David Feldser wrote: ================================================= Question: Where can i find a good source of information about microwave staining technology for the electron mircroscope? ================================================= The most often asked for book in this category, at SPI, is the following:
The Microwave Tool Book Authors: G. R. Login, DMD and A. M. Dvorak, MD http://www.cccbi.chester.pa.us/spi/catalog/books/book29.html
You can see the entire Table of Contents as well as some comments by reviewers, just to make sure this is what you really want.
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 ############################ ==================================================
Hello, I am an undergrad attending the University of Scranton. I am trying to find some beginner information regarding flourescent microscopy. While I have found some web-sites dealing with flourescence, they are all too advanced for me. It is the same situation with the school's library. Most of the information is not geared for the beginner. I am mostly interested in staining neural tissue (CNS).
Thank you very much for any help you can offer. It will be greatly appreciated.
Several list members have requested a summary of the responses to my inquiry after websites with histological images. Thanks again to all who responded, and apologies if I've inadvertently not included your response in this summary. Here goes:
- snip
The following sites contain histological images (including blood):
Scott Henderson, Ph.D. Director of Microscopy, Mount Sinai School of Medicine, Department of Cell Biology & Anatomy
- snip
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.
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
- snip
try at http://www.cba.arizona.edu/histology-lab.html
or http://www.hslib.washington.edu/courses/blood/
or at http://144.92.79.188/histology/histo.html
good luck
Gabriel Adriano Rosa Area Microscopia Electronica, Depto. Cs. Biologicas Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA
- snip
My Histology web page has a collection of links - they ought to get you what you want or at least send you in the correct direction.
} Date: Wed, 01 Oct 1997 11:06:53 -0400 } From: Edgar Voelkl {vog-at-ornl.gov} } X-Sender: vog-at-cosmail1.ctd.ornl.gov } To: "Dr. Larry F. Allard" {allardlfjr-at-ornl.gov} } MIME-version: 1.0 } Status: O } } Dear microscopists, } } I regret to inform you that Professor Gottfried Moellenstedt passed away } on September } 11th, close to midnight. As the father of the electron biprism he led } developments in the area of electron holography -- as well as } many other areas ! -- and contributed strongly to the reputation of the } University of Tuebingen, Germany, in the international community. } Professor } Moellenstedt will be missed by his many students and long-time } collaborators and colleagues. } } } } Dr. Edgar Voelkl } ORNL } Bldg 4515 } 1 Bethel Valley Road } P.O. Box 2008 } Oak Ridge, TN 37831-6064 } } Tel.: (423) 574-8181 } Fax: (423) 574-4913 } email: vog-at-ornl.gov }
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Hello to all, especially dear Rosey (AUSTRALIA) and Dr. GARBER (SPI, = USA), thank you two for your suggestions and greetings (espec. Rosey, for your = hello to the "lippizaners" (for all those not knowing what they are: = those being totally white colored, only male horses, serving for the = famous "Spanische Hofreitschule"- i.e. "Spanish k.k. royal riding = school" - in VIENNA/Austria since about 200 years now) but for all:answering in short for elucidation of my problem: 1) (for Dr. GARBER/SPI:) I meant to say: EPON 81 5 (eight fifteen); it = is a remnant of resin given to me without costs from Polysciences at the = trinational EM-congress at REGENSBURG (FRG, in Sept. 1997) 2) Rosey: thank you very much for sending me a recipe of your mixture = (find below my mixture) and the LUFT-reference.: do you use any left = lot(s) of originally EPON 812 or a substitute of this? If this is true, = would you=20 (or anyone, who uses or has in stock remainders of the original MONSANTO = / SHELL resin component) be so kind to send at a maximum of about 100 g = of that "old", original Epon 812 by gratitude to my Lab (adress given = below)???? 3) The problem, generally adressed, is: The production of the original resin component EPON 812 (Trade mark = of MONSANTO/SHELL) has been cancelled. No one -as to my knowledge- can order EPON 812 any = more. The substance EPON 812 has been replaced by chemically very = similar components (like SERVA/Bioproducts, Boehringer/Ingelheim: = Glycidether 100, which I am using now). A lot of Trade names have been = established by several supplying companies: EMBED 812, POLARBED 812, = SPI-Pon 812 ("exact direct replacement"). I have developed for use in our Lab about 10 years ago a simple, = polychromatic two-step staining for semithin sections, which was devised = for our needs with original EPON 812 resin embeddings. Since 1995 the = intended and standardized staining procedure failed to produce same = results in staining intensity like before. A long time I tried to = overcome that failing in looking forward to "unusual specimen = processing" (which was standardized at that time, too). Nothing happened = until I remembered as the only alteration in our standardized processing = the changing of resin component ORIGINAL Epon 812 (which I had in stock = for about 2 years more than being produced or available from suppliers) = to the replacement by "glycid ether 100 (SERVA)", which is, by = certificate, a 1,2,3-Propanetriol glycidyl ether with a MW of ~ 306.0 = and an epoxy equivalent (g/mol) of 150. Viscosity is certified as 196 = mPa.s at 25 degr. centigrade. I had to change and newly standardize the staining procedure in a very = hard labor, now including 2 short steps more, to get same results as = before. As an explanation for that I asssume the polymerizing character = of the new, replaced substance to be altered, leading to altered = staining properties of the sections too. This interpretation at the = moment is the only, because I had seen and noted an unusual changing of = resin color when mixing up the first three components (intensive red = color!; see below), which fades only to the (with old EPON 812) = "formerly seen yellowbrown color unless mixing components at least for, = say, 20 min, oxygen access provided (if overlayed by freon gas, to = shield from an increased humidity, this will last at least ~ 60 min!). = This was the only difference in my processing I could find! But there is = no difference in using a "red" or "yellowbrown" resin mixture (let me = assume it to be "unoxidized"/"oxidized" resin components): The routine = staining never achieved the same results without adding the two steps = "more". As I am going to prepare a manuscript for a paper on subject = "polychromatic staining of semithin epoxide ("EPON") sections", I should = be prepared to answer the question: "why the staining procedure formerly = did it without, and why now you need unalterable 2 steps more". So I am = trying to lock my hypothesis on altered polymerization properties of my = resin in staining "old fashioned EPON 812"-semithin sections, as = compared to variable EPON replacements. 3a) Hope, that you, Dr. Garber could supply me with any small amounts=20 (up to, say maximum 50 grams) of your SPI -Epon Replacement resin(s) = (kits?) to be able to perform my tests.
4) Rosey: My EPON-(Glycidether 100) mixture=20 ( based on the formerly used EPON 812, WPE ~ 146-150, according to the = A, B-formulations of LUFT 1961 ) is (measurement by weight):
45,23 g Glycidether 100 (SERVA =3D Bioproducts, Boehringer/Ingelheim, = FRG) + 13,29g DDSA (SERVA) + 28,68g MNA/NMA (SERVA) poured together, and according to GLAUERT: warmed up in an oven to 45 = degrees centigrade, then mixing with a special, selfmade glass stirrer, = after some minutes "red color" will appear, mixing as long as the = yello-brown color appears, then add your accelerator (DMP-30, SERVA) as e.g 1.35% (w/w) or as 1.75% = (w/w), respectively, according to Luft=B4s recommendation, to add 1-2% = of accelerator DMP-30. The color of the resin during and after mixing shall be yellow-brown; = polymerization in our lab is classically: 37, 45, 60-64 degrees C, ~ = 12-24 h (over night) each. If we have to perform "fast and hot" = polymerization, we infiltrate tissue specimens (up to 3 times for at = least 1 h/each) in and embed then specimens into a resin mixture = containing 2% (w/w) DMP-30 (acc. to LUFT). This modified resin recipe essentially is a "recalculated" w/w-mixture = of LUFT=B4s A:B-composition, in a relation of A:B =3D 3:7, which yields = polymerized blocks of a higher hardness, suited for human pathological = specimens (also large specimen cutting areas up to 4 x 5 mm) as we = process them now for about 15 years without problems. 5) To Dr. GARBER: we have met several times (I think) at congresses in = Europe as well as in the USA. Unfortunately I don=B4t know anything = about an "MCEM 98 meeting next week at Portoroz, YU", you mentioned. A = copy "directions for the use of SPI-Pon 812" I greatly should appreciate = either via this e-mail server or via my fax indicated below. Thank you = also for the informations on how to order saving 30% of the costs... Thank you very much for responding and your help,=20 other opinions or informations by any colleague out there on the subject =
(anyone out there, performing the two-step polychromatic staining = procedure of HUMPHREY and PITTMAN 1974: Azure II-Methyleneblue-basic = fuchsin?? on Epon 812 or EPON 812-replacement resins?? handling, = results, satisfaction?) are warmest welcome.
best regards=20 Wolfgang MUSS EM-Lab of Dept. Pathology, 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 END of message
Although I have commercial interest in this subject, I am mentioning it because the discussion of archiving systems is already out there, with specific systems listed...I mention our system as an alternative to those being discussed.
BUEHLER, LTD. also sells an image capture/archiving system; the M.A.R.S.(TM) System. M.A.R.S. comes complete with a camera and pentium computer. An optional high resolution (three chip) camera is available.
Features include image capture and enhancement; complete image annotation and editing; measurement tools including: Point to point, Parallel beam, Perimeter/ Area of a polygon, Radius, Angle, and Weld bead angle bisection; Hardness test measurement and Vickers/ Knoop calculation; Image comparison (5 images overlap for grain size comparison, etc.; and automated report generation through a WORD(R) interface. WORD templates come standard, but customized report templates are easily created. Web site info is available at http://www.buehlerltd.com
For more info, you can contact your local salesman or call (800)323-9330 or (847)295-6500.
Scott D. Holt BUEHLER, LTD. PO Box 1 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-6500 http://www.buehlerltd.com
Dear Reader: Is any one know how to fix and process 17 and 18 days mouse embryo in paraffin embedding? I have no problem to cut 16 days mouse embryo paraffin sections. However, not 17 and 18 days, those tissues are too mushy to cut. They looks like unfixed and wax cann't get into tissue. Thanks.
Dorothy Zhang Harvard School of public Health Building 2, CVLAB 677 Huntington Ave, Boston, MA 02115 Phone# 617-432-2970
} } http://www.lumen.luc.edu/lumen/MedEd/Histo/frames/histo_frames.html } http://www.usc.edu/hsc/med-sch/images/images.html } http://www.kumc.edu/instruction/medicine/anatomy/histoweb/ } } Links to these sites (and others) may be found at: } } http://www.pharmacy.arizona.edu/centers/tox_center/swehsc/exp_path/w3-hist } o.htm The last link does not work! Other may try as well out site which has many teaching modules and = link to other pathological sites. http://www1.omi.tulane.edu/classware/pathology/medical_pathology/overvi= ew.html. My own site has lots of inner ear stuff and it is listed as virtual = resource by the Association for Research in Otolaryngology at site = http://www.aro.org/
*Disclaimer: Whatever... is not Tulane =B9s opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web: http://www.tmc.tulane.edu/ferminlab, Internet: = fermin-at-tmc.tulane.edu 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
THE NEW ENGLAND SOCIETY FOR MICROSCOPY (NESM) will hold its October Meeti= ng at M/A-COM, Incorporated 1011 Pawtucket Boulevard, Lowell, MA, (508) 442-4400 on Wednesday, Octobe= r =
22, 1997 at 5:00 PM.
