sci, 73:33-48.
Loud, A.V., W.C. Barany, and B.A. Pack 1965 Quantitative evaluation of cytoplasmic structures in electron micrographs. Lab Invest, 14:258-270.
Loud, A.V. and P. Anversa 1984 Biology of disease: Morphometric analysis of biologic processes. Lab Invest, 50:250-261.
Maser, M.D. 1990 An overview of morphometry and stereology. Tx A&M Univ EM Views, Issue No. 4:3-8,15-16.
Mathieu, O., L.-M. Cruz-Orive, H. Hoppeler, and E.R. Weibel 1981 Measuring error and sampling variation in stereology: comparison of the efficiency of various methods for planar image analysis. J Microsc, 121:75-88.
Mayhew, T.M. and M.A. Williams 1971 A comparison of two sampling procedures for stereological analysis of cell pellets. J Microsc, 94:195-204.
Mayhew, T.M. and L.-M. Cruz 1973 Stereological correction procedures for estimating true volume proportions from biased samples. J Microsc, 99:287-299.
Mayhew, T.M. and L.-M. Cruz-Orive 1974 Caveat on the use of the Delesse principle of areal analysis for estimating component volume densities. J Microsc, 102:195-207.
Mayhew, T.M. 1979 Basic stereological relationships for quantitative microscopical anatomy - a simple systematic approach. J Anat, 129:95-105.
Mayhew, T.M. and F.H. White 1980 Ultrastructural morphometry of isolated cells: methods, models and applications. Pathol Res Pract, 166:239-259.
Mayhew, T.M. 1981 On the relative efficiencies of alternative ratio estimators for morphometric analysis of cell membrane surface features. J Microsc, 122:7-14.
Nazeran, H., F. Rice, W. Moran, and J. Skinner 1995 Biomedical image processing in pathology: a review. Australas Phys Eng Sci Med, 18:26-38.
Peachey, L.D. 1982 A simple digital morphometry system for electron microscopy. Ultramicrosc, 8:253-262.
Pesce, C.M. 1985 Morphometric studies need review by statisticians. Am J Clin Pathol, 83:258
Pesce, C.M. 1987 Biology of disease. Defining and interpreting diseases through morphometry. Lab Invest, 56:568-575.
Schmid-Sch”nbein, G.W. and S. Chien 1989 Morphometry of human leukocyte granules. Biorheol, 26:331-343.
Sokal, R.R. and F.J. Rohlf 1981 Biometry: the principles and practice of statistics in biological research. Freeman, San Francisco,
Sokol, R.J., G. Hudson, J. Wales, and N.T. James 1988 Morphometry of human blood leukocyte ultrastructure: Its potential value in haematology. Haematol, 21:129-139.
Sokol, R.J., G. Hudson, J. Wales, and N.T. James 1991 Ultrastructural morphometry of human leucocytes in health and disease. Electron Microsc Rev, 4:179-195.
Sterio, D.C. 1986 The unbiased estimation of number and sizes of arbitary particles using the disector. J Microsc, 134:127-136.
Stoeber, W. 1965 Statistical size distribution analysis. Lab Invest, 14:892-908.
Underwood, E.E. 1970 Quantitative stereology. Addison-Wesley, Reading,
Vedel Jensen, E.B. and H.J.G. Gundersen 1992 The rotator. J Microsc, 170:35-44.
Webster, P. and G. Griffiths 1994 A novel method for mean cell volume estimation. J Microsc, 174:85-92.
Weibel, E.R., G.S. Kistler, and W.F. Scherle 1966 Practical stereological methods for morphometric cytology. J Cell Biol, 30:23-38.
Weibel, E.R. 1969 Stereological principles for morphometry in electron microscopic cytology. Int Rev Cytol, 26:235-302.
Weibel, E.R. 1972 The value of stereology in analysing structure and function of cells and organs. J Microsc, 95:3-13.
Weibel, E.R. and R.P. Bolender 1973 Stereological techniques for electron microscopic morphometry. In: Principles and techniques of electron microscopy: Biological applications. Vol. 3. M.A. Hayat, ed. Van Nostrand Reinhold Co. New York, pp. 237-296.
Weibel, E.R. 1974 Selection of the best method in stereology. J Microsc, 100:261-269.
Weibel, E.R. 1975 Quantitation in morphology: possibilities and limits. Beitr Pathol, 155:1-17.
Weibel, E.R. 1981 Stereological methods in cell biology: where are we--where are we going? J Histochem Cytochem, 29:1043-1052.
Weibel, E.R. 1982 Biomorphometry in physiological and pathological research. Acta Med Pol, 23:115-125.
Weibel, E.R. 1989 Measuring through the microscope: development and evolution of stereological methods. J Microsc, 155:393-403.
Weibel, E.R. 1991 Fractal geometry: a design principle for living organisms. Am J Physiol, 261:L361-9.
------------------------------------- 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
Alcan International Limited, Kingston Research and Development Centre, is seeking a Materials Characterization Technologist to work in the area of metallography and electron optics (SEM, TEM, Electron Microprobe) support.
You will be involved in a wide range of activities aimed primarily at: (a) characterizing the microstructures of Alcan's automotive and packaging alloys which, in turn, support the Company's product and process development work, and (b) determining the factors which influence the performance of Alcan's sheet products. You will also play a key role in the ongoing development of the laboratory's materials characterization techniques.
We are seeking an individual skilled in metallographic specimen preparation and the operation and maintenance of both optical and electron microscopes. As the perfect candidate, you have a college diploma or university degree in a materials science discipline, or the equivalent, coupled with experience in both optical and electron metallography and in the use of PC-based data acquisition and analysis software. In addition, you must have the ability to cope with numerous activities simultaneously and adapt to changing priorities.
We offer an excellent compensation package commensurate with experience and a first-class working environment.
Interested candidates may send a resume by 18 September 1997 to: Personnel Administrator, Alcan International Limited, Kingston Research and Development Centre, Box 8400, Kingston, Ontario K7L 5L9. Fax: (613) 541-2308
".....CdS nano-particles, at least the ones we have seen, those that would be thin enough to "see" through, are smaller than the smallest holes we have ever seen anyone able to make in a "lacy" film. So I think that there might have been some misinformation accidently posted about this awhile back (see Aug. 11 posting DUNNTEM-at-aol.com working for Ted Pella, Inc.) If I am wrong about this please correct me and set the record straight."
Our posting on this subject stated that nono-particles have a tendency to adhere to the inner surface of the holes in the lacey films - an ideal location for EM imaging. Obviously, since the holes in the lacey film are usually from 1 micron up in diameter, a 5nm particle will not lie across a hole.
Charles Garber went on to say:
"This simple reality is often times missed by people worried about film thickness (which in fact ought to not even be relavent in the case of a lacey or holey film, because after all, you are getting your information through the holes, not through the "lacy" areas). So the whole need to worry about "thickness" per se really ought to be, in the case of lacey films, a non-issue!"
We made no reference to the thickness of the lacey film. Obviously it is a "non-issue". In fact, we do not see that anyone brought that subject up except Charles Garber.
We feel that it is generous of the creator of this list to allow vendors to post information which includes the products they have that might solve a technical problem. It is, we think, misuse of the list to suggest that a vendor is posting misinformation or to promote one's own products by criticizing another's products.
Let's keep competitive sales techniques out of the microscopy list.
We have a CM12 TEM/STEM that is shut down at the moment because the +24 volt supply (located in the remote rack) does not want to start up. This causes the microscope to shut down immediately after you press the main power button. Remote bench testing of this supply with all the safety circuits in place does not help in our diagnosis. We checked the setup with the +5 volt supply, which is almost identical and it works.
If anyone has had similiar problems to the startup sequence of this supply which prevents the microscope from powering up I would be very interested in an email message from you. ie. before we spend $4500 (Can) for a new/rebuilt supply from Philips.
email directly if you desire: eoptics-at-mcmaster.ca
Thanks in advance
Fred
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
} Several experts have advised me that we should give up the } dependence upon the standardless analysis of EDS, although the others } showed strong evidence to prove that it is working well.
Yes, but why not standardize? You are lucky to have a couple of extremely good EDS systems, give them a chance to work properly.
} My former colleague, Bruce, suggested that oily detector window } might cause the problem for softer radiation such as Al-Ka. This message } struck a spark in my poor memory. I really saw somebody who dared to wipe } oil off the thin Be-window of a TN-5400 EDS using a cotton ball with a } drop of acetone. That was ten years ago. } Let's go to the question: Have you ever cleaned the Be window of } your EDS system?
I use EDS for quantitative analysis of minerals, I get results which are comparable to WDS (provocative statement, which I'm happy to defend), I have an old probe with a fairly dirty vacuum system, and I clean my window when the response to Na drops by about 50%, which takes about 6 months. Al drops, too, but not so much. I clean it by gently dribbling a stream of Freon over the whole end of the detector so that the stream doesn't play onto the window directly, but runs down over it. I use Freon because it is a wonderful solvent for oil but is pretty non-aggressive towards epoxies etc which may be used in the construction of the window. I wouldn't use acetone, too aggressive. When I run out of Freon I'll use light petroleum spirit ("ligroin", "petroleum ether").
And DO NOT physically touch the window with ANYTHING at all!!!!!!!!!
But why don't you just follow the recommendations of your detector manufacturer?
In fact, this leads me to a question which often comes to mind when I read some of the postings --- they contain questions which could so easily be answered by the manufacturer, such as the one a couple of days ago about sources of replacement detector windows.
Am I just lucky in the quality of support that I get from Oxford Australia (take a bow, Keith, Julie, and John, for an unsolicited testimonial and thank-you)?
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
Barbara Foster wrote: } } Jennifer T. Morse wrote: } } } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------.} } } I was recently given the following homework assignment: Actual } } resolution as determined by measuring the smallest distance between two } } distinguishable points in a suitable specimen (resolution standard), is } } usually greater than the linear resolution of the microscope. Describe } } the factors that contribute to this discrepency. Find and list suitable } } good references. Can you give me any help in solving this problem? Do } } you know of any good sources to speak to? Do you know the answer? Thank } } you for your help, Fred Meisenkothen } } Jennifer & Fred, } } Two of the best, upper level resources on this type of thing are: } 1. Advanced Light Microscopy by Maksymilian Pluta (Elsevier - 3 volume } set; volume 1 = ISBM 0-444-98939-0) } 2. Progress in Microscopy, M. Francon (Row, Peterson & Co./Pergamon } Press) } } Born & Wolf's book on the Physics of Optics is also a good reference. I } have no info on publisher, etc. } } Let me know what you find. } } Best regards, } Barbara Foster }
I am looking to convert an old Nikon Diaphot TMD with a 80/20 side prism, to a 100/100% side port output. All that is involved is changing the side prism. We have the 80/20 kind, which we would gladly exchange at our expense with anyone in possesion of the 100/100 prism.
If interested please contact me to arrange the particulars.
Two requests. Firstly, would anyone know of a how to guide, including download sites for bringing spectra files from a TN-5500 into a PC? We need cabling, some sort of FTP on both sides and hopefully a spectra manipulation program. I can't help but think there are hundreds of microscopists who would find this useful.
Secondly, I am assembling a list of shareware useful to microscopists. I would like to assemble a list of programs, descriptions and download sites, which will be posted to the listserver (or sent out directly to anyone who requests it, if it gets too long). Should replies to this second request be posted, or come to me directly? I think if the shareware is generally useful to microscopists, a brief description, review and URL would be appropriate material for the listserver. How about it Nestor?
I am send to you information from "Morphology Digest" 1997, issue 5, from 8 september 1997 (Nl) about creation new sterelogy list in Calgary. I think that will be interesting for all.
---------- Forwarded message ----------
I have to prepare some yeast for SEM. The person who requested this work wants some pretty pictures of his yeast, Schizosaccharomyces pombe, for use in seminars. I have read up on some techniques to use but have two main queries: 1. To collect the yeast onto filter paper ( a method used in several publications), do I just drop a suspension of the yeast onto the filter paper? What sort of filter paper do I use? Is there a "better" way of collecting these cells such as settling them onto poly-l-lysine coverslips? 2. Is there a preferred fixative that works? The literature suggests a plethora of fixative cocktails! For TEM, I slam the cells onto a liquid nitrogen cooled copper mirror and process them via substitution in methanol and embed them in lowicryl HM20. We do not have an ESEM so please don't suggest I view them unfixed. Thanking you all in advance, this really is a great way of learning and sharing information! Sarah Ellis
We are interested in measuring the thermal conductivity of micron and submicron phases in a composite.Could any one tell me if SPM (Scanning Probe Microscopy) can be used to measure thermal conductivity and if this is a quantitative or purely qualitative technique ? Are the results relative or absolute ?
Your question is very broad so you will have to narrow the following list to your interests. Enjoy the search!:
Bibliography
Aherne, W.A. and M.S. Dunnill 1982 Morphometry. Edward Arnold (Publishers) Ltd. London,-205.
Bertram, J.F. and R.P. Bolender 1990 Counting cells with stereology: Random versus serial sectioning. J Electron Microsc Techn, 14:32-38.
Bianciardi, G., P. Tanganelli, and G. Weber 1993 Blood cell activation: New perspectives from ultrastructural morphometry. Semin Thromb Hemost, 19:108-114.
Bolender, R.P. 1978 Correlation of morphometry and stereology with biochemical analysis of cell fractions. Int Rev Cytol, 55:247-289.
Bolender, R.P. 1981 Stereology: applications to pharmacology. Annu Rev Pharmacol Toxicol, 21:549-573.
Bolender, R.P. 1982 Stereology and its uses in cell biology. Ann NY Acad Sci, 383:1-16.
Bolender, R.P. 1992a Quantitative morphology for biologists and computer scientists: I. Computer-aided tutorial for biological stereology (version 1.0). Microsc Res Techn, 21:338-346.
Cruz-Orive, L.-M. 1980 On the estimation of particle number. J Microsc, 120:15-27.
Cruz-Orive, L.-M. and E.R. Weibel 1981 Sampling designs for stereology. J Microsc, 122:235-257.
Cruz-Orive, L.-M. 1987 Particle number can be estimated using a disector of unknown thickness: the selector. J Microsc, 145:121-142.
Cruz-Orive, L.-M. and E.R. Weibel 1990 Recent stereological methods for cell biology: a brief survey. Am J Physiol, 258:L148-56.
Dardick, I. and D. Caldwell 1985 Reproducibility of morphometric image analysis. Hum Pathol, 16:1178
Freedman, L.S. 1974 A note on Aherne's method of counting tissue components in relatively thick sections. J Microsc, 100:219-225.
Gundersen, H.J.G. 1977 Notes on the estimation of the numerical density of arbitrary profiles: the edge effect. J Microsc, 111:219-223.
Gundersen, H.J.G., M. Boysen, and A. Reith 1981 Comparison of semiautomatic digitizer-tablet and simple point counting performance in morphometry. Virchows Arch B Cell Pathol, 37:317-325.
Gundersen, H.J.G. 1986 Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William R. Thompson. J Microsc, 143:3-45.
Gundersen, H.J.G. and E.B. Jensen 1987 The efficiency of systematic sampling
in stereology and its prediction. J Microsc, 147(3):229-263.
Gundersen, H.J.G. 1988 The nucleator. J Microsc, 151:3-21.
Hammel, I., D. Lagunoff, M. Bauza, and E. Chi 1983 Periodic, multimodal distribution of granule volumes in mast cells. Cell Tissue Res, 228:51-59.
Henderson, W.R. and E.Y. Chi 1985 Ultrastructural characterization and morphometric analysis of human eosinophil degranulation. J Cell Sci, 73:33-48.
Loud, A.V., W.C. Barany, and B.A. Pack 1965 Quantitative evaluation of cytoplasmic structures in electron micrographs. Lab Invest, 14:258-270.
Loud, A.V. and P. Anversa 1984 Biology of disease: Morphometric analysis of biologic processes. Lab Invest, 50:250-261.
Maser, M.D. 1990 An overview of morphometry and stereology. Tx A&M Univ EM Views, Issue No. 4:3-8,15-16.
Mathieu, O., L.-M. Cruz-Orive, H. Hoppeler, and E.R. Weibel 1981 Measuring error and sampling variation in stereology: comparison of the efficiency of various methods for planar image analysis. J Microsc, 121:75-88.
Mayhew, T.M. and M.A. Williams 1971 A comparison of two sampling procedures for stereological analysis of cell pellets. J Microsc, 94:195-204.
Mayhew, T.M. and L.-M. Cruz 1973 Stereological correction procedures for estimating true volume proportions from biased samples. J Microsc, 99:287-299.
Mayhew, T.M. and L.-M. Cruz-Orive 1974 Caveat on the use of the Delesse principle of areal analysis for estimating component volume densities. J Microsc, 102:195-207.
Mayhew, T.M. 1979 Basic stereological relationships for quantitative microscopical anatomy - a simple systematic approach. J Anat, 129:95-105.
Mayhew, T.M. and F.H. White 1980 Ultrastructural morphometry of isolated cells: methods, models and applications. Pathol Res Pract, 166:239-259.
Mayhew, T.M. 1981 On the relative efficiencies of alternative ratio estimators for morphometric analysis of cell membrane surface features. J Microsc, 122:7-14.
Nazeran, H., F. Rice, W. Moran, and J. Skinner 1995 Biomedical image processing in pathology: a review. Australas Phys Eng Sci Med, 18:26-38.
Peachey, L.D. 1982 A simple digital morphometry system for electron microscopy. Ultramicrosc, 8:253-262.
Pesce, C.M. 1985 Morphometric studies need review by statisticians. Am J Clin Pathol, 83:258
Pesce, C.M. 1987 Biology of disease. Defining and interpreting diseases through morphometry. Lab Invest, 56:568-575.
Schmid-Sch”nbein, G.W. and S. Chien 1989 Morphometry of human leukocyte granules. Biorheol, 26:331-343.
Sokal, R.R. and F.J. Rohlf 1981 Biometry: the principles and practice of statistics in biological research. Freeman, San Francisco,
Sokol, R.J., G. Hudson, J. Wales, and N.T. James 1988 Morphometry of human blood leukocyte ultrastructure: Its potential value in haematology. Haematol, 21:129-139.
Sokol, R.J., G. Hudson, J. Wales, and N.T. James 1991 Ultrastructural morphometry of human leucocytes in health and disease. Electron Microsc Rev, 4:179-195.
Sterio, D.C. 1986 The unbiased estimation of number and sizes of arbitary particles using the disector. J Microsc, 134:127-136.
Stoeber, W. 1965 Statistical size distribution analysis. Lab Invest, 14:892-908.
Underwood, E.E. 1970 Quantitative stereology. Addison-Wesley, Reading,
Vedel Jensen, E.B. and H.J.G. Gundersen 1992 The rotator. J Microsc, 170:35-44.
Webster, P. and G. Griffiths 1994 A novel method for mean cell volume estimation. J Microsc, 174:85-92.
Weibel, E.R., G.S. Kistler, and W.F. Scherle 1966 Practical stereological methods for morphometric cytology. J Cell Biol, 30:23-38.
Weibel, E.R. 1969 Stereological principles for morphometry in electron microscopic cytology. Int Rev Cytol, 26:235-302.
Weibel, E.R. 1972 The value of stereology in analysing structure and function of cells and organs. J Microsc, 95:3-13.
Weibel, E.R. and R.P. Bolender 1973 Stereological techniques for electron microscopic morphometry. In: Principles and techniques of electron microscopy: Biological applications. Vol. 3. M.A. Hayat, ed. Van Nostrand Reinhold Co. New York, pp. 237-296.
Weibel, E.R. 1974 Selection of the best method in stereology. J Microsc, 100:261-269.
Weibel, E.R. 1975 Quantitation in morphology: possibilities and limits. Beitr Pathol, 155:1-17.
Weibel, E.R. 1981 Stereological methods in cell biology: where are we--where are we going? J Histochem Cytochem, 29:1043-1052.
Weibel, E.R. 1982 Biomorphometry in physiological and pathological research.
Acta Med Pol, 23:115-125.
Weibel, E.R. 1989 Measuring through the microscope: development and evolution of stereological methods. J Microsc, 155:393-403.
Weibel, E.R. 1991 Fractal geometry: a design principle for living organisms. Am J Physiol, 261:L361-9.
------------------------------------- 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
Cambridge Healthtech Institute's Advances in Cellular Imaging November 13-14, 1997 Westin Hotel Horton Plaza San Diego, California
TECHNICAL TRENDS AND ADVANCES Multiphoton Excitation Imaging and Photochemistry in Cells and Tissue Dr. Warren Zipfel, Cornell University Highly Resolved Cell and Tissue Optical Imaging in Real Time Dr. Daniel Farkas, Carnegie Mellon University Combined Fluorescent and Gold Cluster Probes: "Simultaneous" Labeling for Light and Electron Microscopy Dr. Richard Powell, NANOPROBES, Inc. Novel Magnetic Messenger Labeling System Mr. Lonnie Adelman, Ericomp Inc.
VIEWING REAL-TIME CELLULAR CHANGES Imaging Drug Uptake and Metabolism in Living Intestinal Tissue with Confocal and Two-Photon Microscopy Dr. Marshall Montrose, Johns Hopkins University School of Medicine Dynamic Changes in Intracellular pH and Ca2+ Dr Randi Silver, Cornell University Medical College (invited) Smart Magnetic Resonance Contrast Agents: A New Generation of Image Enhancement Media Dr. Tom Meade, California Institute of Technology Multiple Fluorescent Proteins and Fluorescence Microscopy to Monitor Live Cell Activity or Screen for Protein Localization Dr. Neal Gliksman, Universal Imaging Corporation Multiphoton Laser Scanning Fluorescence Microscopy Dr. Victoria Centonze Frohlich, University of Wisconsin- Madison
IMAGE ANALYSIS AND INTERPRETATION Quantitative Molecular Image Analysis Dr. Branko Palcic, British Columbia Cancer Research Centre (invited) Image Analysis Tools Dr. Paul Goodwin, Fred Hutchinson Cancer Research Institute (invited) Quantitative Automated Microscopy Dr. Frans Nauwelaers, Becton Dickinson Cellular Imaging Systems (invited) Fluorescence Imaging MicroSpectrophotometer (FIMS) Dr. Douglas Youvan, KAIROS Scientific Inc.
SCREENING AND DRUG DEVELOPMENT High-Content, Cell-Based Screening: Easing the Bottlenecks of Target Validation and Optimization of Lead Compounds Dr. Kenneth Giuliano, BioDx, Inc. Applications of the Fluorometric Imaging Plate Reader (FLIPR) Technology to High-Throughput Screening Dr. Simon Pitchford, Molecular Devices Corporation Use of Fluorescence Polarization in Drug Screening Dr. Michael Jolley, Jolley Consulting and Research Inc. (invited) Fluorescence-Based Screening of Cellular Changes in Ion Concentrations for Drug Development Dr. Carla Suto, SIBIA Neuroscience, Inc. (invited) Imaging Requirements for Faster Drug Development: Screening with Higher Density Formats Dr. Al Kolb, Packard Instrument Company (tentative)
Improved technology for imaging of cells and related targets is having a dramatic impact on pharmaceutical research and development, driven in part by the demand for greater speed, precision, and automation. Novel strategies for labels, better software for image enhancement and analysis, and progress integrating imaging with other laboratory functions are being applied to a growing range of applications. Advantages in such diverse segments as microscopy, cytology, and cellular analysis, as well as more applied uses such as assessment of gels and drug development screening, will be discussed. All of these activities share a similar goal of rapidly and correctly translating images into data that can be stored, used, compared, and manipulated with as much ease and as little human intervention as possible. These developments promise to have a dramatic impact on laboratory productivity, and any research manager involved in these segments should consider participating.
HOTEL INFORMATION Westin Hotel Horton Plaza Reservations made after the cut-off 910 Broadway Circle date will be accepted on a space and San Diego, CA 92101 rate availability basis. Available rooms are limited, so please book T: 619-239-2200 early. F: 619-239-0509 Please identify yourself as a Cut-off Date: October 30, 1997 Cambridge Healthtech Institute Room Rate: $135 Single/Double conference attendee to receive the reduced room rate.
TRAVEL INFORMATION TRAVELWORLD T: 717-288-9311 or 800-828-6033 601 Market Street F: 717-288-4693 Kingston, PA 18704
Exclusive airline discounts are available on American Airlines as well as other specific airlines when tickets are purchased through TRAVELWORLD at least 14 days prior to the meeting date. Some restrictions apply.
CALL FOR POSTERS Cambridge Healthtech Institute encourages attendees to gain further exposure by presenting their work in the poster sessions. Please fill out the registration form, with the poster title and primary author. To ensure inclusion in the conference binder, a one-page summary must be submitted by October 3, 1997.
CALL FOR EXHIBITORS Space is available for companies interested in exhibiting products and services related to cellular imaging. This meeting should attract up to several hundred senior researchers and managers representing a broad range of disciplines and perspectives. Please contact Jim MacNeil of Cambridge Healthtech Institute at 617-630-1341 to obtain an exhibitor package or to inquire about offering a workshop during the meeting. Exhibit space is limited so call now to reserve a space at this premier event.
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Program and speakers are subject to change.
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Advance Registration (by October 3, 1997) |__| $795 Commercial |__| $395 Academic, Government, Hospital-Affiliated On-site or Late Registration (after October 3, 1997) |__| $895 Commercial |__| $445 Academic, Government, Hospital-Affiliated FIRST NAME:______________________________________________________ LAST NAME:_______________________________________________________ TITLE:___________________________________________________________ DIV./DEPT.:______________________________________________________ COMPANY:_________________________________________________________ ADDRESS:_________________________________________________________ City/State/ZIP:__________________________________________________ COUNTRY:_________________________________________________________ TELEPHONE:____________________________ Fax:______________________ E-MAIL:__________________________________________________________ |__| Please send information on exhibiting and opportunities to present workshops. |__| Enclosed is a check or money order payable to Cambridge Healthtech Institute, drawn on a U.S. bank, in U.S. currency. |__| Please charge: |__| AMEX (15 digits) |__| Visa (13 to 16 digits) |__| MasterCard (16 digits) Card #:___________________________________________________________ Exp. Date:________________________________________________________ Cardholder's Name:________________________________________________ Signature:________________________________________________________ Cardholder's Address (if different from above):___________________ __________________________________________________________________ |__| Reserve with credit card information listed above and invoice me. (Invoices must be paid in full by the deadline to retain registration discount. Invoices unpaid one week prior to conference will be billed to credit card at full registration rate.) If you plan to register on site, please check with CHI beforehand for space availability. |__| I am interested in presenting a poster at ADVANCES IN CELLULAR IMAGING and will provide an abstract by October 17, 1997. Poster title:______________________________________________________ ___________________________________________________________________
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In a message dated 97-09-09 03:38:32 EDT, jbest-at-vicon.net (John Best) writes:
{ { Firstly, would anyone know of a how to guide, including download sites for bringing spectra files from a TN-5500 into a PC? We need cabling, some sort of FTP on both sides and hopefully a spectra manipulation program. I can't help but think there are hundreds of microscopists who would find this useful. } }
Greeting John,
We have just the program for you.
The software is called VIDX X-ray Microanalysis. It is a Windows 95 and NT software. (It is regularly sold to function with our PC based X-ray microanalysis and digital Imaging hardware)
The software will open fthe most popular file formats. We can perform both offline and on-line connections to many older vintage X-ray analyzers such as Edax, Kevex, Link, Noran, PGT, Tracor. That means you don't have to go and buy a brand new x-ray analyzer.
Prices are affordable.
For more information contact us at
Evex Analytical 857 State Road Princeton, NJ 08540 609-252-9192
Collect the yeast on to membrane filters with nice circular pores (e.g. Nucleopore), not a torturous-path filter like filter paper (or e.g. Millipore). This will give a nice smooth background against which to view the yeast. Filters with 0.22 micron holes are maybe best, although 0.45 micron will work. The pores will also give an *approximate!* size standard for the yeast cells.
*Before* collecting the yeast, coat both sides of the filters in a sputter coater. This gives better conductivity for viewing. If you're rich, use silver filters instead.
For fixation, I'd use the recipe the article(s) that show SEMs of yeast most like what you're trying to achieve (including 'scope kV), and is the simplest.
Phil
} I have to prepare some yeast for SEM. The person who requested this } work wants some pretty pictures of his yeast, Schizosaccharomyces pombe, } for use in seminars. I have read up on some techniques to use but have } two main queries: } 1. To collect the yeast onto filter paper ( a method used in } several publications), do I just drop a suspension of the yeast onto the } filter paper? What sort of filter paper do I use? Is there a "better" } way of collecting these cells such as settling them onto poly-l-lysine } coverslips? } 2. Is there a preferred fixative that works? The literature } suggests a plethora of fixative cocktails! For TEM, I slam the cells } onto a liquid nitrogen cooled copper mirror and process them via } substitution in methanol and embed them in lowicryl HM20. } We do not have an ESEM so please don't suggest I view them unfixed. } Thanking you all in advance, this really is a great way of learning and } sharing information! } Sarah Ellis
} 1. To collect the yeast onto filter paper ( a method used in } several publications), do I just drop a suspension of the yeast onto the } filter paper? What sort of filter paper do I use? Is there a "better" } way of collecting these cells such as settling them onto poly-l-lysine } coverslips?
You can drop the cells onto poly-l-lysine coated coverslips after osmium fixation. Then process it as usual for dehydration and critical point dry.
Best regards,
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Hello, Here is info on our procedures. We are a "Skin LAB".
PROCESSING OF SKIN TISSUE FOR LIGHT AND ELECTRON MICROSCOPY EMBEDDING IN EPON 812 1. Place newly received tissue* in 1/2 Karnovskys fixative overnight. 2. Take EPON 812 mixture out of refrigerator (let it sit under the hood for at least 2 hours before opening the container). This mixture is a combination of DDSA, NMA, and EMBED 812. (48% of 812, 31% of DDSA, 21% of NMA) 3. Rinse biopsy in 0.1 M sodium cacodylate buffer x2, 15 minutes each rinse. 4. Post-fix in 1% osmium tetroxide, 1 and 1/2 hours. (mix 1:1, 2% OsO4 with 0.2 M sodium cacodylate buffer). 5. Rinse in dH2O x 2, 15 minutes each rinse. 6. En-bloc stain with 1% Uranyl Acetate for 1 and 1/2 hours. 7. Dehydrate through an ascending ETOH series 35% x2 (15 minutes each) 70% x2 (15 minutes each) 95% x2 (15 minutes each) 100% x2 (30 minutes each) 8. Add the catalyst to the EPON 812 mixture (.2ml of DMP30 per 10 ml resin). Stir slowly for 10 minutes. 9. Clear biopsy in Propylene oxide x2, 15 minutes each rinse. 10. Infiltrate by placing biopsy into a 3:1 mixture of Propylene oxide:EPON for 3-4 hours 2:1 mixture for 12-16 hours (overnight) with caps off 1:1 mixture for 12-16 hours (overnight) with caps off 11. Place biopsy into an embedding mold with fresh 100% EPON for 8 hours, then place mold into 60 oC oven for curing (24-48 hours).
EPON 812 can be substituted with either EMBED 812 (EMS), PolyBed 812 (Polysciences), Medcast or Eponate 12 (Ted Pella). All have the ingredients DDSA, NMA, and DMP 30.
EMBEDDING PROCEDURE FOR SKIN BIOPSIES
ROUTINE RAPID
1/2 KARNOVSKYS overnight 2hr to overnight
0.1M NaCaco 15minx2 5-10min 1%OsO4/.1M Na Caco 11/2 hrs 20min
bake for 24-48 hours in 60oC oven DMP .2ml/10ml resin
Note: EPON 812 (which was discontinued in 1979) can be substituted with EMBED 812 (EMS), Polybed 812 (Polysciences), Eponate 12 or Medcast (TedPella). All have the ingredients DDSA, NMA, and DMP-30, plus one 'company specific ingredient'.
I pasted this in from Word, I hope it makes some sense.
Bob Underwood Morphology Core Univ. of Washington -
On Sun, 7 Sep 1997, maria lucia ribeiro caldas wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am having technical problems in EM skin biopsies. I would like to have } a detailed protocol for processing skin biopsies. }
Your question is very broad so you will have to narrow the following list to your interests. Enjoy the search!:
Bibliography
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Bertram, J.F. and R.P. Bolender 1990 Counting cells with stereology: Random versus serial sectioning. J Electron Microsc Techn, 14:32-38.
Bianciardi, G., P. Tanganelli, and G. Weber 1993 Blood cell activation: New perspectives from ultrastructural morphometry. Semin Thromb Hemost, 19:108-114.
Bolender, R.P. 1978 Correlation of morphometry and stereology with biochemical analysis of cell fractions. Int Rev Cytol, 55:247-289.
Bolender, R.P. 1981 Stereology: applications to pharmacology. Annu Rev Pharmacol Toxicol, 21:549-573.