PROGRAM
5:00 pm Registration
5:10 pm Tours Note: 40-minute, escorted tours of the M/A-COM labs will=
start from the NESM registration area, leaving every 20 minutes (or as groups of 10 arrive) until around 6 pm. =
=
6:00 pm Buffet Dinner =
7:15 pm "How to Find Out Why" (Case studies illustrating the use of vario= us tools, including microscopy tools, to determine how and why microelectron= ic components fail) presented by Dana Crowe, Manager of Corporate Reliabilit= y Engineering, M/A-COM, Inc. =
=
8:00 pm "Endocytosis in Alveolar Epithelial Type II Cells" (A discussion = of potential pathways by which ultrafine particles cross the lung epithelium=
and enter the pulmonary interstitium) presented by Rebecca Stearns, Dept.=
of Environmental Health, Harvard School of Public Health =
=
NEW MEMBERS WELCOME!
To register, contact L. Kirstein at tel: 508-473-9673 or E-mail:104365.3522-at-compuserve.com. =
Cesar & others, Actually the URL was missing the "l" on .html part and I always have problems when my URLs are sent via email because the URLs that they allow me at the College of Pharmacy are so doggone long that they wrap to the next line [yes I've tried to get them to alias the sites, but it falls on deaf ears (sorry Cesar ;-) ]. Try using http://www.pharmacy.arizona.edu/exp_path.html and "drilling down" to get to the site.
BTW Cesar, the Tulane site you referenced looks like a good one.
Doug Cromey
At 12:03 PM 10/1/97 +0000, you wrote: } } http://www.lumen.luc.edu/lumen/MedEd/Histo/frames/histo_frames.html } } http://www.usc.edu/hsc/med-sch/images/images.html } } http://www.kumc.edu/instruction/medicine/anatomy/histoweb/ } } } } Links to these sites (and others) may be found at: } } } } http://www.pharmacy.arizona.edu/centers/tox_center/swehsc/exp_path/w3-hist } } o.htm } The last link does not work! } Other may try as well out site which has many teaching modules and link to other pathological sites.=20 } http://www1.omi.tulane.edu/classware/pathology/medical_pathology/overview.h tml. } My own site has lots of inner ear stuff and it is listed as virtual resource by the Association for Research in Otolaryngology at site http://www.aro.org/ } } *Disclaimer: Whatever... is not Tulane =B9s opinion! } Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology } web: http://www.tmc.tulane.edu/ferminlab, Internet: fermin-at-tmc.tulane.edu = =20 } 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 } Fax 504 587-7389, Voice 584-2521, Main Office 588-5224 } } ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
In light of Alwyn's upcoming departure from the University of Illinois a Web Page has been prepared for Alwyn on which you can record messages and regards for those of you who are unable to participate in person. The address is http://ginny.mrl.uiuc.edu/alwyn/ .
Roxanne Luesse Administrative Secretary Materials Research Laboratory 104 S. Goodwin Urbana, Illinois 61801 Phone: 217-333-1370 Fax: 217-244-2278
I published a paper on a modification of the TOTO technique which works well on plants. The reference is: FOOD STRUCTURE, Vol.12 (1993) pp.475-482. If you wish to talk to me about it, feel free to contact me directly.
Bill
William R. McManus Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305 1-801-797-1920 billEMac-at-cc.usu.edu
Wolfgang Muss wrote: ================================================ Unfortunately I donīt know anything about an "MCEM 98 meeting next week at Portoroz, YU ================================================ Acutually it is Portoroz, Slovenia, the dates are October 5-8, and information about it can be found by looking at the SPI website under "Hot Meetings". There is still time to register, contact person is Dr. Miram Ceh, E:mail {mcem97-at-ijs.si} .
Let me give a sales pitch for this meeting, my only "benefit" will be that maybe you might visit our SPI exhibit stand and say "hello":
1] You will be able to hear an outstanding lecture by "our own" Nestor Zaluzec on the topic of "Analytical Electron Microscopy and Materials Research at ANL". It is also my understanding that there is going to be some kind of round table discussion on the "future" in terms of internet communications between scientists in microscopy.
In fact, the quality of the scientific program is outstanding. Another "Plenary lecture" will be "Pigments of the Gastrointestinal Tract.A Saga of Botched Histochemistry" by Feroze N. Ghadially. There is an impressive number of poster presentations on virtually all areas where one uses EM.
2] You really should check out the MCEM website, this is a great meeting about to happen! And if you have have never been to that part of the world, I am told that there is nothing like visiting the old port city of Portoroz!
3] Portoroz is not more than a day's drive from many of the major capitals of Europe!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I believe you are refering to the OTO method described in the article "Thiocarbohydrazide-mediated osmium binding: a technique for protecting soft biological specimens in the scanning electron microscope" by Robert O Kelly, etal. in Principles and Techniques of Scanning Electron Microscopy. vol 4. 1975. ed by M.A.Hayat. ISBN 0-442-25686-8. This method enhances osmium binding and therefore reduces charging and heating of difficult to gold -coat structures. I am not sure how well this would work with plant material. Of course you may be refering to another method.
Hank Adams New Mexico State University Las Cruces, NM
My specimen holder in our Polaron E3000 CPD is not permitting the intermediate fluid to drain out the bottom, ie the tiny hole is blocked. I have removed the retaining screw and spring, but the slider rod is stuck. I have tried gently tapping it from bothh ends, but it will not budge.
Before I really try to sledgehammer it, I thought I would ask if it comes out of one end preferentially, ie from the end withh the retaining spring or the opposite end where it fits over the peg in the CPD itself. I'm assuming it is of uniform diameter with no t shoulders, but don't want to risk ruining it totally. I am soaking it in WD 40 overnight to see if that loosens it any.
You might prefer to email me directly. Thanks
Carolyn J. Emerson email: cemerson-at-plato.ucs.mun.ca
Biology Department Memorial University St. John's, NF A1B 3X9 Tel: (709) 737-7515 Fax: (709) 737-3018
We have had a rash of incorrect postings to the listserver, lately. Most fall into the following 3 categories...
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I have a little problem with a selective colour revealing the microstructure of the Ti6Al4V alloy - this is a wax-lost cast. The gas bubbles appear when etching by imersion a specimen in a water solution of the NH4F.HF reagent so a thin film of the reaction products cannot deposite on a ground face of a specimen. Finaly I receive the black-and-white results of etching.Do you know the reason of these bubbles creation or maybe you know some other etchants withoght NH4F.HF reagent to tint a microstructure of Ti-Al-V alloys ?
Best regards for all
Janina Radzikowska e-mail: jradz-at-iod.krakow.pl Foudry Research Institute Krakow, Poland
I have a little problem with a selective colour revealing the microstructure of the Ti6Al4V alloy - this is a wax-lost cast. The gas bubbles appear when etching by imersion a specimen in a water solution of the NH4F.HF reagent so a thin film of the reaction products cannot deposite on a ground face of a specimen. Finaly I receive the black-and-white results of etching.Do you know the reason of these bubbles creation or maybe you know some other etchants withoght NH4F.HF reagent to tint a microstructure of Ti-Al-V alloys ?
Best regards for all
Janina Radzikowska e-mail: jradz-at-iod.krakow.pl Foudry Research Institute Krakow, Poland
Janina.Radzikowska wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello! } } I have a little problem with a selective colour revealing the } microstructure of the Ti6Al4V alloy - this is a wax-lost cast. } The gas bubbles appear when etching by imersion a specimen in a } water solution of the NH4F.HF reagent so a thin film of the reaction } products cannot deposite on a ground face of a specimen. Finaly I receive } the black-and-white results of etching.Do you know the reason of these } bubbles creation or maybe you know some other etchants withoght NH4F.HF } reagent to tint a microstructure of Ti-Al-V alloys ? } } Best regards for all } } Janina Radzikowska e-mail: jradz-at-iod.krakow.pl } Foudry Research Institute } Krakow, Poland
Janina,
Here is some etchants for color etching of titanium and alloys (from http://www.kaker.com/etch/demo/etch.html):
Material: Titanium alloys (Ti) Type: Microetching Method: Physical etching Etchnant: Thermal etching Procedure: The specimen is polished and tinting removed from bakelite holder. Heating is done in a stainless steel or tungsten-filament basket in air. Basket temperature 1200 C (2192 F) approx. Specimen temperature approx. 400-600 C (752-1112 F). Time approx. 15-60 s. Air blowing improves both oxidation rate and time and specimen's temperature control. Remarks: Color etching. Different coloring of the matrix and the various phases is obtained. In titanium-aluminides-based alloys, the TiAl matrix usually appears yellow and brown. The Ti3Al phase usually appears blue or green. Reference: E. Beraha and B. Sphigler, Color Metallography, American Society for Metals (ASM), Metals Park, Ohio 440073, USA, 1977. p. 111.
Material: Titanium alloys (Ti) Type: Microetching Method: Chemical etching Etchant (electrolyte): 3 g ammonium bifluoridie and 4 ml hydrochloric acid (25 %) in 100 ml distilled water. Procedure: Immersion at room temperature for a few sonds. (To obtain good coloring, last polishing stage should be carried out in, or with, saturated solution of oxalic acid.). Remarks: Color etching. Alpha titanium grains are differently colored. sondary alpha and alpha-prime and various intermetallic phases (such as Ti2Cu) remain white (uncolored) or are evident by coloring. Reference: E. Beraha and B. Sphigler, Color Metallography, American Society for Metals (ASM), Metals Park, Ohio 440073, USA, 1977. p. 111.
Material: Titanium alloys (Ti) Type: Microetching Method: Chemical etching Etchant (electrolyte): 5 g ammonium bifluoride in 100 ml distilled water. Procedure: For pure titanium, immersion at room temperature for a few sonds. Longer etching time required for titanium alloys. Remarks: Color etching. Titanium alpha grains and twins are differently colored according to their crystallographic orientation. sondary phases are evident by coloring. Reference: E. Beraha and B. Sphigler, Color Metallography, American Society for Metals (ASM), Metals Park, Ohio 440073, USA, 1977. p. 111.
Material: Titanium alloys (Ti) Type: Microetching Method: Chemical etching Etchant (electrolyte): 2-3 g sodium molybdate, 5 ml hydrochloric acid (35 %) and 1-2 g ammonium bifluoride in 100 ml destilled water. Procedure: Immersion at room temperature until specimen surface occurs. Remarks: Color etching. Etching of as-cast titanium alloys.Titanium alpha matrix is colored blue or green. TiC is colored yellow or dark brown. Reference: E. Beraha and B. Sphigler, Color Metallography, American Society for Metals (ASM), Metals Park, Ohio 440073, USA, 1977. p. 111.
Henrik
-- Henrik Kaker SEM-EDS Laboratory, Metal Ravne d.o.o. Koroska c.14, 2390 Ravne, Slovenia Tel: +386-602-21-131, Fax: +386-602-20-436 SEM-EDS Laboratory Web Site: http://www2.arnes.si/guest/sgszmera1/index.html Microscopy Vendors Database: http://www.kaker.com/mvd/vendors.html Kaker.Com: http://www.kaker.com
} Let me give a sales pitch for this meeting, my only "benefit" will be that } maybe you might visit our SPI exhibit stand and say "hello": } } In fact, the quality of the scientific program is outstanding. Another } "Plenary lecture" will be "Pigments of the Gastrointestinal Tract.A Saga of } Botched Histochemistry" by Feroze N. Ghadially. There is an impressive } number of poster presentations on virtually all areas where one uses EM. } } Chuck }
Also, revised versions of papers presented at MCEM 97 will appear as a special issue of Journal of Computer Assisted Microscopy, early next year.
} } Is any one know how to fix and process 17 and 18 days mouse embryo in paraffin embedding? I have no problem to cut 16 days mouse embryo paraffin sections. However, not 17 and 18 days, those tissues are too mushy to cut. They looks like unfixed and wax cann't get into tissue. Thanks. { {
It could be fixation but it sounds like a combination of both inadequate fixation and processing. Some specifics would help ie: -Type of fixative, length of time in fixative -Are the specimens bisected prior to fixation and processing -What type of tissue processor is being used -What is your processing protocol, reagents, time in reagents, vacuum and temperature applied to which stations in the processor
Feel free to either e-mail me or call.
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
The magnetic field in our laboratory is too high for the new high resolutio= n=20 SEM we are going to install. I know of one active magnetic field=20 cancellation system (Oxfords). Can anyone tell me if there are other simila= r=20 systems in the market?
Best regards Lars Oestensson Graenges Technology=20
For years we have "airfuged" our fluorescent antibodies at 95 000 rpm before use to eliminate any aggregates which may have formed during storage. Does anyone know if this can (or should) be done with gold conjugated antibodies?
TIA,
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
This is a multi-part message in MIME format. --------------E32E6228270A2877B26D0CC7 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
Carolyn Emerson (by way of Nestor J. Zaluzec) wrote: } } } My specimen holder in our Polaron E3000 CPD is not permitting } the intermediate fluid to drain out the bottom, ie the tiny } hole is blocked. I have removed the retaining screw and spring, } but the slider rod is stuck. I have tried gently tapping it } from bothh ends, but it will not budge. } } Before I really try to sledgehammer it, I thought I would ask if } it comes out of one end preferentially, ie from the end withh } the retaining spring or the opposite end where it fits over the peg } in the CPD itself. I'm assuming it is of uniform diameter with } no t shoulders, but don't want to risk ruining it totally. I am } soaking it in WD 40 overnight to see if that loosens it any. } Carolyn, DO NOT TRY TO REMOVE IT FROM THE END WITHOUT THE SCREW. The rod has shoulders on it. Remove it from the screw end. I have the Jumbo model and the holders should be similar. The brass rod is most likely corroded. Good luck Gregory Rudomen S.U.N.Y. Stony Brook University Microscopy imaging center --------------E32E6228270A2877B26D0CC7 Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Rudomen, Greg Content-Disposition: attachment; filename="vcard.vcf"
Here are some replies taken from the "Tips & Tricks" archive I maintain. If you would like to check out the whole thread, got to the web address at the end of this message and follow the "Tips & Tricks" link. From there go to "SEM" and look fo the link dealing with fields. Good luck.
The guy I had contact with for the field compensation system at
Linear Reasearch Associates (it's a small company) is Curt Dunnam
(crd4-at-cornell.edu). He's the chief engineer, and will likley be happy to
talk.
Ben Simkin (simkin-at-egr.msu.edu)
simkin-at-egr.msu.edu
I've had similar problem with our microscope, and if it's truly fields,
I'd recommend just living with not using the outlet. AC fields can be activly
supressed if they are from an external source (we have purchased a system
from Linear Research Associates; they run ads in Microscopy Today (I don't
have the address with me right now)), but this sounds much more like a ground
loop, and the solutions to that (so I've been told) include either sinking
your own dedicated common ground (anywhere from easy to nearly impossible), or
disconnecting the ground connection between your asscessories and your SEM,
and running a "floating ground". I've discussed this as one solution with
our SEM service technician, and he says it sometimes works (assuming the
noise it adds to your instrument signal from the differance in "ground"
potentials is acceptably low).
Ben Simkin (simkin-at-egr.msu.edu)
Dept. Mat. Sci. and Mech.
Michigan State University
Incidentally, there was a series of articles in Microscopy Today (Don Grimes,
Ed., MicroToday-at-aol.com) recently on magnetic fields, their causes and
cures. It was written by Curt Dunnam of Linear Research Associates,
Trumansburg, NY 14886 (Fax: 770-368-8256). LRA apparently specializes in
diagnosing and dealing with magnetic fields in EM labs. You might get useful
help from them. I know there are other companies that do the same, but I
don't happen to have names and addresses readily available.
Wil Bigelow
Wil_Bigelow-at-mse.engin.umich.edu
Hello all,
thanks to all of you who have answered my questions. Here is the relevant
information:
1.1. Two addresse are quoted, maybe the first one is more recent:
THE DINDIMA GROUP P/L
10 Argent Place
RINGWOOG, Victoria, 3134
Australia
Telephone Number +61 3 9873 4455
AND/OR
Post Office Box 106
VERMONT,
VIC 3133
AUSTRALIA
PHONE;
+613-9873-4455
FAX:
+613-9873-4749
1.2. Email is: 100241.3642-at-compuserve.com.au
1.3. If you are in USA, better deal with Chuck (cgarber-at-2spi.com), he has
already done for you the custom process and distributes Arlunya products.
1.4. If you are in Pakistan or nearby country then it might be more useful
2. "Lupe" (actually binocular, sorry for the mistake) I have got several
interesting answers and will answer them privately.
At 02:38 PM 10/2/97 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
There is company based in USA that can provide an active field cancellation system, that may suit your needs.
address:
Integrated Dynamics Engineering Inc. 150-P New Boston Street Woburn MA 01801 USA
tel: (617) 938-5120 fax: (617) 938-5122
Disclaimer: I have no affiliation with this company, just a satisfied customer.
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
******************************************************** On Thu, 2 Oct 1997 lars.oestensson-at-techno.graenges.se wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } The magnetic field in our laboratory is too high for the new high resolution } SEM we are going to install. I know of one active magnetic field } cancellation system (Oxfords). Can anyone tell me if there are other similar } systems in the market? } } Best regards } Lars Oestensson } Graenges Technology }
We are a lab that does muscle and nerve biopsies, both histology and TEM. We would like to know if there are any atlas-type reference books available of TEM for muscle and nerve (possibly both normal and disease-state). We are interested in using it for teaching purposes (both lab personnel and MD residents).
Thanks in advance, Susan Danielson Medical College of WI Muscle/Nerve lab Milwaukee 414-259-3836
At 09:34 AM 10/2/97 -0700, Pat Hales wrote: } For years we have "airfuged" our fluorescent antibodies at 95 000 rpm before } use to eliminate any aggregates which may have formed during storage. Does } anyone know if this can (or should) be done with gold conjugated antibodies?
Unfortunately, there is no simple answer to this question. Our BioSite gold conjugates are prepared in such a way as to almost totally eliminate clusters (95% of the particles are guaranteed singlets), and centrifuging will actually create, rather than reduce, clumping. However, other gold colloids, produced by other manufacturers, are said to improve with centrifuging. I suggest that you ask the supplier of your gold conjugates for their recommendation.
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 following is being forwarded to the list as a courtesy. Do Not use E-mail reply function. Thank you. -Karen
Please respond by mail to: Dr. Douglas C. Youvan, CSO KAIROS Scientific Inc. 3350 Scott Blvd., Bldg 62 Santa Clara, CA 95054 USA
dyouvan-at-kairos-scientific.com wrote: } [Below is an]... announcement regarding staff positions that are currently open at KAIROS. The corresponding ad will appear in the October 10, 1997 issue of Science magazine. } } For the software enginnering position, we are especially interested in identifying graduate students or postdocs who are just finishing their studies and who have experience in C++ programming. } } Thanks, } } Doug } } ***************************************************************** ************************** } BASIC + APPLIED R&D POSITIONS } } KAIROS is integrating optical design and software engineering with molecular genetics to develop novel instrumentation, reagents, and methodologies in the fields of biotechnology, microscopy, medicine, and materials science. } } Successful candidates should relate their training and work experience to the content of our website: } } www.kairos-scientific.com } } KAIROS has three immediate openings for scientists and engineers trained in one or more of the following fields: } } Software Development (C++/MFC) } Optical Spectroscopy and Microscopy } Protein Engineering and Cell Biology } } Curriculum Vitate and names of references should be mailed to: } } Dr. Douglas C. Youvan, CSO } KAIROS Scientific Inc. } 3350 Scott Blvd., Bldg 62 } Santa Clara, CA 95054 USA } } Please do NOT respond via e-mail. } ***************************************************************** ************************** } -- } Douglas C. Youvan, Ph.D. } Chief Scientific Officer } KAIROS Scientific Inc. } Bldg. 62, 3350 Scott Blvd. } Santa Clara, CA 95054 } } T: 408-567-0400 x11 } F: 408-567-0440 } W: http://www.kairos-scientific.com } } Editor, Biotechnology et alia http://www.et-al.com
You are dealing with a whole set of interrelated, complicated problems. Briefly- Avoid useing the old Epon from Shell. Noone has it anymore, and if they do, they will not use it because of its unreliablilty between batches. (It was known to contain up to 13% of chlorine left over from the manufacturing process at one time). Noone will be able to duplicate your results, if you use the old Shell Epon in your publication. Use the replacements which are known to be the purified chemical equivalents and are not mixtures of Araldite, Epon, various dilutents (as some of them tend to be). Use LX-112, or Eponate 812.
The interaction between methylene blues and azures, etc. and epon sections are far more complicated than suspected. The coloration depends on osmium penetration (to some extent), on the percentage of unpolymerized monomers left in your tissue, etc. If you would send me your address via e-mail, I will send you a copy of my standard stain (methylene blue, azure, basic fuchsin) method which was a handout at an MSA meeting some years ago. Also you must pay close attention to your stains - are you buying the correct CI number? Have you changed suppliers?
I am short on time right now, but if you contact me with specific questions I would be happy to tell you what I know.
We are trying to do TEM (thin sections) on large liposomes containing cholesterol and triolein (triglyceride) droplets, fixing in solution at room temp then using freeze-substitution & Lowicryl HM23 to minimize extraction of lipids. Ruthenium tetroxide seemed to give better preservation of membrane structure than osmium did, but I don't see the triolein droplets any more. With osmium the triolein droplets are a nice uniform grey. With ruthenium there are dark structures that may be triolein droplets, but they are not homogeneous grey, rather they look like myelin figures, i.e. look like tightly wrapped layers of onion skin (phospholipid membrane?), dark with many concentric thin white lines (railroad tracks?). Particularly odd is that the thin white lines in these dark structures always look sharp as if the membranes were cut perpendicularly, but there is just no way we could always be cutting them perpendicularly.
Does anyone have experience with ruthenium and triglyceride? Is this what you would expect, or does triglyceride stained with ruthenium normally look similar to TG stained with osmium? (We are not experienced at processing liposomes, and these in particular are very sensitive to the process used for EM, in case you have any suggestions.)
I have some color photographic prints of blue fluorescence and yellow fluorescence. Does anyone know how well B&W photos of these prints will turn out? Are there any special filters/tricks that I could try?
TIA
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu
You can get the same effect as that provided by the large, bulky, and expensive revolving darkroom doors rather more simply by using two doors, separated by a small (3 ft) entry way chamber. Instead of solid doors, however, use heavy black curtains. For each of the doorways use two curtain panels that are fastened at the top and along opposite sides of the door frame, but which overlap by about 18 inches down the middle of the doorway. With this arrangement, you can walk through the curtain panels of the first doorway into the entry chamber. The curtain panels of the first doorway will close behind you, and then you simply walk through the panels on the sedcond doorway into or out of the darkroom.
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
From my experience with moving color photos over to B&W, the best way is to bring them into something like Adobe Photoshop and manipulate the grey scale. Blue and Yellow should respond well to this. Photoshop has a control called "Levels" which gives a histogram of grey levels then lets you set where you want the black, white and the midpoint to be within that histogram. I don't know what your pictures look like but if you had a print of a blue image on a black background, for instance, you could even adjust this so that the blue shows up white against the black, both being "pure" black and white if you desire (e.g. 0 and 256). Sometimes this is better than simply adjusting the "brightness" and "contrast" controls. There is also a similar adjustment called "curves" but to be honest, I never seem to get what I want out of that control (most likely due to ignorance on my part).