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Cruz-Orive, L.-M. and E.R. Weibel 1990 Recent stereological methods for cell
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Gundersen, H.J.G., M. Boysen, and A. Reith 1981 Comparison of semiautomatic digitizer-tablet and simple point counting performance in morphometry. Virchows Arch B Cell Pathol, 37:317-325.
Gundersen, H.J.G. 1986 Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William R. Thompson. J Microsc, 143:3-45.
Gundersen, H.J.G. and E.B. Jensen 1987 The efficiency of systematic sampling
in stereology and its prediction. J Microsc, 147(3):229-263.
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Henderson, W.R. and E.Y. Chi 1985 Ultrastructural characterization and morphometric analysis of human eosinophil degranulation. J Cell Sci, 73:33-48.
Loud, A.V., W.C. Barany, and B.A. Pack 1965 Quantitative evaluation of cytoplasmic structures in electron micrographs. Lab Invest, 14:258-270.
Loud, A.V. and P. Anversa 1984 Biology of disease: Morphometric analysis of biologic processes. Lab Invest, 50:250-261.
Maser, M.D. 1990 An overview of morphometry and stereology. Tx A&M Univ EM Views, Issue No. 4:3-8,15-16.
Mathieu, O., L.-M. Cruz-Orive, H. Hoppeler, and E.R. Weibel 1981 Measuring error and sampling variation in stereology: comparison of the efficiency of various methods for planar image analysis. J Microsc, 121:75-88.
Mayhew, T.M. and M.A. Williams 1971 A comparison of two sampling procedures for stereological analysis of cell pellets. J Microsc, 94:195-204.
Mayhew, T.M. and L.-M. Cruz 1973 Stereological correction procedures for estimating true volume proportions from biased samples. J Microsc, 99:287-299.
Mayhew, T.M. and L.-M. Cruz-Orive 1974 Caveat on the use of the Delesse principle of areal analysis for estimating component volume densities. J Microsc, 102:195-207.
Mayhew, T.M. 1979 Basic stereological relationships for quantitative microscopical anatomy - a simple systematic approach. J Anat, 129:95-105.
Mayhew, T.M. and F.H. White 1980 Ultrastructural morphometry of isolated cells: methods, models and applications. Pathol Res Pract, 166:239-259.
Mayhew, T.M. 1981 On the relative efficiencies of alternative ratio estimators for morphometric analysis of cell membrane surface features. J Microsc, 122:7-14.
Nazeran, H., F. Rice, W. Moran, and J. Skinner 1995 Biomedical image processing in pathology: a review. Australas Phys Eng Sci Med, 18:26-38.
Peachey, L.D. 1982 A simple digital morphometry system for electron microscopy. Ultramicrosc, 8:253-262.
Pesce, C.M. 1985 Morphometric studies need review by statisticians. Am J Clin Pathol, 83:258
Pesce, C.M. 1987 Biology of disease. Defining and interpreting diseases through morphometry. Lab Invest, 56:568-575.
Schmid-Sch”nbein, G.W. and S. Chien 1989 Morphometry of human leukocyte granules. Biorheol, 26:331-343.
Sokal, R.R. and F.J. Rohlf 1981 Biometry: the principles and practice of statistics in biological research. Freeman, San Francisco,
Sokol, R.J., G. Hudson, J. Wales, and N.T. James 1988 Morphometry of human blood leukocyte ultrastructure: Its potential value in haematology. Haematol, 21:129-139.
Sokol, R.J., G. Hudson, J. Wales, and N.T. James 1991 Ultrastructural morphometry of human leucocytes in health and disease. Electron Microsc Rev, 4:179-195.
Sterio, D.C. 1986 The unbiased estimation of number and sizes of arbitary particles using the disector. J Microsc, 134:127-136.
Stoeber, W. 1965 Statistical size distribution analysis. Lab Invest, 14:892-908.
Underwood, E.E. 1970 Quantitative stereology. Addison-Wesley, Reading,
Vedel Jensen, E.B. and H.J.G. Gundersen 1992 The rotator. J Microsc, 170:35-44.
Webster, P. and G. Griffiths 1994 A novel method for mean cell volume estimation. J Microsc, 174:85-92.
Weibel, E.R., G.S. Kistler, and W.F. Scherle 1966 Practical stereological methods for morphometric cytology. J Cell Biol, 30:23-38.
Weibel, E.R. 1969 Stereological principles for morphometry in electron microscopic cytology. Int Rev Cytol, 26:235-302.
Weibel, E.R. 1972 The value of stereology in analysing structure and function of cells and organs. J Microsc, 95:3-13.
Weibel, E.R. and R.P. Bolender 1973 Stereological techniques for electron microscopic morphometry. In: Principles and techniques of electron microscopy: Biological applications. Vol. 3. M.A. Hayat, ed. Van Nostrand Reinhold Co. New York, pp. 237-296.
Weibel, E.R. 1974 Selection of the best method in stereology. J Microsc, 100:261-269.
Weibel, E.R. 1975 Quantitation in morphology: possibilities and limits. Beitr Pathol, 155:1-17.
Weibel, E.R. 1981 Stereological methods in cell biology: where are we--where are we going? J Histochem Cytochem, 29:1043-1052.
Weibel, E.R. 1982 Biomorphometry in physiological and pathological research. Acta Med Pol, 23:115-125.
Weibel, E.R. 1989 Measuring through the microscope: development and evolution of stereological methods. J Microsc, 155:393-403.
Weibel, E.R. 1991 Fractal geometry: a design principle for living organisms. Am J Physiol, 261:L361-9. ------------------------------------- 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
Someone recently commented about having a glassblower grind down a bell jar that has a chipped rim. We have had this done a number of times with good success. It is indeed a feasible solution, provided you can find a glassblower willing to undertake the task. On the other hand, you could probably do it yourself, if you are willing to devote the time and energy required, because all one glassblower I observed did was to spread a slurry of silicon carbide abrasive on a piece of window glass, and sit and rub the bell jar around over it.
Alternatively, we have been successful in some instances in 'patching' the chips in bell jars using a stiff epoxy (e.g. Torr Seal). Clean all grease and oil off the bell jar (try using Tilex Soap Scum Remover), then fill the chipped hole with the epoxy, and set the bell jar on a flat, smooth surface covered with waxed paper (so that the surface comes out flat and smooth) while the epoxy cures. Good luck,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Please define your problems more clearly. I have a lot of experience with skin biopsies, (2 published papers also) but I need to know what the problems are - embedding, cutting, staining for LM, wrinkles, etc. I would be happy to help if I had the details. Also, the size of the biopsy which you are required to handle will have a great influence on your final protocol. My handout at FORUM booth at the MSA meeting was an exact protocol which we use in our laboratory. I would be happy to send it to you, if I knew it would be of use in solving your particular problem. (Need your address).
} ... } Someone recently commented about having a glassblower grind down a bell jar } that has a chipped rim. We have had this done a number of times with good } success. ...
I consider a bell jar with any defect to be a considerable risk for an implosion.If your users don't use a implosion gaurd with 100% consistency then replace the bell jar ...
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/
} ... } Someone recently commented about having a glassblower grind down a bell jar } that has a chipped rim. We have had this done a number of times with good } success. ...
I consider a bell jar with any defect to be a considerable risk for an implosion.If your users don't use a implosion gaurd with 100% consistency then replace the bell jar ...
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/
Maria, We have worked with skin from a variety of species for over 20 years. We primarily work with human, pig, and in vitro equivalents. We do SEM, TEM, Immuno EM, enzyme histochemistry and basic light microscopy. We will be happy to share any of our techniques with you. What type of problem are you having? We use half strength Karnovsky's for fixation and embed in Spurr. Along time ago we used EPON 812 and then switched to Polybed and now Spurr. Sectioning with a diamond knife greatly enhances your sections.Please let us know your specific problems. Good Luck!!! NAMR
Nancy A. Monteiro-Riviere,Ph.D.,DABFE,DABFM Professor of Investigative Dermatology/Toxicology North Carolina State University College of Veterinary Medicine Cutaneous Pharmacology and Toxicology Center 4700 Hillsborough St. Raleigh, NC 27606 Telephone:(919)829-4426 FAX:(919)829-4358 email: Nancy_Monteiro-at-ncsu.edu CTPC Homepage:http://cptc.ncsu.edu
Allow me to second this repair method. I used it to repair a seriously chipped Denton 502A bell jar (it would work on any), and after overnight curing (room temperature), the unit pulled as good a vacuum as quickly as it did before the chip.
Phil
} Alternatively, we have been successful in some instances in 'patching' the } chips in bell jars using a stiff epoxy (e.g. Torr Seal). Clean all grease } and oil off the bell jar (try using Tilex Soap Scum Remover), then fill the } chipped hole with the epoxy, and set the bell jar on a flat, smooth surface } covered with waxed paper (so that the surface comes out flat and smooth) } while the epoxy cures. } Good luck, } } Wilbur C. Bigelow, Prof. Emeritus } Materials Sci. & Engr., University of Michigan } Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; } Fx:313-763-4788; Ph:313-764-3321
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Someone recently commented about having a glassblower grind down a bell jar } that has a chipped rim. We have had this done a number of times with good } success. It is indeed a feasible solution, provided you can find a } glassblower willing to undertake the task. On the other hand, you could } probably do it yourself, if you are willing to devote the time and energy } required, because all one glassblower I observed did was to spread a slurry } of silicon carbide abrasive on a piece of window glass, and sit and rub the } bell jar around over it. } } Alternatively, we have been successful in some instances in 'patching' the } chips in bell jars using a stiff epoxy (e.g. Torr Seal). Clean all grease } and oil off the bell jar (try using Tilex Soap Scum Remover), then fill the } chipped hole with the epoxy, and set the bell jar on a flat, smooth surface } covered with waxed paper (so that the surface comes out flat and smooth) } while the epoxy cures. } Good luck, } } Wilbur C. Bigelow, Prof. Emeritus } Materials Sci. & Engr., University of Michigan } Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; } Fx:313-763-4788; Ph:313-764-3321 } } We have repaired our Bell Jars 10-15 times using Vacuum epoxy, such as Bell Torr or Torr Seal. We do it a bit different than above. We actually file and sand the surface smooth. Basically, I have come to the conclusion that for the level of vacuum that Denton Coaters use, the only time a new bell needs to be bought is if it cracks. Chipping is easy to take care of.
oh...The reason I replied to the above is that if you use the wax paper technique, make sure you get the epoxy thick enough in the chip (ie. as thick as the glass)
In behalf of the "boss" of the EM core, I'll ask you for help... We like to know if there is somebody around who used LR white resins in an AFS from Leica. What are your conditoins, problems encountered etc... Thanks a lot for your help
Marc
PS Thank you Nestor for your work...
------------------------------ SCHMUTZ Marc PhD IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
I am a user of an old Coates&Welter/Nanometrics FEG-SEM and am interested in producing Electron Channeling Patterns. I need to rock the beam about a point on the sample for this but my microscope doesn't have this capability.
Are there any users (probably ex-users!) of this venerable machine who can tell me whether a beam deflection system was ever produced for the C&W or who have experience of ECPs with it? If so, where can i get hold of the necessary electronics?
Any help would be much appreciated.
ANGUS BEWICK Physics Dept. University of Bristol UK phab-at-siva.bris.ac.uk
This is a multi-part message in MIME format. --------------3F23D1509D80037EE77529EF Content-Type: text/plain; charset=us-ascii Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit Content-Transfer-Encoding: 7bit
Barbara Foster wrote:
} ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ----------------------------- } -----------------------------------------. } Born & Wolf's book on the Physics of Optics is also a good reference. } I have no info on publisher, etc. } Let me know what you find. } } Best regards, } Barbara Foster } Microscopy/Microscopy Education } 53 Eton Street } Springfield, MA 01108 } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Barbara:
I think you may mean:
Principles of Optics : Electromagnetic Theory of Propagation, Interference and Diffraction of Light by M. Born, E. Wolf Sixth Edition (Paperback) Published by Pergamon Press Publication date: June 1981 ISBN: 0080264816
-------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com The Web is:"Vaster than empires and more slow" Andrew Marvell (1621-1678)
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We are replacing our old SEM + EDS and would like dispose of it ourselves since the manufacturer of the new unit is offering only a nominal sum in a trade in. Can anyone suggest effective places to advertise this equipment? (We are located in NE USA, and would like to make room for the new system in 3-4 mo.)
A follow up question that I have is: what is a reasonable price to ask? ( The SEM is a 16 y.o. AMRAY 1600 turbo LaB6/W, secondary & Link backscatter, 2 CRT's, vibration isolation table, in good condition & under factory service contract. The EDS is 3 y.o. Oxford Isis thin window 136eV spec., 126eV MnKa calibrated, with beam control and image capture, under factory service contract. )
I will greatly appreciate any & all suggestions | comments.
Many thanks for your numerous and energetic inquiries about "Optimizing Light Microscopy for Biological and Clinical Labs"! Since our web site is not quite ready, in answer to those requests, we have put together a short order form (see below) as well as the description sent in an earlier email "re: infinity optics".
For those of you in the Northeast, you can also get a complimentary copy of the book by attending the one-day lecture-demo, "Optimizing Light Microscopy". Dates: Oct 3 (Providence College, Providence RI), Nov 3 (NYC), Nov 5 (Springfield, MA), Nov 7 (Boston, Tufts Med School MRC) Email for details.
Book Description: "Optimizing Light Microscopy for Biological and Clinical Laboratories" is available from MME as well as through the American Society for Clinical Laboratory Sciences and the publisher, Kendall-Hunt. Approximately 200 pages with over 100 diagrams, illustrations, and micrographs. Peppered with short experiments which are geared to help practicing microscopists learn more about how their light microscopes work. There are also introductory chapters on video microscopy and other microscopy techniques, ranging from Confocal to EM to microspectrometry.
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Barbara Foster Consortium President MME -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-Microscopy/Microscopy Education is a consortium of 24 consultants who specialize in customized on-site training in all areas of microscopy, sample preparation, and image analysis. Our goal: to help you use your microscope more effectively. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
We are in the process of rebuilding our darkroom from the ground up, and were wondering if anyone had any information on suppliers of the revolving, light-tight darkroom doors and/or other darkroom fixtures.
Thanks.
Craig Lending Department of Biology SUNY Brockport Brockport, NY 14420
Torr Seasl is an epoxy compound especially formulated for use in vacuum systems that was sintroduced by Varian Associates, Vacuum Products Division, 121 Hartwell Ave, Lexington, MA 02173-3133 (800-882-7426) a number of years ago. It is stated to be good at pressures below 10-9Torr, and is bakeable at temperarures up to 120C. It adheres to most clean materials (glass, metals, ceramics) and holds up well over long term service. Some companies that handle EM supplies also handle it (e.g. I find it listed in the Ladd catalog).
Other similar products are also sold by other companies. For example, Duniway Stockroom Corp. (800-446-8811; info-at-duniway.com) sells a product called 'Epoxy Patch' that is stated to be equivalent to Torr seal
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
------------------------------------------------------- Dan Edwards Structural Materials Research Section Battelle Pacific Northwest National Laboratory P.O. Box 999, MSIN P8-15 Richland, WA 99352
Hello, We have a problem fixing imaginal discs (third instar - Drosophila). We are following a protocol used by Andrew Tomlinson, 1985- "The cellular dynamics of pattern formation in the eye of Drosophila" in J. Embryol. exp. Morph. 89, 313-331. The fixative used is a combined cold glutaraldehyde/osmium followed by an osmium post-fix. We're seeing pore fixation of membranes including inner cristae of mitochondria and some clear areas within cytoplasm. Thanks in advance for any suggestions. Rosemary
#################################################### Rosemary Walsh Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:rw9-at-psu.edu ####################################################
We supply Chromium Targets. If you would like to give us a call at 800/444-3137 and let us know what type of system you have, we will be happy to help you.
At least once over the history of this listserver there has been extensive discussion about osmium "pepper" and other artifactual inclusions seen in embedded, sectioned TEM samples. Does anyone have a compilation of replies/discussion or remember what conclusions were drawn? Answer directly please, to avoid boring others with a rediscussion, OK? Many thanks. Grace P.S. Was it related especially to phosphate buffered osmium solutions?? I just had a major disaster with some and would like to solve the problem quickly.....
The semiconductor industry uses "wafer" tweezers. They are used to pick up wafers from the side. The contact is not as minimal as the triceps, but they may work. There are some listed at the following URL. I am not sure where the ones we use around here came from. URL: http://www.ebsciences.com/labsupply/tweezers.htm
We have a student in our lab who is having a problem with her protoplasts that have been embedded in Epon/Araldite pulling away from the resin when she sections. This causes a lot of problems, to say the least. Do any of you have suggestions as to how to keep this from happening? She has heard that adding tannic acid to the fix helps prevent the problem. Has anyone done that? She's also having a problem with the inclusion bodies falling out of her sections when she cuts. Any suggestions about that? We recommended she reduce her cutting speed. All suggestions will be gratefully passed along to her.
Always eager to learn new things,
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I have sent a repeated requests to resubscribe but without any results. I should be most grateful if you can put me on the list. If there are any problems, Please let me know.
Can anyone recomend a good, user friendly LM IM analysis software program? Our current one is useless (hate to say). We will be demoing a Noesis pacckage soon. I have experience with Optimas, and personally do not care for it. Any other shareware besides NIH? All comments etc welcome.
I posted the below earlier this week but the email bounced back. I believe that the the previously advanced concept of 'quantitative' analysis does require examination. JD.
I thank Garratt-Reed for corroberating my previous posting. John Bazzola had asked for a rough guide (he has confirmed that since) on detection limits of EDS versus WDS techniques. Anybody who has any significant experience with microprobe analysis knows that there is no single line correct answer. However, it is imperative for analysts to remember a few general figures so they can advise on appropriate instrumentation and techniques.
John B's initial inquiry deserved a reply and when none was given, I posted mine more than a day later. I believe that my posting is a useful guide for non-specialist analysts. Nothing that GR writes makes nonsence of my posting.
Nobody had asked about other differences between the techniques eg. resolution or simultaneous acquisition. I am pleased that G-R has supplied some information on those topics and on new, very high count-rate acquisition facilities for EDS. G-R is proud of his 0.6% "quantitative analysis" of Cr at an accuracy of +/- 0.1wt%.
I suggested that in EDS the presence of 1% of an element is the approxiamte lower limit for quantitative analysis. G-R has lowered that limit by some 40% - or has he? +/- 0.1% is good when 20% of the element is present, as it represents an accuracy of 0.5%. +/- 0.1% when 0.6% is present is about +/- 16% accuracy; I call that qualitative or at best semi-quantitative.
Another correspondent emailed me and noted that it was President Trueman who had been looking for a one handed adviser. But that was for an economist and not a scientist as I, apparently wrongly, remembered. The correspondent could see my point though; thanks to Brian Demczyk. I think that Trueman had an excellent idea but he should have extended that search to a scientist as well. Certainly microscopists and economists share disciplines which combine art and science. And I should add require 'good judgement'.
Why now was I abused with that opening: "Jim Darley's posting is hardly furthering good science". Am I to believe that good science is advanced by quarelsome nitpicking and that all broadbanding is verboten? I plead not guilty. 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, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} } Subject: Re: detection limits x-ray } } Date: Friday, 5 September 1997 6:21 } } } } Jim Darley's posting is hardly furthering good science. } } } } 1: Detection limits (John's original question). Modern EDX systems are } } easily capable of quantitative analysis in the sub-1wt% range. See, for } } example http://prism.mit.edu/facltis/stem/stmexam.htm (the second } } illustration on that page) where I was getting analyses for Cr in steel } of } } the order of 0.6+/-0.1 wt%. I don't know about you, but I consider this } } quantitative. This, mind you, was in a measurement where I was } attempting } } to optimize spatial resolution rather than sensitivity, and the } acquisition } } time was 60sec. per data point. This example, of course, was from a } STEM. } } There are many more examples in the literature. } } } } Using beam gating techniques such as those developed by Charlie Lyman and } } colleagues, EDX data can readily be acquired at 20,000-40,000 counts per } } real second, if spatial resolution is sacrificed. Combine this with an } } acquisition time of 1,000 seconds (by no means unrealistic for an } important } } measurement) and the detection limit is in the rage of, or better than, } 0.01wt%. } } } } Of course, in the SEM, which may have been the point of John's original } } question, the situation is not the same, the beam voltage is lower } } (resulting in poorer peak/bremmstrahlung ratios) and the emission of } x-rays } } from a solid sample is different from that in a thin foil, but these are } } some of the variables that Michael quite rightly pointed out must be } considered. } } } } 2: Comparison between EDX and WDX. } } } } This is like comparing apples and oranges, because the instruments } designed } } with them are generally intended for different purposes. } } } } WDX has a much better peak resolution than WDX, which results in better } } measured P/B ratios (and hence improved statistics), as well as much } better } } capability in resolving nearby x-ray lines. Also, because the x-ray } } counting and wavelength analysis are different functions in the crystal } } spectrometer, available countrates have traditionally been higher in WDX } } than EDX (although modern EDX detectors are an order of magnitude faster } } than they were fifteen years ago). Against this must be set the fact } that } } WDX is inherently a serial technique (although multiple spectrometers } help } } here), while EDX is a parallel technique (compare the advantages of PEELS } } over SEELS - although this is not a totally fair comparison). Anyway, } the } } advantage of WDX (ignoring the capability of resolving peak overlaps) is } } improved statistical precision. Is this useful? } } } } Well, maybe. } } } } By far the most significant parameter which affects electron-induced } x-ray } } emission spectra is sample geometry. Two extreme cases are where the } sample } } is a thin foil (as in the STEM) where, to a first approximation, the } x-ray } } spectrum recorded by the detector is the same as that emitted, and to a } } second order, it is possible to derive a reasonable thickness correction } for } } cases where the error is small. Alternatively, when the sample has } } dimensions large compared with the volume irradiated by the electron } beam, } } and has an accurately known geometry compared to the incident beam and } the } } detector (such as a polished flat sample in the microprobe), correction } } programs such as ZAF can do a reasonable job of extracting a quantitative } } analysis. } } } } What about where the sample geometry is unknown (for example, a rough } } surface such as might be examined in the SEM)? In that case the } uncertainty } } in the analysis is far, far worse than any uncertainty caused by the poor } } statistics of the EDX spectrum, so there is no point or advantage } whatever } } in using WDX to try to improve things, because it won't. This is why } } typically an SEM has an EDX detector - it is cheaper and gives just as } good } } an analysis (except for the somewhat poorer detection limit). In the } } microprobe, great care is taken in polishing and mounting the sample, so } the } } advantage of the WDX detector can be realised. } } } } Where does this leave us? The advantage of a WDX detector over an EDX } } detector *ON THE SAME SAMPLE* is limited - perhaps an order of magnitude } in } } detection limit, and, on a flat, polished sample, also perhaps } approaching } } an order of magnitude in precision. The WDX detector can also resolve } many } } cases where peaks would overlap in EDX. On general rough SEM samples, } the } } only advantages of WDX are a small improvement in detection limits and } the } } ability to resolve overlaps (which could be important if a trace element } } peak overlaps a major peak in the EXD spectrum. } } } } The President's science advisors were right - it all depends. I seem to } } remember that one of Jim Darley's Prime Ministers (Gough Whitlam?) had to } be } } dismissed because Parliament would not pass his budget - but does that } have } } anything to do with Science? } } } } Tony Garratt-Reed } } } } } } } Hi John and Michael et al: } } } I recall that one of your Presidents (Kennedy?) was looking for a one } } } handed science adviser; the ones he had always prevaricated "on the one } } } hand and on the other hand". } } } } } } Michael is quite right, detection limits vary greatly depending on } endless } } } factors. However, it is useful to have some figures as guidelines: } } } Considering say the 10 elements following sodium. Detection of these is } } } about best. } } } For these in EDX the limit for quantitative analysis is about 1%. } Detection } } } limit is about 0.1%. Increasing counts and counting times beyond the } } } customary 100 seconds at perhaps 2000cps will scarcely improve either } } } limit. } } } } } } WDX is near quantitative to its detection limit and that is at least two } } } orders of magnitude greater than is EDX. } } } } } } I'll enter correspondence only when it concerns errors in excess of five } } } orders of magnitude. } } } 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, 400+ Links, MSDS } } } ************************ http://www.proscitech.com.au } } } } John J. Bozzola wrote: } } } } } } } } } } Under ideal (and reasonably attainable conditions), what is the } } } } } detection } } } } } limit (in grams) for EDX and WDX? Thanks. } } } } } } } } The question you ask is not specific enough ... to many factors to } be } } } } considered with regard to which elements you are interested in, and } } } } (e.g.) ... how sensitive your specimen is to long count times and high } } } } beam currents ... ... ask again ... } } } } cheerios, shAf } } Anthony J. Garratt-Reed } } MIT Room 13-1027 } } 77 Massachusetts Avenue } } Cambridge, MA 02139-4307 } } United States of America } } } } Ph: 617-253-4622 } } Fax: 617-258-6478
Would anyone else like to help this soul. Send mail to his address (mluiselli-at-davinci.cnart.mx) not mine. Thanks
} Return-Path: {mluiselli-at-davinci.cnart.mx} } Date: Wed, 10 Sep 1997 18:33:42 -0700 } From: "Lic. Mariana Luiselli" {mluiselli-at-davinci.cnart.mx} } Organization: C N C A } To: sdw-at-biotech.ufl.edu } Subject: can you help me to find out what is K=F6eler lighting... } X-URL: http://www.biotech.ufl.edu/~emcl/tips.html } } In my homework they ask me, what is k=F6eler lighting, but i don=B4t know= =20 } what it is. Can you help me to find it out? } Thank you so very much, } mariana luiselli } }
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { 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 "
Thanks to all who responded to my CM12 +24 volt power supply problem.
The answer to the problem is to change C104 and C106, 47 ufd. 40 volts capacitors to 47 ufd. 63 or 100 volts. These capacitors are in the auxiliary power supply for startup, within the unit.
After the repair, I powered up the microscope and it worked the first time.
Thanks again Fred
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
Hello, I=B4m interested in staining protoplast with DAPI, but I don=B4t know how d= o=20 it.Is necessary a fixation?. I would like know a protocol to make this=20 staining. Best regards, M.D.Gomez email: gomezm-at-plantas.ibmcp.upv.es =20
If at all possible, could compilations of replies/discussion regarding osmium pepper be posted on the list server. I know that would serve useful for myself. Thanks.
Dan Caruso Biological Technician Medjet-at-worldnet.att.net
---------- } From: Grace Kennedy {kennedy-at-nsi.edu} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: osmium pepper } Date: Wednesday, September 10, 1997 7:52 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } At least once over the history of this listserver there has been extensive } discussion about osmium "pepper" and other artifactual inclusions seen in } embedded, sectioned TEM samples. Does anyone have a compilation of } replies/discussion or remember what conclusions were drawn? Answer } directly please, to avoid boring others with a rediscussion, OK? Many } thanks. Grace P.S. Was it related especially to phosphate buffered } osmium solutions?? I just had a major disaster with some and would like to } solve the problem quickly..... }
At 04:13 PM 9/10/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Consolidated International Corp. 4501 South Western Blvd Chicago, IL 60609 800/621-3680 312/376-5600 312/376-5835 FAX
I don't know if there are other suppliers. Be very careful with the installation. We've had difficulties with ours primarily due to an incompetent installation.
} Can anyone recomend a good, user friendly LM IM analysis software program? } Our current one is useless (hate to say).
Which one is it?
} We will be demoing a Noesis pacckage soon.
Superb but not particularly friendly. Steep learning curve.
} I have experience with Optimas, and personally do not care } for it. Any other shareware besides NIH? All comments etc welcome.
ImagePro is probably the most friendly and is fairly powerful. For shareware, I strongly suggest you look at ImageTools (http://ddsdx.uthscsa.edu/dig/itdesc.html). An alternative which I liked quite a bit less but which seems powerful is Osiris (http://www.expasy.ch/www/UIN/html1/projects/osiris/ReadmeOsiris.html).
I want to supply Karnovsky (modified to 2.5% G and 2.5% "paraformaldehyde") to a lab to collect 5mm biopsies over a period of time. In the past I have used Karnovsky for up to a week after producing it but I am not sure about storing it for 1-2 months. Should I store at 4 deg C or consider freezing some (it will be in caodylate with 2.5mM CaCl2)?
Malcolm Haswell Electron Microscopy University of Sunderland
"Another correspondent emailed me and noted that it was President Trueman who had been looking for a one handed adviser. But that was for an economist and not a scientist as I, apparently wrongly, remembered. The correspondent could see my point though; thanks to Brian Demczyk. I think that Trueman had an"
I think this must have been a Freudian slip, and at certain times he might have even been called President Bluntman, but his name was "Truman".
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 617-275-4695 FAX: 617-271-0252
I've been using the BioQuant system for about 5 years now and love it. It's sold by R&M Biometrics Inc. Phone 615-350-7866.
It's a Windows based program that is very versatile program, color recognition as well as grey scale. It can do sterography, cell counts (I automated my cell proliferation studies with it), etc.
the usual discliamer :) I have no connection with this Co. except as a statisfied customer. -- Begin original message --
} From: Microls-at-aol.com } } Can anyone recomend a good, user friendly LM IM analysis software program? } Our current one is useless (hate to say). We will be demoing a Noesis } pacckage soon. I have experience with Optimas, and personally do not care } for it. Any other shareware besides NIH? All comments etc welcome. } } Sincerely: } Lou Solebello } JM Huber Corp. } } }
-- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
I have just returned from holidays and have found a number of responses related to the question of gelatinized glutaraldehyde I had posed just before I left. Thanks to everyone for their interest. Oh by the way, I have now been able to entice one of my chemist colleagues to look at some of these samples to see whether she can discover a reason for this phenomenon as a result of differences in berry chemistry. If anyone is interested in the results, please contact me offline, but it will be awhile before the chemical analyses are done.
Thanks again.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia, Canada B4N 1J5
Dear Craig: Try Arkay Corporation, 228 South First Street, Milwaukee, Wisconsin Phone #1-800-TO-ARKAY; 414-276-9196. My catalogue shows that they carry both. Don Gantz Boston Univ. Med School
I would be interested to hear from Philips XL30 & XL40 users regarding how long their tungsten filaments last.
Thanks for your input. Nancy R. Smith Director of Operations Microscope And Graphical Imaging Center California State University, Hayward http://www.csuhayward.edu/SCI/sem
Wil Bigelow wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Torr Seasl is an epoxy compound especially formulated for use in vacuum } systems that was sintroduced by Varian Associates, Vacuum Products } Division, 121 Hartwell Ave, Lexington, MA 02173-3133 (800-882-7426) a } number of years ago. It is stated to be good at pressures below 10-9Torr, } and is bakeable at temperarures up to 120C. It adheres to most clean } materials (glass, metals, ceramics) and holds up well over long term } service. Some companies that handle EM supplies also handle it (e.g. I } find it listed in the LADD catalog). } } Other similar products are also sold by other companies. For example, } Duniway Stockroom Corp. (800-446-8811; info-at-duniway.com) sells a product } called 'Epoxy Patch' that is stated to be equivalent to Torr seal } } 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
As a manufacturer of vacuum evaperators we have been plagued by chipping of bell jars for many years. We have used (and sold) Torr Seal and several other epoxies over the years and they are satisfactory for small chips. However, for large chips we have found that epoxy-putty GAPOX10(tm) works extremely well. It is inexpensive and cures in one hour for sanding and smoothing. We can pull a vacuum of 10(-7) and have yet to encounter a problem.
Thanks to all who responded to my osmium pepper question. I now know not to osmicate in phosphate but haven't decided what I should use for a buffer-I don't think phosphate and cacodylate are compatible so a simple switch may be out. Has anyone out there used Dalton's dichromate/osmium? I cannot find any reference to anything by Dalton. I do have a formula and have used it but am curious to know if anyone else has played with it. Tx Grace
} Thanks to all who responded to my osmium pepper question. I now know not } to osmicate in phosphate but haven't decided what I should use for a } buffer-I don't think phosphate and cacodylate are compatible so a simple } switch may be out. Has anyone out there used Dalton's dichromate/osmium? I } cannot find any reference to anything by Dalton. I do have a formula and } have used it but am curious to know if anyone else has played with it. Tx } Grace
In a former life, I used Dalton's fixative for immersion fixation of vertebrate retina. I don't remember all the details, but the results were very good. I don't remember this fix being widely used, though.
Hi All, does anyone know of a portable IR microscope vendor? A collleague of mine would like to purchase one to be able to look at sub surface features on silicon crystals as-grown. Since the crystals are rather large, it is easier to take the microscope to the crystal then vice-versa if possible.