I hope this helps. Feel free to contact me off line if you need additional info.
Cheers,
John V. Pacific Northwest National Laboratory Richland, WA USA js_vetrano-at-pnl.gov (509)372-0724
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I have some color photographic prints of blue fluorescence and yellow fluorescence. Does anyone know how well B&W photos of these prints will turn out? Are there any special filters/tricks that I could try?
TIA
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu
Dear Bob: The biggest problem is to have the greys represent, light areas as fluorescing. I expect that the yellow will be no problem but the blue will probably be too dark. I would rephotograph the prints in B&W film using colour filters. If you remember the Oswald colour circle which has complimentary - opposite colours one can easily determine which colours will result in lighter or darker greys. So blue, photographed using a blue filter will result in a lighter grey. Orange in a darker. You can achieve the same with digital imaging and colour replacements. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au } } I have some color photographic prints of blue fluorescence and yellow } fluorescence. Does anyone know how well B&W photos of these prints will } turn out? Are there any special filters/tricks that I could try? } } TIA } } Bob } } Dr. Robert R. Wise } Department of Biology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu } }
} The magnetic field in our laboratory is too high for the new high resolution } SEM we are going to install. I know of one active magnetic field } cancellation system (Oxfords). Can anyone tell me if there are other similar } systems in the market? } } Best regards } Lars Oestensson } Graenges Technology
Hi Lars,
Besides the systems already mentioned by Scott and Fred, there is a system named EMF-1 produced by Advanced Research Systems in Illinois. You can find information about the system on the web-address:
http://www.mcs.net/~ars/ars/emf-1.htm
I have no experience with the system - I just came across this webpage some time ago.
Best regards, Joergen.
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
Dear Susan, one reference for that: W.J.Kenneth CUMMING, FULTHORPE J., HUDGSON P., MAHON M (Eds): Color Atlas of MUSCLE PATHOLOGY MOSBY-WOLFE (London) 1994, ISBN 0 7234 2016 5, iii-vi, 1- 202 (incl. = subj. index), ~ 140.- US-Dollars is, as to my knowledge, one of the most recently published color atlas, = dealing with Histology, Histochemistry, EM and a bit of Molecular = Biology in Muscle (+/- nerve/nerves not representatively included). For Nerve interpretation/concerning musculature you could look for:=20 a) RICHARDSON E. P. Jr., DeGIROLAMI U. (Eds) Pathology of the Peripheral = Nerve, (+/- TEM, B/W, also histology); Vol. 32 in the Series: Major Problems in Pathology (LiVOLSI Virginia, Consulting Editor). W.B. SAUNDERS (Philadelphia, London...) 1995, 1-164, incl. subj.index; = ISBN0-7216-3298-X (ordered from HARCOURT-BRACE-IOVANOVICH, London, = GB-Pounds ~46.- =3D ~58-60.- US-Dollars) b)VITAL C., VALLAT J.-M.(Eds) Ultrastructural Study of the Human = Diseased Peripheral Nerve, 2nd Ed., ELSEVIER, N.Y.,1987, 1- 286, incl. = subj. index; ISBN 0-444-01136-6, ~165.- US-Dollars
All three books are (in my opinion) worth their price; they are used = also for interpreting muscle cases. Hope this helps, best wishes and have a good day Wolfgang MUSS, A-5020 SALZBURG/Austria e-mail: W.Muss-at-lkasbg.gv.at.
I would be interested to hear from other SEM users with the same (JOEL JSM 5400) or a similar SEM what sort of filament life you are achieving ?
I fear that we may have a vacuum leak since the filament life is characteristically low and tarnishing is usually visible on the filament holder. The latter I am told may be an indication of a vacuum leak in the system.
3rd Oct.97 Dear all, today I got the data sheets (in English, as well as contact adress of = the selling company in USA) on the silicone rubber product I use for = long time. So I am able to send within the next 3-4 days my infos on how to produce = home-made silicone rubber embedding molds to all colleagues out there = who wanted to get that informations.=20 For those who wish/wished only e-mail info on the technique: I shall = contact you directly by e-mail within the next days.=20 A short version of how to plan/to do it, what type of silicone rubber I = use, technical hints and considerations, especially for posting in the = archives (to Scott WHITTAKER) is considered to be prepared next week.
Wish good luck to all=20 (hopefully my technique helps to solve problems. I should be glad = receiving a feedback, if anyone will try the whole.
Note added: I am/was producing and selling (at prime cost + 10% + postage) such = molds for some european as well US- colleagues who don=B4t/didn=B4t want = to fabricate their own negative-forms (which is the most expensive and = time-consuming part remembering that you should be able to produce not = only one or two, but many molds from them, but, a very nice part for = "left-brains") for conventional TEM-embeddings of = epoxide-resins/polyester-resins in following types: - cubic blocks, numbered 1-48 - cubic blocks, numbered 1-12, dito, numbered 1-10 - pyramidal, size + 0 - mold for specimen infiltration consisting of 7 by 4 rounded cavities, = ~ 1.5 ml resin / each cavity. If you want more information on that, feel free to mail to me your = questions or queries. =20 Disclaimer:=20 I have no financial interest in nor am affiliated to the company selling = the silicone rubber product and only speak as a satisfied customer.
Wolfgang MUSS, PhD. Dept. Pathol., LKA, EM-Lab Muellner Hauptstrasse 48 A-5020 SALZBURG, AUSTRIA phone: ++43++662+4482-4720 Ext. fax: ++43++662+4482-882 Ext (c/o W.Muss) e-mail: W.Muss-at-lkasbg.gv.at (character next right to -at- is a "small" L)
Jurgen Paetz wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Dear fellow SEM users } } I would be interested to hear from other SEM users with the same (JOEL } JSM 5400) or a similar SEM what sort of filament life you are achieving } ? } } I fear that we may have a vacuum leak since the filament life is } characteristically low and tarnishing is usually visible on the filament } holder. The latter I am told may be an indication of a vacuum leak in } the system. } } Regards } } J. Paetz (Senior mineralogist) } } Amplats Research Center } Republic of South Africa
Dear Jurgen Paetz,
You are correct that a discolored base is usually a sign of a poor vacuum. A normal burnout of a filament should have a bubble on top of the broken wire. If your filament burns out this way and the base is discolored it is probably a bad vacuum. If the filament is craked , no bubble, then their could be a flaw in the wire or a slight crack was made by human handling. We do not use a JEOL ourselves, but we do sell filaments for all the different scopes and our customers seem to feel you should get 50 - 200 hours out of a filament.
I have the same machine as yours with selected operation for low vacuum chamber (JSM-5400LV). For high vacuum operation, more than 100 hours, sometime 130 hours of filament life can be achieved.
****************************************** Zhiyu Wang Electron Microscope Lab and Imaging Center Western Kentucky University(WKU) Bowling Green KY 42101
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear fellow SEM users } } I would be interested to hear from other SEM users with the same (JOEL } JSM 5400) or a similar SEM what sort of filament life you are achieving } ? } } I fear that we may have a vacuum leak since the filament life is } characteristically low and tarnishing is usually visible on the filament } holder. The latter I am told may be an indication of a vacuum leak in } the system. } } Regards } } J. Paetz (Senior mineralogist) } } Amplats Research Center } Republic of South Africa } } }
We have a position open for an EM technician.(see below) Informal enquiries can be made to me cgilpin-at-man.ac.uk
UNIVERSITY OF MANCHESTER SCHOOL OF BIOLOGICAL SCIENCES
RESEARCH TECHNICIAN, GRADE C (ELECTRON MICROSCOPY)
Applications are invited for the position of electron microscope technician within the School of Biological Sciences' Electron Microscope, Graphics and Photography Unit. The post will involve working closely within a team of technicians within the Unit in providing assistance with electron microscope operation, sample preparation, image processing, image archiving, data interpretation, networking and computer software management and maintenance. Applicants should have working experience in electron microscopy and preferably a working knowledge of computers. The salary for this post is stlg11,365 p.a.
Application forms are available from Mr. A. Nicholas, School of Biological Sciences, University of Manchester, 2.205 Stopford Building, Oxford Road, Manchester M13 9PT UK. arthur.nicholas-at-man.ac.uk The deadline for applications is October 31, 1997.
The University of Manchester is an equal opportunities employer. 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
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I have had all of my nerve and muscle questions answered in; Myology by Andrew G. Engel and Betty Banker, Vol I and II, McGraw-Hill. I have no idea of the year of the latest edition or the price. My edition (1986) has 2106 pages. Kate Connolly
We are looking for a Reichert-Jung FC4E cryochamber and its control unit for our Reichert-Jung Ultracut E ultramicrotome. If you have an FC4E that you'd like to sell, please contact me.
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
Bob: The trick is to use Kodak Panalure II Repro RC paper which is designed for making B&W from color negatives. Tom
} I have some color photographic prints of blue fluorescence and yellow } fluorescence. Does anyone know how well B&W photos of these prints will } turn out? Are there any special filters/tricks that I could try?
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 some color photographic prints of blue fluorescence and yellow } fluorescence. Does anyone know how well B&W photos of these prints will } turn out? Are there any special filters/tricks that I could try? } } TIA } } Bob } Bob,
They should turn out fine. I take it you're meaning to take copy-stand photos of the prints? Just use the filters as if you're taking B&W negatives of the original preps. Yellow 12 filter to lighten the yellow and darken the blue, a blue filter for the reverse, a red to really darken the blue.
Or no filter. If the blue areas are nicely a blue (not light blue) they should naturally have a darker grey value. Try shooting a roll of a few prints, doing each one no filter, Yellow 12, blue (I forget the number). Maybe even a red.
I assume you're shooting with T Max? Another choice would be to use mumbletymumble, some orthochromatic B&W. This will have a reduced sensitivity to blue, and will shoot as if you were using a yellow filter (more-or-less).
The only other trick is the usual: watch for reflections off of the prints, but if you're using a copystand, this should be moot.
} I would be interested to hear from other SEM users with the same (JOEL } JSM 5400) or a similar SEM what sort of filament life you are achieving } I fear that we may have a vacuum leak since the filament life is } characteristically low and tarnishing is usually visible on the filament } holder. The latter I am told may be an indication of a vacuum leak in } the system.
How long is your filament lifetime?
I work on a CamScan SEM/EDX and have lifetimes from 90-240 hours, depending on how much EDX (20kV, SEM: 15kV) I made. The lifetime of the filament does not seem to depend on the sort of the filamnet, cheap ones have similar lifetimes to more expensive ones.
Our SEM has no lock, so we have to ventilate (with N2) the whole column, including the filament.
Greeings, O.Rother -- Oliver Rother Institute for Geology and Paleontology Scanning Electron Microscope (SEM) University of Kiel, Germany Tel. +49 431 35021 Fax: +49 431 35262
Reference to the problem with stray magnetic field. We have installed a Field Cancellation System by Spicer Consulting which we purchased from Agar Scientific,Stanstead, Essex, UK in all the labs in which we have high resolution instruents. They cost (3years ago) about 8000 Uk pounds each. They work VERY well.