I appreciate any help.
cheers
Lucio
Dr.Lucio Mule'Stagno MEMC Electronic Materials Inc University of Missouri -St.Louis Silicon Materials Research Group Physics Dept., 501 Pearl Dr., 8001 Natural Bridge rd., St.Peters, St.Louis MO 63376 MO 63121 tel: 314 279 5338 314 516 5931/3 fax: 314 279 5363 314 516 6152 email: lmulestagno-at-memc.com luciom-at-newton.umsl.edu
} Can anyone recomend a good, user friendly LM IM analysis software } program? Any other shareware besides NIH? All comments etc welcome. } } Sincerely: } Lou Solebello } JM Huber Corp.
ImageTool is a free image processing and analysis program for Windows 95/NT from the University of Texas Health Science Center at San Antonio.
http://ddsdx.uthscsa.edu/dig/itdesc.html
This list needs a FAQ.
-------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com The Web is:"Vaster than empires and more slow" Andrew Marvell (1621-1678)
jss wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. }
} The Sparc5. Microscopy Station Master } unsubscribe
} Thanks to all who responded to my osmium pepper question. I now know not } to osmicate in phosphate but haven't decided what I should use for a } buffer-I don't think phosphate and cacodylate are compatible so a simple } switch may be out. Has anyone out there used Dalton's dichromate/osmium? I } cannot find any reference to anything by Dalton. I do have a formula and } have used it but am curious to know if anyone else has played with it. Tx
Grace, In our experience, osmium pepper is not caused by phosphate problems but by using aldehydes that have partially polymerized (e.g., old glutaraldehyde or formaldehyde solutions). The polymers are small enough to get into the cell but once attached to proteins, can not be removed. The polymers then vigorously reduce osmium which then shows up as pepper. I have routinely used phosphate buffered osmium and have never had the pepper problem. In fact, however, for osmication you really don't need to buffer at all. Distilled water is fine.
I used Dalton's chrome osmium many years ago for tissue culture cells and it worked fine. Now we prefer to use osimum ferrocyanide which gives better contrast. You can prep the solution using cacodylate. We fix in either cacodylate OR phosphate buffered glut/form, rinse extensively (overnight) in cacodylate buffer and then into the osmium.
If you need more info, contact me again as I can fax you the info for Dalton's.
John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
It's been cute sitting in the corner, but now this prof. needs the space. Does anyone want a Philips 75 TEM, 1960s vintage? It's small, not too heavy, and actually ran until it blew a filament sometime back. I'm not sure which decade that was... I have the manuals and tools as well.
You would, of course, have to pay packing and shipping.
For more info, drop me a line at tina-at-pbrc.hawaii.edu
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* ****************************************************************************
When taking a biopsy, careful attention to the following points will ensure that the histopathologist is quite unable to give any useful information.
1. Paint the skin with a strong antiseptic solution, preferably one that is deeply coloured (after all, the tissue has to be stained sooner or later).
2. Rub on the antiseptic vigorously to ensure that the surface layers of the epidermis are so disturbed as to be histologically uninterpretable.
3. Inject a considerable volume of local anaesthetic solution into the middle of the area to be biopsied and preferably inject the fluid rapidly. This will produce a satisfactory degree of tissue distension and distortion.
4. Seize the area to be biopsied with artery forceps, making sure that the forceps are clamped firmly enough to crush the tissue. Should the biopsy be too large to be effectively be dealt with by one pair of forceps then do not hesitate to use several pairs.
5. The tissue excised should be from the centre of the lesion and should not include any of the adjacent normal tissue. It is making things too easy for the Pathologist if the specimen allows the relationship of the lesion to the adjacent tissue to be seen.
6. When making the excision, two alternative and equally effective techniques may be considered. Either the biopsy can be only a fraction of a millimetre thick (thus saving the trouble of cutting sections) or a large piece of tissue can be excised. In this case make a number of tentative cuts so that the biopsy is partly cut through in several places. This will help to make the specimen impossible to orientate for proper sectioning.
7. If you think that the lesion may be an invasion tumour, always remove it piecemeal with a curette. The Pathologist will then probably not be able to tell whether it is invasive or not.
8. Before placing the tissue into fixative, ensure that is adequately covered with blood. This should be done so effectively that after fixation the specimen is entirely concealed within clot. This ensures that the Pathologist is kept busy trying to find the specimen and provides a reasonable chance that every section will be covered with red blood cells dislodged from the clots during the sectioning process.
9. If the biopsy is to be really uninterpretable, care must now be exercised in the selection of the container and the fixative. The container should be very small so that there is room for a minimum of fixative. Plastic sequestrene bottles (pink label) are admirable for this purpose. It is quite surprising how much tissue can be compressed into one of these. Alternatively, use a container with a very narrow neck. A mass of tissue is often quite soft when freshly excised and can be forced through a small hole. When fixed the tissue can not be removed from the bottle without breaking the glass. With luck fragments of glass will then be driven into the tissue. This lends excitement to the process of section cutting.
10. The fluid into which the specimen is placed should on no account be a conventional histological fixative, such as 10% neutral-buffered formol saline. Plain water or physiological saline will ensure adequate breakdown of the cells. If available, Stuart's transport medium is even more effective in producing putrefaction. If such a culture medium is used, the specimen should be kept on a radiator until dispatch and should be sent to the laboratory by post (this is especially effective during the summer months).
11. On no account should the container be labelled with the patient's name or any other mark of identification. If you can arrange for several unlabelled containers to arrive in the laboratory on the same day so much the better. There is an excellent chance that the specimens from different patients will be confused. An alternative, and equally effective, measure is to place more than one biopsy in the same container. If they are from the same patient make sure the pieces of tissue are all of a similar shape and size. Thus, if one biopsy shows a neoplasm no-one will know which of the biopsy sites contains the tumour.
12. In the accompanying request form on no account should you provide the Pathologist with the name of either the patient, yourself or your address. Otherwise, there will be some means of indexing the specimen and of knowing where to send the report when eventually prepared.
13. So that the Pathologist shall not be influence or biased in interpretation, avoid giving any information regarding the age of the patient, or the site of the biopsy, or the duration and appearance of the lesion. Above all, never mention your own differential diagnosis.
Finally, one or more of the following stratagems should be used in selected cases:
(a) Ask for serial specimens, bearing in mind that a specimen 5 mm thick will yield about a 1000 sections.
(b) Telephone the laboratory frequently for the report. If possible arrange for the first two calls to reach the laboratory before the specimens.
(C) Ask for rapid-frozen sections especially on large heavily-calcified masses that have been present for several years.
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
HASWELL Malcolm wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I want to supply Karnovsky (modified to 2.5% G and 2.5% "paraformaldehyde") } to a lab to collect 5mm biopsies over a period of time. } In the past I have used Karnovsky for up to a week after producing it but I } am not sure about storing it for 1-2 months. Should I store at 4 deg C or } consider freezing some (it will be in caodylate with 2.5mM CaCl2)? } } Malcolm Haswell } Electron Microscopy } University of Sunderland
Malcolm:
I have stored this fixative in glass at room temperature for extended periods (1-2 months) and have had no problems. Once you see some bottom ppt. or cloudiness, discard with copious amounts of water. Also be careful of the formalin conc. in the air. Best regards, Jerome.
Dalton's dichromate-osmium fixative brings back some nostalgic memories. I used it to fix rat testicular Leydig cells for my PhD thesis research (about 1956-58, Harvard Biology), and the results suggested that the smooth endoplasmic reticulum was tubular, rather than than vesicular, as was commonly believed at the time. In my subsequent postdoctoral work with Don Fawcett, opossum Leydig cells fixed with Dalton's dichromate osmium exhibited tubular smooth ER, while the same material fixed with osmium-acetate veronal, commonly used at the time, had vesicular SER. This work was published in 1961 (Christensen, Fawcett 1961, J Biophys Biochem Cytol 9:653-670).
The reference for the fixative is an abstract: Dalton, A.J. 1955, "A chrome-osmium fixative for electron microscopy," Anat Rec 121:281. I knew Jack Dalton, who was at the National Cancer Institute, at NIH in Bethesda. He and Marie Felix were the first to show the Golgi complex by EM (1954, as I remember).
Kent
A. Kent Christensen Department of Anatomy and Cell Biology Medical Sciences II Building University of Michigan Medical School Ann Arbor, MI 48109-0616 akc-at-umich.edu Tel (313) 763-1287 http://www-personal.umich.edu/~akc/
----------------------------
On Thu, 11 Sep 1997, Grace Kennedy wrote:
} Thanks to all who responded to my osmium pepper question. I now know not } to osmicate in phosphate but haven't decided what I should use for a } buffer-I don't think phosphate and cacodylate are compatible so a simple } switch may be out. Has anyone out there used Dalton's dichromate/osmium? I } cannot find any reference to anything by Dalton. I do have a formula and } have used it but am curious to know if anyone else has played with it. Tx } Grace
Our lab is using the fluorescent antibody technique to detect and count Cryptosporidium and Giardia in water samples. One of the biggest interference?s with this is the auto-fluorescence of algal cells which is often brighter than the FITC stained cells we are looking for. This is particularly inconvenient as we are attempting to automate the microscopy with the use of Image analysis. Part of our efforts are going into producing a cleaner final concentrate but it is not always possible to eliminate all the algae. In view of this has any one come across a product/method for quenching auto-fluorescence that does not affect the FITC stain.
Phil Dobson Australian Water Quality Centre Hodgson Rd, Bolivar South Australia 5118 Phone 61 8 8259 0341 Fax 61 8 8259 0228 Email phil.dobson-at-sawater.sa.gov.au
I'm translating a microscope manual from Japanese into English and constantly running into sentences that go like "part of the field of view may be cut off" and "image cut-off in peripheral part may occur."
Could anyone tell me how to put them correctly? Should I instead say "the field of view may be vignetted" and "image vignetting may occur" which sound more technical to me but I'm not sure about.
Any suggestion is welcomed. Thanks in advance.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
have you considered measuring the emission at two wavelengths? The autofluorescence of algae should be pretty red, whereas fluorescein is predominantly green. Subtracting a scaled red image from the corresponding green image should leave you with only fluorescein fluorescence, presuming that all of the algae have the same fluorescence spectrum. Alternatively, if the cells remain discrete in the image, it would be possible to get your image analysis package to ignore the algae on the basis of size and/or create a mask in the red image that prevents the algae being counted in the green one.
Ray
At 3:39 pm +0930 12/9/97, Phil Dobson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ray Hicks ________________________________________________________________________ |University of Cambridge |Tel 01223 330149 | |Department of Medicine |Fax 01223 336846 | |Level 5, Addenbrookes Hospital |e-mail {rh208-at-cus.cam.ac.uk} | |Hills Road Cambridge |Web http://facsmac.med.cam.ac.uk | |CB2 |ftp server ftp://131.111.80.78 | |UK | | |_________________________________|_____________________________________|
We currently have the first version of the Buehler image analysis system and I have nor problems with it. It is a very powerful package and for all of our Materials analysis needs it performs very well. We have not upgraded to the latest version wich is supposed to be even better due to lack of funds. They will be happy to demo it for you if you give them a call.
Roberto Garcia EMF Manager Wright State University
Sorry about keeping you waiting. Bernies Photo Center can be reached at 800 346-8884. They will have verything you need and if I could make a suggestion splurge on a sodium vapor light, it is worth it. No more stumbling around the darkroom. They also have the best prices around on bulk 4X5 film. Good luck with the darkroom.
Roberto Garcia EMF Manager Wright State University Dayton, OH
In a message dated 97-09-12 08:45:19 EDT, Chiba writes: { { sentences that go like "part of the field of view may be cut off" and "image cut-off in peripheral part may occur." Could anyone tell me how to put them correctly? Should I instead say "the field of view may be vignetted" and "image vignetting may occur" which sound more technical to me but I'm not sure about. } }
The phrase "part of the field of view may be cut off" sounds ok to me. Without an illustration, I do not know what it means. My mental image is that there is a black part within a photographic image taken with the microscope from a flash/shutter timing problem, or from blocking of the field of view by an accessory that is inserted in the optical path, such as the quarter wavelength plate typically used in a polarizing microscope.
The phrase "image cut-off in peripheral part may occur." could be reworded as "...the peripheral part of the image may be cut-off." As above, I do no know really what it means. But I imagine that it means the effect from closing down the sub-stage iris. I do not think that is really vingetting, a gradual shading, but it is close to it. If the effect being discussed is from gross misalignment of the bulb in the illuminator housing, then it could be vignetting.
Steve Stokowski Stone Products Consultants Concrete Petrographers http://members.aol.com/CrushStone/index.htm
} Sorry for my layman's question. } I'm translating a microscope manual from Japanese into } English and constantly running into sentences that goes } like "part of the field of view may be cut off" and } "image cut-off in peripheral part may occur." } } Could anyone tell me how to put them correctly? } Should I instead say "the field of view may be } vignetted" and "image vignetting may occur" which sounds } more technical to me but I'm not sure about. }
I would not use the term "vignette" myself. It is a technical term that is used in lens design to mean that you have blocked part of a bundle of rays (hopefully on-purpose to kill the wild rays that would have not been well focussed, but sometimes by mistake because a lens element was too small). Although the word could be used for your purpose, few people know what it means, and some of them may be confused because of the technical use. I would just say "the field of view may be reduced," and "part of the field of view may be cut off." Making it sound more technical is helpful only for people with trouble falling asleep :)
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
} Sorry for my layman's question. } I'm translating a microscope manual from Japanese into } English and constantly running into sentences that goes } like "part of the field of view may be cut off" and } "image cut-off in peripheral part may occur." } } Could anyone tell me how to put them correctly? } Should I instead say "the field of view may be } vignetted" and "image vignetting may occur" which sounds } more technical to me but I'm not sure about. }
I would not use the term "vignette" myself. It is a technical term that is used in lens design to mean that you have blocked part of a bundle of rays (hopefully on-purpose to kill the wild rays that would have not been well focussed, but sometimes by mistake because a lens element was too small). Although the word could be used for your purpose, few people know what it means, and some of them may be confused because of the technical use. I would just say "the field of view may be reduced," and "part of the field of view may be cut off." Making it sound more technical is helpful only for people with trouble falling asleep :)
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
I have been trying several stains on fixed monolayer fibroblasts. The cells have been fixed by either formalin, formalin fumes, or gluteraldehyde. None of the stains are penetrating the cells. Does anyone have a suggestion?
We just bought a new Olympus inverted microscope for epi-fluorescence. We were very surprised to find that our 1.2NA water immersion objective gave BRIGHTER pictures and SHARPER pictures than our 1.4 NA oil immersion objective. We thought it was a problem with the 1.4 NA objective so the sales representative brought by:
1) another 1.4 NA oil immersion objective: whose pictures were as poor as the previous objective 2) a 1.25 NA oil immersion UV objective: whose bright spots were brighter, whose black regions were darker, and whose contrast was sharper than on the 1.4 NA objective. (The 1.25 NA objective is also much more affordable objective).
Does anyone have an explanation? One possibility was that the 1.4 NA objective is NOT a UV objective: we thought that some UV light may be coming through our 460-490 nm excitation filter causing auto-fluorescence in the objective. So we put another very sharp 488 nm filter in front of it and it did not resolve the problem.
In comparing the 1.4 NA oil to the 1.2 NA water objective, shouldn't the signal be 85% brighter? - on the excitation side: (1.4/1.2)2=1.36, or 36% more transmission of excitation - on the emission side: (1.4/1.2)2=1.36, or 36% more collection of emission in total: (1.4/1.2)**4=1.85.
So how could the lower NA objectives give brighter sharper signals?
We are collecting our images with a Hamamatsu Cooled-CCD camera. Both the shutter on the excitation and the collection are under software control, so we know that data from both objectives are collected under the same condition. We've used a series of test slides so each objected could be used on the same image. Any suggestions are welcome.
Thanks,
Sanford M. Simon Laboratory of Cellular Biophysics Box 304 Rockefeller University 1230 York Avenue New York, N.Y. 10021 212-327-8130 (voice) 212-327-8022 (fax) simon-at-rockvax.rockefeller.edu (e-mail)
In regards to (i) EM immunocytochemical staining on primary antibodies and collodial gold and (ii) uranyl acetate and Pb citrate, I would like to poll the list to determine how many people either float their EM grids or immerse them in droplets so that both sides are stained. I have always floated on a 20 ul drop but have started wondering if I am missing half the action. Comments about advantages and disadvantages welcomed!
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
I have a couple of questions and will appreciate any replies and suggestions.
1) What is the best sample to test the resolution of a TEM at a tilt angle (} 30degree)?
I am testing the resolution of a CM300FEG microscope. I used oriented gold, evaperated gold particles, graphitized carbon and so on. There is no problem of those samples at zero tilt. But after tilt the specimen to 30 degree or higher, I got only the good resolution (2.3A in case of evaperated gold particle) in one direction, along the tilt axis. I believed it is not because of the focus gradient that I could not get the same resolution in both directions at this tilt angle. I think the samples (evaporated gold particles and graphitized carbon) may not be the suitable samples for the resolution test of the high tilt angles (higher than 45 degree and up to 60 degree). I wonder if any of you has done such test before and has any suggestions on which specimen should be used in this test and where to get it?
2) Is there any published documentation about the line resolution of Kodak SO-163 film?
I am writing an paper about the performance of the CM300FEG at low magnificaiton, and need to know the line resolution of Kodak SO-163 (something I can cite for). But I could not find any published documentation. Of course, I called Kodak technique support and of course they didn't give me a clue. There is a web site called "Bibliography on EM Imaging and Related Technologies" (http://www.uct.ac.za/depts/emu/imaging/papers.htm), but I can not find any thing there either about SO-163. The only other related information I got was from a similar discussion in this board a year ago. But the data about line resolution found in that discussion were mostly estimated but not from any published documentation. I wonder if any of you know there is any kind of published documentation including research paper, technique report or so which gives the line resolution of Kodak SO-163. (BTW, the topic about the line resolution has been discussed a year ago and I am sorry to ask it again.)
I appreciate any information or suggestion on my request.
Yifan Cheng
BTW, I am not sure if my email address is already on the list or not. So that I appreciate also that when you reply this email, send a reply to me directly as well as to the board. Thanks again.
-- ********************************************************************** * Dr. Yifan Cheng * Phone: +1-850-644-4104 (Office)* * Institute of Molecular Biophysics * or: +1-850-644-9769 (Lab) * * Florida State University * Fax: +1-850-561-1406 * * Tallahassee, FL 32306-4380 * Email: ycheng-at-sb.fsu.edu * * U.S.A. * http://www.sb.fsu.edu/~ycheng * **********************************************************************
We have a Zeiss DSM-960 with LaB6. This system has a long history of problems with the column isolation valve (manually operated) leaking during specimen exchanges. Numerous fixes have been tried including slightly larger and softer o-rings and metal shims to change the height/pressure which the gate cams to when closed, all to no avail. Has any one experienced similar problems on a Zeiss or on other instruments and was a fix found.
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-uimrl7.mrl.uiuc.edu {that's the letter l} _____________________________________________________
} HASWELL Malcolm wrote: } } } } I want to supply Karnovsky (modified to 2.5% G and 2.5% "paraformaldehyde") } } to a lab to collect 5mm biopsies over a period of time. } } In the past I have used Karnovsky for up to a week after producing it but I } } am not sure about storing it for 1-2 months. Should I store at 4 deg C or } } consider freezing some (it will be in caodylate with 2.5mM CaCl2)? } } } } Malcolm Haswell } } Electron Microscopy } } University of Sunderland } } Malcolm: } } I have stored this fixative in glass at room temperature for extended } periods (1-2 months) and have had no problems. Once you see some bottom } ppt. or cloudiness, discard with copious amounts of water. Also be } careful of the formalin conc. in the air. Best regards, Jerome.
I would recommend purging the vials with nitrogen gas prior to sealing. Use teflon lined caps, if possible and store at 4C.
cheers
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 http://www.personal.psu.edu/ejb11/
} } We will be demoing a Noesis pacckage soon. } } Superb but not particularly friendly. Steep learning curve.
Kalman, my records indicate you are using our old version Visilog4.1.3. The new release is much more powerfull and easier to use. Give me a call and I'll upgrade you at no cost.
Regards,
---------------------------------------------------------------------------- -------- 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 ---------------------------------------------------------------------------- --------
NANCY SMITH wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I would be interested to hear from Philips XL30 & XL40 users } regarding how long their tungsten filaments last. } } Thanks for your input. } Nancy R. Smith } Director of Operations } Microscope And Graphical Imaging Center } California State University, Hayward } http://www.csuhayward.edu/SCI/sem
Nancy,
Although we do not use a Philips XL30 or XL40 in our own lab, we have supplied filaments for years. So based on our experience and comments from our customers, let me make the following observations:
Some Factors Affecting Tungsten Filament Life
1) quality of vacuum 2) single or multiple user instrument 3) high or low KV 4) height setting of filament 5) oversaturation 6) quality of the Tungsten
Some of our customers report life spans of 50 to 200 hours and beyond. A normal burnout is when the filament has a bubble on the top of the broken wire. Look for these indicators when the filament burns out too quickly:
a) normal burnout where the base is clean could mean there is a good vacuum but an incorrect height setting or oversaturation. b) a discolered base could mean a bad vacuum c) a cracked filament (no bubble) could be a flaw in the wire or a slight crack in the wire during installation
line resolution of film: films were compared by Downing and Grano (1982) Ultra- microscopy 7:381-404 the Kodak 4463, as used in this study, is now the SO163, as given in a data sheet No P-252 by Kodak. There is no simple figure for the resolution, but they measured the modulation transfer function for different frequencies. Check the paper, it is worth reading. Regards, Reinhard
I'm translating a microscope manual from Japanese into English and constantly r unning into sentences that go like "part of the field of view may be cut off" a nd "image cut-off in peripheral part may occur."
Could anyone tell me how to put them correctly? Should I instead say "the field of view may be vignetted" and "image vignetting may occur" which sound more te chnical to me but I'm not sure about.
Any suggestion is welcomed. Thanks in advance.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
Chiba Atsushi wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Sorry for my layman's question. } } I'm translating a microscope manual from Japanese into English and constantly running into sentences that go like "part of the field of view may be cut off" an } } Could anyone tell me how to put them correctly? Should I instead say "the field of view may be vignetted" and "image vignetting may occur" which sound more tec } } Any suggestion is welcomed. Thanks in advance. } } Chiba Atsushi [(Mr.) -- *Chiba* is my surname] } Voice: (+81) 010-045-9451Chiba, I would go with the simpler language. While vignetting is the appropriate term, it is less explicit.
MARK YOUR CALENDARS NOW!!! EVERYONE IS WELCOME TO ATTEND!
The Fall meeting of the Midwest Microscopy and Microanalysis Society will be held on Friday, November 7, 1997, at Eli Lilly & Co. in Greenfield, Indiana (near Indianapolis). The meeting is hosted by Jeffrey Horn of the Toxicology Research Laboratories at Lilly.
The schedule for the meeting is:
8:30 Welcome and Brief Overview of EM Lab Functions at Lilly - J. Horn, M.S., Eli Lilly and Company
9:00 Electron Microscopy in Pharmaceutical Drug Development - V. Meador, DVM/PhD, Eli Lilly and Company
9:30 Everything You Always Wanted to Know About Molecular Morphology (But Were Afraid to Ask) - the Basics of In Situ Labeling Methods - J. Fagerland, Ph.D., Abbott Laboratories
10:00 Coffee Break
10:30 Ultrastructural Evidence in Legal Cases, N. Cheville, DVM/PhD, Iowa State University
11:30 Lunch (Provided)
12:45 Tour Eli Lilly & Co. Toxicology Facility
2:00 Use of SEM for Selecting Therapeutic Candidate in Various in vivo Thrombosis Models - G. Sandusky, DVM/PhD Indiana University
2:30 Diagnostic Electron Microscopy in the Hospital Setting - M. Goheen, M.S. Indiana University Hospitals
3:00 Coffee Break
3:30 Digital Imaging for Dummies - J. Gagne, M.S., Abbott Laboratories
4:15 Conclusion
Meeting announcements, maps, and registration instructions will be sent to members of MMMS in U.S. Mail soon. Anyone else can receive them by contacting me at (847) 935-0104 or via email at jane.a.fagerland-at-abbott.com.
Sanford, first are you speaking of the differences with objectives as seen through eyepieces or with the CCD camera? if the problem is with the CCD camera one problem is probably IR leakage onto CCD with one objective and not the other. The NA of the lens should be taken as a measure of resolution capability not amount of light transmission, though with two objectivesof exact same type of glass and design then the NA can be used to see who collects more light. But when you start to compare a PlanApo to PlanNeofluar then the glass and lens design insdie objective may have drastically different spectral properties, especially outside visible range (Up in IR } 850nm) this is what will cause a low contrast soft image.
Another likely possiblity is that the lower the NA the more depth of field the objective will have and thereby when looking at a 3-d object have a sharper edge as the fluorecesence is coming from above and below focal plane (This is why confocal looks so good and why most quality microscopes will offer, and you should be purchasing an objective with an iris diaphram -- or have body of scope that has it built into body--aperture diaphramfor epi path-- to cut down on NA to cut down on bloom in sample. (your flourescent cell)
To check yourself put both objective on upright scope with high resolution condenser and look at a very thin {5micoron sectionsof H&E stain tissue. at this point the higher NA should be clearer and sharper then the 1.25NA . But other than BF you can see that nore NA is not always better.
Scott E. Berman Advanced Imaging Concepts, Inc. (609) 921-3629 x26 Princeton, NJ scotte57-at-aol.com
Sanford, first are you speaking of the differences with objectives as seen through eyepieces or with the CCD camera? if the problem is with the CCD camera one problem is probably IR leakage onto CCD with one objective and not the other. The NA of the lens should be taken as a measure of resolution capability not amount of light transmission, though with two objectivesof exact same type of glass and design then the NA can be used to see who collects more light. But when you start to compare a PlanApo to PlanNeofluar then the glass and lens design insdie objective may have drastically different spectral properties, especially outside visible range (Up in IR } 850nm) this is what will cause a low contrast soft image.
Another likely possiblity is that the lower the NA the more depth of field the objective will have and thereby when looking at a 3-d object have a sharper edge as the fluorecesence is coming from above and below focal plane (This is why confocal looks so good and why most quality microscopes will offer, and you should be purchasing an objective with an iris diaphram -- or have body of scope that has it built into body--aperture diaphramfor epi path-- to cut down on NA to cut down on bloom in sample. (your flourescent cell)
To check yourself put both objective on upright scope with high resolution condenser and look at a very thin {5micoron sectionsof H&E stain tissue. at this point the higher NA should be clearer and sharper then the 1.25NA . But other than BF you can see that nore NA is not always better.
Scott E. Berman Advanced Imaging Concepts, Inc. (609) 921-3629 x26 Princeton, NJ scotte57-at-aol.com
Carlos E. Barbosa wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Does someone can say me what type of lamp have I to use for performing } UV micropscopy of thin-sections of rocks. } Thank you in advance! } } Carlos BarbosaDear Carlos,
Both high pressure mercury arcs and xenon arcs show good emission in the UV. One caution: most conventional optics do not transmit below about 365-380 nm. Talk to your microscope vendor about quartz optics for all relevant components. I assume that you will be doing fluorescence, since humans cannot see into the UV (although I am told that those of us who have had cataract operations see further into the near UV than those of us with convention eyes) *and* of course, direct viewing of UV light can literally fry the delicate tissue in the eye. With UV fluorescence from a thin section, the job is a bit easier, but you will need quartz optics throughout the illumination system , including the objective. You may also want to test the glue which is holding your thin section to its substrate, to make sure that it is not autofluorescent, creating undesirable background.
Let me know how things work out.
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
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Thomas, As I understand for this type of staining, it depends on whether the grids are 'formvar' coated or not. If the grids that you are using are coated in some way, even if you did immerse them into solutions, the sections will only be stained on one side anyway. So there is no advantage in immersing the grids.
However, if the sections are on uncoated grids, then immersion into the Ab sols will give you greater staining, (both sides!)
Hope this helps,
Rich.
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
=46or both immuno staining and U acetate/Pb citrate, I've always immersed. In 25 =B5l drops for the former and in a multigrid staining thing for the latter. THe immersion for immuno was to pick up as much stain as possible on a structure that was about half as thick as an EM section. Oh, I was using uncoated grids, of course.
Rosemary White Department of Ecology and Evolutionary Biology Monash University, Melbourne, Victoria 3168, Australia phone 61-3-9905 5670 email r.g.white-at-sci.monash.edu.au fax 61-3-9905 5613 or rgwhite-at-vaxc.cc.monash.edu.au
For reasons best known to ourselves, we are attempting to resurrect an OMAR SPC 900/EX critical Point Dryer. It has not been used in some time and none of us can recall using it, let alone how it was used!! Of course, the manuals have long since disappeared (presumably to the same place that odd socks and pens go) and we need to get it going. Does anybody out there have a manual that they could fax us or even an idea of the pressures and times we should be using with this little beast? Any help would be gratefully accepted!
Many thanks,
Colin V.
Colin J. Veitch Instrumentation Scientist CSIRO Division of Wool Technology PO Box 21, BELMONT, Vic. 3216. Australia.
I work with a LaB6 Auger in a university environment and have been given a project to identify contaminants on and just below the surface of a leadframe (Au on epoxy). I can overcome the charging prior to depth profiling by dropping to low voltage (2Kv or Less). Once I start ion sputtering for short times the charging is enormous and continuous. Any suggestions would be appreciated.
Sincerely,
Tamara E. Bloomer Assistant Scientist Ames Laboratory 137 Wilhelm Hall Ames, IA 50011 (515) 294-2564
The text "Advanced Scanning Electron Microscopy and X-Ray Microanalysis" by Newbury, Joy, Echlin, Fiori, and Goldstein, 1986, Plenum, has a chapter on characterization of semiconductors which covers EBIC and voltage contrast.
regards,
Dave Audette Olin Research Center Cheshire, CT 06410 USA (203)- 271-4272 deaudette-at-corp.olin.com
colin.veitch-at-dwt.csiro.au wrote: } } Hi All, } } For reasons best known to ourselves, we are attempting to resurrect an } OMAR SPC 900/EX critical Point Dryer. It has not been used in some time } and none of us can recall using it, let alone how it was used!! Of } course, the manuals have long since disappeared (presumably to the same } place that odd socks and pens go) and we need to get it going. Does } anybody out there have a manual that they could fax us or even an idea } of the pressures and times we should be using with this little beast? } Any help would be gratefully accepted! } Hi Colin: Times have to be determined by trial and error. It depends upon the sample. As to temperature, the chamber must be cooled to between 15-20C in order for there to be liquid CO2. The solvent used to dehydrate the sample must be removed and this is done with repeated flushing of the chamber. When this is done the temp is raised to above 32C(36C just to be sure) pressure will be around 1200lbs. At this point the critical point has been reached and passed and pressure can be slowly lowered. Another option is Hexamethlydisilazane(HMDS). This is a liquid that can be used in place of CPD. After dehydration HMDS is subistuted with several changes and then allowed to air dry. It gives good results. You will have to try it on your sample
Hope this helps. Good luck. THE OPINIONS HERE ARE NOT THOES OF THE FACILITY Greg Rudomen Greg-at-umic.sunysb.edu University Microscopy Imaging Center S.U.N.Y. Stony Brook Stoy Brook, NY 11794-8088
I think it is O.K. to follow phosphate with cacodylate. We do so routinely, and don't seem to have any problems. Lesley Weston.
On Thu, 11 Sep 1997, John Chandler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Thanks to all who responded to my osmium pepper question. I now know not } } to osmicate in phosphate but haven't decided what I should use for a } } buffer-I don't think phosphate and cacodylate are compatible so a simple } } switch may be out. Has anyone out there used Dalton's dichromate/osmium? I } } cannot find any reference to anything by Dalton. I do have a formula and } } have used it but am curious to know if anyone else has played with it. Tx } } Grace } } In a former life, I used Dalton's fixative for immersion fixation of } vertebrate retina. I don't remember all the details, but the results were } very good. I don't remember this fix being widely used, though. } } John } chandler-at-lamar.ColoState.EDU } } }
I need a TPR 010 Piranni gauge for my Balzers BAF 301 Freeze etcher. Techno Trade does have them for $250.00 each...which is ok on an unlimited budget. Has anyone else gotten these elsewhere? Is there an aftermarket brand that I might use? And yes I have cleaned many a TPR 010 {I have a real good technique...} but after around four cleanings they act very strange. Any help, advise, or berations are welcome.
John Grazul Rutgers University Electron Imaging Facility
All manuals should follow the "KISS" principle! ie: Keep It Stupidly Simple!!!! Forget about whether it sounds "technical" or not, just make sure the directions are abundantly clear to a 5 year old child and researchers around the world will applaud you.
} } Sorry for my layman's question. } } I'm translating a microscope manual from Japanese into English and } constantly running into sentences that go like "part of the field of view } may be cut off" and "image cut-off in peripheral part may occur." } } Could anyone tell me how to put them correctly? Should I instead say "the } field of view may be vignetted" and "image vignetting may occur" which } sound more technical to me but I'm not sure about. } } Any suggestion is welcomed. Thanks in advance. } } } Chiba Atsushi [(Mr.) -- *Chiba* is my surname] } Voice: (+81) 010-045-9451
Cheers :) George Department of Earth and Atmospheric Sciences University of Alberta Edmonton, Alberta T6G 2E3 Canada ph: 403-492-5746 fax: 403-492-2030
Anyone aware of a "nearly" conventional photographic process for development of prints made from enlarged negatives? Examples: stabilization processors that use heat to develop sensitized papers OR (less desirably) concentrated chems as in the Kodak Ektamatic-like scheme.
Unfortunately, the heat-developed papers use mercury and silver and will be phased out shortly while the Ektamatic-type stabilization processors generate liquid wastes. The ideal situation would involve conventional negs enlarged onto a paper that could be processed using some sort of dry (or nearly so process) AND yielding photographic quality images.
Now, digital imaging people please don't jump all over me, because sometimes digital images and prints just do not offer adequate rendition of the information -- without spending big bucks. Right now, we are working with dense, highly contrasted negatives (difraction patterns, high contrast and thick specimens) and the quality of the digital images (direct image capture or scanning of TEM negs) is inadequate. My own idea would be to have a system akin to a xerographic process wherein one would enlarge a negative onto a high quality paper that would then be "developed" in a manner similar to a dry copier probably using toner powders. I believe all of these components currently exist but have not been put together - to my knowledge.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
O.k., I know this has been discussed MANY times before, but I I didn't save all the relavent messages.
Looking for two things:
(1) Refurbished / replacement screen for a JEOL 100-S (hey it still works fine)
(2) Looking for places to recoat our old 100s screen. I believe that SPI still does it, and will call soon (I did save your message Chuck). Grant Sci Corp doesn't seem to exist any more. Any others to consider? Any comments god or bad?
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
We have 10 years of blocks filed and stored in small plastic boxes from Althor Products. Our last purchase order was returned: no forwarding address. Does anyone know how to contact these people or of another source for those 1 3/4 x 7/8 x 3/4" boxes with snap-on lids? Thanks Joyce Craig Chicago State University
We inherited an old embedding oven that is caked in resin. Does anyone ever clean out their embedding oven? If so, what is used to do this?
I am not so concerned about the cleanliness of the oven, but rather the problems it seems to be causing when things start sticking to the oven. There is a plastic petri dish with old desiccant in it that has stuck itself onto the bottom of the oven, with no way to easily dislodge it.
Any suggestions or comments would be greatly appreciated.
Susan
Susan Carbyn Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
I am making TEM samples of modern and fossil brachiopod (a bivalved marine invertebrate) shells composed of calcite. I was intending to use an ion milling device (argon) to thin the samples. However, it has been suggested that a tripod polisher would be less time consuming and provide a larger "viewing" area.
Has anyone had experience using a tripod polisher on carbonate materials? Would tripod polishing produce more defects in the crystallographic structure than ion thining? My intent, if possible, is to evaluate diagenetic alteration by observing crystallographic defects.
Thanks.
Nancy Buening
Nancy Buening E-mail: buening-at-geology.ucdavis.edu Department of Geology Phone: (916) 752-0350 University of California Fax: 916 752-0951 Davis, CA 95616
Julian, I didn't expect the reply so quick. I certainly try that. Thanks so much. Regarding serial sectioning, you didn't mention what kind of knife you used for GMA sectioning. Is tungsten or glass knife? Which is better? Regular Oil Red O staining procedure won't work with GMA section. Why? The reason we choosing GMA is that the morphology is better than frozen tissue. Do you have suggestion about this? Dorothy
We recoat screens in our lab. Please contact us for pricing.
Dianne Ernest F. Fullam, Inc.
O.k., I know this has been discussed MANY times before, but I I } didn't save all the relavent messages. } } Looking for two things: } } (1) Refurbished / replacement screen for a JEOL 100-S (hey it still } works fine) } } (2) Looking for places to recoat our old 100s screen. I believe } that SPI still does it, and will call soon (I did save your message } Chuck). Grant Sci Corp doesn't seem to exist any more. Any others } to consider? Any comments god or bad? } } Thanks. } } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } } "640K ought to be enough for anybody." } -- Bill Gates, 1981 } } Ernest F. Fullam, Inc. Phone: (518) 785-5533 FAX: (518) 785-8647 E-Mail: pdf-at-fullam.com
I apologize to Grant Scientific Corp for passing apparent misinformation, for indeed Grant Scientific does exist (as it has since 1975 I have been informed). I have spoken with the good folks at Grant, and they have been able to provide me some very useful information regarding TEM Screen re-coating.
I wish to make it all known to the listserver goupr that Grant does exist and can be contacted at: 803-829-2841.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
I am replying to my subscription request. All information is correct.
Nancy Buening
Nancy Buening E-mail: buening-at-geology.ucdavis.edu Department of Geology Phone: (916) 752-0350 University of California Fax: 916 752-0951 Davis, CA 95616
} We inherited an old embedding oven that is caked in resin. Does anyone } ever clean out their embedding oven? If so, what is used to do this?
I used to chisel away at the stuff, and try to use various solvents to clean off the glass door without dissolving my gloves, with limited success. One day I came into the lab and, lo! the oven was clean. Turns out someone had cleaned it out with our putty knife WHILE IT WAS HOT! Duh. She said it was pretty easy. Be careful of all surfaces. Wear gloves. Worry about toxicity. Disregard me if anyone says this is not recommended.
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* ****************************************************************************
} } We inherited an old embedding oven that is caked in resin. Does anyone } ever clean out their embedding oven? If so, what is used to do this? }
I am not sure what kind of resin is involved, but once I had to clean out a 5 litre three-neck flask that had been used to synthesize alkyd resins - these are basically phthalic polyesters crosslinked through polyunsaturated fatty acids. There was a caked-on film that would not yield to concentrated sulphuric acid, tetrahydrofuran, or any of the usual things. I put some .880 Ammonia in the bottom of the flask, blocked the necks with cotton wool, and left it overnight, and next morning the film had swollen and fallen away and could be yanked out with tongs or whatever.
Ammonia vapour is very effective with polyester based resins because of (a) it basic nature and (b) most important, its small molar volume.
If, on the other hand, your resin is an epoxy, it might be better to put a dish of methylene chloride (dichloromethane) in the bottom, seal the oven, and go away overnight. Methylene chloride is the basis of most commercial paint strippers.
The use of vapour technique does make for much less messy operation. Once the film is loosened, strong detergent should be good enough for scrubbing.
However, that PLASTIC PETRI DISH would probably turn into a gooey mess with the methylene chloride vapour, so try the ammonia first. (Also consider, does your oven seal with an O-ring?)
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I have a procedure for Oil Red O staining of GMA sections which was sent to me by Bob Schoonhoven. I haven't tried it myself.
Stain: Oil Red O Indication: Demonstrate lipids Solutions: 1) 60% aqueous triethyl phosphate 2) 0.5% Oil Red O solution 0.5g Oil Red O (CI 26125) 100 ml 60% aqueous triethyl phosphate Filter before use 3) Celestin Blue 0.5g celestin blue B 100 ml 5% aqueous ferric ammonium sulfate Boil gently 2-3 minutes; cool to room temperature; filter; and add 12 ml glycerol Filter before use Procedure: 1) Rinse briefly in 60% triethyl phosphate 2) Stain 5-20 minutes in Oil Red O solution 3) Rinse 1-2 seconds in 60% triethyl phosphate 4) Rinse well in distilled water 5) Counterstain in Celestin Blue 15 minutes 6) Rinse well in distilled water 7) Mount in glycerin jelly or other water-soluble mount Results: lipids: red-orange nuclei: blue Reference: Feldman, A.T. and Dapson, R.W., "Relative Effectiveness of Various Solvents for Oil Red O," _Medical Laboratory Technology_, Vol. 31:335-341, 1974.
Disclaimer: Energy Beam Sciences manufactures the JB-4 and JB-4A microtomes for sevtioning plastic-embedded tissue, and sells GMA kits.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
The vignetting I most frequently see is a cut-off of the image at the corners due to the microscope's camera tube. This can commonly be caused by using an adapter (for example, a C-mount adapter for the digital or video microscope camera) with too wide a field of view. The descriptions you cite in the Japanese text could be describing this or other phenomena. You'd need to know more from the original author(s) to be sure. Brooks Corl Senior Applications Manager, Polaroid Corporation E-mail: corlb-at-polaroid.com
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In a message dated 97-09-12 08:45:19 EDT, Chiba writes: { { sentences that go like "part of the field of view may be cut off" and "image cut-off in peripheral part may occur." Could anyone tell me how to put them correctly? Should I instead say "the field of view may be vignetted" and "image vignetting may occur" which sound more technical to me but I'm not sure about. } }
The phrase "part of the field of view may be cut off" sounds ok to me. Without an illustration, I do not know what it means. My mental image is that there is a black part within a photographic image taken with the microscope from a flash/shutter timing problem, or from blocking of the field of view by an accessory that is inserted in the optical path, such as the quarter wavelength plate typically used in a polarizing microscope.
The phrase "image cut-off in peripheral part may occur." could be reworded as "...the peripheral part of the image may be cut-off." As above, I do no know really what it means. But I imagine that it means the effect from closing down the sub-stage iris. I do not think that is really vingetting, a gradual shading, but it is close to it. If the effect being discussed is from gross misalignment of the bulb in the illuminator housing, then it could be vignetting.
Steve Stokowski Stone Products Consultants Concrete Petrographers http://members.aol.com/CrushStone/index.htm ---------------------------------- Forwarded ----------------------------------
Are you into digital? Please help to reevaluate digital image processing for the development of a program for next year's MSA-98 meeting (Follow-up and Summary will be placed here):
"Applied Image Processing: What Can It Do For Digital Imaging"
If you like to help, type your reply directly into this message and send it to me at
Klaus-Ruediger Peters {Peters-at-bsac.uchc.edu}
Thanks for your help and voicing your opinion. Klaus ****************************************************************************
What is important, what are you using, what deserves more attention, what has "practical value" and may represent one of the topics? What is of your interest? What can't you do but like to? Please, hit "REPLY" now, then "PASTE" my address into "TO:" and just fill in your Spontaneous thoughts on:
-----------------------------------Being Digital--------------------------- Digital image handling (Shifting data, formatting, labeling, annotating, 8-bit to 16-bit/per channel)
----------------------------------Trying Digital----------------------------- Digital darkroom (Printing images as hard copies on paper, overheads and slides)
----------------------------------Doing Digital----------------------------- Digital image display and "enhancement" (Presenting the desired information)
---------------------------------Using Digital------------------------------ Data quantification (Finding and measuring the desired information)
----------------------------Other topic of interest?------------------------
Images are as diverse as their usages. But, what are the common concepts and how good do they apply to the common daily imaging tasks? Any possible contributions from yourself to this MSA 1998 program?
Thanks for your initiative and for your help. Klaus
The Minnesota Microscopy Society FALL BUFFET DINNER & TALK will take place Tommorrow, SEPTEMBER 18, 1997 from 5:30 - 8:00 PM at The Campus Club, University of Minnesota Minneapolis, East Bank Campus
SPEAKER: William P. Wergin Agricultural Research Service,U.S. Dept. of Agriculture, Beltsville, MD
TOPIC: "THE MICROSCOPY OF SNOW:" "The 3-D Structure and Metamorphoses of Snow and Ice Crystals as Revealed by Low Temperature SEM". For more details see our website at http://resolution.umn.edu/MMS/
We hope to provide a pleasant evening during which microscopists will be moved to renew or begin memberships in MMS, MSA and/or MAS.
Program 5:30-6:00 Wine, Cider & Cheese Social 6:00-7:00 Buffet Dinner. 7:00-8:00 Talk The Buffet Dinner is $12 per MMS member, payable at the door. (non-member fee is $22, includes new membership, to attend without becoming member, $15.) STUDENTS: NEW FEATURE THIS YEAR: Current student members or new student members - $5.00 membership fee payable at the door- will receive a complimentary buffet dinner courtesy of MMS and sponsoring vendors.
Please make an advance reservation by contacting:
Mike Coscio (612)569-1331, 569-1284 FAX, mike.coscio-at-medtronic.com Stuart McKernan, (612)626-7942, 626-7530 FAX, stuartm-at-maroon.tc.umn.edu
Parking is available behind the Union in the East River Road Ramp (connected by walkway to Union) for $2.50 (per day rate), at the Radisson Ramp on Washington Ave. S.E., a block east of the Union, and at other Minneapolis Campus locations
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
A collegue contacted me asking for information on labeling of a small protein in potato tuber. Apparently, she is having problems with the potato starch disintegrating in the electron beam. Do any of you have any suggestions about how to retain antigenicity and, at the same time, attain good (or reasonable) fixation. If so, please contact her offline at the email address below as she is not yet a member of this list.
Her name is Bonnie Compas: email address is: becompas-at-csupomona.edu
Thank you,
De Wood
*****************************************
Delilah F. Wood United States Department of Agriculture Western Regional Research Center 800 Buchanan Street Albany, CA 94710
} Did anybody know the size of large and small viewing screen of CM120 } and CM200/FEG. Your information will be greatly appreciated! } gary } } } +--------------------------------------------------------------------+ } | Gary Gang REN, Ph.D. | } | Department of Cell Biology #MB219 Email: gangren-at-scripps.edu | } | Mail Stop MB21 Tel: (619) 784 9815 (O) | } | The Scripps Research Institute (619) 546 1585 (H) | } | 10550 N. Torrey Pines Road Fax: (619) 784 9927 | } | La Jolla, CA 92037, USA http://leonardo.scripps.edu/ren | } +--------------------------------------------------------------------+ } } }
I am looking for a person to fill a 1 year surface chemist position at a petroleum company in Beacon, NY (Dutchess County). The person must have Ultra High Vacuum, X-Ray Spectoscopy (XPS) and Scanning Electron Microscopy (SEM)or AES experience. The chemist will be doing sample prep and intro, analysis, and running data. It is an applied research, product related position. Please e-mail me at kwagscha-at-aerotek.com or call 1-800-973-1518 ext. 1045 if interested. Kristen Wagschall
I am posting this for a friend who is not on the listserver. Please reply=
directly to Trinity College.
Thank you!
David Henriks South Bay Technology, Inc.
************************ PLEASE POST ************************
Department of Human Resources Trinity College Hartford, Connecticut 06106
Position Announcement
Electron Microscopy Laboratory Manager/ Technician
Trinity College has an immediate opening for an experienced electron microscopist to work in a newly established Electron Microscopy Center. A= s this facility is designed to serve both the physical and biological sciences, familiarity with both disciplines is highly preferred. Primary responsibilities include assisting faculty and students in the use of the=
facility for teaching and research, and overseeing the upkeep and use of TEM, STEM and specimen preparation facilities, including the analytical E= M EDX and EELS). A B.S. degree with advanced research training and/or extensive experience with TEM are essential; master's degree with requisi= te experience is preferred.
This position is a full-time, 12-month appointment with full College benefits. Applications will be reviewed upon receipt; search will continu= e until position is filled. Please respond with a resume, cover letter stating salary expectatons, and the names, telephone numbers and addresses of three professional references to: Prof. Daniel Blackburn, c= /o Human Resources, 300 Summit Street, Trinity College, Hartford, CT 06106.=
(Resumes also may be faxed: (860) 297-5140, or sent via the internet: Sandra.Magee-at-Mail.Trincoll.edu).
Trinity College is an Equal Opportunity/Affirmative Action Employer. Wome= n and mminorities are encouraged to apply. Applicants with disabilities should request any needed accommodation in order to participate in the application process.
The Duniway Stockroom Corp. ((800-446-8811) deals in new, used, and rebuilt vacuum devices, including all types of vacuum gauges. They might be able to help you with a replacement gauge, or possibly they could rebuild yours.
Good luck,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
I have received a couple of inquiries about what Torr Seal is and where to get it.
Torr Seasl is an epoxy compound especially formulated for use in vacuum systems that was introduced by Varian Associates, Vacuum Products Division, 121 Hartwell Ave, Lexington, MA 02173-3133 (800-882-7426) a number of years ago. It is stated to contain no solvents, and therefore to be good at pressures below 10-9Torr. It is bakeable at temperarures up to 120C. It adheres to most clean materials (glass, metals, ceramics), and holds up well over long term service. Some companies that handle EM supplies also handle it (e.g. I find it listed in the Ladd catalog, probably SPI handles it too).
Other similar products are also sold by other companies. For example, Duniway Stockroom Corp. (800-446-8811; info-at-duniway.com) sells a product called 'Epoxy Patch' that is stated to be equivalent to Torr seal
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
Dear Microscopists, I am forwarding this message from SAFETY listserver. Marek Malecki.
On Sep 17, 8:56am, Dr. Gary Gang REN wrote: } Subject: screen size } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Did anybody know the size of large and small viewing screen of CM120 } } and CM200/FEG. Your information will be greatly appreciated! } } gary } } } } } } +--------------------------------------------------------------------+ } } | Gary Gang REN, Ph.D. | } } | Department of Cell Biology #MB219 Email: gangren-at-scripps.edu | } } | Mail Stop MB21 Tel: (619) 784 9815 (O) | } } | The Scripps Research Institute (619) 546 1585 (H) | } } | 10550 N. Torrey Pines Road Fax: (619) 784 9927 | } } | La Jolla, CA 92037, USA http://leonardo.scripps.edu/ren | } } +--------------------------------------------------------------------+ } } } } } } } } } -- End of excerpt from Dr. Gary Gang REN
You can measure it quite accurately by using the measuring function of the CM scope. Just move any thing you can identify from one side of the screen to the other side of the screen.
Yifan Cheng
-- ********************************************************************** * Dr. Yifan Cheng * Phone: +1-850-644-4104 (Office)* * Institute of Molecular Biophysics * or: +1-850-644-9769 (Lab) * * Florida State University * Fax: +1-850-561-1406 * * Tallahassee, FL 32306-4380 * Email: ycheng-at-sb.fsu.edu * * U.S.A. * http://www.sb.fsu.edu/~ycheng * **********************************************************************
Dear Microscopists, To evaluate effects of various cryoprotectants on the cells' viability, we would like to image cultured cells being frozen. Is anybody aware of a cryo-stage (below -100deg.C) for light microscopy? Any information will be greatly appreciated. Vendors welcome. Sincerely, Marek Malecki.
FOR SALE: Brand New 6.2 mm Dupont diamond knife. Never used. Current retail for a knife of this length is approximately $6000+. I would like to sell the knife for $3200 or best offer. Ron Kalil
Ronald Kalil Center for Neuroscience University of Wisconsin 1300 University Ave. Madison, WI 53706
Has anyone ever heard of something called the "Arkograf electrical metal etching pen?" Or more importantly, the name of the manufacturer and where they are located? It is a device for making permanent markings on metal surfaces for identification purposes.
Thanks.
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 ############################ ==================================================
We have an Electric Discharge Machine Mark III, Ser 338854; It came originally from Concept EDM Ltd, Maidenhead, Berkshire England. I checked the UK yellow pages but they seem no longer to exist.
Our problem is that the Electric Discharge Machine won't work and we would like the circuit diagrams (schematics) before trying to fix it.
If anyone out there has knowledge of the current address etc. of the original suppliers, OR has a circuit diagram they could copy for us, we would be most grateful.
Thanks,
Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
My belated thanks for responding to my query into "vignetting" I posted on the Microscopy list. Your comment was very helpful and enable me to get the job done.
Thank you very much again.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
My belated thanks for responding to my query into "vignetting" I posted on the Microscopy list. Your comment was very helpful and enable me to get the job done.
Thank you very much again.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
My belated thanks for responding to my query into "vignetting" I posted on the Microscopy list. Your comment was very helpful and enable me to get the job done.
Thank you very much again.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
My belated thanks for responding to my query into "vignetting" I posted on the Microscopy list. Your comment was very helpful and enable me to get the job done.
Thank you very much again.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
} Dear Microscopists, } I am forwarding this message from SAFETY listserver. } Marek Malecki. } } Date: Wed, 17 Sep 1997 13:25:29 -0400 } From: "Ralph Stuart, Vermont SIRI" {rstuart-at-esf.uvm.edu} } } A Tiny Drop Of Mercury Shatters Lives And Science } } A9 1997 The Associated Press } } LYME, N.H. (September 13, 1997 6:45 p.m. EDT) - } snips
} "She loved her work," he says. "It made her happy." } } She couldn't have known the risks. She couldn't have known how bad the bad } stuff really was. Truth is, no one knew. } } Just a tiny drop of liquid. Sweet-smelling. Dense. Deadly. } } By HELEN O'NEILL, The Associated Press
While not wishing to diminish the dangers of mercury, I really don't see that this sort of journalistic hype helps anybody. It makes a good story, I suppose, for the sort of newspaper that runs 'Elvis Lives' stories, but it isn't science and even the courts regard hearsay as highly unreliable.
} Dear Microscopists, } I am forwarding this message from SAFETY listserver. } Marek Malecki. } } Date: Wed, 17 Sep 1997 13:25:29 -0400 } From: "Ralph Stuart, Vermont SIRI" {rstuart-at-esf.uvm.edu} } } A Tiny Drop Of Mercury Shatters Lives And Science } } A9 1997 The Associated Press } } LYME, N.H. (September 13, 1997 6:45 p.m. EDT) - } snips
} "She loved her work," he says. "It made her happy." } } She couldn't have known the risks. She couldn't have known how bad the bad } stuff really was. Truth is, no one knew. } } Just a tiny drop of liquid. Sweet-smelling. Dense. Deadly. } } By HELEN O'NEILL, The Associated Press
While not wishing to diminish the dangers of mercury, I really don't see that this sort of journalistic hype helps anybody. It makes a good story, I suppose, for the sort of newspaper that runs 'Elvis Lives' stories, but it isn't science and even the courts regard hearsay as highly unreliable.
Terribly embarrassed... Please accept my apology for the 5 or 6 private thank-yous I just posted. I just kept hitting reply without realizing it was a mailing list.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
The procedure that Steven posted does indeed work very nicly.....Provided that no alcohol was used in the "dehydration" process. If you wish to look for lipids in GMA embedded tissues you must use a series of graded (with water) monomer solutions instead of alcohols. Somewhere ????? I have the procedure written but I can't put my hands on it right now (ie: I haven't got a clue as to where I 'filed' it). :(
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
My thoughts exactly, see the August? issue of Scientific American for a better tribute to Karen Wetterhahn and a less subjective discussion of the risks associated with chemical handling.
} } While not wishing to diminish the dangers of mercury, I really don't see } that this sort of journalistic hype helps anybody. It makes a good story, I } suppose, for the sort of newspaper that runs 'Elvis Lives' stories, but it } isn't science and even the courts regard hearsay as highly unreliable. } } Regards, } } Larry Stoter
================= C. John Runions Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
Evex Analytical, manufacture of the VIDX X-ray Analyzer and the VIDX = Scan Digital Imaging system for SEM/STEM and TEM, would like your feed = back.
We are now engineering Version 3.0 of our VIDX X-ray analyzer software. = We would like to ask you for your wish list. Your wish list may include = functions, routines, macros, etc. which you would like to see = implemented in your ideal x-ray analyzer, This inquiry is open to every = one.
Please respond via e-mail. You may foward drawing also.
Thank you
Peter Tarquinio Evex Analytical www.evex.com 609-252-9192 Tel 609-252-9091 Fax
Linkam make a very nice heating/cooling stage for light micrscopes with a temperature range of -196 to 600 C. They are distributed locally by Fryer Co. Phone 847-669-2000.
Regards,
Joe Neilly Abbott Laboratories Microscopy and Microanalysis 200 Abbott Park Rd. Abbott Park, IL 60064
I am posting this message for a friend who would like to have the e-mail address of Bill McManus, Utah State Univ., Dept . Biology, Logan, Utah. Please reply to :
Kalabm-at-em.agr.ca
Ann Fook Yang EM Unit, ECORC, Agriculture and Agri-Food Canada, Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
If anyone is over in UK at the end of February please bear in mind the fol= lowing meeting....
************************************************************* Microscopy of Internal Interfaces: Determination of= Nanochemistry
Institute of Physic= s, 76 Portland Place, London W1N 4AA,= UK.
Friday 27 February 1998=
10.00am - 5.30pm
Organizer: Dr Rik Brydson, School of Process, Environmental and Materials = Engineering, University of Leeds, Leeds LS2 9JT, UK.
EMAG Committee of Institute of Physics Joint with Royal Microscopical Society: Materials Section.
The chemistry of internal interfaces in both structural and functional mat= erials is often a critical factor affecting resultant physical properties, sometimes overrid= ing or controlling the effects of interface structure. The majority of interfaces have a spatial = extent, in at least one dimension, of a few nanometres. Clearly it is desirable to be able to dete= rmine accurately interfacial chemistry and bonding. With this in mind, a one day IOP meetin= g will be held in London at the end of February 1998 on the techniques for interfacial micro= analysis and relevant case studies in this rapidly expanding field. The day will be div= ided into fundmentals and applications sessions.
Confirmed Invited Speakers: oProf. Mick Brown (Cambridge) "Grain boundary chemistry in metals and all= oys" (Sponsored by RMS). o Dr Rik Brydson (Leeds) "Interfacial bonding determined by EELS" o Dr David Jefferson (Cambridge) "Determination of surface chemistry using= HREM" o Prof. John Titchmarsh (Sheffield Hallam) "Determining Interfacial Segreg= ation using EDX" o Dr John Watts (Surrey) "Surface Analysis applied to Interfaces" o Prof Bruce Hamilton/ Dr Uschi Bangert (UMIST) "STM/STEM of cleaved semic= onductor multilayers" o Dr Paul J Warren (Oxford) "Atom Probe Field Ion Microscopy of Internal I= nterfaces".
Other topics will include: o Interfacial Chemistry and HREM o STM on cross-sections o Theoretical aspects of interface chemistry o Metal-support interactions in Supported Catalysts o Semiconductor interfaces o Biomaterial interfaces o Coatings o Corrosion Films
Additional POSTER contributions are extremely welcome.
For further details about registration etc. please contact IOP conference desk Tel: 0171 470 4800 Fax: 0171 470 4848 Email: physics-at-i= op.org or Rik Brydson Tel: 0113 233 2369 Fax: 0113 242 2531 Email: mtlrmdb-at-leeds.ac= .uk _____________________________ Dr. Rik Brydson, University Research Fellow, Electron Optical Unit, Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
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As part of the group involved in the postmortem examination of the death of Karen Wetterhahn I was grateful for all the information and warnings I could lay my hands on before I dealt with any material. The event ,however reported, serves as a reminder to all of us of the dangers of our materials and the need to keep up with the information involved in our safety. This institution has completely reexamined it's glove policy.
Hey Larry- how many electron microscopists have you met who's hand trembles and shakes as they reach out to shake your hand, or dumps the food from their fork or plate as they struggle to eat lunch? I believe they, their family and friends don't look at this as trash journalism, we should all remember SAFETY FIRST. -Mike Rock
On Thu, 18 Sep 1997, Larry Stoter wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear Microscopists, } } I am forwarding this message from SAFETY listserver. } } Marek Malecki. } } } } Date: Wed, 17 Sep 1997 13:25:29 -0400 } } From: "Ralph Stuart, Vermont SIRI" {rstuart-at-esf.uvm.edu} } } } } A Tiny Drop Of Mercury Shatters Lives And Science } } } } A9 1997 The Associated Press } } } } LYME, N.H. (September 13, 1997 6:45 p.m. EDT) - } } } snips } } } "She loved her work," he says. "It made her happy." } } } } She couldn't have known the risks. She couldn't have known how bad the bad } } stuff really was. Truth is, no one knew. } } } } Just a tiny drop of liquid. Sweet-smelling. Dense. Deadly. } } } } By HELEN O'NEILL, The Associated Press } } While not wishing to diminish the dangers of mercury, I really don't see } that this sort of journalistic hype helps anybody. It makes a good story, I } suppose, for the sort of newspaper that runs 'Elvis Lives' stories, but it } isn't science and even the courts regard hearsay as highly unreliable. } } Regards, } } Larry Stoter } } }
I have recently bought several different types of bulbs from Lamp Technology (Bohemia, NY) at 800-533-7548. There prices are a lot better that anything I have ever found through a microscope dealer. One of my dealers told me that I was getting a better price than he was wholesale. I recommend you give them a call. They have a web page but I can't remember the address - use a search engine if you are interested. I have no interest in the company except as a very happy user.
} Does anyone know of a source for the HBO 100W/2 100 watt bulbs } used for fluorescence light microscopy? } } Thanks, } } Dennis Shubitowski } University of Michigan } School of Dentistry } dshubito-at-umich.edu
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
They were rated a favorite last year by Beth Richardson (UGA) who was on a similar quest.
(No affiliation blah, blah, blah.....)
cheers ed
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 http://www.personal.psu.edu/ejb11/
Advanced MicroBeam, Inc. would like to announce its new web presence to t= he microscopy community. We can be found at http://www.advancedmicrobeam.co= m. Please take a look at how we may be of service to you.
Thanks Dave
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D David A. Stanford Advanced MicroBeam, Inc. 4217C King Graves Rd PO Box 610 Vienna OH 44473-0610
-------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com The Web is:"Vaster than empires and more slow" Andrew Marvell (1621-1678)
Without in any way downplaying the tragedy for the people involved, the dose-response relationship postulated in the story is not very plausible. Mercury toxicity has been studied extensively for at least half a century. It is a cumulative poison which does cause the type of neurological effects reported, but a "single tiny drop", through a glove and quickly washed off, being lethal seems suspect. Associated Press is not peer reviewed (although maybe it depends on your definition of "peer").
In 1996, the EPA published a 7-volume report on environmental mercury. For some data from recent animal studies (rodent and primate) and the current EPA and FDA reference exposure limits, try the following Web pages.
An excellent sample to test resolution at high tilt angles is a single crystal silicon wafer. If mounted as a cross-sectional sample (electron beam parallel to a {011} zone axis), there are several advantages. In th= is initial orientation, the (111) lattice spacings of 3.14 A, (200) spacings=
of 2.715A and (220) spacings of 1.92A are all available for resolution tests. Being a cubic single crystal, it is a simple procedure to follow t= he Kikuchi bands to any other cubic zone axis for the additional resolution checks you mentioned.The resolution at high tilt angles can be checked by=
mounting the cross-sectional silicon sample with the sample's glue line either parallel or perpendicular to the sample holder rod. With the glue=
line parallel to the sample holder rod, {100} zone axes are accessible by=
rotating the sample holder 45 degrees in either direction. The {100} zone=
includes (220) spacings (200) spacings for resolution tests. Similarly, = if the sample is mounted with the glue line perpendicular to the sample hold= er rod, the second tilt attachment can be used to tilt the sample 45 degrees=
to the {100} zone axis in either direction about the second tilt axis. = If your goniometer has a limited tilt capability, there are many other choce= s of zones at lower (and higher) angles for you to choose from.
While you are performing these resolution tests, there are several other calibrations that are easy to perform and can be done with the same sampl= e. For example, the complete magnification range of the TEM (the supplied values from the manufacturer can be 5-10% off), the camera constant calibration, and the imge/diffraction patter rotation calibration. All o= f these tests and calibrations are extensively documented for the MAG*I*CAL=
calibration sample, which is a single crystal of silicon upon which are precisely grown calibration marks. =
PLEASE NOTE: South Bay Technology, Inc. does offer the MAG*I*CAL Calibration Standard and so we have a finiancial interest in promoting it= s use. For more inforamtion on the MAG*I*CAL, please contact me off-line.
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. I have a couple of questions and will appreciate any replies and =
suggestions.
1) What is the best sample to test the resolution of a TEM at a tilt angle (} 30degree)?
I am testing the resolution of a CM300FEG microscope. I used oriented gold, evaperated gold particles, graphitized carbon and so on. There is no =
problem of those samples at zero tilt. But after tilt the specimen to 30 degree or =
higher, I got only the good resolution (2.3A in case of evaperated gold particle) =
in one direction, along the tilt axis. I believed it is not because of the =
focus gradient that I could not get the same resolution in both directions at =
this tilt angle. I think the samples (evaporated gold particles and graphitized carbon) may not be the suitable samples for the resolution test of the high tilt angles (higher than 45 degree and up to 60 degree). I wonder if any of you has done such test before and has any suggestions on which specimen should =
be used in this test and where to get it?
2) Is there any published documentation about the line resolution of Kodak SO-163 film?
I am writing an paper about the performance of the CM300FEG at low magnificaiton, and need to know the line resolution of Kodak SO-163 =
(something I can cite for). But I could not find any published documentation. Of =
course, I called Kodak technique support and of course they didn't give me a clue. There is a web site called "Bibliography on EM Imaging and Related Technologies" (http://www.uct.ac.za/depts/emu/imaging/papers.htm), but I can not find any thing there either about SO-163. The only other related information I got =
was from a similar discussion in this board a year ago. But the data about line resolution found in that discussion were mostly estimated but not from any published documentation. I wonder if any of you know there is any kind of published documentation including research paper, technique report or so which gives the line resolution of Kodak SO-163. (BTW, the topic about the line resolution has been discussed a year ago and I am sorry to ask it again.)