Parick Echlin Multi-Imaging Centre University of Cambridge
On Thu, 2 Oct 1997 lars.oestensson-at-techno.graenges.se wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } The magnetic field in our laboratory is too high for the new high resolution } SEM we are going to install. I know of one active magnetic field } cancellation system (Oxfords). Can anyone tell me if there are other similar } systems in the market? } } Best regards } Lars Oestensson } Graenges Technology }
Rinse grids easily this way: Find a source of freshly, freshly, (not a typo) distilled water. (The collecting jars for the water must be maintained cleanly, as well as the tubing, and the water must not "sit" for days in the jars). If you must use deionized water be aware that it may not be as clean. That is, deionized water may be filtered through a 3 micrometer filter and then through a 1 micrometer filter. If you then apply a 0.22micrometer filter before use, the water may still not be really clean enough for TEM. Please be aware of all these possibilities if you see miscellaneous dirt on your sections. What most of us do not realize graphically is that a particle which passes through a 0o.22 micrometer filter may be really huge at a 15x mag. At any rate, fill a clean syringe with good quality water. Attach a 0.22 micrometer filter which you have cleaned by running through it (at a previous time) about 15cc of boiling water. (some filters contain dust aquired during the manufacturing process). When rinsing grids allow genrous quantities of water to run down the forceps and over the grids. Blot with dustless filter paper. If we have a lot of grids to do, we use the Hiroka Staining Kit. This works really well and is easy to use, especially if one only loads the center three or four rows of the pad. Most important - always pay attention to your water! It has to be as clean as you can get it. Bye, Hildy
How are the lines applied to TEM fluorescent screens? I have new screen that is blank and would like to apply at least a recognizable spot at the center, maybe a frame outlining the photo area. I'd think that I could use a drafting pen. True?
TIA, Owen
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
Since I have archived a discussion on how to make your own TEM screens, I would appreciate if you would be so kind as to send me the replies to this thread. It would make a nice addition. Thanks
At 11:41 AM 10/3/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
Chris, Terribly sorry chap, but I'm rather confused about the salary you offered in your posting for the technician position. Would you mind translating {stlg11,365 p.a.} into English, American English that is! ;-);-);-) ------------------------------------- Name: Winston W Wiggins E-mail: wwiggins-at-carolinas.org
Fellow Histoneters that responded to running controls on immunofluorescent (IF) studies. The response was not great. Maybe 6. The responses ranged from not running controls at all; to using tonsils, which seemed to appeas the inspectors; to being unrealistic "request that your pathologist or urolgist give you removed kidneys with tumors that can be negative for everything except IgG." As of 9/29/97 our lab had a positive (renal needle bx) lupus, in which I sectioned several blank slides, cold acetone fixed, and am storing in a -30oC. Upon receiving another IF case and tagging as per usual with IgG, IgM, IgA, C3, C1q, K & L, one of the known lupus slide will be tagged with Polyvalent (IgA, IgM, IgG, Kappa, Lambda). Thank you everyone for allyour imput and valuable help. If anyone else has a realistic suggestion, please e-mail me. Teresa
Richard Thrift asked about preservation of liposomes in embedded tissue. For traditional ethanol-dehydrated, epon-embedded preparations for TEM, check out reference by Angermuller and Fahimi from 1982 in Histochemical Journal 14:823-835 on imidazole-buffered osmium tetroxide. They obtained excellent preservation and staining of lipid droplets and lipoproteins with this technique applied to rat liver. Was particularly effective in preserving lipids with unsaturated fatty acids such has the oleic acid in triolein. We had good results in preserving emulsions composed of lecithin, cholesterol, and triolein. Don Gantz Boston Univ. School of Medicine gantz-at-med-biophd.bu.edu
We are in need of some extremely low power objectives (1X, 2.5X and 5X) lens for a Leica light microscope (all brands will be considered). The lenses should have flat field and extremely good resolution cababilities.
#################################################################### 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 ####################################################################
The National Institute of Standards & Technology has many Post Doctoral Positions open. These are offered competitively through the National Research Council. Within microscopy/microanalysis research areas we have several possible openings described at the following sites:
http://rap.nas.edu/lab/NIST/50837106.html This opportunity highlights our analytical research using Analytical Electron Microscopy/Compositional Mapping
http://rap.nas.edu/lab/NIST/50837109.html This opportunity highlights our Submicroscopic Chemical and Physical Characterization of Materials and Particles
http://rap.nas.edu/lab/NIST/50837110.html This opportunity highlights our Electron-Probe Microanalysis/Scanning Electron Microscopy research.
These are very general descriptions of broad areas of research. If you have research ideas that are related to these analytical approaches and are looking for a Post Doc opportunity, please contact me soon.
The NIST/NRC program offers a two year post doc at an annual salary of $45,500. The applications are due to the NRC in January 1998. This includes a technical proposal and several recommendations.
A candidate must be a U.S. citizen that receives their PhD and starts work at NIST by Jan 15, 1999 (You can start as early as July 1, 1998). So, this is the perfect opportunity for those of you that are graduating this spring through next fall. PLEASE NOTE that NIST/NRC only accepts applications ONCE a year, unlike some other institutions.
Eric B. Steel, Leader e-mail: eric.steel-at-nist.gov Microanalysis Research Group Office: 301-975-3902 N.I.S.T. FAX: 301-417-1321 Bldg. 222/Rm A113 Gaithersburg MD 20899 http://www-sims.nist.gov/Division/MicroGroup.html
I have used both techniques with varying results. CPD often caused artifacts, but also produced more attractive images than freeze drying. Many cells collaps with FD. My best results were obtained in cryo or with fresh hydrated samples (you get about 30 min. observation time before the cells collaps; depending on what your specimens are, of course). If you have access to an ESEM this may be the best bet.
best wishes
Sincerely +----------------------------------------------------------------- |Dr Stephan Helfer, SSO |Senior Mycologist - MSc Course Director | |Royal Botanic Garden Edinburgh, Inverleith Row, EDINBURGH EH3 5LR, |Scotland UK | |http://www.rbge.org.uk | |phone: +44 (0)131 552 7171 ext 280 | or +44 (0)131 459 0446-280 (direct digital VoiceMail line) |fax: +44 (0)131 552 0382 +------------------------------------------------------------------
"What is the best commercial source of consistently high-quality holey grids?"
This is a seductive question for a vendor to answer?
I am associated with Ted Pella Inc. in Redding, Northern California and would venture to say that our holey films are of consistently high quality.
I am not clear from your question whether you are looking for holey films for astigmatism correction or holey films for specimen application. If the latter - we supply a product we call NetMesh Grids, Lacey films. These contain many holes of different sizes in a netlike pattern and are very strong.
Please contact us at 1(800)237-3526 or 1(808)573-8945.
To everyone in my e-mail address file (sorry if you have this information already): please note that I am taking a new job and moving in about ten days.
ALWYN EADES
(full name: John Alwyn Eades)
IS MOVING TO LEHIGH UNIVERSITY IN OCTOBER 1997
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 yet activated ** 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
Filament life is a function of many factors. The presence of tarnishing usually signifies some sort of oxidation, implying a vacuum leak. I must say, if your SEM is a new one, that filament life usually improves over the first year of life. I think this is a result of outgassing the whole system. BTW, if you have a "bubble" at the end of your burnt-out filament, this is a result of over-saturating the filament. Remember to check the satuation level every hour for the first six hours of a new filament's life, as the saturation level drops fairly quickly, then levels off. After the second year, a filament lasts me a month.
I do not have a JEOL 5400, these are just general W-filament comments.
You wrote: } Dear fellow SEM users } } I would be interested to hear from other SEM users with the same (JOEL } JSM 5400) or a similar SEM what sort of filament life you are achieving } ? } } I fear that we may have a vacuum leak since the filament life is } characteristically low and tarnishing is usually visible on the filament } holder. The latter I am told may be an indication of a vacuum leak in } the system. } } Regards } } J. Paetz (Senior mineralogist) } } Amplats Research Center } Republic of South Africa } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I am impressed with, and use, holey carbon films prepared by Structure Probe. Dozens of grids with these films have been uniformly excellent. Recently I had a chance to use a couple of holey grids made by Pella, and they were excellent also. I don't recall the pricing comparison, but both are cheaper than I can make them myself (and probably better :-) ).
Larry
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Vachik Hacopian wrote: =================================================== What is the best commercial source of consistently high-quality holey grids? =================================================== As a long time manufacturer of filmed grids, including "holey" and "lacey" types, my answer would not be exactly that of an independent third party.
However, just remember one thing: The "making" of filmed grids is easy to describe, however the "art" of making a superb grid is not. But at the end of the day, no one knows for sure what they have made unless they have their own in-house TEM facilities to inspect the quality of their products. Be certain your intended vendor has their own facilities to check themselves what they are getting ready to send out their door.
Otherwise you, the customer, will be the QC department for that vendor yourself! Anyone who has been unhappy with purchased filmed grids in the past will know exactly what I am talking about. Customers are always welcome to visit our production facility and meet our staff members responsible for the in-house production and TEM inspection of our coated grids.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I have just come back from working in the USA and on reading my e-mail I saw your request and some of the answers provided.
Perhaps I have been in this game too long! I just had to sit down and tr= y to help you out as I feel the answers that I saw were not exactly correct= ? =
We are talking about Tungsten, the life of LaB6 is another question!
1. I can not speak directly for the instruments that you mention but =
ALL SEM are the same when it comes to filament life 1.1 Filament life is inversely proportional to the current you wish to=
draw from the gun, emission current measured in micro amps. 1.2 Typically Japanese instruments need ~100uA for good quality high resolution images, Philips use a lower current than other instruments usually between 30 and 50 uA at saturation 1.3 The position of the filament in the cathode and the bias setting decide the total current that you may pull from the gun. The nearer the filament to the cap the higher the current that you may attain. 1.4 To check to see if you have the "optimum - high performance" filame= nt position run up to 40,000X (working distance 10 to 15mm) with a nice specimen which gives lots of signal and see if you obtain an image (even = if its noisy) through the whole of the spot size (often called probe current= ) range. If you do the filament is fine if you do not the filament is in a= n "economy position" which will reduce current but increase filament life. =
If you need to work at high resolution you will need the filament in the optimum position, for less than 20,000X and for EDX this is not required.=
2. Filament life is also very dependant upon gun vacuum, it is possible = to have a poor gun vacuum even if the vacuum gauge says the vacuum is good. =
The gun will be very smelly in a poor vacuum environment, the chamber tending to be orange in colour and the filament base will also tend towar= ds an orange colour. Filament bases will ALWAYS show a colour, pale blue =3D=
low heating level low current use, dark blue =3D higher heating and more=
normal if the instrument is being used for highish performance, orange-brown =3D contamination through poor vacuum 3. Filament life also depends on how carefully the filament is saturated=
i.e. use a wave form and make sure you fully saturate but do not overheat=
the filament. 4. You should check saturation and alignment every time you switch the k= V on and every time you change the kV. As the filament ages and thins it will require a different current to attain saturation. When you change t= he kV the gun becomes more (up) or less (down) efficient and the saturation will change, on many instruments so will the emission current. 5. Due to the higher currents used in the SEM 90% of filaments failing under normal use will have small "melt" blobs at the failure point, this = is normal. A large melt blob usually means oversaturation. In the TEM the currents drawn are far lower and the usual break is between two tapered ends, blobs are more rare unless oversaturated.