I appreciate any information or suggestion on my request.
Yifan Cheng
BTW, I am not sure if my email address is already on the list or not. So that I appreciate also that when you reply this email, send a reply to me directly as well as to the board. Thanks again.
-- ********************************************************************** * Dr. Yifan Cheng * Phone: +1-850-644-4104 (Office)* * Institute of Molecular Biophysics * or: +1-850-644-9769 (Lab) * * Florida State University * Fax: +1-850-561-1406 * * Tallahassee, FL 32306-4380 * Email: ycheng-at-sb.fsu.edu * * U.S.A. * http://www.sb.fsu.edu/~ycheng * ********************************************************************** =
Dear reader, I tried to detect mRNA and protien in the same slide. My insitu hybridyzation and immunohistochemistry both worked. but not working when I put them together. I tried insitu first then IHC. Is there anyone tell me some tricks about it? Thanks. Dorothy
Kate, would you share with us the outcome of your institution's re-examination of its glove policy? Thanks. /Vachik Hacopian
} As part of the group involved in the postmortem examination of the death of } Karen Wetterhahn I was grateful for all the information and warnings I could } lay my hands on before I dealt with any material. The event ,however reported, } serves as a reminder to all of us of the dangers of our materials and the } need to keep up with the information involved in our safety. } This institution has completely reexamined it's glove policy. } } Kate Connolly
Dorothy, You may want to visit the WWW site by Dr. Gwen Childs. It contains the detailed hands on protocols whuch you need and the excellent examples of labelings:ISH and IL. http://cellbio.utmb.edu/childs/childs.htm
Sincerely, Marek Malecki.
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************************ PLEASE POST ************************
Department of Human Resources Trinity College Hartford, Connecticut 06106
Position Announcement
Electron Microscopy Laboratory Manager/ Technician
Trinity College has an immediate opening for an experienced electron microscopist to work in a newly established Electron Microscopy Center. As this facility is designed to serve both the physical and biological sciences, familiarity with both disciplines is highly preferred. Primary responsibilities include assisting faculty and students in the use of the facility for teaching and research, and overseeing the upkeep and use of TEM, STEM and specimen preparation facilities, including the analytical EM EDX and EELS). A B.S. degree with advanced research training and/or extensive experience with TEM are essential; master's degree with requisite experience is preferred.
This position is a full-time, 12-month appointment with full College benefits. Applications will be reviewed upon receipt; search will continue until position is filled. Please respond with a resume, cover letter stating salary expectatons, and the names, telephone numbers and addresses of three professional references to: Prof. Daniel Blackburn, c/o Human Resources, 300 Summit Street, Trinity College, Hartford, CT 06106. (Resumes also may be faxed: (860) 297-5140, or sent via the internet: Sandra.Magee-at-Mail.Trincoll.edu).
Trinity College is an Equal Opportunity/Affirmative Action Employer. Women and minorities are encouraged to apply. Applicants with disabilities should request any needed accommodation in order to participate in the application process.
*****************
Christine Caragianis Broadbridge, Assi. Prof. Trinity College, Department of Engineering 300 Summit Steet Hartford, CT 06106-3100
Forwarded Email please reply direclty to fischg98-at-providence.edu
Below is the result of your feedback form. It was submitted by (fischg98-at-providence.edu) on Tuesday, September 16, 1997 at 11:34:39 ---------------------------------------------------------------------------
Email: fischg98-at-providence.edu Name: Gia Fischetti
School: Providence College
State: RI
Zip: 02918
Question: I am currently doing research on Leishmania enrietti, and I am now tring to prepare specimines for our TEM. I have been unsuccessful at finding a "recipe" for the preparation of this organism. I am reluctant to just try Karnovsky's fixative, and would appreciate any help that you could give me on this matter. Thank you very much. I hope to hear from you soon. Gia Fischetti
If you read the obituary etc., she was clearly quite a scientist. A moral here is check that your gloves are good for what you are using, we have toxicologists/occasional electron microscopists using tributyl tin which is also not nice.
Rick, While the AP is not peer reviewed Science is. Suggest that you check out the August issue. Karen was a collaberator with several people in our lab. Also suggest that you look up "dimethylmercury". One drop (50ul) is equal to about 300X the occupational exposure limit. -- Begin original message --
} } } Without in any way downplaying the tragedy for the people involved, the } dose-response relationship postulated in the story is not very plausible. } Mercury toxicity has been studied extensively for at least half a century. } It is a cumulative poison which does cause the type of neurological effects } reported, but a "single tiny drop", through a glove and quickly washed off, } being lethal seems suspect. Associated Press is not peer reviewed (although } maybe it depends on your definition of "peer"). }
} Rick Mott } rick-at-pgt.com } } (These are personal views having nothing to do with my employer) }
-- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
I'm doing automated particle analysis (SEM/EDX) of suspended particulate material from drinking water reservoirs. Particles are filtered onto nuclepore membranes and carbon coated. Features are located using a thresholded bse image. Diatoms (some species) are so thin that that they don't image very well. Does anybody know of a 'staining' method to increase the atomic number contrast for amorphous silica?
reply to jptmvl-at-mailbox.syr.edu thanx, dave johnson
I would appreciate any information about the software QUANTITEM for quantitative determination of interfacial roughness and composition mappings in HREM images. Thanks in advance.
F. Peiro *******************************+ Francesca Peiro
EME, Enginyeria i Materials Electronics Dpt. Fisica Aplicada i Electronica Universitat de Barcelona Avda. Diagonal 645-647 08028 Barcelona, Spain
The story of the death of Dartmouth College chemistry professor Karen Wetterhahn is unfortunately very true and not greatly exaggerated. She was using very small amounts of dimethylmercury as an NMR standard. The amount reported is "one to a few drops". This is not a large exposure and the exposure occured only once. She was reported to be a very careful chemist. The latex gloves she was wearing did not stop the absorption of the dimethylmercury through her skin. Treatment for mercury poisoning was unsuccessful in saving her life.
Everyone needs to minimize their exposure to toxic materials and microscopists are well aware of this as is noted in this listserver. The composition of gloves worn in the laboratory is very important and each application should be evaluated according to need. It serves no purpose to downplay the potential for tragedy in the lab from toxic materials. Remember also that the greatest potential for problems is the long term exposure to small amounts of materials that the lab person has become accustomed to using - familiarity breeds contempt. Two good articles about Karen Wetterhahn appear in: Chemical and Engineering News, June 16, 1997, p. 11, and Scientific American, September, 1997, p. 20.
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The Dartmouth College Biosafety Office in combination with OSHA has produced a comprehensive study /report on the characteristics and abilities of available gloves. On a smaller scale , anyday, I expect a poster for all the labs summarizing the chief points of this work. Anyone wishing information on gloves should contact Michael Blayney at: Michael.Blayney-at-dartmouth.edu
I cannot believe this discussion. No doubt mercury is a nasty cumulative toxin. But blaming a faulty glove and a single drop passing onto the skin is pure nonsense. In the good ol days I cleaned a number of times mercury diffusion pumps and purified the mercury by shaking the stuff with nitric acid and solvents in a separating funnel. No gloves! I have no trace of Minnemata disease, steadier hands than most people and I am "approximately normal". I knew a man who had active mercury poisoning. This was contracted in a large but closed room after a large industrial thermometer fractured on a very hot oven. The man inhaled fumes for several hours. Mercury poisoning most frequently is caused by ingestion and inhalation. I do not advocate bathing in mercury, but that story I take with a drop of something else! 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, 400+ Links, MSDS ************************ http://www.proscitech.com.au } } Kate, would you share with us the outcome of your institution's } re-examination of its glove policy? Thanks. /Vachik Hacopian } } } } } As part of the group involved in the postmortem examination of the death of } } Karen Wetterhahn I was grateful for all the information and warnings I could } } lay my hands on before I dealt with any material. The event ,however reported, } } serves as a reminder to all of us of the dangers of our materials and the } } need to keep up with the information involved in our safety. } } This institution has completely reexamined it's glove policy. } } } } Kate Connolly
If I recall correctly, the compound in question was dimethyl mercury, which turned out to pass quickly through her gloves, in contrast to metallic mercury.
} Without in any way downplaying the tragedy for the people involved, the } dose-response relationship postulated in the story is not very plausible. } Mercury toxicity has been studied extensively for at least half a century. } It is a cumulative poison which does cause the type of neurological effects } reported, but a "single tiny drop", through a glove and quickly washed off, } being lethal seems suspect. Associated Press is not peer reviewed (although } maybe it depends on your definition of "peer").
Scott Schwinge Friday Harbor Labs University of Washington
Can we move the Hg discussion to www.Hgtox.com, the gloves discussion to the handwear listserv, and get on to death of more important people (Di)?
Seriously, Prof. Wetterhahn and Di were interesting individuals who have received their eulogies in other more appropriate forums. Whether one drop of diMeHg causes death is debatable, but not here (are any of you microscopists using it?). Gloves get holes and have unsealed wrists (usually). Enough OK? Rob Palmer CEB/UT
If you can still excite the x-ray lines of interest, try lowering the incident beam potential. Woody ______________________________ Reply Separator _________________________________
I'm doing automated particle analysis (SEM/EDX) of suspended particulate material from drinking water reservoirs. Particles are filtered onto nuclepore membranes and carbon coated. Features are located using a thresholded bse image. Diatoms (some species) are so thin that that they don't image very well. Does anybody know of a 'staining' method to increase the atomic number contrast for amorphous silica?
reply to jptmvl-at-mailbox.syr.edu thanx, dave johnson
I get my Osram HBO 100W/2 for a price of $128.00 from
Elkay P.O. Box 1105 La Jolla CA 92038-1105
Larry Kuritzky 619-454-5742
} Does anyone know of a source for the HBO 100W/2 100 watt bulbs } used for fluorescence light microscopy? } } Thanks, } } Dennis Shubitowski } University of Michigan } School of Dentistry } dshubito-at-umich.edu
Dr. Steven Barlow, Associate Director EM Facility/Biology Department 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/EM_Facility
Would anyone know of a used tripod polisher I could buy? I need it to prepare TEM sections of modern and fossil shells (brachiopods) made of calcite.
Thanks.
Nancy Buening
Nancy Buening E-mail: buening-at-geology.ucdavis.edu Department of Geology Phone: (916) 752-0350 University of California Fax: 916 752-0951 Davis, CA 95616
I believe you have missed the point: dimethyl mercury is different from the metallic mercury that you are addressing.
} I cannot believe this discussion. No doubt mercury is a nasty cumulative } toxin. But blaming a faulty glove and a single drop passing onto the skin } is pure } nonsense. } In the good ol days I cleaned a number of times mercury diffusion pumps and } purified the mercury by shaking the stuff with nitric acid and solvents in } a separating funnel. } No gloves! I have no trace of Minnemata disease, steadier hands than most } people } and I am "approximately normal". } I knew a man who had active mercury poisoning. This was contracted in a } large } but closed room after a large industrial thermometer fractured on a very } hot oven. The man inhaled fumes for several hours. } Mercury poisoning most frequently is caused by ingestion and inhalation. I } do not advocate bathing in mercury, but that story I take with a drop of } something else!
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Can we move the Hg discussion to www.Hgtox.com, the gloves discussion to } the handwear listserv, and get on to death of more important people (Di)? } } Seriously, Prof. Wetterhahn and Di were interesting individuals who have } received their eulogies in other more appropriate forums. Whether one drop } of diMeHg causes death is debatable, but not here (are any of you } microscopists using it?). Gloves get holes and have unsealed wrists } (usually). Enough OK? } Rob Palmer } CEB/UT } } }
If you have the option, I would try low kV secondary imaging to do your image analysis, using kV's of 5-10, or even lower. Then later, go to backscatter and increase your accelerating voltage for EDX.
The problem with stains that increase your atomic number contrast, it seems to me, is that they would also greatly affect your EDX results.
Also, you could try doing your EDX first, then sputter-coating your samples for imaging purposes. Of course, then you lose the capability of going back to recheck your x-ray data.
If you must find a particular particle and image and do x-ray on it at the same time, then let me know what you find out from others, because that's a problem I've faced, too. The only thing that comes to mind immediately is image it at low kV, then reconfigure your scope for higher kV backscatter and EDX and hope you don't lose the particle in the process.
Best wishes, Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Moving the mercury story after the "air " time it has had I can see.
But , I do see a need to discuss gloves on this listserve, we work with too many nasty chemicals. Especially with chemicals such as lowicryl.
Does anyone know of a WWW site for glove permiability information? Perhaps even with microscopy specific chemicals?
I would like to make it a bookmark, or put an anchor to it on our web pages.
Thanks,
Lou Ann
} Can we move the Hg discussion to www.Hgtox.com, the gloves discussion to } the handwear listserv, and get on to death of more important people (Di)? } } Seriously, Prof. Wetterhahn and Di were interesting individuals who have } received their eulogies in other more appropriate forums. Whether one drop } of diMeHg causes death is debatable, but not here (are any of you } microscopists using it?). Gloves get holes and have unsealed wrists } (usually). Enough OK? } Rob Palmer } CEB/UT
*************************** Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine University of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lamiller-at-ux1.cso.uiuc.edu
Microscopy Home Page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Central States Microscopy Society http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/CSMS/csms.html
Personal Home Page: http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/LAM.html
Whether it is proper or not to discuss the death of a scientist in this list, I do not know.
BUT, I do not think a SCIENTIST's death is less valued than those CELEBILITIES' or Di. Without scientists, our world will less progress but without those "celebrities", our world has no impact or even better !!
We are scientists and engineers, we should respect ourselves and be proud of it.
I am not a chemist, but have received quite extensive training on hazardous materials here at work. I have learned that materials not normally absorbed through the skin can be readily absorbed when they are carried by a material that is absorbed. I believe that it may amplify the toxic effects in some cases.
It is dangerous to assume that a material that presents a minor hazard in one state, is not hazardous in another!
David L Johnson wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I'm doing automated particle analysis (SEM/EDX) of suspended particulate } material from drinking water reservoirs. Particles are filtered onto } nuclepore membranes and carbon coated. Features are located using a } thresholded bse image. Diatoms (some species) are so thin that that they } don't image very well. Does anybody know of a 'staining' method to } increase the atomic number contrast for amorphous silica? } } reply to jptmvl-at-mailbox.syr.edu thanx, dave johnsonDave,
I don't know of an EM approach but there is a very old technique called "Rheinberg Illumination" which works a treat with diatoms. Basically, the technique is called "optical staining" and involves selectively filtering the background versus the scattered light. One of my favorites produces a blue background and yellow diatoms .... a combination which should be pretty effective for automated image analysis systems. We've used them for years on many non-stained specimens, ranging from diatoms to mold in ketsup.
The filters, originally available from KODAK, worked really well with 10x objectives. When they discontinued producing these filters, they were kind enough to give us the originals. We can make a set of about 20 different filters available to you for a modest price. Contact me directly and I can provide you with the details for both how to use these filters and pricing.
Plase let us know how you make out.
Best regards,
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
What size are the particles you are trying to image?
For the first time I am going to add my two cents worth. In regards to the posting and discussion about dimethylmercury, I would rather read about safety problems and concerns than be subjected to the bickering between list members. This is not the first occasion that some list members have used this public forum to deliver jibes. That the occassion revolves around someone's death makes it all the more objectionable to me. Although Nestor has reminded the entire list of the rules of the list, and the objectionable traffic has abated, I'm seeing it again. Please respect the published purpose and rules of this list. I speak for myself, but perhaps I am not alone in feeling this way.
Sincerely, Maureen Petersen
************************************************************************ Maureen Petersen Department of Plant Pathology 1453 Fifield Hall University of Florida
Dear David, A light gold or gold-palladium sputter coat will increase BSE response and will interfere minimally with EDX identification. Diatoms are pure Si anyway, so small Au and Pd peaks will not interfere. I always use Au-Pd when I image by BSE, unless there are heavy elements already present.
David Johnson wrote: } I'm doing automated particle analysis (SEM/EDX) of suspended particulate } material from drinking water reservoirs. Particles are filtered onto } nuclepore membranes and carbon coated. Features are located using a } thresholded bse image. Diatoms (some species) are so thin that that they } don't image very well. Does anybody know of a 'staining' method to } increase the atomic number contrast for amorphous silica? } } reply to jptmvl-at-mailbox.syr.edu thanx, dave johnson 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 keep Paraplast Plus melted in a bath more or less continuously, so it is always ready to use, even though it may be one or two months between uses. Lately my students have been having some sectioning problems (with freshly sharpened blades) and before I suggest that maybe they have done something wrong during embedding, is it possible that the wax has "gone bad?"
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
I always immerse the grids for IEM-colloidial gold expt's. That is if I am using only 1 primary antibody. If using 2 primary antibody, then I float the grids, one side per antibody/gold. As a side note, I also immerse the grids when staining with UA and Pb.
Best of Luck Ed Calomeni Medical College of Ohio Dept Pathology Toledo, OH 43614 emlab-at-opus.mco.ecu
Colleagues , I can't help this guy can any of you? Reply back to him not the listserver...
Nestor Your Friendly Neighborhood SysOp
Below is the result of your feedback form. It was submitted by (dmatthew-at-providence.edu) on Sunday, September 14, 1997 at 20:46:01 ---------------------------------------------------------------------------
Question: Hello. I am an undergrad student at PC just beginning a class using the EM. For a project I plan on examaning the amebocytes of limulus polyphemus (horshoe crab). I am intersted in any information on fixation techniques of the cells for the TEM. I had planned on centrifuging blood and then processing the pellet. I've begun a literature search for specific techniques, but getting my hands on the more obscure journals can be difficult. Any help would be appreciated. Thanx so much.
We got our target from Goodfellow Metals in Cambridge UK Tel: +44-1223-568068 Fax: +44-1223-420639. They have high purity chromium foils of various thicknesses and will laser cut to what ever size is needed. We ordered a 1mm thick target for the Denton Magnetron Sputter Coater we have on our Oxford Instruments CM1500 cryopreparation unit. It works very well. Get Goodfellows to send you their very informative catalogue.
Patrick Echlin Director, Multi-Image Centre University of Cambridge
On Wed, 10 Sep 1997, T. Graham wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } I need to get ahold of a chromium sputtering target. Does anyone know } which company I may purchase this from? } } Tom Graham } } }
Does anyone know of the Histoclear and Histomount clearing and mounting media? I'd like to know the name of the manufacturer and where they are located? Thanks,
Imre Kovacs M.D. kimre-at-comser.szote.u-szeged.hu
Alzheimer's Disease Research Center Albert Szent-Gyorgyi Medical University H-6720 Szeged, Somogyi u. 4. Hungary
Dear colleagues, I have formvar powder and choloform at my disposal to prepare holey carbon films. Could you give me the method to use or the references of some literature where I can find it? Thank you for your help! Nathalie Bozzolo _________________________________________________
Nathalie Bozzolo Laboratoire d'Analyse des Materiaux Centre de Recherche Public - Centre Universitaire 162a, avenue de la Faiencerie L-1511 Luxembourg tel : (352)46 66 44 402 fax : (352)46 66 44 400 _________________________________________________
Many thanks to Phil Dahlstrom for sorting out the "mercury" confusion. It reminds me of the time when I was an inexperienced young scientist at the Paint Research Association (and dinosaurs walked the earth!). I had thought of using tetraethyl lead as an (electron? X-ray?) dense material, and was forcefully warned off by one of the senior staff there. Simply because it was found in petrol did not mean that it was OK to use: in fact, at the "lead" factory there would be paraffin (kerosene) showers, and on the slightest contact the victim would be shoved under, clothes and all - speed is of the essence.
More recently, I was looking for ways of putting heavy metal into specimens either for TEM density or for backscattering SEM, and I discovered recent work in the literature, suggesting triphenyl bismuth as a relatively low toxicity material - bismuth compounds are apparently remarkably innocuous compared to its neighbours in the periodic table. I didn't get so far with that, as we found another way of imaging our specimens - water trees in electric cables - but if anybody is interested, here are a couple of references they could follow up:
(1) TI: THERMOMECHANICAL INVESTIGATION OF POLY(METHYLMETHACRYLATE) CONTAINING AN ORGANOBISMUTH RADIOPACIFYING ADDITIVE AU: RAWLS_HR, GRANIER_RJ, SMID_J, CABASSO_I JN: JOURNAL OF BIOMEDICAL MATERIALS RESEARCH, 1996, Vol.31, No.3, pp.339-343
(2) TI: Radiopaque copolymers of styryldiphenylbismuth vinylbenzylphosphonate and methyl methacrylate AU: Tamber_H, Smid_J, Cabasso_I JN: CHEMISTRY OF MATERIALS, 1997, Vol.9, No.6, pp.1335-1341
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
} -----Original Message----- } From: Vladimir Dusevich [SMTP:dusevich-at-astro.ocis.temple.edu] } Sent: Friday, September 19, 1997 2:56 PM } To: Robert J. Palmer Jr. } Cc: Microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Hg, gloves, death } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } 100% agree!!! } } On Fri, 19 Sep 1997, Robert J. Palmer Jr. wrote: } } } } ---------------------------------------------------------------------- } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } ---------------------------------------------------------------------- } -. } } } } Can we move the Hg discussion to www.Hgtox.com, the gloves } discussion to } } the handwear listserv, and get on to death of more important people } (Di)? } } } } Seriously, Prof. Wetterhahn and Di were interesting individuals who } have } } received their eulogies in other more appropriate forums. Whether } one drop } } of diMeHg causes death is debatable, but not here (are any of you } } microscopists using it?). Gloves get holes and have unsealed wrists } } (usually). Enough OK? } } Rob Palmer } } CEB/UT } } } } } }
INVITATION TO SUBMIT PROPOSALS FOR 1) PROGRAM PARTICIPATION and 2) 1998 FACULTY FELLOWSHIPS
The Shared Research Equipment (SHaRE) User Facility and Program at the Oak Ridge National Laboratory (ORNL) provides access to a variety of advanced instrumentation for collaborative materials science research. Through the SHaRE User Program, materials scientists from universities, industries, or other government laboratories may access the SHaRE Facility as well as other instrumentation within ORNL's Metals and Ceramics (M&C) Division. Facility instrumentation includes a variety of electron microscopes, atom probe field-ion microscopes, and mechanical properties microprobes.
SHaRE Program Participation Proposals are being solicited at this time for facility use during fiscal year 1998 (October 1, 1997 - September 30, 1998). The program is intended to support collaborations between M&C staff members and researchers external to ORNL. Therefore, proposals must identify at least one staff member and one non-ORNL participant who will act as a co-principal investigator and share responsibility for the project. Proposals will be reviewed by a committee and evaluated with regard to scientific excellence, relevance to the interests of the U.S. Department of Energy, Division of Materials Sciences, and the likelihood of project success. Additionally, principal investigators and graduate students from U.S. accredited universities are eligible to receive funds to defray certain program-related travel and subsistence expenses.
The SHaRE Facility instrumentation and the related User Program are described in detail at http://www.ornl.gov/share. In the past, only letter proposals were submitted for program participation. However, application for program participation and travel support is now made by using a downloaded form located at http://www.ornl.gov/share/pdf/proposal98.pdf. The form will be mailed to potential applicants upon request.
1998 Faculty Fellowships Faculty fellowships provide outstanding university faculty extended access to the SHaRE User Facility at ORNL. It is anticipated that at least one junior and one senior university faculty member will be appointed as fellows. Information regarding the fellowships, including eligibility requirements, length of appointment, stipends, and application guidelines may be found at http://www.ornl.gov/share. The guidelines will be mailed to potential applicants upon request.
SHaRE will accept and review proposals for projects and fellowships at any time during the fiscal year, but allocates the majority of funds during the month of October. Proposals for review during the October meeting should be received at the address below by October 10, 1997. Proposals will be reviewed, and travel awards announced, in mid-October. Fellowship applications received after the October date will be reviewed during February 1998.
Proposals submitted (5 copies) should be sent to:
SHaRE Proposals Education and Training Division, MS-36 Oak Ridge Institute for Science and Education P.O. Box 117 Oak Ridge, Tennessee 37831-0117
SHaRE is jointly administered for the U.S. Department of Energy by ORNL and the Oak Ridge Institute for Science and Education (ORISE). For additional information or clarification on proposal submissions, please contact me directly.
Regards,
Neal
Dr. Neal D. Evans voice: (423) 576-4427 Shared Research Equipment Program facsimile: (423) 574-0641 Oak Ridge National Laboratory email: evansnd-at-ornl.gov Building 5500, MS 6376 Oak Ridge, TN 37831-6376
Greetings! I am looking for a site where I can find microscopic pictures of various kinds of protists for my fifth grade science students. I have drawings, but we do not have access to slides of the real thing. Perhaps someone out there can direct me to a useful website that will interst my students. Thanks! Reply to: jpolak-at-esu6.esu6.k12.ne.us Julie Polak Exeter Public Schools Exeter, Nebraska
} I have to prepare some yeast for SEM. The person who requested this } work wants some pretty pictures of his yeast, Schizosaccharomyces pombe, } for use in seminars. I have read up on some techniques to use but have } two main queries: } 1. To collect the yeast onto filter paper ( a method used in } several publications), do I just drop a suspension of the yeast onto the } filter paper? What sort of filter paper do I use? Is there a "better" } way of collecting these cells such as settling them onto poly-l-lysine } coverslips?
We use regular microscope slides, cleaned well (detergent, acid or alcohol): coat with l mg/ml aqueous solution of poly-L-lysine for several minutes, rinse in distilled wate.
Take a (preferably) aqueous suspension of the yeast and place onto the microscope slide and allow the cells to settle for about one hour at RT. Carefully tip the slide and allow the unattached cells to flow off. Now, gently dropper some fixative (2% glut/4% formald in buffer of choice) onto the slide to completely cover the smear. Allow to set undisturbed overnight at RT in a humid chamber (petri dish with filter paper or Tupperware container with moist paper towels). Transfer the slide into a rinse solution (Coplin jar of distilled water or petri of distilled water). With some yeasts, the aldehyde fix is adequate to preserve the integrity of the cells (others may collapse without osmium post-fixation). The slides are then slowly dehydrated in ethanol (20-40-60-80-100-100%) for 10 min each. Critical point dry using liquid carbon dioxide as transitional solvent. You will need to break the slide into 1 inch squares to fit into the CPD device. Do this by scoring the slide with a diamond marker pen and pressing down gently onto an applicator stick. Mark an "X" the back side with the diamond marker to help identify the good side later. Freeze drying from the ethanol should also work well.
IF osmium is needed place slide/specimen into 2% aqueous osmium tetroxide overnight - take care to avoid evaporation by either immersion of the slide into a shallow container of osmium or by vapor fixation of the slide/specimen. Vapor fixation (CAUTION WITH OS FUMES - DO IN PROPERLY OPERATING FUME HOOD) may be accomplished by placing the wet slide/specimen into a plastic petri dish and then placing a small volume (4-5 ml) of 2% osmium solution nearby in the dish. Rinse in distilled water and dehydrate and CPD as described above.
Caveats: do not overload the slides with culture since you do not want a heaped up mess but isolated cells. A suspension that is quite turbid should work well - not a paste or milky/opaque solution.
Contact me if you have any other questions. I have done a lot of imaging of yeasties by TEM and SEM.
Cheers,
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I would like to ask you for your advise on cryo-sectioning of plant material in possibly low temperature. We tried to section the frozen piece of leaf using Cryocut E Reichert, playing with different embedding media but the results were not satisfactory. We found difficulties in cutting thick sections (5-30 microns) even of very young leaf. Morphological structure was not well preserved.
I really would appreciate your advise, suggestions, tips and technical tricks in this matter. What equipment do you use and what can you recommend?
Thank you in advance for all your time and assistance
With best regards
Jolanta Mesjasz-Przybylowicz ************************************************************************ Dr Jolanta Mesjasz-Przybylowicz National Accelerator Centre P.O. Box 72 Faure 7131 South Africa tel: 27-21-8433820 fax: 27-21-8433543 Internet: MESJASZ-at-nac.ac.za ************************************************************************
I have used both with excellent results, for 10 micron and 100-200 micron sections. Histomount *really* likes to shrink, so it needs to be watched while drying. Also, it makes zillions of air bubbles if you try to dry it with heating like is done with Permount. Dry finished slides at room temp. Excess Histomount cleans up with Histoclear and a razor blade.
Both are made/sold by National Diagnostics in New Jersey, US, but I don't have the contact information. (This is a year old--I assume the information hasn't changed.)
Fisher sells a similar (identical?) product to Histoclear, but I don't remember if they have a Histomount analog.
Phil
} Does anyone know of the Histoclear and Histomount clearing and } mounting media? I'd like to know the name of the manufacturer and } where they are located? } Thanks, } } } Imre Kovacs M.D. } kimre-at-comser.szote.u-szeged.hu } } Alzheimer's Disease } Research Center } Albert Szent-Gyorgyi } Medical University } H-6720 Szeged, } Somogyi u. 4. } Hungary
1. Add 4 drops of solution b to solution a, and shake vigorously for 30 seconds. 2. Pour on clean glass slides and dry. (ie: dip the slides into a staining dish.) 3. Breath on slide several times. 4. Float layer off the slides. 5. On the floating layers, gently place grids, dull side down. 6. Pick up layer with parafilm, letting the film stick to the parafilm. 7. Let dry for 30 minutes. 8. Put in petri dish with filter paper saturated with 50% methanol for 20 minutes. 9. Take each grid off mesh and dip in 50% methanol and place on filter paper to dry. 10. Coat with carbon in a vacuum evaporator. 11. Immerse each grid in chloroform briefly.
} ---------- } From: bozzolo-at-crpcu.lu[SMTP:bozzolo-at-crpcu.lu] } Sent: 22 September, 1997 06:11 } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: TEM - holey carbon films } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi The company is:National Diagnostics 404-699-2121
bob
On Mon, 22 Sep 1997, IMRE KOVACS M.D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopists, } } Does anyone know of the Histoclear and Histomount clearing and } mounting media? I'd like to know the name of the manufacturer and } where they are located? } Thanks, } } } Imre Kovacs M.D. } kimre-at-comser.szote.u-szeged.hu } } Alzheimer's Disease } Research Center } Albert Szent-Gyorgyi } Medical University } H-6720 Szeged, } Somogyi u. 4. } Hungary } }
} Rick, } While the AP is not peer reviewed Science is. Suggest that you check out } the August issue. Karen } was a collaberator with several people in our lab. Also suggest that } you look up } "dimethylmercury". One drop (50ul) is equal to about 300X the } occupational exposure limit. } -- Begin original message -- } } Without in any way downplaying the tragedy for the people involved, the } } dose-response relationship postulated in the story is not very plausible. } } Mercury toxicity has been studied extensively for at least half a century. } } It is a cumulative poison which does cause the type of neurological effects } } reported, but a "single tiny drop", through a glove and quickly washed off, } } being lethal seems suspect. Associated Press is not peer reviewed (although } } maybe it depends on your definition of "peer"). } } } } Rick Mott } } rick-at-pgt.com } } } } (These are personal views having nothing to do with my employer) } } } } -- End original message -- } } } regards, } Bob } Robert Schoonhoven
Of course, one of the problems is that the original newspaper article that started this thread refered to 'mercury'. Which as many have pointed out, as elemental metallic mercury while dangerous is essentially a long term, cumulative poison and there is no possibility that single drop on even unprotected skin will cause the slightest harm.
On the other hand, dimethylmercury is an entirely different kettle of fish (apologies for colloquialism). This is a classic example on the confusion that arises when the scientifically ignorant start talking about things they don't understand.
It reminds me of the debate in the British parliament a number of years ago on the merits of fluoridation of water - the principle case was made by a scientifically ignorant MP who based his argument against fluoridation on a dictionary definition of the chemical and biological properties of fluorine. One wonders if he had ever looked up the same information for chlorine and took a similar stand on his use of salt on his food.
While there are many arguments in science, particularly in areas relating to ethical issues, which may be open to the general public, there are equally many that aren't. It is simply a case that if you haven't studied and learnt the basic facts, you are ignorant and aren't qualified to comment. And notions of democracy, and free speech don't change that.
The Division of Nephrology in the University of Florida College of Medicine is seeking a full-time electron microscopy technician. This individual will be one of two full-time electron microscopy technicians staffing the Division's Electron Microscopy Facility, which serves as a research facility for the members of the Division of Nephrology and their collaborators. The research conducted primarily strives to determine correlations between renal ultrastructure and function using transmission and scanning electron microscopy, morphometric analysis, and immunogold and immunoperoxidase cytochemistry. Equipment housed within the facility include 2 Zeiss EM-10A transmission electron microscopes, a Topcon DS130C scanning electron microscope, 2 LKB Nova ultramicrotomes, a Leica Automatic Freeze Substitution unit, and all related support equipment. The candidate's duties will include tissue processing, ultramicrotomy, immunocytochemistry, routine maintenance of the electron microscopes, viewing samples, photographic processing, and related support functions. The candidate may be hired as an Electron Microscopy Technician of Senior Electron Microscopy Technician depending on experience.
Reply to Dr. Jill Verlander verlandr-at-medicine.ufl.edu ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Our Ohio company is seeking engineering / scientist support for microelectromechanical systems research & development. Work will involve new materials and surface treatments to control the friction and wear of MEMS devices; Friction & wear measurement of MEMS systems / materials.
It is important that candidates have capabilities in cross-section TEM, analytical TEM of tribological materials; especially wear tracks; creation of unique microstructures; and understand friction & wear on a fundamental level.
Contact Ronald Decker - deckerrc-at-utcdayton.com
Ronald C Decker Program Manager Universal Technology Corporation 1321 Research Park Drive, Suite 100 Dayton OH 45432-2817 Voice (937) 426-8530, Fax (937) 426-7753
Dear All: I inherited a used Jeol 100ZX. It is in excellent working condition and about 20 years old, located in the Los Angeles area. If you are in need of a TEM or want it for spare parts please make me an offer. Peter Jordan, EMSI 909 694-1939
This information is correct as of March 95 when we last ordered. A European source of Hystomount (and I think also Hystoclear) is:
Hughes and Hughes Ltd. Unit 1F, Lowmoor Industrial Estate, Tonedale, Wellinton, Somerset TA21 0AZ England Phone +44 823 660222 FAX +44 823 660186
As regards to Hystoclear, there are other companies which sell a limonene-based xylene substitute. As an example, Fisher calls its product Hemo-De. They are different than xylene in more than just the toxicity. Thus I would ask for a small sample from your supplier to try before you buy. In my experience these products are normally sold in large quantities. (Fisher lists it smallest size as 1 gallon).
Good luck, Azriel Gorski
On Mon, 22 Sep 1997, IMRE KOVACS M.D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopists, } } Does anyone know of the Histoclear and Histomount clearing and } mounting media? I'd like to know the name of the manufacturer and } where they are located? } Thanks, } } } Imre Kovacs M.D. } kimre-at-comser.szote.u-szeged.hu } } Alzheimer's Disease } Research Center } Albert Szent-Gyorgyi } Medical University } H-6720 Szeged, } Somogyi u. 4. } Hungary }
We need a Zeiss Axioplan microscope for transmitted light and fluorescence work. This would be the original Axioplan, not the new Axioplan 2 model. If you own this microscope, and are interested in selling or trading your instrument, please send me details of the configuration you have. Part numbers would be helpful.
Patrick
-------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com
I want to attach a video system to my light microscope. I just bought the camera (Sony ccd 370) and the corresponding adaptor and lens, although now I'm having trouble with the board. I bought a miro dc30 board but I only get a small image (I want the image on the entire screen), and, besides, I'm not having good printings. I'm working with and HP 133 Mhz, 48 MB ram, and my printer is an HP 870. I guess that this are good working conditions, thus, I believe that the board is not correct. Can anyone help me? Thank you in advance!
I would check out the adapter and lens system before the board.
We have a Pixera camera that we have tried to replace our old RS-170 camera with. The lens/adapter for the RS-170 camera screws right in to the forn of our Pixera, however, the field of view is only about one third the size it was with our RS-170 camera. I came to find out that that should not be a surprise. The chip on the Pixera is only 1/3" across while it is 1" across on the RS-170. We have yet to get the right adapter, but have looked at items from Optem, Diagnostic Instruments, and Edmund Scientific. We should be able to get much closer.
There are others out there that have gone through the same exercise. Maybe they will speak up too.
At 08:21 AM 9/23/97 -0300, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear microscopists: We have very good deal of barely used, looks like new NORAN confocal and RMC ultramicrotome and their accesories for sale. Please contact Dorothy at 617-432-2970. Thanks.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I want to attach a video system to my light microscope. I just bought } the camera (Sony ccd 370) and the corresponding adaptor and lens, } although now I'm having trouble with the board. I bought a miro dc30 } board but I only get a small image (I want the image on the entire } screen),
Not necessarily. I am not familiar with your camera or board but the problem could be that the relay lens for the camera has too large a field of view. Have you tried connecting the camera directly to a monitor? Is the image acceptable on that?
Thanks to all who responded to my question about my Denton not sucking. The problem turned out to be a broken rough pump bellows. Ida (the parts lady) at Denton was great! Even though there was a long backorder for the part, she searched all over Denton and found a bellows for me. She did this out of the kindness of her heart. Thanks to her we were able to repair my machine lickety-split. I just thought I'd let everybody know how much I appreciate them, the BBer's for advice and the Denton folks for being so helpful. My Denton now sucks like you wouldn't believe. It hasn't worked this well in a long time. It can now pump down to 2 X 10-5 without any LN2 added to the trap. I guess the moral of the story is...It always pays to ask those who know.
Happily carbon coating in Berkeley,
Paula = )
p.s. I have no financial interest in Denton, etc, etc, etc.
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I am not familiar with the Sony system, but if it is NTSC video, then you will be limited to about 500 lines of vertical resolution. Horizontal resolution will depend on the Sony and the capture card. It is probably about 400 lines... This will equate to 500x~400 pixel image. If your computer screen is set to - say 1024x768 pixel display, the unmodified image would fill about 1/2 the CRT. There may not be a convenient way to increase the Sony resolution, but you could fill the screen by reducing the CRT resolution to 640x480. Better yet, (although "hollow" magnification) would be to increase the image pixel array size (software manipulation) after capture to fill the screen. Increasing the pixel array for the same size hard copy may help produce better halftone printer output also.
For work like this, I would prefer higher resolution, digital still cameras. The obvious drawback is lack of "real-time" output.
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I want to attach a video system to my light microscope. I just bought the camera (Sony ccd 370) and the corresponding adaptor and lens, although now I'm having trouble with the board. I bought a miro dc30 board but I only get a small image (I want the image on the entire screen), and, besides, I'm not having good printings. I'm working with and HP 133 Mhz, 48 MB ram, and my printer is an HP 870. I guess that this are good working conditions, thus, I believe that the board is not correct. Can anyone help me? Thank you in advance!
Our lab is in the transition of doing immunogold tagging in HRLM-TEM.But we continue to do immunofluorescent techniques received in Michels transport media. Is there anyone out there still doing immunofluorescent techniques (direct or indirect immunofluorescent techniques) on renal, skin, muscle or nerve biopsies? If yes, is the biopsy snap-frozen in liquid nitrogen as soon after removal as possible (precooling a metal chuck for about a minute. Placing a small volume of saline or water on top of the colled chuck, immediately placing the renal or skin bx into the water or saline drop. As the water and the bx begin to freeze, the chuck is turned down to eliminate excess water or saline. The chuck is placed in liquid nitrogen for approx one minute to snap-freeze the bx or tissue. When nitrogen stops bubling bx is adequately frozen and bx is ready to be stored at -70oC or to be sectioned). If not possible to do technique is the tissue being storred in Michels' transport media also sold commercially as Zeus. Are the renal biopsies being tagged with IgG, IgM, IgA, C3, C1q, Kappa & Lambda? Skin, muscle and nerve biopsies tagged with IgG, IgM, IgA, C3 and Fibrinogen? Please respond. Thanx Teresa
Our lab is in the transition of doing immunogold tagging in HRLM-TEM.But we continue to do immunofluorescent techniques received in Michels transport media. Is there anyone out there still doing immunofluorescent techniques (direct or indirect immunofluorescent techniques) on renal, skin, muscle or nerve biopsies? If yes, is the biopsy snap-frozen in liquid nitrogen as soon after removal as possible (precooling a metal chuck for about a minute. Placing a small volume of saline or water on top of the colled chuck, immediately placing the renal or skin bx into the water or saline drop. As the water and the bx begin to freeze, the chuck is turned down to eliminate excess water or saline. The chuck is placed in liquid nitrogen for approx one minute to snap-freeze the bx or tissue. When nitrogen stops bubling bx is adequately frozen and bx is ready to be stored at -70oC or to be sectioned). If not possible to do technique is the tissue being storred in Michels' transport media also sold commercially as Zeus. Are the renal biopsies being tagged with IgG, IgM, IgA, C3, C1q, Kappa & Lambda? Skin, muscle and nerve biopsies tagged with IgG, IgM, IgA, C3 and Fibrinogen? Please respond. Thanx Teresa
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } I would check out the adapter and lens system before the board.
Agreed.
} We have a Pixera camera that we have tried to replace our old RS-170 camera } with. The lens/adapter for the RS-170 camera screws right in to the forn of } our Pixera, however, the field of view is only about one third the size it } was with our RS-170 camera. I came to find out that that should not be a } surprise. The chip on the Pixera is only 1/3" across while it is 1" across } on the RS-170. We have yet to get the right adapter, but have looked at } items from Optem, Diagnostic Instruments, and Edmund Scientific. We should } be able to get much closer.
I am not sure which particular kind of microscopy will suit my needs, and would very much appreciate it if somebody could help me in this regard.
My application is as follows: I need to look at molecular-level changes occurring at a gas-liquid interface with respect to some parameters. The liquid will have, among other components, proteins in it. I want to look at the larger molecules which are affected by an interface. For this, I plan to take a liquid volume and disperse bubbles in them. Then I plan to freeze a part of the liquid rapidly and carry out imaging at various sections. I don't know what kind of technique or microscopy is the best for this. I would appreciate comments and suggestions on this. Also, if somebody knows of a paper or book that has this kind work in it, then that would give me some leads. I have searched some of the indexes with keywords like surface, bubble, modification, microscopy, protein etc., but could not find what I wanted. Am I missing something here? We have some generic metrology equipment like Surface profilers (contact and Optical) , SEM, AFM, Light Microscopes (not near-field). I would also be interested in knowing if any of this can be adapted for my needs. I have NOT done any of the following: biological tissues examining/sectioning, staining, fixing, cryomicroscopy, AFM etc. My experience is with microfabricated structures and examination under the SEM, light microscope and surface profilers. However, I am very willing to learn a new field, and can work towards obtaining a new piece of equipment, or try to have some arrangement with interested commercial/non-commercial parties.
Thanks very much.
Ashok Institute for Micromanufacturing Louisiana Tech University E-mail: krishnan-at-engr.latech.edu ph: (318) 251-8110, fax: (318) 257 5104
"Smaller, lighter, more functional and less expensive consumer products, industrial machines, instruments...possibilities limited only by man's imagination"
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ms. Polak: Please look at the website list that is section V of the Project MICRO bibliography (address below). And if you're really into protozoa, I recommend the recent book by Anderson & Druger listed in section IIB.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
The Analytical Microscopy Core facility at Hospital for Special Surgery, New York, NY, is searching for a full time electron microscopy technician. The Core provides TEM and SEM service to about 50 clinical and basic scientists, whose major interests are in orthopedics and connective tissue diseases. About 2 days per week would be used to provide technical assistance to a senior researcher, which would involve TEM, SEM, immuno and histochemistry, and development of image analysis techniques. This individual should be familar with basic TEM and SEM techniques but will be trained to work with connective tissues and calcified cartilage and bone. The job can be classified as EM Technician or Senior Technician, depending on qualifications. Interested persons please respond to Steve Doty at address below:
Stephen B. Doty, PhD. Phone: (212) 606-1417 Director, Analytical Microscopy Core Fax: (212) 717-1192 Hospital for Special Surgery email: dotys-at-hss.edu 535 E. 70th Street, NY, NY 10021
Dear All: I made two mistakes on my posting yesterday, first the Jeol is a 100C and not a 100ZX and my phone number is 909 694-1839 and not 1939. The thing I got right was that it is 20 years old and it is still for sale. Sorry, I'll never write an e-mail letter at midnight. Peter Jordan, EMSI
Dear All: A few months ago there was a posting about a Zeiss 9 TEM given away for free. If this is still available or if you know who did the posting please let me know. If I remember right it was in the San Diego area. Thank you. Peter Jordan
Greetings again! I want to thank all of you terrific people out there who have so graciously sent me information about protist pictures!!!! I took my 5th graders to several of the sites and they said the protists were "AWESOME!!!" (I concurred!) We especially liked the "zoo." Several of you sent personal messages encouraging my students and me, and also sent links and other interesting information. Thanks again for all your help! I think I've even made a couple of new net-friends! Many thanks, Julie Polak
At 10:32 AM 9/24/97 +0200, you wrote: } I have a problem with an import the file with an extension *.sp ( spectrum ) } and put it to the document in word 6 or amipro } } Malgorzata Warmuzek
As a rule, you must have a program that supports OLE (object linking and embedding) for the type of file that you have at hand before you can import that file into another program. I suspect that you have no such program for your SP files.
You will probably have to export the file to some form that can be read by your spreadsheet or graphing program, prepare your graph there, and then copy that graph into your word processor. I haven't worked much with Microsoft's MSGRAPH that comes with Word, but it might do what you need in a rudimentary sort of way without invoking a spreadsheet program. ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering, or 270 Metals Development Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
We had so much fun yesterday at the Microbe Zoo and at Kunkel's Gallery, that I want to share our four favorite sites with the rest of the world (as it were). I think these would be helpful to other upper elementary/middle school teachers and students.
I use Thumbs Plus (approx. $60 US if I remember correctly) Cerious Software, Inc. 1515 Mockingbird Lande Suite 209 Charlotte, NC 28209 http://www.cerious.com
I have no financial interest in this company other than doing my part as a very satisfied customer.
} ------------------------------------------------ } 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} } } }
We are in the process of resurrecting Cambridge 250 Mk3 SEM and we are looking for the source of spare parts. Any feedback on the subject will be greatly appreciated.
As far a s cryosections of leaves are concerned. It ain't easy because of all the internal air spaces. Cryofracturing is no problem and with a kittle care you can cryoplane the leaf ti get a nice smooth surface. I think I know what you want to do Jolanta (are you still doing ion beam microscopy ?) May be freeze substitution is the best approach. If all else fails read my book
Best wishes
Patrick Echlin University of Cambridge
On Mon, 22 Sep 1997, Jolanta Mesjasz-Przybylowicz wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } } Dear Microscopists, } } } I would like to ask you for your advise on cryo-sectioning of plant } material in possibly low temperature. We tried to section the frozen } piece of leaf using Cryocut E Reichert, playing with different embedding } media but the results were not satisfactory. We found difficulties in } cutting thick sections (5-30 microns) even of very young leaf. } Morphological structure was not well preserved. } } I really would appreciate your advise, suggestions, tips and } technical tricks in this matter. } What equipment do you use and what can you recommend? } } Thank you in advance for all your time and assistance } } With best regards } } Jolanta Mesjasz-Przybylowicz } ************************************************************************ } Dr Jolanta Mesjasz-Przybylowicz } National Accelerator Centre } P.O. Box 72 } Faure 7131 } South Africa } tel: 27-21-8433820 } fax: 27-21-8433543 } Internet: MESJASZ-at-nac.ac.za } ************************************************************************ }
Thumbs plus by Cerius Software works very well for us as an image filing tool. It provides a thumbnail of all images in a directory.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
On Wed, 24 Sep 1997 mark_munro-at-bio-rad.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } } Dear all, } does anyone know of any shareware/inexpensive image database and } archiving software. } } Thanks in advance, } } Mark Munro } } }
mark_munro-at-bio-rad.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear all, } does anyone know of any shareware/inexpensive image database and } archiving software. } } Thanks in advance, } } Mark Munro
Mark,
Company Cerious Software Inc. http://cerious.catalogue.com/index.html has a great software ThumbPlus 3.0 for 60 US$.
Henrik Kaker SEM-EDS Laboratory Metal Ravne d.o.o. Slovenia
We are looking for any information we can find on the microscopy/identification of particles from automobile tires. Seems we have a case where we need to try to match particles from a pair of tennis shoes to a particular set of automobile tires(best case), or at least identify particles as possibly coming from automobile tires (more likely). (I'll let your imagination fill in the details on this one!)
If anybody has done anything like this, please let us know. In the meantime, we'll be hitting the indexes/abstracts and databases.
Never a dull moment.... Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
ThumbsPlus is very good. I've used a single site license. In my new position, I've just gotten 5 concurrent site licenses for myself and my team. I don't know how good their Mac version is. I believe that it is still in beta testing. It is shareware and Cerious software has a website where you can download the shareware version. Once you are registered, there are some things that are available, but the unregistered version is very good. I tried it out that way after it was suggested previously on the Microscopy Listserver.
-Scott Walck
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
We do direct immunofluorescence on renal and skin biopsies. For the most part, the specimens are received in Michels transport media (which we make ourselves). Upon arrival, the specimens are washed in 3 changes of transport buffer to flush out the excess salts of the michels media (3 washes, 10 minutes each, room temp). The specimen is then lightly blotted, oriented in a cryomold, covered in OCT, and frozen. To freeze, we cool iso-pentane (2-methylbutane) in liquid nitrogen, and lower the cryomold into the cooled iso-pentane for a few seconds. We then attach a chuck in the cryostat, and cut. When the specimen is received fresh, we skip the washing part, and proceed as above. On the renal biopsies, we run: IgG, IgA, IgM, C3, C1q, Fibrinogen, Kappa, Lambda, and Albumin. On the skin biopsies, we run: IgG, IgA, IgM, C3, C1q, Fibrinogen, and Albumin.
Hope this answers your question.
Phyllis Davie Immunocytochemistry Laboratory University of Washington Medical Center pdavie-at-u.washington.edu
On Tue, 23 Sep 1997, Flores, Teresa wrote:
} Our lab is in the transition of doing immunogold tagging in HRLM-TEM.But we } continue to do immunofluorescent techniques received in Michels transport } media. Is there anyone out there still doing immunofluorescent techniques } (direct or indirect immunofluorescent techniques) on renal, skin, muscle or } nerve biopsies? If yes, is the biopsy snap-frozen in liquid nitrogen as } soon after removal as possible (precooling a metal chuck for about a } minute. Placing a small volume of saline or water on top of the colled } chuck, immediately placing the renal or skin bx into the water or saline } drop. As the water and the bx begin to freeze, the chuck is turned down to } eliminate excess water or saline. The chuck is placed in liquid nitrogen } for approx one minute to snap-freeze the bx or tissue. When nitrogen stops } bubling bx is adequately frozen and bx is ready to be stored at -70oC or to } be sectioned). } If not possible to do technique is the tissue being storred in Michels' } transport media also sold commercially as Zeus. } Are the renal biopsies being tagged with IgG, IgM, IgA, C3, C1q, Kappa & Lambda? } Skin, muscle and nerve biopsies tagged with IgG, IgM, IgA, C3 and Fibrinogen? } Please respond. Thanx Teresa } } } }
You can capture the spectrum with a software, such as Paint Shop Pro we are using, and save it as TIFF or Bitmap file. If you have the software, I can show you the details.
With the best wishes, Charlie Kong kong-at-t-rex.materials.unsw.edu.au
On Wed, 24 Sep 1997, Malgorzata.Warmuzek wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } } I have a problem with an import the file with an extension *.sp ( spectrum ) } and put it to the document in word 6 or amipro } } Malgorzata Warmuzek } Foundry Research Institute } Research Materials Department } Structural and Physical Research Laboratory } } Zakopianska 73 Call +48 12 2605022 ext. 317 } 30-418 KRAKOW - POLAND Fax +48 12 2665478, +48 12 2660870 } } }
I don't have access to a nice workshop, where they can construct a glow discharger, where in Europe can I buy one? Or, is it out there someone, who has an old mashine you don't need anymore?
TIA
Gunnel Karlsson
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Gunnel Karlsson E-mail Gunnel.Karlsson-at-oorg2.lth.se Biomicroscopy Unit Tel +46 222 8229 Inorganic Chemistry 2 Fax +46 222 4012 Box 124 S-221 00 LUND, Sweden ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ This message was sent by Eudora with recycled electrons
Malgorzata, There should be some function on your Link system for outputting the data of a *.sp file onto a DOS disk. This can then be read in to Microsoft Excel, plotted as a graph and the graph imported into MS word. We have a different Link system to you so the details may vary on how you get the Link system to output onto a DOS disk but I would be happy to send you a copy of the procedure that works for us if that would help.
Yours sincerely
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ 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. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
I have your book as a Bible on my desk. I remember our discussions concerning this problem as well but I do hope that I will find someone around the world who get closer to solve it. May be you know or heard about such person?
In cryofracturing we will be not able to obtain one layer of cells, that is a reason why I was trying cryo-sectioning. Freeze-substitution can be an option, but one should be careful about redistribution of ions which is our main worry in preparation for X-ray microanalysis.
Best regards
Jolanta
} As far a s cryosections of leaves are concerned. It ain't easy because } of all the internal air spaces. Cryofracturing is no problem and with a } kittle care you can cryoplane the leaf ti get a nice smooth surface. I } think I know what you want to do Jolanta (are you still doing ion beam } microscopy ?) May be freeze substitution is the best approach. If all } else fails read my book } } Best wishes } } Patrick Echlin } University of Cambridge
************************************************************************ Dr Jolanta Mesjasz-Przybylowicz National Accelerator Centre P.O. Box 72 Faure 7131 South Africa tel: 27-21-8433820 fax: 27-21-8433543 Internet: MESJASZ-at-nac.ac.za ************************************************************************
For that matter, if you are running under Windows, you can use the PrintScreen or Alt-PrintScreen keys to capture the screen and paste it into Word or whereever. You can get fancier by using the Format Picture item in Word to trim the bitmap to the area you want and to size it. Or you can get fancier still and first paste the bitmap into a picture editor (e.g., LView Pro or MS Imager or even MS Paint which come with Windows). Then you can crop or annotate the image and save it to a file or copy it from there to your word processor.
Note that the difference between PrintScreen and Alt-PrintScreen is that the Alt form copies only the active window instead of the whole screen. Thus you can size your spectrum (or any other application) as you want before snapping a copy.
This may not be as nice as importing the spectrum in a form in all its detail, but it will convey the information.
At 02:40 PM 9/25/97 +1000, you wrote: } Malgorzata, } } You can capture the spectrum with a software, such as Paint Shop } Pro we are using, and save it as TIFF or Bitmap file. If you have the } software, I can show you the details. } } With the best wishes, } Charlie Kong } kong-at-t-rex.materials.unsw.edu.au } } } On Wed, 24 Sep 1997, Malgorzata.Warmuzek wrote: } } } I have a problem with an import the file with an extension *.sp ( spectrum ) } } and put it to the document in word 6 or amipro } } } } Malgorzata Warmuzek } } Foundry Research Institute } } Research Materials Department } } Structural and Physical Research Laboratory } } } } Zakopianska 73 Call +48 12 2605022 ext. 317 } } 30-418 KRAKOW - POLAND Fax +48 12 2665478, +48 12 2660870 ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering, or 270 Metals Development Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
To make a glow discharge all you need is a poor vacuum ( ~ 100 mTorr) and a high voltage power supply. You can use a vacuum bell jar with an electrical feedthru. Install a leak value in the system and rough pump it out, then put in a controlled leaked of what ever gas you want, air will work, but Argon is nice and be careful with Oxygen.
Arrange your electrode to be in the bell jar (insultate it up to the point where you want the "glow" to start.) Then slowly crank up the kV. Depending on your geometry, gas pressure etc.. you should get something by the time you reach ~ 1 kV.
The more important question is what do you want to do with the discharge?
Have you tried the (Walter) McCrone Institute in Chicago? I don't have the address handy, but maybe someone else does. They have worked on everything from the Shroud of Turin to ancient paints.
------------------------------------------------ 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 --
Greetings All, After each step of the UA stain and the LC stain, I rinse the copper grids by immersion. I have four changes of water and "dip" the grid about 40 times in each change. I am interested in changing that to the procedure that rinses the grid by "flooding" it using a syringe or whatever. Can someone share with me exactly how that is done ... what type of water ... etc. Also ... do the sections ever wash off the grid when rinsing that way??
Thanks in advance, Sharron Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
09/25/97 Hi everywhere/everyone, as a brand-new member of this listservice I want to add a notice on = self-fabrication of silicone rubber molds for epoxide-resin-embedding, = which Lonie Kerr asked for 10/28/96 (only for the case, someone has = similar problems now). There are articles (in English) on self fabrication of moulds for = epoxide-embedding, at least 2 of them are: 1) MUSS W., SIMONSBERGER P. (1983): Home-made Silicone Rubber Embedding = moulds for Electron Microscopy, Mikroskopie (Vienna) 40, 207-209 (simple version for the unexperienced) 2) MUSS W.(1984): Self fabrication of Silicone Rubber embedding Moulds = for use in Electron Microscopy; Mikroskopie (Vienna) 41, 34-39 (sophisticated version for the unexperienced including silicone rubber = material to be used). I am producing all my moulds (types as shown in article 2) like cubic, = flat, "beem capsule"- type w/round or pyramidal block face, with = engraved numbers for identification and others as requested) by myself. = They are needed in our routine diagnostic EM-Lab and fabricated since = 1984 without problems, with high quality in shearing force and = unmoulding properties as well as very low costs (quantity needed a year = for about 1300 to 1600 specimen blocks ~ 5 moulds 48 specimen = cabinets/each; costs/mould ~ US-Dollars 5-15.-, depending on amount of = silicone rubber mass needed). Fabrication is simple, if negative moulds of sufficient quality are at = hand and several general rules in working up the silicone mass are = strictly followed.=20 Silicone rubber moulds of this type are o.k. for use with epoxide = resins, use for hydrophilic resins like LR White, Lowicryls not tested = yet. If interested in how to produce such moulds efficiently and interested = in which kind/quality of silicone rubber one should use, please send = e-mail request to:
Wolfgang MUSS PhD Head of EM-Lab at Pathology Department LKA Muellner Hauptstrasse 48 A-5020 SALZBURG/AUSTRIA/Europe phone: ++43++662+4482-4720 Ext. fax: ++43++662+4482-882 Ext (c/o W. MUSS) e-mail: W.Muss-at-lkasbg.gv.at.
Good luck, hope this helps somebody. END of Message, no attachment added.
} I don't have access to a nice workshop, where they can construct a glow } discharger, where in Europe can I buy one? Or, is it out there someone, who } has an old mashine you don't need anymore? } } TIA } } Gunnel Karlsson
You don't need a workshop to make a usable system. Get a small, cheap handheld Tesla coil of the sort used for physics demonstrations. Identify an unused current feedthru in your vacuum evaporator, and attach a wire fitting to it to use as a "docking port" for the Tesla coil. Discharge the coil into the vacuum during rough (mechanical) pump. Take care to not discharge the coil in air; the ozone produced damages the cilia in your respiratory system.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Dear Randy, } } We are looking for any information we can find on the } microscopy/identification of particles from automobile tires. Seems we } have a case where we need to try to match particles from a pair of tennis } shoes to a particular set of automobile tires(best case), or at least } identify particles as possibly coming from automobile tires (more likely). } (I'll let your imagination fill in the details on this one!) } } If anybody has done anything like this, please let us know. In the } meantime, we'll be hitting the indexes/abstracts and databases. } We have looked at some polymer blends for which the components can be differentially stained, and I can give you the name of our user who could possibly tell you whether this can be done for tire particles. The HVEM can be used to look at the structures of such particles if they are some few micrometers thick. We can also do EDX (and have done so on a few forensic specimens). Yours, Bill Tivol
See the following article for an easy to build glow discharge unit. I built a similar one several years ago for ionizing grids, and it works very well. All you need is a drill, a hack saw, a drill bit, and some epoxy. You will also need a plastic desciccator, a few feet of wire, a few machine and sheet metal screws, and some sheet aluminum about 1/8 to 1/16 of an inch thick. Mine is simpler than the one described in the article, consisting of only an aluminum disk screwed into the cut-off end of the high voltage generator, a larger disk placed inside the desciccator (with a wire going to an outside ground). You will also need a mechanical vacuum pump.
Aebi U, 1987 [See Related Articles] A glow discharge unit to render electron microscope grids and other surfaces hydrophilic. J Electron Microsc Tech 7(1), 29-33 (1987) ---------------------- Doug Keene DRK-at-shcc.org
See the following article for an easy to build glow discharge unit. I built a similar one several years ago for ionizing grids, and it works very well. All you need is a drill, a hack saw, a drill bit, and some epoxy. You will also need a plastic desciccator, a few feet of wire, a few machine and sheet metal screws, and some sheet aluminum about 1/8 to 1/16 of an inch thick. Mine is simpler than the one described in the article, consisting of only an aluminum disk screwed into the cut-off end of the high voltage generator, a larger disk placed inside the desciccator (with a wire going to an outside ground). You will also need a mechanical vacuum pump.
Aebi U, 1987 [See Related Articles] A glow discharge unit to render electron microscope grids and other surfaces hydrophilic. J Electron Microsc Tech 7(1), 29-33 (1987) ---------------------- Doug Keene DRK-at-shcc.org
Wolfgang: I , for one, am very interested in learning more about this technique. I started trying to do this last month without a lot of success. I just looked for your publication but unfortunately our somewhat mediocre library doesn't carry Mikroskopie . So it would be a great help to me if you could outline your procedure. I am especially interested in your formulation of silicon rubber and where you buy the components. Thanks in advance. Tom Phillips
-------------------------------------. } } 09/25/97 } Hi everywhere/everyone, } as a brand-new member of this listservice I want to add a notice on } self-fabrication of silicone rubber molds for epoxide-resin-embedding, } which Lonie Kerr asked for 10/28/96 (only for the case, someone has } similar problems now). } There are articles (in English) on self fabrication of moulds for } epoxide-embedding, at least 2 of them are: } 1) MUSS W., SIMONSBERGER P. (1983): Home-made Silicone Rubber Embedding } moulds for Electron Microscopy, Mikroskopie (Vienna) 40, 207-209 } (simple version for the unexperienced) } 2) MUSS W.(1984): Self fabrication of Silicone Rubber embedding Moulds for } use in Electron Microscopy; Mikroskopie (Vienna) 41, 34-39 } (sophisticated version for the unexperienced including silicone rubber } material to be used). } I am producing all my moulds (types as shown in article 2) like cubic, } flat, "beem capsule"- type w/round or pyramidal block face, with engraved } numbers for identification and others as requested) by myself. They are } needed in our routine diagnostic EM-Lab and fabricated since 1984 without } problems, with high quality in shearing force and unmoulding properties as } well as very low costs (quantity needed a year for about 1300 to 1600 } specimen blocks ~ 5 moulds 48 specimen cabinets/each; costs/mould ~ } US-Dollars 5-15.-, depending on amount of silicone rubber mass needed). } Fabrication is simple, if negative moulds of sufficient quality are at } hand and several general rules in working up the silicone mass are } strictly followed. } Silicone rubber moulds of this type are o.k. for use with epoxide resins, } use for hydrophilic resins like LR White, Lowicryls not tested yet. } If interested in how to produce such moulds efficiently and interested in } which kind/quality of silicone rubber one should use, please send e-mail } request to: } } Wolfgang MUSS PhD } Head of EM-Lab at Pathology Department LKA } Muellner Hauptstrasse 48 } A-5020 SALZBURG/AUSTRIA/Europe } phone: ++43++662+4482-4720 Ext. } fax: ++43++662+4482-882 Ext (c/o W. MUSS) } e-mail: W.Muss-at-lkasbg.gv.at. } } Good luck, hope this helps somebody. } END of Message, no attachment added.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
I published an article entiled as "A Modified Autowasher Device for Rapidly Washing Large Numbers of EM-grids" in Microscopy Researdh and Technique vol 26:177-179 (1993). Here I just copy the summary part for you:
This device consists of a siphon system and 5 or 10 grid disks, modified from the previous mode (Chen, 1973), for large numbers of grids with ultrathin sections. This method improves the ease of assembling grids onto the grids disk and also requires much less stain solution. This system only takes 5 min for one single stain washing, at a maximum of 100 grids, and also avoids stain contamination. The grid disk can also be used for immunocytochemistry work and for critical point drying of grids with biological specimens.
You may go to SPI Supplies website at http://www.cccbi.chester.pa.us/spi/new/stanwash.html/ for details.