So after all that how long should they last? Well 30 to 50 hours is OK i= f you are running in a normal lab environment. If you are pushing the resolution expect 15 to 30 hours. Aiming at better resolution than the manufacturer claims, then you must expect to have {10 hours filament life=
and you will end up with a very dirty gun and first condenser system; but=
you are getting more than you paid for! If you get well in excess of 50 hours my personal belief is that the instrument is not being used to its best effect, there is far more in a SEM than using it as a super light microscope! Of course it is horses for courses, unfortunately too few people realize just how much performance they could really pull from thei= r SEM if they only asked the instrument in the correct way for more performance. =
Electron microscope operators must realize that the filament is a consumable item! As I travel the world I really believe that the life of=
the filament in most laboratories takes priority over the quality of the image. Every laboratory I visit has the filament in the long life econom= y position and they almost all complain that the instrument will not do wha= t they want. You cannot get good high resolution or low kV results with ou= t plenty of beam current and that starts by setting the gun up correctly an= d in that case you must sacrifice filament life. Claims of 100 hours plus = in a SEM must mean the gun is under run, probably the operator is using the first peak - too big for imaging at any level of performance. Nothing wrong with this but any complaints of poor performance need to be rectifi= ed by first looking at filament position and saturation.
In my books "Working With a SEM" and "Maintaining and Monitoring the Electron Microscope" I discuss filament breaks and the colour of the base=
under different conditions.
Sorry to rabbit on but the problem of being in this game for 33 years is that you have seen it all before! It is so frustrating that SEM are considered by many people to be instruments to look at flies eyes or bee'= s knees, when they are really a scientific instrument with a very complex imaging system and tremendous potential when used correctly. You should = be able to run even a 10 year old instrument at 50,000X if it is set up correctly, we do time after time.
I am still looking for a TEM for checking quality of the em products I produce. Are their any free or low-priced Zeiss 9s or similar instruments out there. I do not need particularly high resolution but rather a machine that is relatively easy to maintain. I would be most grateful for any help.
The corollary to this argument is that only items made by the supplier can be trusted. Frankly, I would worry about a supplier who makes WDS standards, grids, apertures, refilaments and more. Nothing wrong with an in-house facility to film grids, but what is wrong with the manufacturer maintaining standards? I know of three (not counting SPI) EM consumable suppliers who have their grids filmed by suppliers with TEMs. Presumably all such coated grids are made with TEM access and checking. This 'contribution' to the server by SPI is, all things considered, another blatant advertisement and a waste of subscribers time. 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, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } From: Garber, Charles A. {cgarber-at-2spi.com} } Vachik Hacopian wrote: } =================================================== } What is the best commercial source of consistently high-quality holey grids? } =================================================== } As a long time manufacturer of filmed grids, including "holey" and "lacey" } types, my answer would not be exactly that of an independent third party. } } However, just remember one thing: The "making" of filmed grids is easy to } describe, however the "art" of making a superb grid is not. But at the end } of the day, no one knows for sure what they have made unless they have their } own in-house TEM facilities to inspect the quality of their products. Be } certain your intended vendor has their own facilities to check themselves } what they are getting ready to send out their door. } } Otherwise you, the customer, will be the QC department for that vendor } yourself! Anyone who has been unhappy with purchased filmed grids in the } past will know exactly what I am talking about. Customers are always } welcome to visit our production facility and meet our staff members } responsible for the in-house production and TEM inspection of our coated } grids. } } 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're looking for an apparatus for preparing vitrified TEM specimens, such as the Reichert KF80, which is not produced anymore. Does anybody have something like standing around?
Philip
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
Dear colleagues, 35 mm.color slides are still a relevant media for us. We have been addressing the problem of PC/slide interface. We have obtained a slide scanner, so the problem of putting our large slide inventory on disk is solved. The slide printers we are aware of are too expensive (US$ 5,000 range?) I am considering setting up a good quality flat screen monitor with a 35 mm. camera and appropriate lens, dedicated to tranforming PC screens to slides. Does anyone have experience with such an arrangement, or a better solution? Thanks for your help, Prof. Walter A. Mannheimer Dept. of Metallurgy and Materiais Eng. Federal University of Rio de Janeiro POBox 68505, 21945 Rio de Janeiro, Brazil Vox (55 21) 590-0579 Fax (55 21) 290-6626 wamann-at-metalmat.ufrj.br
I have been making slides this way for years, it works very well. It works best with slow film (ASA 25 or 64) and long exposures (2 to 10 seconds) so that screen flicker is averaged out. I use a 120mm lens (longer lens yield less distortion), tripod, and cable release. Use as dark of room as you can to avoid glares on the screen. Also, taping matte black paper over the edges of the monitor prevents them from showing up on your slide, giving a much more polished look. Be sure to place the mouse pointer out of the way while taking the picture (I tend not to notice the mouse on the monitor as it looks natural there but on a slide it looks like you are trying to highlight a feature). If color balance is important for your slides then you may have to do quite a bit of fiddling with the monitor. Also, it is important to adjust the image width and height to remove distortions before taking pictures. The easy way to do this is to display an image you know is a circle and adjust the monitor until it really is a circle.
Dan Moore
At 09:38 AM 10/6/97 EST3EDT, you wrote: } Dear colleagues, } 35 mm.color slides are still a relevant media for us. } We have been addressing the problem of PC/slide interface. } We have obtained a slide scanner, so the problem of putting our large } slide inventory on disk is solved. } The slide printers we are aware of are too expensive (US$ 5,000 } range?) } I am considering setting up a good quality flat screen monitor with a } 35 mm. camera and appropriate lens, dedicated to tranforming PC screens } to slides. Does anyone have experience with such an arrangement, } or a better solution? Thanks for your help,
INTERNATIONAL CONFERENCE ON PHYSICAL MESOMECHANICS AND COMPUTER AIDED DESIGN OF ADVANCED MATERIALS AND TECHNOLOGIES - M E S O M E C H A N I C S ' 9 8 (May 31 - June 2, 1998)
and/or at
WORKSHOP ON MICRO- AND MESOMECHANICS ASPECTS OF MATERIAL FAILURE - M E S O F A I L U R E ' 98 (June 3 - 4, 1998)
in Tel Aviv, Israel (one paper per presenter).
What is Mesomechanics? A new science, a branch of the physics and mechanics of deformation and fracture, has recently received its own name - Mesomechanics. The "meso" range of experimental and theoretical analysis of processes related to deformation and fracture of solids is defined as lying between the scale at which continuum mechanics is sufficiently accurate for a description of the events, and the atomistic scale. Within this range, several subscales can be identified, for instance those related to grains, grain boundaries, particles, shear bands, voids, microcracks and dislocations. To study phenomena within the meso range, new tools are often needed, sometimes outside the normal use of classical mechanics. Mesomechanics deals also with phenomena of self-organization in materials under loading.
Conference and Workshop Co-Chairmen: Prof. V.E. Panin, Director of State Research Center Institute of Strength Physics and Materials Science, Siberian Branch of Russian Academy of Sciences, Tomsk, Russia. Prof. R.L. Salganik, Center for Technological Education Holon, Israel. Prof. G.C. Sih, Xi'an Jiaotong University, China. Prof. M.P. Wnuk, University of Wisconsin, Milwaukee, USA
Conference Topics: -Physics and mechanics of heterogeneous media as a basis for computer aided design of advanced materials. -Computer aided design of advanced materials based on metals, ceramics and polymers. -Basic scientific principles of strengthening and surface treatment of materials. -Non-destructive testing based on mesomechanics. -Mesomechanics of fatigue. -Experimental methods of mesomechanics. -Fractals in mesomechanics. -Mathematical models and methods for mesomechanics. -Contact mesomechanics. -Strain localization processes at pre-fracture stage on meso-level of material structure. -Influence of yielding, irreversible deformation and fracture on phase transformations in solids on micro- and meso-levels. -Mesomechanics of time-dependent deformation, damage and fracture. -High rate deformation and fracture processes. -Mesomechanics of highspeed and shock wave deformation. -Mesomechanics of rocks and soils -Engineering applications of mesomechanics.
Some invited key-note lectures that will be presented to the Conference: Prof. K.B. Broberg, "Modelling of Materials in Mesofracture" (University College Dublin, Ireland); Prof. Y.C. Gao,"Fatigue Mechanism of Fiber-Reinforced Materials" (Northern Jiaotong University, China); Prof. J.K. Knowles, "Continuum Modeling of Solid-Solid Phase Transitions" (Caltech, USA); Prof. B.R. Lawn, "Damage Accumulation Beneath Hertzian Contacts in Ceramics and Other Brittle Materials" (National Institute of Standards and Technology, USA); Prof. G.C. Sih, "Transitional and Stability Character of Mesofracture" (Xi'an Jiaotong University, China); Prof. M.P. Wnuk, "Constitutive Modeling of Damage Accumulation and Fracture in Multiphase Materials" (University of Wisconsin, Milwaukee, USA)
A synopsis of one page should be sent or faxed and also sent by e-mail (if possible) to the Conference and Workshop Coordinator not later than October 31, 1997: see the Application and Information Request below.
Please inform your Colleagues to submit tentative paper titles immediately, thanks.
For more information please see
http://www.cteh.ac.il/happen/meso98.html
or address:
Prof. R.L. Salganik - the Conference and the Workshop Coordinator. Center for Technological Education Holon P.O. Box 305, Holon 58102. Israel
****************************************** INTERNATIONAL CONFERENCE: "MESOMECHANICS'98" May 31. - June 2, 1998 WORKSHOP: "MESOFAILURE'98" June 3 - 4, 1998 Tel Aviv, Israel
APPLICATION AND INFORMATION REQUEST
Please mail this form to: Prof. R.L. Salganik - the Conference and Workshop Coordinator. Center for Technological Education Holon, Affiliated with Tel Aviv University P.O. Box 305, Holon 58102, Israel
I intend to: =C9 attend the Conference/Workshop (Yes/No), =C9 present an oral/poster paper to the Conference (Yes/No), =C9 present a paper to the Workshop (Yes/No) entitled:______________________________________________________ ___________________________________________________________ ___________________________________________________________ ___________________________________________________________ ___________________________________________________________ ___________________________________________________________
A synopsis of one page should be sent or faxed and also sent by e-mail (if possible) to the Conference and Workshop Coordinator not later than October 31, 1997.
=C9My organization intends to participate in the Exhibition (Yes/No).
} } Dear colleagues, } 35 mm.color slides are still a relevant media for us. } We have been addressing the problem of PC/slide interface. } We have obtained a slide scanner, so the problem of putting our large } slide inventory on disk is solved. } The slide printers we are aware of are too expensive (US$ 5,000 } range?) } I am considering setting up a good quality flat screen monitor with a } 35 mm. camera and appropriate lens, dedicated to tranforming PC screens } to slides. Does anyone have experience with such an arrangement, } or a better solution? Thanks for your help, } Prof. Walter A. Mannheimer } Dept. of Metallurgy and Materiais Eng. } Federal University of Rio de Janeiro } POBox 68505, 21945 Rio de Janeiro, Brazil } Vox (55 21) 590-0579 Fax (55 21) 290-6626 } wamann-at-metalmat.ufrj.br
Walter, I've been doing this for several years and am quite happy with the results. We have a slide making service here that accepts PC disks and charges too much money for slide production. With my tripod, macro lens and color slide film I can shoot and process an entire role of 36 exposures for less than $10US.
A good quality monitor is important. I meter directly off of the screen and use a mid range f stop 8 - 11. For text with a lot of light background, I bracket the exposure times. Turn off room lights to avoid reflections. I've compared slides from my monitor to those produced by a slide film recorder and yes, the resolution is better from the expensive dedicated slide film recorder. But not that much better.
My .02 worth.
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/
Has anyone performed image processing and analysis on nerve? Following is the question posed to me, I am clueless on what I would be measuring/doing regarding nerve analysis. I do not have any IA experience with nerves.