Good luck,
Ming
On Wed, 24 Sep 1997, Chism, Sharron wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Greetings All, } After each step of the UA stain and the LC stain, I rinse the } copper grids by immersion. I have four changes of water and "dip" the } grid about 40 times in each change. I am interested in changing that to } the procedure that rinses the grid by "flooding" it using a syringe or } whatever. Can someone share with me exactly how that is done ... what } type of water ... etc. Also ... do the sections ever wash off the grid } when rinsing that way?? } } Thanks in advance, } Sharron Chism HT (ASCP) } Electron Microscopy Lab } Harris Methodist Hospital } Fort Worth, Texas }
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Mark, I've been using a product called imageAXS CE v3.0. This is a free version of a slightly more flexible commercial program. It automatically finds image files and creates a MS Access database which you can edit and use for searching and indexing images. the url is:
http://www.dascorp.com
Hope this helps glenn
Glenn Poirier Tel (514) 398 -6774 Electron Microprobe Laboratory Fax (514) 398 4680 Earth and Planetary Sciences glennp-at-stoner.eps.mcgill.ca McGill University http://castaing.eps.mcgill.ca 3450 University St. Montreal, Qc H3A 2A7
There are three sides to every story: Your side, My Side and the truth
Dear Tom, I have been casting my own silicon rubber molds for some time now. These are one-inch molds for epoxy metallurgical mounts. The silicon rubber comes from Dow Corning and they have several strengths and hardnesses. First, I have a kit consisting of several 100 ml tri-pour (plastic) beakers with a hole drilled in the bottom to fit a short screw. A block of aluminum, one inch in diameter and about 3/4 inch high, is tapped to receive the screw. You screw the aluminum block securely into the bottom of the beaker, mix the SiRubber (according to the directions) in a disposable cup, outgas the SiRubber in a vacuum desicator, then pour the rubber into the beaker. Outgas again. Leave it overnight to set, then remove the set rubber in the morning. It is a bit of a struggle to remove: I take out the screw, then push up on the Al block with a sharp point. Any flaps can be cut off with a razor blade. You can cast these things in any shape you can imagine.
You wrote: } Wolfgang: I , for one, am very interested in learning more about this } technique. I started trying to do this last month without a lot of } success. I just looked for your publication but unfortunately our somewhat } mediocre library doesn't carry Mikroskopie . So it would be a great help } to me if you could outline your procedure. I am especially interested in } your formulation of silicon rubber and where you buy the components. } Thanks in advance. Tom Phillips } } -------------------------------------. 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
What are some of the problems associated with making a photograph of a single atom? and What methods are used for getting around this problem? please send response to :
We currently use Potssium ferrocyanide reduced OsO4 as our 'routine' post fixative, primarily for enhanced membrane staining. When it works, tissue stains up beautifully! However, every now and then, we get tissue which appears to not have infiltrated properly ater using this post fixative. Nine times out of ten, it is a result of the potassium ferrocyanide (as repeat tests with new/no Pot Ferro show). According to the literature, (I think even to the original pot ferro paper) they do mention that extra care must be taken when infiltrating into epoxy resin when using this stuff. It only seems to be intermittent, even with fresh Pot ferro made up.
Does anybody know what is the pot ferro stock solution shelf life is? (we use stuff between 4 and 8 weeks old after 'brewing' for the first four) What does the pot ferro react with in the tissue to prevent good infiltration? Any other ideas/suggestions?
I'd appreciate any help on this,
Thanks
Rich.
P.S. Thanks for all the replies to an earlier request (4 - 5 weeks!) for info re: charging policies for EM Units too! :-)
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
Hello all, Has anyone been successful in labelling Substance-p for TEM? I am trying to tag it in endothelial cells, and have been having as much luck as a snowball would have in hell. Any help would be appreciated, as the old PhD may hinge on this some day! While I am here, if anyone has used DiI (3,3'-dioctadecyloxacarbocyanine percholate) or DiO (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine percholate) for neuronal tract tracing, please email me? ...................................................................... ....................................... Alex Black Department of Anatomy National University of Ireland Galway
alexander.black-at-ucg.ie
Alex Black BSc, MMedSc Department of Anatomy National University of Ireland, Galway Galway, Ireland
RENAL BIOPSY - IMMUNOFLUORESCENCE CONTROLS } Anyone doing immunofluorescenst techniques in renal biopsies running a control? } No.
One reason is that there is not enough tissue to go around. It is very difficult to get control sections from a positive case and there are not enough positive cases that are archived to pproduce the material.
Any suggestions? |--------------------------------| | Karlene Hewan-Lowe, M.B., B.S. | | Department of Pathology | | Emory University | | Phone: 404-686-2926 | | Fax: 404-6864978 | |--------------------------------|
Dear colleagues, thank you for replies to my topic from yesterday and your interest. = Lucky to see that there are some out there in need for such a procedure. = God bless you! To handle the problem via e-mail I think would go to far, concerning = spaceneeds of informations. Unfortunately I do have neither a homepage = (for the future there is planned one) nor a scanner unit, so I am not = able to reproduce those papers for you by e-mail. So I shall send ASAP = to all of you (see below) written infos to you. Unfortunately I don=B4t have original reprints of that papers mentioned = in my information.So you will get copies, as an attachment I shall send = Instructions which summarize the most important things in my opinion to = do the job as optimal as possible.=20 So my message to all who responded till now and others maybe following: Thank you all very much for your interest/greetings (Keith! welcome = greetings too!): - Jerzy BOHDANOWICZ, GDANSK/Poland, - Julian P.S. SMITH III, ROCK-HILL, SC/USA - Phil OSHEL, CHAMPAIGN, IL/USA (will contact you separately with = respect to "Microscopy today") - Tom PHILLIPS, COLUMBIA, MO/USA - Ann Fook YANG, OTTAWA, ONT/CANADA - Scott WHITTAKER, GAINESVILLE, FL/USA - Gabriel Adriano ROSA, BUENOS AIRES/ARGENTINA=20 - John J. BOZZOLA, CARBONDALE, IL/USA you will get written/copied information ASAP. Sincerely yours Wolfgang MUSS Dept. Pathology LKA (Gen.County Hospital), EM-Lab, Muellner Hauptstrasse = 48, A-5020 SALZBURG/AUSTRIA-Europe, phone: ++43++662+4482+4720 Ext, = Fax:++43++662+4482+882 Ext ("c/o W.MUSS") End of message, no attachment added.
Re to Sharron CHISM, Fort Worth Tx; 09/26/97 Dear Sharron, whatever type of aid (apparatus) you will use for staining your = ultrathins: concerning water quality: if possible, use UHQ (ultrahigh-purified = water) or at least bi- (better triple-)distilled water from a = quartz-glass distilling apparatus (most conveniently in your own lab!). = My/our story: our quartz-glass bi-distilling apparatus which at that time had a = life-span of nearly 12 years (in which time we got no bigger problems at = all) had broken down (heating wire burned through). Therefore we thought = to overcome the problem in ordering } } A.bidest.sterile, "non pyrogenic" = { { via our hospital pharmacy (+/- every time freshly produced, etc.). In = fact we got worsest stainings of our sections due to precipitates never = seen before in such an amount and shapes. We thought this to be a = possible source with respect to our handling in washing the grids, = unclean glass ware, syringes, etc.... or just more dust in the lab or = else; nothing of that all: despite using same methods for mixing up our = staining solutions as usual, we were not able to locate this source of = junk on the grids! When I asked at our pharmacy, how they produce their = water, they showed me & told me about their apparatus: it was/is a still = producing vapours by means of copper plates! An analytic chemist told me = some days after, that their central analytic lab got their own = distilling (UHQ) machine because of too high copper-contents of the = bidistilled water of the former source. Since that info I used only = their UHQ (coming along in clean glassware, most preferably quartz = glass) with no precipitate-problems any more, hoping to get my old = quartzglass still as soon as possible from repairing. Hope this adds another aspect in "hunting our elephantine precipitates", Best regards Wolfgang MUSS SALZBURG/Austria, Europe
Sharron, I also stain with Uranyl Acetate 5 min and Lead Citrate 5 min. I place copper grids, section side down, on drops of UA and LC. Inbetween stain and after I grasp edge of copper grid and flood by dripping from top of forcep for approx 10 sec each grid. I use Distilled, Sterile water to rinse. any reason for change. I know several techs that rinse as you do, but felt there needed to be changed. I've been staining as above for over 22 years. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to point out another source of contamination in stills: the tubing. Not the obvious crud, but breakdown products. We were using manufacturer-recommended Tygon tubing on our Barnsted still, and we getting some mysterious scum in the distillate. The micro-analytical lab couldn't identify it, other than as some complex organic kind of thing. Process of elimination ended at the tubing. The distillate was coming off at below, but not much below, the breakdown temperature of the tubing. The tubing was changed for higher-temperature rated silicone tubing. This seemed to solve the "new contamination" problem, but the previous contamination had coated the inside of the glass (not sure if it was quartz) carbouy and wouldn't clean off (not even will Tilex Scum Remover).
Phil } } Re to Sharron CHISM, Fort Worth Tx; 09/26/97 } Dear Sharron, } whatever type of aid (apparatus) you will use for staining your ultrathins: } concerning water quality: if possible, use UHQ (ultrahigh-purified water) } or at least bi- (better triple-)distilled water from a quartz-glass } distilling apparatus (most conveniently in your own lab!). My/our story: } our quartz-glass bi-distilling apparatus which at that time had a } life-span of nearly 12 years (in which time we got no bigger problems at } all) had broken down (heating wire burned through). Therefore we thought } to overcome the problem in ordering } } A.bidest.sterile, "non pyrogenic" { { } via our hospital pharmacy (+/- every time freshly produced, etc.). In fact } we got worsest stainings of our sections due to precipitates never seen } before in such an amount and shapes. We thought this to be a possible } source with respect to our handling in washing the grids, unclean glass } ware, syringes, etc.... or just more dust in the lab or else; nothing of } that all: despite using same methods for mixing up our staining solutions } as usual, we were not able to locate this source of junk on the grids! } When I asked at our pharmacy, how they produce their water, they showed me } & told me about their apparatus: it was/is a still producing vapours by } means of copper plates! An analytic chemist told me some days after, that } their central analytic lab got their own distilling (UHQ) machine because } of too high copper-contents of the bidistilled water of the former source. } Since that info I used only their UHQ (coming along in clean glassware, } most preferably quartz glass) with no precipitate-problems any more, } hoping to get my old quartzglass still as soon as possible from repairing. } Hope this adds another aspect in "hunting our elephantine precipitates", } Best regards } Wolfgang MUSS } SALZBURG/Austria, Europe
Among the awards to be presented next year is the "Morton D. Maser Distinguished Service Award," to be presented to an individual to ..."recognize outstanding volunteer service to the Society, ... and who has served the Society for many years with great dedication."
The nomination is to include a letter (this e-mail) from the primary nominator and supplemental letters (e-mails) of support from others.
It is with great personal pleasure that I nominate Nestor Zaluzec for this award.
Nestor is our friendly SYSOP, has organized the Argonne e.m. computer resources on behalf of microscopists everywhere, has organized the computer workshop/software exchanges at our meetings, served as MSA Program Chair for the Minneapolis meeting, etc., etc.
If you would like to provide a supplemental e-mail of support please direct your contribution to Gracie Burke, MSA Awards Committee. Deadline for input is December 31, 1997.
mgburke-at-pitt.edu
Thank you.
Ron Anderson
p.s. Please DO NOT send or copy your letters to the listserver, it's bad enough that Gracie will never speak to me again!
What is probably happening is that the paraffin wax is seperating from the monomers that have been added to it. This can happen under several circunstances. The most common being that the temperature on the paraffin dispencer is set too high for the wax being used. Another reason could be that the wax has been sittting unused (but hot) for too long a time (several weeks or more).
-- Begin original message --
} From: Gary Radice {gradice-at-richmond.edu}
} } I keep Paraplast Plus melted in a bath more or less continuously, so it is } always ready to use, even though it may be one or two months between uses. } Lately my students have been having some sectioning problems (with freshly } sharpened blades) and before I suggest that maybe they have done something } wrong during embedding, is it possible that the wax has "gone bad?" } } Gary Radice, Associate Professor gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond VA 23173 804-289-8233 (FAX) -- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
} } Malgorzata Warmuzek wrote: } } I have a problem with an import the file with an extension *.sp ( spectrum ) and put it to the } } document in word 6 or amipro
Dear Malgorzata,
I use the ISIS suite 3.0 and there is an easy way to copy spectra to any other windows program, maybe it works also with your version. Use the command 'Buttons' 'Print' and a new window will appear called 'Spectrum printing'. There it is possible to activate the command 'Edit' 'Copy'. The Spectrum will now be transfered to the clipboard in form of a vector graphic.
Kind regards
Rainer ------------------------------------------- Rainer Ziel Akzo Nobel Central Reasearch 63784 Obernburg - Germany
It's possible to make excellent moulds from the material that dentists use to make dental impressions. This is usually polyvinyl siloxane, though there is also a polyether material that is more awkward to work with. The polyviynl siloxane (Perfourm, Reprosil etc.) comes in tubes (cheaper) or in cartridges which give a much better result. We have been using these materials in our lab for many years. They are available from any dental supplier. Hope this helps. Lesley Weston.
On Thu, 25 Sep 1997, Wolfgang Muss wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } 09/25/97 } Hi everywhere/everyone, } as a brand-new member of this listservice I want to add a notice on self-fabrication of silicone rubber molds for epoxide-resin-embedding, which Lonie Kerr asked for 10/28/96 (only for the case, someone has similar problems now). } There are articles (in English) on self fabrication of moulds for epoxide-embedding, at least 2 of them are: } 1) MUSS W., SIMONSBERGER P. (1983): Home-made Silicone Rubber Embedding moulds for Electron Microscopy, Mikroskopie (Vienna) 40, 207-209 } (simple version for the unexperienced) } 2) MUSS W.(1984): Self fabrication of Silicone Rubber embedding Moulds for use in Electron Microscopy; Mikroskopie (Vienna) 41, 34-39 } (sophisticated version for the unexperienced including silicone rubber material to be used). } I am producing all my moulds (types as shown in article 2) like cubic, flat, "beem capsule"- type w/round or pyramidal block face, with engraved numbers for identification and others as requested) by myself. They are needed in our routine diagnostic EM- Lab and fabricated since 1984 without problems, with high quality in shearing force and unmoulding properties as well as very low costs (quantity needed a year for about 1300 to 1600 specimen blocks ~ 5 moulds 48 specimen cabinets/each; costs/mould ~ US- Dollars 5-15.-, depending on amount of silicone rubber mass needed). } Fabrication is simple, if negative moulds of sufficient quality are at hand and several general rules in working up the silicone mass are strictly followed. } Silicone rubber moulds of this type are o.k. for use with epoxide resins, use for hydrophilic resins like LR White, Lowicryls not tested yet. } If interested in how to produce such moulds efficiently and interested in which kind/quality of silicone rubber one should use, please send e-mail request to: } } Wolfgang MUSS PhD } Head of EM-Lab at Pathology Department LKA } Muellner Hauptstrasse 48 } A-5020 SALZBURG/AUSTRIA/Europe } phone: ++43++662+4482-4720 Ext. } fax: ++43++662+4482-882 Ext (c/o W. MUSS) } e-mail: W.Muss-at-lkasbg.gv.at. } } Good luck, hope this helps somebody. } END of Message, no attachment added. } }
We have a problem obtaining frozen ultra-thin sections of cultured cells. We have tried both 2.3 M sucrose and 20% PVP/1.8 M sucrose to infiltrate the cells before freezing without success. Briefly, after fixing the cells and rinsing with buffer, the plates are scraped and the cells are microfuged briefly to pellet the cells. Then we add a small amount (approximately 1X the volume of the cell pellet) of sucrose or PVP/sucrose to the pellet of cells, and tap the tube to resuspend the cells. The next day a small amount of the cell suspension is placed on a metal peg and frozen in liquid nitrogen. When sectioning we obtain snow-like flakes on the knife edge. Tissues processed in a similar manner cut well. We suspect the problem to be too much buffer associated with the cell pellet, even though we try to remove as much buffer from the pellet as possible. Any suggestions would be appreciated.
Christine Roy and David Hall Albert Einstein College of Medicine Bronx, NY
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Considerable! If you or anybody else can image a single atom in an SEM, then you're edging into Nobel Prize territory (although it's relatively easy in a dedicated STEM). Under the correct circumstances, it's not too difficult to do it in a TEM either. For example, isolated uranium atoms on a thin carbon film support are relatively easy. If you can manage a single sulphur atom on a carbon support in a SEM, then I guess you'll be off to Sweden. On the other hand, there are a number of 'probe' instruments - STM, AFM, etc - around which can visualise individual atoms without any difficulty. However, if you want to understand what the image really means, then it starts to get difficult:)
This seems to me to be another case of where the real answer to the question is a visit to your local library. I know they aren't necessarily 'high-tech', but there still isn't much around electronicaly to beat a visit to a good library, sitting down, going through the journals, indexes, citation catalogues, etc. It takes time but it will give you what you want - refereed papers in reputable journals.
Sharron, if you don't like the dip method try the dilution method, because any flooding method with the syringe or whatever may destroy your sections, try allowing the grid to float to the bottom of a small beaker (10-20 ml) repeat 3-5 times, pour off the excess water to retrieve the grid, and rember to pick it up by the edge of the grid. -MR
On Wed, 24 Sep 1997, Chism, Sharron wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Greetings All, } After each step of the UA stain and the LC stain, I rinse the } copper grids by immersion. I have four changes of water and "dip" the } grid about 40 times in each change. I am interested in changing that to } the procedure that rinses the grid by "flooding" it using a syringe or } whatever. Can someone share with me exactly how that is done ... what } type of water ... etc. Also ... do the sections ever wash off the grid } when rinsing that way?? } } Thanks in advance, } Sharron Chism HT (ASCP) } Electron Microscopy Lab } Harris Methodist Hospital } Fort Worth, Texas }
Sharron, if you don't like the dip method try the dilution method, because any flooding method with the syringe or whatever may destroy your sections, try allowing the grid to float to the bottom of a small beaker (10-20 ml) repeat 3-5 times, pour off the excess water to retrieve the grid, and rember to pick it up by the edge of the grid. -MR
On Wed, 24 Sep 1997, Chism, Sharron wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Greetings All, } After each step of the UA stain and the LC stain, I rinse the } copper grids by immersion. I have four changes of water and "dip" the } grid about 40 times in each change. I am interested in changing that to } the procedure that rinses the grid by "flooding" it using a syringe or } whatever. Can someone share with me exactly how that is done ... what } type of water ... etc. Also ... do the sections ever wash off the grid } when rinsing that way?? } } Thanks in advance, } Sharron Chism HT (ASCP) } Electron Microscopy Lab } Harris Methodist Hospital } Fort Worth, Texas }
Hello, all- Are any of you interested in a Denton DFE-3 Freeze Etch Unit that fits on a Denton DV-502 vacuum evaporator? Please don't respond unless you are reasonably sure you know what this is, and think you might want it, anyway! It seems to be in excellent condition.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I am finally starting to prepare some TEM samples of glass. Does anyone know whether a light coat of carbon should be applied to both sides of the sample or is just one side (I assume the top side in the microscope) good enough?
-Scott
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I have been asked to prepare some delicate fungal hyphae samples. I have a Denton critical point dryer and a freeze dryer. I'm more comfortable using the freeze dryer. Has anyone used a freeze drying technique on similar samples? Or does anyone in Alberta or British Columbia have a Denton critical point dryer that I might be able to come and see how to properly use. I'm not convinced that I'm using this one properly. Any suggestions appreciated.
It is indeed possible there may be excess buffer with the cells. Do you get *anything* in the way of sections that you can observe in the scope? If so, what do you see?
1. Do the fixed cells stay nicely as a pellet? After you fix and wash the cells, perhaps you could transitionally embed the pellet in low-melt agarose or 1.5% agar to hold the cells together. Do this only if the fixed pellet does not hold together on its own. Some fixed cells will form a nice pellet and stay that way. Others tend to go back into suspension.
2. *Definitely* increase the amount of cryoprotectant, well in excess of the volume of the pelletted cells. Try a trick which we used to use for some tissues: Assuming you have maybe a 50-100 microliter packed cell volume, fill a small 1-2 ml centrifuge tube, Eppendorf (TM) or similar, with the cryoprotectant. Then gently layer the cell pellet, embedded if necessary as in #1, above, on top of the cryoprotectant and very gently push the pellet *just under* the surface of the cryoprotectant. Allow the pellet to sink to the bottom of the tube on its own accord. This may take overnight. Infiltration with the cryoprotectant is complete when the pellet reaches the bottom of the tube. There is usually no need to leave the pellet in the cryoprotectant for longer periods of time. You can remove it and mount on a pin immediately after it reaches the bottom of the tube.
Good Luck! Let us know the secret after you get the problem solved.
Imaging atoms? Carefully consider the affects of potential probe size on the image, electron beam(and descrete potential electron optic subcomponents) and AFM tip size. SPM(AFM) and EM may be producing images heavily convoluted by tip or e-beam probe size contour. Atoms or multiple probe images?
---------- } From: Larry Stoter {LPS-at-teknesis.demon.co.uk} } To: mskittee-at-swbell.net; Microscopy-at-sparc5.microscopy.com } Subject: Re: SEM photography } Date: Friday, September 26, 1997 12:35 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } What are some of the problems associated with making a photograph of a } } single atom? and What methods are used for getting around this problem? } } please send response to : } } } } mskittee-at-swbell.net } } Considerable! If you or anybody else can image a single atom in an SEM, } then you're edging into Nobel Prize territory (although it's relatively } easy in a dedicated STEM). Under the correct circumstances, it's not too } difficult to do it in a TEM either. For example, isolated uranium atoms on } a thin carbon film support are relatively easy. If you can manage a single } sulphur atom on a carbon support in a SEM, then I guess you'll be off to } Sweden. On the other hand, there are a number of 'probe' instruments - STM, } AFM, etc - around which can visualise individual atoms without any } difficulty. However, if you want to understand what the image really means, } then it starts to get difficult:) } } This seems to me to be another case of where the real answer to the } question is a visit to your local library. I know they aren't necessarily } 'high-tech', but there still isn't much around electronicaly to beat a } visit to a good library, sitting down, going through the journals, indexes, } citation catalogues, etc. It takes time but it will give you what you want } - refereed papers in reputable journals. } } Regards, } Larry Stoter } } ------=_NextPart_000_01BCCADB.8B1B4500 Content-Type: text/html; charset=ISO-8859-1 Content-Transfer-Encoding: quoted-printable
{html} {head} {/head} {BODY bgcolor=3D"#FFFFFF"} {p} {font size=3D2 = color=3D"#000000" face=3D"Arial"} Imaging atoms? Carefully consider the = affects of potential probe size on the image, electron beam(and descrete = potential electron optic subcomponents) and AFM tip size. SPM(AFM) = and EM may be producing images heavily convoluted by tip or e-beam = probe size contour. Atoms or multiple probe images? {br} {font size=3D2 = color=3D"#008080"} {br} {br} {font color=3D"#000000"} ---------- {br} > =
} } I am finally starting to prepare some TEM samples of glass. Does anyone } know whether a light coat of carbon should be applied to both sides of the } sample or is just one side (I assume the top side in the microscope) good } enough? } -Scott
One side carbon coating is sufficient. It can be on the top or the down side in the microscope, though you should get a better image if you place it on top, actually. If you do not coat, be sure you will charge and explode the sample immediately.
} Date: Fri, 26 Sep 1997 15:16:43 -0400 } From: Dr. David Hall {hall-at-aecom.yu.edu} } To: Microscopy Forum {microscopy-at-sparc5.microscopy.com} } Subject: frozen thin sections of isolated cells } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We have a problem obtaining frozen ultra-thin sections of cultured cells. } We have tried both 2.3 M sucrose and 20% PVP/1.8 M sucrose to infiltrate } the cells before freezing without success. Briefly, after fixing the cells } and rinsing with buffer, the plates are scraped and the cells are } microfuged briefly to pellet the cells. Then we add a small amount } (approximately 1X the volume of the cell pellet) of sucrose or PVP/sucrose } to the pellet of cells, and tap the tube to resuspend the cells. The next } day a small amount of the cell suspension is placed on a metal peg and } frozen in liquid nitrogen. When sectioning we obtain snow-like flakes on } the knife edge. Tissues processed in a similar manner cut well. We } suspect the problem to be too much buffer associated with the cell pellet, } even though we try to remove as much buffer from the pellet as possible. } Any suggestions would be appreciated. } } Christine Roy and David Hall } Albert Einstein College of Medicine } Bronx, NY } Sounds to me as if your sections are sucrose (the "snow" or white stuff). I suspect your cells are thinly dispersed in the sucrose, and not stuck together enough to make a "section."
We add paraformaldehyde to the monolayers, swirl for just a few min (~5), scrape with a rubber policeman and pellet into a small tipped tube:
| | | | | | | | | | | | | | | | | | | | \ / H U almost exactly this size.
We pellet in a swinging bucket centrifuge and then microfuge to pack the cells. We let them fix for another hour or two and cut off the very bottom and again just above the cells, forming a log with the cells in the center that can be pushed out with a paper clip. If they stick together, fine, proceed. If not, push them into small piles (~0.5-1 mm), on a piece of Parafilm, drain them with filter paper cut into pie-shaped wedges using the very tip to touch the pellet gently, and coat them with cooled, still molter 1% agar. Cut away any excess agar. Inflitrate with 3 changes of sucrose (2.3M) over about 30-60 min. Place onto stubs and flash freeze. This keeps the cells together, not dispersed thinly in your sucrose.
If you fix very long before pelleting, your cells will not like to stick together, and will disperse in the sucrose. The consistency of the cell pellet should be like cooked oatmeal. (I could make some "snotty" comment about consistencies of other substances). If they are too wet, they will disperse, and you'll have to hunt all over your grid for them. If they're too dry, you could alter the ultrastructure.
Good luck. S
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Below is the result of your feedback form. It was submitted by (dmatthew-at-providence.edu) on Saturday, September 27, 1997 at 16:01:50 ---------------------------------------------------------------------------
Email: dmatthew-at-providence.edu Name: Douglas Matthews
School: Providence College
State: RI Question: Can anyone help me find a recipe for Millonig's phosphate buffer?
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Thanks!
Gracie Burke
Dr. M.G. Burke Westinghouse Electric Corp. Bettis Laboratory West Mifflin, PA 15122 tel: (412) 476-5883; fax: (412) 476-5151 e-mail: mgburke-at-pitt.edu
Dear Douglas, 09/29/97 original recipe for MILLONIG=B4s buffer was published as follows: MILLONIG G: Advantages of a Phosphate Buffer for OsO4-Solutions in = Fixation. Abstract B 26, publ. on behalf of the EMSA in: Journal of Applied Physics (LANCASTER =3D Procs of the ann. Meetings of = EMSA) 32, p.1637 (1961) Original Text of this abstract:
} } Since the sodium-mono- and diphosphate exists as an effective buffer = system in the body fluids of animals, it has seemed reasonable to test = it as a vehicle for OsO4 in fixation of biological tissues. An isotonic = (delta=3D -0.56 degrees centigrade) solution at pH 7.3 of the following = composition was tested: sol. a) 2,26% NaH2PO4 sol. b) 2.52% NaOH sol. c) 5.4% Glucose (about 5 ml added to an end volume of OsO4-soln. = of 50 ml.)
[sol. d): 41,5 ml sol. a) + 8.5 ml sol. b) [fixative: 45 ml sol. d) + 5 ml sol. c) + 0.5g OsO4 (as crystals)] [It has been found that this solution is stable for several weeks, if = stored in the refrigerator in a clean bottle. In comparative studies, = mainly with the veronal-acetate buffer + Osmium and on different animal = tissuies, the phosphate buffer seems to prevent extraction of the = cytoplasmic matrix, preserves the glycogen and fibrillar elements and = gives an uniform fixation at different levels in the tissue block { {] --------End of original abstract text of Millonig himself.---------
Recipe for convenient lab-volumes therefore: for an isotonic buffer solution ( ~ 300 mosmol, pH ~ 7.2 - 7.3 ) u = should mix the following: 5.65 g NaH2PO4.H2O ad 250 ml of A.dest ( =3D 33.9 g ad 1500 ml) 1.26 g NaOH-pastilles ad 51 ml A.bidest ( =3D 7.76 g ad 307 ml) makes: ~300 ml ( ~ 1800 ml)
You should measure pH, as well as osmolality: in my experience osmolality is a little bit lower than 300 mosmols therefore I have to add about 0.1 to 0.2 (w/v) of D-Glucose (~0.5 g, or = ~3.3g respectively, for endvolume of 1800 ml) to end up with 300 = mosmols.
MILLONIG=B4s 0.13 M sodium phosphate buffer (isoosmotic with mammalian = blood) (from: MILLONIG G. (ed): Laboratory Manual of Biological Electron = Microscopy; published and distributed by Mario SAVIOLO-Editore, = VERCELLI/ITALIA, copyright by G.Millonig, 1976 , 67 pages) Solution A: 0.164 M monosodium phosphate ( =3D 2.26% NaH2PO4.H2O, =3D 2.56% NaH2PO4 .2H2O) Solution B: 0.63 N sodium hydroxide (=3D 2.52% NaOH)
pH 6.0 6.2 6.4 6.8 7.0 7.2 = 7.4 7.6 7.8 ml A 96.2 94.7 92.5 87.9 85.8 83.9 82.5 = 81.6 80.8 ml B 3.8 5.3 7.5 12.1 14.2 16.1 = 17.5 18.4 19.2=20
The Centre for Microscopy and Microanalysis at the University of Queensland has the following specimen holders surplus. All holders to be sold in working order, the cold stage controller is not included. Interested parties should contact,
Assoc. Prof. Alasdair W. McDowall Deputy Director: Center for Microscopy & Microanalysis University of Queensland, Brisbane. QLD 4072 Phone: (07) 3365-4211 International: 61 (7) 3365-4211 Facsimile: (07) 3365-4422 International: 61 (7) 3365-4422 WWW: http://www.uq.oz.au/nanoworld/nanohome.html
EXCESS SPECIMEN HOLDERS DUE TO JEOL 4000FX-4010 UPGRADE JEOL 4000FX SPECIMEN HOLDERS (a) GATAN (D8020106-1) Normal double tilt, motorised, low background. Holder has had routine use. (b) GATAN (D8020105) Tilt holder, motorised rotation. Holder has not been used. (c) JEOL standard single tilt holder. Holder has had minimum use. Also Hitachi 200kV TEM H 800 Specimen Holder (d) GATAN ( D003130 ) LN2 holder. Holder has had minimum use.
Cost: Aus$ 5,000.00 per holder plus shipping.
-- ************************************************************ Duncan Waddell (BSc) Centre for Microscopy and Microanalysis The University of Queensland. St. Lucia. Qld. 4072 Telephone: +61-7-3365-4216 Facsimile: +61-7-3365-2199 WWW: http://www.uq.edu.au/nanoworld/nanohome.html ************************************************************ Any opinion expressed is that of the writer, and not necessarily that of CMM or of the University. ************************************************************
Following the thread about coating of a glass sample, Jacky Larnould added off line that when the objective aperture is set (above the sample) the "charging effect" disappears, and the sample does no break. While I have noticed this phenomenon for a while I have never come with a satisfactory explanation. Has anyone found a good explanation to this?
} I am finally starting to prepare some TEM samples of glass. Does anyone } know whether a light coat of carbon should be applied to both sides of the } sample or is just one side (I assume the top side in the microscope) good } enough? }
We only ever coat one side. We usually try to put the coated side towards the electron source, but not always. It has always worked fine in preventing charging. I've often wondered why this should be, but it certainly works, on either ceramics or polymers!
Tony.
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
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Reply to: RE} frozen thin sections of isolated cells
Dear Christine and David, We routinely prepare culture cell pellets for ultra-thin cryosectioning but we tend to put the resuspend the cells in gelatin then pellet this to form a nice gelled block of cells which can be treated the same as tissue for cryosectioning. Here is a copy of our protocol. If you have any questions please feel free to call me at 203-785-3646. You may also want to try adjusting the temperature and thickness you are cutting your cells. This may alleviate the "snow" effect. Linda Chicoine Center for Cell Imaging yale Univ. School of Medicine linda.iadarola-at-yale.edu EMBEDDING CELLS IN GELATIN/SUCROSE
1. Fix cells in buffered fixative directly in the culture dish. Can fix for 30' to overnight at 4C.
2. Remove fixative and replace with PBS/10%FCS to cover monolayer.
3. Scrape cells off culture dish with teflon , use glass pippette to transfer cells to an eppendorf tube.
4. Centrifuge gently to form a loose pellet.
5. Remove half the PBS/FCS and replace with 10% gelatin at a 1:1 dilution with PBS/FCS still in the tube. Final concentration 5% gelatin. Resuspend cells in this and re-centrifuge gently to form a pellet again.
6. Place tube in ice bucket for 15-30' or in fridge for longer, until gelatin has hardened.
7. Cut bottom of tube with a razor blade. Cut straight through tube to get the gelled pellet in the bottom piece of the tube.
8. In a petri dish of buffer or sucrose sitting on ice, remove the cell pellet from the bottom of the tube using a shaved wooden stick. Under a dissecting microscope, cut the pellet into small triangular or rectangular pieces.
9. Place peices in eppendorf tube filled with sucrose. Leave pieces for 30' or longer to infiltrate.
10. Pour sucrose into small petri dish, on ice,under disecting microscope. Remove a piece using capillary action of a fine point forceps and place the piece on a specimen nail. Check for excess sucrose around the pellet. Remove excess sucrose with a wedge of filter paper.
11. Immediately plunge the nail with the specimen into liquid nitrogen. Swirl the specimen somewhat vigorously and when completely frozen place the specimen in a nunc tube on an aluminum storage cane. Specimens can remain on the cane in liquid nitrogen for several years.
--------------------------------------
We have a problem obtaining frozen ultra-thin sections of cultured cells. We have tried both 2.3 M sucrose and 20% PVP/1.8 M sucrose to infiltrate the cells before freezing without success. Briefly, after fixing the cells and rinsing with buffer, the plates are scraped and the cells are microfuged briefly to pellet the cells. Then we add a small amount (approximately 1X the volume of the cell pellet) of sucrose or PVP/sucrose to the pellet of cells, and tap the tube to resuspend the cells. The next day a small amount of the cell suspension is placed on a metal peg and frozen in liquid nitrogen. When sectioning we obtain snow-like flakes on the knife edge. Tissues processed in a similar manner cut well. We suspect the problem to be too much buffer associated with the cell pellet, even though we try to remove as much buffer from the pellet as possible. Any suggestions would be appreciated.
Christine Roy and David Hall Albert Einstein College of Medicine Bronx, NY
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Hi Folks !
Does anyone of you know of some specific staining technique for Titanium, and where to find a cook-book for it? We need it for recovery of the new type of Gun Shot Residues.
Best regards Bengt Bengt Stocklassa , Managing Director Cox Analytical Systems AB | Phone: +46 31 7725300 House of Innovations, CTH | Fax: +46 31 7725600 412 96 - Gothenburg, SWEDEN | E-mail: bengt-at-xco.se
Hi Jill, I've done some preliminary work using Leica's FD unit on fungal material. I've FD Bergamot (Monarda didyma) leaves with a powdery mildew fungal infection and yeast (Sacc. cer). The leaves came out fine but the fungal hyphae were collapsed. The yeast came out very nicely. I ran both fixed (Karnovsky's) and unfixed pieces. All were cryo prepped by propane plunge freezing. All were examined at 2.0 kV uncoated. Overall I was pleased with the results, I haven't had a chance to go over all of the samples but the chemical fixation seemed to be better for maintaining the fungus. I would run both unfixed and fixed as I believe the mechanical agitation of adding a liquid solution may have washed away some fungal material. Aldehyde and/or Osmium vapor fixation would likely be less disruptive.
My FD time/temp schedule was: 48hrs-at--80C 36hrs-at--60C 24hrs-at--40C 12hrs-at--30C 12hrs-at--15C 24hrs-at--5C 6hrs-at-+10C 30hrs-at-+20C Vacuum was maintained with a cryo sorption pump.
I can post some images to my web page by the end of the week, if you are interested. The unfixed yeast pics are there already. Follow the links from Research Projects to Sample prep for SEM... http://www.personal.psu.edu/ejb11
cheers ed
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 http://www.personal.psu.edu/ejb11/
Let me modify my earlier suggestion for using the PrintScreen copying method to concur with Rainer's suggestion below. The PrintScreen trick still works, but other methods are probably more suitable when available. And this package does allow for copying the spectrum to the Windows clipboard.
You should be advised that you will probably have to use the Paste Special function to select the spectrum "picture" for pasting instead of the sample id "text". The text will come up as default.
At 08:51 PM 9/26/97 +0200, Rainer wrote: } } } Malgorzata Warmuzek wrote: } } } I have a problem with an import the file with an extension *.sp ( spectrum ) } and put it to the } } document in word 6 or amipro } } Dear Malgorzata, } } I use the ISIS suite 3.0 and there is an easy way to copy spectra to any other } windows program, maybe it works also with your version. Use the command } 'Buttons' 'Print' and a new window will appear called 'Spectrum printing'. } There it is possible to activate the command 'Edit' 'Copy'. The Spectrum will } now be transfered to the clipboard in form of a vector graphic. } } Kind regards } } Rainer ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering, or 270 Metals Development Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
Dear Colleagues I'm trying to cut 60 um parafin sections, but I've found it very difficult. It's rare to botain a good section There is some procedure to facilitate this work
Francisco Javier Hernandez Blazquez Centre commun de Quantimetrie, 8 avenue Rockefeller 69373 LYON CEDEX 08. France. Tel : (33) 4 78 77 75 19.
Responding to the message of {v03007804b05554e7eb94-at-[206.69.208.21]} from Yves Maniette {yves-at-giga.sct.ub.es} (by way of Nestor J. Zaluzec): } Following the thread about coating of a glass sample, Jacky Larnould added } off line that when the objective aperture is set (above the sample) the } "charging effect" disappears, and the sample does no break. While I have } noticed this phenomenon for a while I have never come with a satisfactory } explanation. Has anyone found a good explanation to this? } } Yves Maniette } We looked at this some time ago. JEOL machines work better than Philips, unless you install a really large aperture (we use 800 micron) and move to that aperture rather than moving the apertures out of the beam. The lack of symmetry induced by moving the aperture rod to the side rather than moving to an empty hole prevents the system from effectively stabilizing the charge, and the beam is displaced from the specimen.
The beneficial effect of the aperture is probably due to some capacitative charge dissipation from the sample by the very close proximity of the aperture. It is important to have everything properly aligned - beam and aperture, or the lack of radial symmetry will again deflect the beam away from the area of interest.
Different specimens respond differently - we have had very good success with single crystal alumina, but less success with polycrystalline specimens.
Good luck.
Stuart
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
We are trying to calculate the surface area of gap junctions. Has anyone done something similar to this? Are the gap junctional complexes (groups of connexons) arranged in a circular profile? If you could provide me with a reference or any information, I would appreciate it.
With regard to the following: ================================================= We are looking for any information we can find on the } microscopy/identification of particles from automobile tires. Seems we } have a case where we need to try to match particles from a pair of tennis shoes to a particular set of automobile tires(best case), or at least identify particles as possibly coming from automobile tires (more likely). ================================================= Assuming what you are saying is you have a need to demonstrate "common origin", which is not all that of an uncommon request, we have found the best approach is to use thin section TEM on the particulates. Tire-origin particulates have a nice characteristic dispersion of carbon black plus other inorganics.
The elastomeric system found in tennis shoes also has inorganic additions present. Generally speaking (based on our own in-house experience) the additive particles are larger than would be the carbon black and other particles found in a tire. Hence to differentiate between tires vs. tennis shoes, this should not be a major problem. If you wanted to show that the particulates came from a specific tire or a specific pair of tennis shoes, this would be a much higher level question, and considerable additional work (and the running of far more samples) would be indicaed.
The reason why this is more of a TEM than SEM request is that the particulates, especially the carbon black, is really below the practical limit of SEMs in terms of resolution and furthermore, the TEM view of this kind of sample is far easier to understand and interpret. And when EDS does have to be done, that data also is far easier to interpret and understand (because you don't have to deal with "depth" effects).
All thin sectioning on these kinds of samples must be done with the use of a good ultramicrotome, using a cryo stage, and using diamond knives, and we would always use a "Materials Science" diamond knife in order to not unnecessarily balloon up the cost of doing this kind of work.
Disclaimer: Our Structure Probe laboratories offer this kind of laboratory analytical service for clients needing to have done this kind of work. We are also a major provider of "materials science" diamond knives for persons wanting to do this kind thin sectioning.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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Charlie Kong wrote: } } Rainer, } } Have you found that the spectrum you got it by copying become a low resolution (287x287 pixels) graph? } } I have tried the method you described in the internet, and felt the difficulty to see the lebels of axis... } } p.s. I pasted the spectrum in Paint Shop Pro. } } Charlie Kong kong-at-t-rex.materials.unsw.edu.au
Dear Charlie,
The spectrum is copied to the clipboard in form of a vector graphic. PaintShopPro is a pixel graphic program. So the spectrum is converted when it is imported. You can import the graphic directly to your Word processor. In the german version of Winword it is done by the command 'Einfugen' 'Inhalte Einfugen' 'Graphic'. Afterwards it is possible to double click the graphic and to change e.g. labels.
Kind regards
Rainer
------------------------------------------- Rainer Ziel Akzo Nobel Central Reasearch 63784 Obernburg - Germany
On Mon, 29 Sep 1997, Francisco Javier Hernandez Blazquez wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Colleagues } I'm trying to cut 60 um parafin sections, but I've found it very difficult. } It's rare to botain a good section } There is some procedure to facilitate this work
A neat old trick for doing this is to use rubber cement in the mixture. It gives the paraffin the needed tensile strength to prevent crumbling.
David Feldser asked for references on microwave-accelerated staining for electron microscopy. I list 14 references FYI:
1. Estrada JC, Brinn NT, Bossen EH: A rapid method of staining ultrathin sections for surgical pathology TEM with the use of the microwave oven {italic} . {/italic} Am J Clin Pathol 1985, 83:639-641
2. Zondervan PE, de Jong A, Sorber CWJ, Kok LP, de Bruijn WC, van der Kwast TH: Microwave stimulated incubation in immunoelectron microscopy: a qualitative study {italic} . {/italic} Histochem J 1988, 20:359-364
3. Matsutani S, Yamamoto N: Improved methods for immuno-electron microscopy of cultured cells: use of a novel substrate and application of microwave irradiation {italic} . {/italic} Acta Histochem Cytochem 1990, 23:227-236
4. Wouterlood FG, Boon ME, Kok LP: Immunocytochemistry on free-floating sections of rat brain using microwave irradiation during the incubation in the primary antiserum: light and electron microscopy {italic} . {/italic} J Neurosci Methods 1990, 35:133-145
6. van Deuren B, van Reempts J, Borgers M: Microwave-enhanced silver staining of degenerating neuronal processes {italic} . {/italic} Eur J Morphol 1992, 24:597(abstract)
7. Giammara BL, Hopfer RL, Yates PE, Hanker JS: A rapid silver stain for the DNA of microorganisms cultured from AIDS or other immunocompromised patients {italic} . {/italic} Proc Twelfth Int'l Congr Electron Micros 1990, 48:762-763
8. Hanker J, Giammara B: Microwave-accelerated cytochemical stains for the image analysis and the electron microscopic examination of light microscopy diagnostic slides {italic} . {/italic} Scanning 1993, 15:67-80
11. Utsunomiya H, Shan L, Kawano I, Iwasaki A, Ono K, Kobayashi A, Kuma K, Kishikawa S, Kakudo K: Immunolocalization of parathyroid hormone in human parathyroid glands with special references to microwave antigen retrieval {italic} . {/italic} Endocr Pathol 1995, 6:223-227
12. Stirling JW, Graff PS: Antigen unmasking for immunoelectron microscopy: labeling is improved by treating with sodium ethoxide or sodium metaperiodate, then heating on retrieval medium {italic} . {/italic} J Histochem Cytochem 1995, 43:115-123.
13. Login GR, Dvorak AM. Microwave fixation and microwave staining methods for microscopy. In: Hayat MA ed. Immunogold-silver staining: methods and applications. Boca Raton, CRC Press, 1995, pp. 163-182
14. Login GR, Dvorak AM: The Microwave Toolbook. A Practical Guide for Microscopists. Boston, Beth Israel Hospital, 1994, pp. 184
Reminder: "Optimizing Light Microscopy" will be hosted by the Biology Department of Providence College this Friday, October 3. Course cost: $150,includes a copy of "Optimizing Light Microscopy for Biological and Clinical Laboratories". Despite the title of the book, the course has wide application to light microscopy in any venue.
For further information, write, call, or email:
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
Reminder: "Optimizing Light Microscopy" will be hosted by the Biology Department of Providence College this Friday, October 3. Course cost: $150,includes a copy of "Optimizing Light Microscopy for Biological and Clinical Laboratories". Despite the title of the book, the course has wide application to light microscopy in any venue.
For further information, write, call, or email:
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
Reminder: "Inside Fluorescence", a lecture-demonstration on factors affecting fluorescence microscopy, will be hosted this SATURDAY, OCTOBER 4, by Providence College.
Cost of class: $150 which includes a detailed course workbook.
For further information, please contact our office.
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
The best explanation for the effect of the objective aperture was published (I recall) in a Philips bulletin.
The section in a TEM has a positive charge generated in it as the primary beam produces secondary electrons in the section which then exit from it. This charge will destroy the section if it is not neutralised or conducted away through e.g. a carbon coat.
The objective aperture neutralises the positive charge in the specimen by reflecting back backscattered and secondary electrons which re-enter the section.
Mel Dickson Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Dear Yves, In every TEM I've seen, the objective aperture slides in just below the sample. In samples that are likely to break, such as formvar-covered slots, I was told never to look at the sample without the objective aperture in. Having all the high-kV electrons hitting on just the top side would pop the film. If the objective aperture is in, the high-kV electrons scattered back from the aperture below the specimen will balance the flux from above and the film probably won't break. BTW, it doesn' matter which side of the sample you coat, since the electrons go right through, anyway. Otherwise, it wouldn't be TEM. You wrote: } All, } } Following the thread about coating of a glass sample, Jacky Larnould added } off line that when the objective aperture is set (above the sample) the } "charging effect" disappears, and the sample does no break. While I have } noticed this phenomenon for a while I have never come with a satisfactory } explanation. Has anyone found a good explanation to this? } } Yves Maniette } 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
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I am interested in purchasing some time on a commercial ESEM to do some microscopy of fractures. Does anyone know of any ESEMs in CA, AZ, OR or WA that will provide commercial time? If so please email me at HWachob-at-fail.com. Thank You for your assistance.
Problem: I do have a excellent Zeiss stand and a complete series of Leitz planapochromatic objectives. However as you probable know Zeiss microscopes have a 160 mm mechanical tubelength whereas teh Leitz objectives are calculated for 170 mm. What kind of correction lens can I use in between the tube so that I can use the Leitz lenses on the Zeiss stand without loose of optical quality? Can anybody give me a good advice .
feldsdm4-at-juniata.edu wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Email: feldsdm4-at-juniata.edu } Name: David Feldser } } School: Juniata College } } Question: Where can i find a good source of information } about microwave staining technology for the } electron mircroscope? } } ---------------------------------------------------------------------------
Hi David,
this Link might be helpful http://www.rfglobalnet.com
--- ImmunoFluorescence Controls for Renal Biopsies------ From T. Fores: } Anyone doing immunofluorescenst techniques in renal biopsies running a control? } } From: Wanda } Hi Teresa: } } At the present we are not running a control on our immunofluorescent } techniques in renal biopsies. Has anyone replying back doing this? Even if we pooled resources, there would not be enough material to distribute to all laboratories that run this test. One solution is to run a known positive patient sample at the time titre the antibody. Another suggested solution was to run a tonsil control with each batch of patient samples (simplifying solution - the test material in each tissue is equivalent). And what about the control for the complement (C1, C3, C4) and fibrinogen?
Business idea: rats with immune mediated GN, frozen kidney samples on slides ...
The situation regarding immunofluorescence controls (IF) for renal biopsies is an example of the tension that exists between pure science and the practical application of science in clinical laboratories. Manuals are written and regulations are mandated without regard to the practical aspect of running a test in a clinical laboratory. Some rational solution is needed to stop laboratorians from resorting to draconian measures in order to comply with the regulatory agencies.
} Also would you know of anyone who might be interested in serving as a } consultant for EM or an EM Tech interested in relocating? I can consult on Transmission EM of human tissues. |--------------------------------| | Karlene Hewan-Lowe, M.B., B.S. | | Department of Pathology | | Emory University | | Phone: 404-686-2926 | | Fax: 404-6864978 | |--------------------------------|
Message text written by INTERNET:MWIS-at-crf.cuis.edu } microscopy-at-Sparc5.Microscopy.Com {
Check out PAX-it, an excellent image capture, archiving and databasing system. It has pretty powerful report generation software which allows y= ou to effectively replace traditioinal film in the photo documentation process. Going digital seems to pay for itself quickly. The other good thing is that it has network software, so you can access the database of images from your desktop PC's.
------------------------------------------------------------------------ If you work in MATERIALS R&D, SOLID STATE PHYSICS, CRYSTALLOGRAPHY or in a SCIENTIFIC LIBRARY, this is interesting for you.
A true compilation of all world literature data on MATERIALS CONSTITUTION; PHASE DIAGRAMS and RELATED DATA is available in print: the annual "RED BOOK" series . It started with the publication year 1990 and is produced by MSI and the Russian information service VINITI.
This compilation extracts data, diagrams and text from the world literature and arranges the information after alloy systems (not after publications), every year. - No need to browse hundreds of journals + proccedings - All foreign language publications translated into English - Best coverage of "eastern" publications available anywhere. - Uniformly structured detailed summaries of binary, ternary,..., multicomponent systems.
The introduction package, comprises the publication years 1990 to 1995 inclusive. It provides over 5300 summaries, more than 6000 diagrams and more than 1200 tables printed on over 8500 pages. Have a look at sample summaries and order information at
http://www.msiwp.com or inquire at info-at-msiwp.com
NOTE: The very attractive introduction offer expires mid December `97.
------------------------------------------------------------------------- This mailing list will inform you on the global phase diagram evaluation program of MSI and its team MSIT. It will announce how you can access its further products, such as the literature data base, electronic phase diagrams, etc. The traffic will be as low as 1 or 2 mailings per quarter. If this information is of no interest to you, please accept our appologies for the present message and send an e-mail to majordomo-at-msiwp.com with the message in the body: unsubscribe msi-adverts This will remove your address from the list.
Dear Microscopists I'm just beginning to use LR white as embedding medium for TEM- immuno- cytochemistry. For previous embeddings of plant root segments for conventional TEM research with SPURRs resin, I used flat embedding mold of silicon rubber for simple orientation of the segments. Is it possible to use flat embedding mold also with LR white and how is it possible to avoid contact with oxygen during polymerization? What kind of molds are otherwise most suitable for LR white? Furthermore I'm looking for a common embedding procedure of plant roots in LR white for use in TEM immunogold-labelling. I would like to contact in this way a lab with similar interests. Thanks to all comments.
Martin Bartels FB Biologie / AG Pflanzenoekologie C.v.O. University Oldenburg /Germany PO Box 2503 26111 Oldenburg phone ++49 441 798 3436 fax ++49 441 798 3436 e mail: bartels-at-uni-oldenburg.de
Here is a copy of replies I got about teh charging effect. One was sent off line. I made an horrible mistake saying that the aperture is above the sample, which is nonsense. Well let's say that I had a cold yesterday. YM.
All,
Following the thread about coating of a glass sample, Jacky Larnould added off line that when the objective aperture is set (below the sample) the "charging effect" disappears, and the sample does not break. While I have noticed this phenomenon for a while I have never come with a satisfactory explanation. Has anyone found a good explanation to this?
capacitance effect between the sample and aperture - the smaller the aperture the better for diffraction burgers vector analysis. Move the aperture around and note what happens. Also compare Philips CM and Jeol aperture geometries - you should find Philips easier to use with non conducting samples owing to aperture being nearer to specimen. It's been a while since I was doing this, however, when setting up diffraction conditions, it's best to use large objective aperture when locating yourself in the pattern. Removing the aperture entirely, as normally done, causes massive beam tilt in strongly charged samples (e.g. dirty MgO). cheers
We looked at this some time ago. JEOL machines work better than Philips, unless you install a really large aperture (we use 800 micron) and move to that aperture rather than moving the apertures out of the beam. The lack of symmetry induced by moving the aperture rod to the side rather than moving to an empty hole prevents the system from effectively stabilizing the charge, and the beam is displaced from the specimen.
The beneficial effect of the aperture is probably due to some capacitative charge dissipation from the sample by the very close proximity of the aperture. It is important to have everything properly aligned - beam and aperture, or the lack of radial symmetry will again deflect the beam away from the area of interest.
Different specimens respond differently - we have had very good success with single crystal alumina, but less success with polycrystalline specimens.
In every TEM I've seen, the objective aperture slides in just below the sample. In samples that are likely to break, such as formvar-covered slots, I was told never to look at the sample without the objective aperture in. Having all the high-kV electrons hitting on just the top side would pop the film. If the objective aperture is in, the high-kV electrons scattered back from the aperture below the specimen will balance the flux from above and the film probably won't break. BTW, it doesn' matter which side of the sample you coat, since the electrons go right through, anyway. Otherwise, it wouldn't be TEM.
The best explanation for the effect of the objective aperture was published (I recall) in a Philips bulletin.
The section in a TEM has a positive charge generated in it as the primary beam produces secondary electrons in the section which then exit from it. This charge will destroy the section if it is not neutralised or conducted away through e.g. a carbon coat.
The objective aperture neutralises the positive charge in the specimen by reflecting back backscattered and secondary electrons which re-enter the section.
To: Manufacturers of microscopes, imaging and image analysis systems, and sample preparation equipment
ASCB has informed us that there is still space available for MME to conduct market research at the upcoming Cell Biology meeting. If there is enough interest from you, we would be pleased to conduct a multi-client survey, by subscription. This meeting provides a prime venue for testing new ideas for instrument development. Since MME conducted research at this meeting in 1993, there are also select opportunities for long-term trend evaluation.
October 9th will be the deadline for our decision to go. Please note that there will be only one block of questions available per technology area and a total of approximately 25 questions, so space will be limited to first come/first served.
For further information, please respond directly to:
Barbara Foster Consortium President Microscopy/Marketing & Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Message text written by Randy Schnack } Microscopy-at-Sparc5.Microscopy.Com {
The Leco system is the one that most are using. It uses a high res CCD camera to provide film replacement using digital means. The software the= y provide is PAX-it, which does the archiving in an easy cabinet & folder style database. It has a really cool "report generation" tool that links=
all of the data fields with the images in MS Word and gives you a great print-out. =
The budget of 15k sounds about right for a complete computer, capture car= d, software AND high res camera. If the user wants to provide their own pentium, that will save some money as well.
Dear Yves, } } One side carbon coating is sufficient. It can be on the top or the down } side in the microscope, though you should get a better image if you place } it on top,
The image should be the same regardless of which side is coated. Assuming a perfect plane-wave for the incident beam, coating the bottom will cause the image--assumed to be perfect--to be convoluted with the scattering from the carbon; whereas, coating the top will form the image with a beam which has been convoluted with the same scattering. Both resulting images will be the same. Yours, Bill Tivol
Greetings to all, is there any person or EM-Lab out there who/which uses/used a resin = formulation with EPON 815 (Polysciences).=20 Would be glad to receive a mixing recipe which works/worked as I have a = small amount left for testing a staining procedure on semithin sections = made from original Epon embedded blocks. If also a reference could be = added it would be fine. Thanking you in advance Wolfgang MUSS, EM-Lab, Dept. Pathology, A-5020 SALZBURG/AUSTRIA, Europe e-mail: W.Muss-at-lkasbg.gv.at
This is a question for the Noran Voyager 4 users. After I obtain an image of my sample, on the monitor, is it possible to e-mail it from the Voyager to a person on MS Exchange? Does it have to be converted into a particular type of file? Does the person on the other end, the one receiving the image, have to convert it and then view as a certain type of file? Are instructions to do this in the Voyager manuals?
Hi would it be simpler to put a correction collar on the scope that would extend your tube length to 170mm?
Bob
On Mon, 29 Sep 1997, P.M. HOUPT wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } dear microscopists, } } Problem: I do have a excellent Zeiss stand and a complete series of } Leitz planapochromatic objectives. } However as you probable know Zeiss microscopes have a 160 mm mechanical } tubelength whereas teh Leitz objectives are calculated for 170 mm. } What kind of correction lens can I use in between the tube so that I can } use the Leitz lenses on the Zeiss stand without loose of optical } quality? } Can anybody give me a good advice . } } thank you in advance, } } Pieter Houpt } } (the Hague ,the Netherlands) } } }
I flat embed LR White all the time. I got an old Vacume oven out of surplus and hooked up a vacume pump and a dry nitrogen tank. When it is time for polymerization and the temperature is stable at 55C, I purge the chamber 3 times with nitrogen by pumping it down and letting in the nitrogen. On the last perge, I fill the chamber with the nitrogen leaving a slight vacume 1-3 lbs. Just to keep the door sealed. Polymerize for 24-48 hrs.
I use the peel-away polypropylene embedding molds. Dont use polystyrine.
On Wed, 1 Oct 1997, Martin Bartels wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopists } I'm just beginning to use LR white as embedding medium for TEM- immuno- } cytochemistry. For previous embeddings of plant root segments for } conventional } TEM research with SPURRs resin, I used flat embedding mold of silicon } rubber for } simple orientation of the segments. Is it possible to use flat embedding } mold } also with LR white and how is it possible to avoid contact with oxygen } during } polymerization? What kind of molds are otherwise most suitable for LR } white? } Furthermore I'm looking for a common embedding procedure of plant roots in } LR } white for use in TEM immunogold-labelling. I would like to contact in this } way } a lab with similar interests. } Thanks to all comments. } } Martin Bartels } FB Biologie / AG Pflanzenoekologie } C.v.O. University Oldenburg /Germany } PO Box 2503 } 26111 Oldenburg } phone ++49 441 798 3436 fax ++49 441 798 3436 } e mail: bartels-at-uni-oldenburg.de } }
There is a paper from 1987 describing how to make a simple glow-discharge unit. We made one from this design and have been using it successfully for years, slightly modified - the needle-type valve doesn't seem to be necessary, just close off the feed for the argon with a removable clamp. The reference is : Aebi U. and Pollard T.D., A glow discharge unit to render electron microscope grids and other surfaces hydrophilic. J. Electron Microsc.Technique, 7:29-33 (1987). Hope this helps. Lesley Weston.
On Thu, 25 Sep 1997, Gunnel Karlsson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello, } } I don't have access to a nice workshop, where they can construct a glow } discharger, where in Europe can I buy one? Or, is it out there someone, who } has an old mashine you don't need anymore? } } TIA } } Gunnel Karlsson } } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } Gunnel Karlsson E-mail Gunnel.Karlsson-at-oorg2.lth.se } Biomicroscopy Unit Tel +46 222 8229 } Inorganic Chemistry 2 Fax +46 222 4012 } Box 124 } S-221 00 LUND, Sweden } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } This message was sent by Eudora with recycled electrons } } }
There is a paper from 1987 describing how to make a simple glow-discharge unit. We made one from this design and have been using it successfully for years, slightly modified - the needle-type valve doesn't seem to be necessary, just close off the feed for the argon with a removable clamp. The reference is : Aebi U. and Pollard T.D., A glow discharge unit to render electron microscope grids and other surfaces hydrophilic. J. Electron Microsc.Technique, 7:29-33 (1987). Hope this helps. Lesley Weston.
On Thu, 25 Sep 1997, Gunnel Karlsson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello, } } I don't have access to a nice workshop, where they can construct a glow } discharger, where in Europe can I buy one? Or, is it out there someone, who } has an old mashine you don't need anymore? } } TIA } } Gunnel Karlsson } } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } Gunnel Karlsson E-mail Gunnel.Karlsson-at-oorg2.lth.se } Biomicroscopy Unit Tel +46 222 8229 } Inorganic Chemistry 2 Fax +46 222 4012 } Box 124 } S-221 00 LUND, Sweden } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } This message was sent by Eudora with recycled electrons } } }
At 09:17 AM 9/30/97 -0400, James Martin wrote: } Can anyone steer me to a website that has photomicrographs of histological } samples; in particular, blood samples?
There are a number of such sites. There are annotated links to the best of them from "The Histotech's Home Page" (http://www.histology.to). Go to "Peggy's Links" and select the section on images.
Best regards, Steven Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Some years ago our lab studied substance P and did some immuno-EM. There is a nicely detailed experimental procedure in this paper: "A substanceP-like peptide in bullfrog autonomic nerve terminals: anatomy biochemistry and physiology," C.W.Bowers, L.Y. Jan & Y.N. Jan, Neuroscience, vol 19, No 1, pp343-356, 1986. Good luck! Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
O.k., with all the positive comments about the Epson Stylus Photo printer, I have one more question: Since the postscript level 2 emmulation for the Stylus photo adds ~ 20% to the cost of the printer is it worth it? Every other printer I have has postscript capabilities and we generally use it (even though we're Intel PC / Windows based, not Macintosh) any opinions would be great! Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
This WWW site has hundreds of histology images, including several related to blood samples.
Best Regards,
Kirk J. Czymmek University of Delaware kirk-at-udel.edu
On Tue, 30 Sep 1997, James Martin wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Can anyone steer me to a website that has photomicrographs of histological } samples; in particular, blood samples? } } Thanks. } } James Martin } Williamstown Art Conservation Center } } }
Martin Bartels wrote: ================================================ I'm just beginning to use LR white as embedding medium for TEM- immuno- cytochemistry. For previous embeddings of plant root segments for conventional TEM research with SPURRs resin, I used flat embedding mold of silicon rubber for simple orientation of the segments. Is it possible to use flat embedding mold also with LR white and how is it possible to avoid contact with oxygen during polymerization? What kind of molds are otherwise most suitable for LR white? ================================================== One can use flat UV transparent silicone molds for this type of work. However, since the (transparent) silcone, in order to be UV transparent, does not have any of the additives normally incorporated to provide chemical resistance, don't expect long lifetimes, some people report being able to use a cavity (with L. R. White(TM) for example) only once or twice. The problem with oxygen exposure is easily "solved" by over filling the cavity with resin slightly, so there is a positive miniscus, and then placing another identical transparent mold on top, flat side down onto the over- filled cavities. Capillary action ensures that there is a quite adequate seal against the presence of oxygen.
Such molds can be purchased at the main suppliers of accessories of consumables for microscopy laboraotries including SPI, full details about which can be found on the SPI website, given below. These molds do not all "come out of the same source" and are not all the same, so don't assume the result experienced from one brand would be the same for all other brands.
Disclaimer: SPI Supplies manufactures transparent silicone molds for this application and we would obviously have an interest in seeing more people using these transparent molds.
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 ############################ ==================================================
"Electron, X-ray and Ion Spectroscopies-A Primer", an all day educational seminar, will be presented Friday, November 14, 1997, at Argonne National Laboratory. Advance registration is required. For more information contact Chas. Allen at allen-at-aaem.amc.anl.gov or Anita Brandes at g10809-at-email.mot.com including your complete mailing address. We will send you information and a registration form. Registration fee is $30 ($10 for students) which includes lunch and refreshments at breaks.
======================================== Charles W. Allen Electron Microscopy Center-HVEM-Tandem Facility MSD 212/E211 Argonne National Laboratory Argonne. IL 60439 USA
Email:allen-at-aaem.amc.anl.gov Tel: 630-252-4157 Fax:630-252-4798 (Note: On August 3,1996, Area Code changed from 708 to 630) ========================================
I'm trying to find a protocol that describes the TOTO or PATOTO (not potato!) technique for fixing plant specimens for SEM. I've tried doing a literature search and all I get are references to Dorothy and Oz, just kidding. I've read about the procedure but I cannot find a good descriptive protocol. Thanks for helping with this.
I'll get that pretty picture, and it's little dog, too.
Hoping a house doesn't fall on you,
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
To everyone with an e-mail address in my computer (sorry if you have this information already):
ALWYN EADES
(full name: John Alwyn Eades)
IS MOVING TO LEHIGH UNIVERSITY IN OCTOBER 1997 (moving in about ten days)
Present address
Materials Research Laboratory 104 S Goodwin Urbana Illinois 61801-2985
is moving to
Department of Materials Science and Engineering Lehigh University Whitaker Laboratory 5 East Packer Avenue Bethlehem PA 18015-3195
610 758 4231 610 758 4244 FAX jae5-at-lehigh.edu not activated yet. ** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones)
In mid-October this year (1997), I will be moving to Lehigh. The new address will be:
Department of Materials Science and Engineering Lehigh University Whitaker Laboratory 5 East Packer Avenue Bethlehem, PA 18015-3195
Peter: There is a more serious problem which I am sure the Zeiss and Leitz people will quickly point out. That is that the older series of objectives had to be used with a corresponding ocular to reduce optical aberrations, etc. in the objectives, giving you that high optical quality. Really old scopes had a way of adjusting the tube length, but that is the least of your problems in a more modern scope when you switch to another manufacturer's objectives. Of course the latest scopes have "infinity corrected" optics, but I wouldn't switch them either. If you use them, you will not have as good an image, but it would give you an image. The best tack would be to find an appropriate Leitz stand with appropriate oculars. Many older scopes are available, and without the optics are pretty cheap.
Regards, Mike
} } dear microscopists, } } } } Problem: I do have a excellent Zeiss stand and a complete series of } } Leitz planapochromatic objectives. } } However as you probable know Zeiss microscopes have a 160 mm mechanical } } tubelength whereas teh Leitz objectives are calculated for 170 mm. } } What kind of correction lens can I use in between the tube so that I can } } use the Leitz lenses on the Zeiss stand without loose of optical } } quality? } } Can anybody give me a good advice . } } } } thank you in advance, } } } } Pieter Houpt } } } } (the Hague ,the Netherlands) ================================================ Michael L. Boucher Sr. mboucher-at-isd.net 13345 Foliage Avenue Apple Valley, MN 55124-5603 Ph 612-432-8836 ================================================