Input from Experts on neurology would be appreciated.
I suspect that there are probably reports of quantitative approaches to assessing peripheral nerves?? Does anyone have any suggestions?
Project:
I will be working with a diabetic drug, I believe designed to be an insulin "secretagog" (don't quote me on that) which in a previous mouse study caused a peripheral neuropathy morphologically similar to the classic spontaneous ageing neuropathy of mice. The hypothesis is that drug related changes in blood glucose contributed to the pathogenesis and thus it is a pharmacologic effect.
What questions should I be asking Gregory.Argentieri-at-pharma.novartis.com
At 09:00 AM 10/3/97 +0200, Jurgen Paetz wrote: } I would be interested to hear from other SEM users with the same (JOEL } JSM 5400) or a similar SEM what sort of filament life you are achieving? } I fear that we may have a vacuum leak since the filament life is } characteristically low and tarnishing is usually visible on the filament } holder. The latter I am told may be an indication of a vacuum leak in } the system.
I want to add my corroboration to this diagnosis. A combination of short filament life and discolored filament base is a sure indication of a vacuum leak or other source of contamination.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Hello to Prof. Mannheimer and those reading along,
I'm a technician and SEM operator at the Univ. of Penn. I've had very acceptable results in 'screen shooting' PC monitor images using a 35 mm Nikon camera and a slow color slide film. All we did was completely darken the room, (no lights and shaded the windows) to avoid any screen glare from these sources, put the camera on a good tripod with cable shutter release (exposures can be anywhere from 1/2 sec. to 5-10 sec. at full open lens); position the camera so that the screen image fills the field, adjust the screen brightness and contrast for an optimal appearing image (what you see is what you get) and shoot. We use the camera's interior light meter and usually bracket each shot. The bracketing can be reduced, or avoided, once you become comfortable with the film sensitivity you're using and the screen brightness of the image.
Also, Daniel Moore's procedures seem real good. Thanks for the tip on matte black paper.
Hello, I am hoping someone out there has had some experience doing in-situ techniques using semi-thin plastic sections. Frozen sections of the tissue I will be working with does not provide adequate morphology. Any suggestions or references would be greatly appreciated. Thanks in advance, Gary
Hello, I am hoping someone out there has had some experience doing in-situ techniques using semi-thin plastic sections. Frozen sections of the tissue I will be working with does not provide adequate morphology. Any suggestions or references would be greatly appreciated. Thanks in advance, Gary
wamann2-at-METALMAT.UFRJ.BR wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Dear colleagues, } 35 mm.color slides are still a relevant media for us. } We have been addressing the problem of PC/slide interface. } We have obtained a slide scanner, so the problem of putting our large } slide inventory on disk is solved. } The slide printers we are aware of are too expensive (US$ 5,000 } range?) } I am considering setting up a good quality flat screen monitor with a } 35 mm. camera and appropriate lens, dedicated to tranforming PC screens } to slides. Does anyone have experience with such an arrangement, } or a better solution? Thanks for your help, } Prof. Walter A. Mannheimer } Dept. of Metallurgy and Materiais Eng. } Federal University of Rio de Janeiro } POBox 68505, 21945 Rio de Janeiro, Brazil } Vox (55 21) 590-0579 Fax (55 21) 290-6626 } wamann-at-metalmat.ufrj.brDear Dr. Mannheimer,
I have taken a large number of slides in this fashion. I have an old Olynpus OS2N camera with a Vivitar 90 mm Macro/Telephoto lens on it. This sort of lens (prefferably with a zoom), gives you lots of flexibility. As I remember, we used tungsten film and set the shutter ot 1/60th second. Since we had a non-interlaced screen, this worked very well.
Hope this information is helpful. Best of luck!
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 ********** Americas First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
Greg, Here's a publication that I was involved with years ago that did quantitation on peripheral nerve basement membrane thickness.
Johnson PC, Doll SC, Cromey DW (1986) Pathogenesis of diabetic neuropathy. Annals of Neurology 19:450-457.
Dr. Peter C. Johnson (formerly of the UA, and then the Barrow Neurological Institute in Phoenix AZ) and (as I recall) Dr. Peter J. Dyck (Mayo Clinic) were/are big users of morphometric approaches to peripheral nerves. I'd suggest running a MedLine on these two as a place to start for ideas.
Good Luck. Doug
At 05:11 PM 10/6/97 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
} How are the lines applied to TEM fluorescent screens? I have new screen } that is blank and would like to apply at least a recognizable spot at the } center, maybe a frame outlining the photo area. I'd think that I could use } a drafting pen. True? } AEI supplied us with a plastic template which had the proper posi- tions for the corners and centers marked for both "tilt" & "horiz" screen positions. We used to lay this over the screen and stick a probe through the holes in the plastic, then draw corners & crosses. Subsequently, we had our shop scribe the appropriate lines in our screen blanks with their smallest mill bit. When the phosphor is poured onto the blanks, the marks are quite visible. We also added marks for the middle of the film edges. Since there is ~2 mm uncertainty in the screen position from the mount, all these marks are only approximate. Yours, Bill Tivol
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Owen,
Trying to remember how I marked screens that I had cast for the 1MeV microscope in Madison a long time ago. I think that I used an extremely sharp pointed, very soft lead pencil. As I recall, I sharpened the pencil on very fine sandpaper and the surface was hard enough to write on. I would expect that ink from a Rapidograph-type pen would wick and make a wide blotchy mark.
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Good morning,
How are the lines applied to TEM fluorescent screens? I have new screen that is blank and would like to apply at least a recognizable spot at the center, maybe a frame outlining the photo area. I'd think that I could use a drafting pen. True?
TIA, Owen
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 --IMA.Boundary.189261678 Content-Type: text/plain; charset=US-ASCII; name="RFC822 message headers" Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part Content-Disposition: inline; filename="RFC822 message headers"
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REDUCED FEE REGISTRATION DEADLINE January 31, 1998 Workshop on Tripod Polishing
Workshop Objective This course will cover all aspects of pre-thinning and focus on final thinning for TEM via Tripod Polishing. Due to the limited class size an= d the extensive hands-on opportuinities, this course is well suited to novices as well as advanced Tripodders. Attendees will also learn the latest techniques available in ion milling and in plasma cleaning for TEM=
samples. The course will include sections on:
How to do it and why should I? What's really going on and what am I really seeing? How to prepare small, specific area cross-sections. The problem of wildly differing materials (eg tungsten). Rapid preparation of TEM cross-sections. Preparation of a wide range of materials: semiconductors, ceramics, metals,...
Hands-on Opportunity This course will be unique in that it will provide a hands-on opportunity=
for every class participant. Tripod Polishers, Polishing Wheels, and pre-thinning equipment will be made available to all participants and actual samples will be prepared - by the students - as part of the course= =2E =
This is a great opportunity to get your hands dirty and actually learn by=
doing. The instructors will walk you through each step of the process an= d then let you loose on the equipment. This course is designed to teach th= e Tripod Polishing technique. Silicon samples will be provided to the students and used as the basis for the course teaching.
Workshop Location and Dates South Bay Technology - San Clemente, CA Dates: Friday & Saturday - March 13 & 14
Previous Participants (partial list) INTEL, AMD, Motorola, LSI Logic, Conner Peripherals, Univ of Maryland, Un= iv of New Mexico, UNAM (Mexico), LG Electronics (Korea), Battelle, MEMC, MVA=
Inc., Univ of Michigan, U.S. Bureau of Mines, IBM, Naval Research Lab, Purdue Univ, Univ of Alabama, Univ of Arizona, Univ of Colorado, Univ of Wisconsin.
Class Size Due to the intensive hands-on aspects of this course, class size will be strictly limited to 10 participants.
Registration Fee: $795 (includes lunches and Friday night Dinner) $695 if registration fee paid by January 31, 1998=
Registration Deadline: 30 days prior to workshop
For additional Information: Monica Pflaster South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 TEL: 800-728-2233 FAX: 714-492-1499 e-mail: sbt-at-southbaytech.com
ON-LINE Registration available at: http://www.southbaytech.com
Registration Form
To register for the workshop, please fill out this form and send it, with=
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Payment must be made in the form of a check, money order, Visa or MasterCard. Checks must be drawn on a U.S. Bank and made payable to Sout= h Bay Technology, Inc. Credit card orders by FAX may be sent to South Bay Technology at 714-492-1499. Please do not send credit card information v= ia e-mail.
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We are embarking on some bromo-deoxyUridine (BrDU) studies with cell cultures and are wondering if we need to make up fresh stocks each time or can we sterile filter and store at 4 C or -80C. Does anybody have practical experience with its stability? TIA.
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 know this is sort of off the topic of the listserv, but can anyone help a colleague who is:
looking for any information on a company or individual who sells Anatoxin-a(s) [P=O structure]. This is a toxin isolated from the blue-green freshwater algae Anabaena flos-aquae.
Or if anyone knows how the toxicity differs between anatoxin-a(s) and the synthetic anatoxin-a fumarate (produced by Sigma and others).
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Jurgen Paetz wrote: } } } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------.} } } Dear fellow SEM users } } } } I would be interested to hear from other SEM users with the same (JOEL } } JSM 5400) or a similar SEM what sort of filament life you are achieving } } ? } } } } I fear that we may have a vacuum leak since the filament life is } } characteristically low and tarnishing is usually visible on the filament } } holder. The latter I am told may be an indication of a vacuum leak in } } the system. } } } } Regards } } } } J. Paetz (Senior mineralogist) } } } } Amplats Research Center } } Republic of South Africa } } } } Dear Jurgen Paetz, } } You are correct that a discolored base is usually a sign of a poor } vacuum. A normal burnout of a filament should have a bubble on top of } the broken wire. If your filament burns out this way and the base is } discolored it is probably a bad vacuum. If the filament is craked , no } bubble, then their could be a flaw in the wire or a slight crack was } made by human handling. } We do not use a JEOL ourselves, but we do sell filaments for all the } different scopes and our customers seem to feel you should get 50 - 200 } hours out of a filament. } } John Arnott } Ladd Research } We have a JEOL 5800LV. Our filiment life on that machine has been 150-200. We feel that is low. Our JEOL 733 filiments last in the 1000's of hours. The 5800 burnt filiments display the small "ball" (normally only present on one end of the break unless the filiment has been very over-driven). The bases of the filiments are always slightly discolored and we do not have a vacuum leak (that we know of). The bases may get discolored when we work in LV mode (though I have been assured that the vacuum in the gun chamber stays very high). Our JEOL 733 filiment bases are always somewhat discolored, and we watch the gun chamber vacuum very closly, so I know there is no leak.
One thing that may extend the filiment life is allowing the filiment to cool before venting the chamber (I don't know if the 5400 uses an exchange port or just vents the chamber). On our Hitachi S-450 we found that doing this does increase filiment life, but the 5800 is new and we have just started cooling the filiment, so I do not know what the effect will be. I do know that machines that change Acc kV on the fly have shorter filiment lives.
I write this not to contradict John Arnott's above statement, but just to show that there are "extreme" differences in machine filiment lives that are not just based on the type of machine being used but also possibly on the way the machine is used. The number 50-200 hours seems low to me, but most of my experience is on the 733, so maybe I am spoiled. As far as discoloration being a sign of a leak, I bet it is, but how much discoloration is normal should also be a question.
We are doing TEM on cultured Koala lymphocytes. We are having an on going problem with Spurr's resin (we use medium Spurr's). In some cases resin doesn't get polymerised and stays soft even after 3-4 days in sixty degree oven. The interesting point is that it doesn't happen with every sample. Even in a series of different samples which are processed at the same time and embeded in the same batch of resin, some remain soft while the others are quite Ok.
We heard that some components of culture media may interfere with resin polymerisation so we have been careful to wash the cultured cells properly but it did not eliminate the problem. Any advice on how to tackle the problem is highly appreciated. I would also be thankful if you suggest a way to revive those samples which are embeded in soft resin.
Regards
M. Ghoddusi
Majid Ghoddusi Division of Veterinary Pathobiology The University of Queensalnd QLD 4072 Australia
My colleague Neil Hand has recently published a paper entitled:
Non-isotopic in-situ hybridization to detect chick Sox gene mRNA in plastic embedded tissue. Histochemical Journal Vol 29, pp 625-629 (1997)
This may be of help, including other references.
Neil may be contacted at: mpznhand-at-unix.ccc.nottingham.ac.uk
Cheers
Kev
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Kevin J. Randall Dept of Histopathology Queen's Medical Centre University Hospital NHS Trust Nottingham NG7 2UH Tel: 0115 924 9924 x 43725 ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^K
We need to analise Cu, Ag, Fe, etc. as impurities in very small samples of gold alloys. The best technique that we could find is thin foild EDS using an analytical Hitachi 8100 TEM with a Ge detector. To improve accuracy, we would like to use elemental standards, but we did not find a supplier of EDS/TEM standards up to now. I would appreciate to receive your suggestions.
Best regards Rui Vilar DeMAT/IST -- ####################################### Rui Vilar Departamento de Engenharia de Materiais Instituto Superior Ticnico Av. Rovisco Pais, 1096 Lisboa Codex Portugal Tel.: -351-1-8418121 Fax: -351-1-8418121 or -351-1-8418120 Email: pcrvilar-at-alfa.ist.utl.pt #######################################
reidr1-at-uofs.edu wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Email: reidr1-at-uofs.edu } Name: Richard Reid } } School: University of Scranton } } Hello, } I am an undergrad attending the University } of Scranton. I am trying to find some beginner } information regarding flourescent microscopy. } While I have found some web-sites dealing with } flourescence, they are all too advanced for me. } It is the same situation with the school's } library. Most of the information is not geared } for the beginner. I am mostly interested in } staining neural tissue (CNS). } } Thank you very much for any help you can } offer. It will be greatly appreciated. } } Sincerely, } } Richard Reid } } --------------------------------------------------------------------------- Dear Richard,
We occasionally run a class in fluorescence. The best resource for the class has been a set of books by ROST: Fluorescence Microscopy (Vol 1), and Quantitative Fluorescence Microscopy. Publisher: Cambridge Press.
Our new book "Optimizing Light Microscopy for Biological and Clinical Labs" also has a sound chapter on Fluorescence. Email me for details, if you are interested.
Hope these help. Good luck in your studies.
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 ********** Americas First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
Fellow micrsocpist, Has anyone had experience identifying blood residue on obsidian stone tools using sem. These tools are between 800 to 1000 years old. Does anyone know of any references that bases identification of blood on morphology or elemental analysis or has any hints for features that may be apparent?
Hank Adams Electron Microscopy Lab New Mexico State University, :"Home of the worst football team in the country" and proud of it!
EMPLOYMENT ANNOUNCEMENT: ElectroImage has an immediate opening for a technical person who has microscopy, computer, and image analysis skills. We are based on Long Island and distribute digital cameras, software, and other imaging products. If you would be interested in discussing the opportunity, please contact me at Matt-at-electroimage.com or by telephone at 516-773-4305.
Thank you.
Matt Irwin ElectroImage, Inc. 277 Northern Blvd. Suite 101 Great Neck, NY 11021
First of all, please DO NOT "REPLY" to this message, but contact the person listed below, for whom I am posting this message to Microscopy (she is not subscribed). Thanks. Gib Ahlstrand.
Used JEOL 100B TEM, free giveaway, but you pay shipping costs. This scope is in good working order with lots of extras: EDAX brand x-ray microanalaysis system (older model), SED detector, STEM detector, specimen current detector.
Contact: Ms. Barb Clark, University of Minnesota, Dept. of OBGYN, Minneapolis, MN USA 55455.
One of the companies would be M.A.C. in the U.K. E-mail standards-at-dial.pipex.com Fax: +44 1480 462901 they also have a web page i think. htpp://www.macstandards.co.uk/town/street/yr49/ ------------------------------------- Name: Luc Harmsen E-mail: anaspec-at-.icon.co.za
Vachik Hacopian wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } What is the best commercial source of consistently high-quality holey grids? } } Vachik Hacopian
Dr. Hacopian, I am inclined to say that since Ladd has been producing holey film for over forty years that we produce the most consistent and best film , but I suppose all manufacturers would say the same thing or go into a lomg-winded discourse on why theirs is the best. I would suggest that the best course for you is to try some different suppliers who have a long history of supplying high quality products over the years (such as Ladd, Pella Fullam, etc.) and see whose film best suits your application. Since their reputations for quality have existed for such a long period I don't think you would go wrong with any of them.
Developmental Biologist Department of Zoology Miami University Oxford, Ohio
We seek an Assistant Professor for a tenure-track position to begin in August, 1998. Ph.D. in zoology or biology and postdoctoral experience required. Individuals with expertise in any area of Animal Developmental Biology are invited to apply, but preference will be given to those who use electron microscopy as a major research tool. We expect this person to develop an independent research program in developmental biology that will enhance the department's research capabilities.
Teaching responsibilities include: (1) a sophomore level course in developmental biology; (2) participation in a team-taught introductory biology course; and (3) and advanced course in a specialty area. The successful applicant will be expected to seek external funding to support his/her research and to supervise and advise graduate and undergraduate students. Advancement will be based on teaching, research, and professional service, with primary emphasis on teaching and research.
Miami University is a state-assisted institution in SW Ohio. The department has excellent research facilities; the EM facility is well-equipped for SEM, TEM, cryopreservation, and confocal microscopy (see http://www.muohio.edu/~zoocswis for more details about the department and our facilities). The department has strong Ph.D. and M.S. programs, 32 faculty members, several postdoctoral researchers, and 50 graduate students on the Oxford campus.
Interested persons should submit a curriculum vitae, a statement of teaching philosophy, a description of current research and long-term research interests, and should arrange for three letters of recommendation and transcripts of graduate and undergraduate academic work to be sent to:
Dr. Douglas H. Taylor, Chair of Zoology, Miami University, Oxford, OH 45056.
Review of applications will begin on 1 December, 1997, and continue until the position is filled. Miami University offers equal opportunity in employment and education.
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 use a 160 tube length upright Zeiss microscope for plankton research.Because I have to study living cells I want to use DIC (differential interference contrast or Nomarkski). I asked Zeiss in the UK and in the Netherlands , but they told me that this device is not available anymore for that kind of "old" type microscopes. Knows anybody an address where I can get secondhand DIC attachment.
"Filament Life" The life of a filament in an electron probe has been mentioned during the=
discussions of SEM filament life, they cannot be compared!
A filament in a probe is being used to generate x-rays not to produce a high resolution image, the cap design and component geometry is different=
from a high resolution SEM. Those of you who use your SEM for EDX work will know that with the correct geometry you do not need much beam curren= t to generate a good x-ray spectra; people use lower emission currents or much smaller spot sizes to compensate for the SEM overcurrent.
"Filament Base" Filament bases indicate the environment in which that filament is being forced to operate. A good vacuum and economy filament position (TEM styl= e) even in an SEM will give a base lightly coated with tungsten, which is powder blue in colour. Drive the filament harder and the base will gathe= r even more tungsten and become much darker. If the vacuum environment is = of a poor quality the filament base will become yellow to orange in colour, this IS an indication of a poor environment which, in spite of what the vacuum gauges say, indicates a leak or an inefficient vacuum system. =
"Virtual Leaks" Be aware that the practice of situating the diffusion pump or turbo pump=
immediately below the specimen chamber does mean that filament life is related to how dirty the specimen is. A gassy specimen will reduce the pumping efficiency of the gun and thus filament life. A bad vacuum in th= e gun produces discharge and thus instability but most of all you will smel= l the problem when you open the gun; it has an oily-ozone smell!
Thanks to all who responded to my question. Quite a number of people photograph a monitor with good results. The several hints are appreciated. Best regards to all Prof. Walter A. Mannheimer Dept. of Metallurgy and Materiais Eng. Federal University of Rio de Janeiro POBox 68505, 21945 Rio de Janeiro, Brazil Vox (55 21) 590-0579 Fax (55 21) 290-6626 wamann-at-metalmat.ufrj.br
We are currently observing gold foils in TEM, along with EDS analysis. However, in order to perform absortion corrections we need to know the thickness of the sample. Question: how can we determine sample thickness from TEM observations? (We have a double tilt analytical holder).
} } Dear All } } We are doing TEM on cultured Koala lymphocytes. We are having an on going } problem with Spurr's resin (we use medium Spurr's). In some cases resin } doesn't get polymerised and stays soft even after 3-4 days in sixty degree } oven. The interesting point is that it doesn't happen with every sample. } Even in a series of different samples which are processed at the same } time and embeded in the same batch of resin, some remain soft while the } others are quite Ok. } } We heard that some components of culture media may interfere with resin } polymerisation so we have been careful to wash the cultured cells properly } but it did not eliminate the problem. Any advice on how to tackle the } problem is highly appreciated. I would also be thankful if you suggest a } way to revive those samples which are embeded in soft resin. } } Regards } } M. Ghoddusi } } Majid Ghoddusi
Dear Majid Ghoddusi,
Please keep in mind that Ladd is a supplier of all the chemicals dicussed but with that in mind I would like to suggest the following:
1) It is very important to thoroughly mix the complete resin mixture. From your brief description, incomplete mixing would seem a likely reason for your problems. 2) If you are sure that incomplete mixing is not the problem, have you allowed enough time for complete infiltration of complete resin into your cells? 3) Humidity might also be an issue. The resin mixture is hygroscopic thus the resin may be absorbing water which would adversly effect curing and cutting properties. Try curing in a closed BEEM or gelatin capsule.
A solution that sometimes extracts epoxy reins from embedded samples can be prepared as follows: a) Prepare standard solution of KOH in absolute ethanol b) Allow to stand overnight c) The dark-colored supernatant is used as solvent d) Trim areas that contain only epoxy resin and immerse the sample in the solvent e) After epoxy resin is disolved wash 4 or so times in absolute ethanol F) Re-embed CAUTION!!!!!! I can not predict how if this treatment will destroy the vital components of your cells so please test this procedure or a couple of samples.
} } We need to analise Cu, Ag, Fe, etc. as impurities in very small samples } of gold alloys. The best technique that we could find is thin foild EDS } using an analytical Hitachi 8100 TEM with a Ge detector. To improve } accuracy, we would like to use elemental standards, but we did not find } a supplier of EDS/TEM standards up to now. I would appreciate to receive } your suggestions. } } Best regards } Rui Vilar } DeMAT/IST } -- } ####################################### } Rui Vilar } Departamento de Engenharia de Materiais } Instituto Superior Ticnico } Av. Rovisco Pais, 1096 Lisboa Codex } Portugal } Tel.: -351-1-8418121 } Fax: -351-1-8418121 or -351-1-8418120 } Email: pcrvilar-at-alfa.ist.utl.pt } #######################################
Dear Rui Vilar: Because thickness of specimen and standards are very difficult to control within 10 or even 20% of desired, quantitative analysis in TEM is elusive. Thickness (and other factors) change X-ray generation greatly. There are always exceptions, for instance just comparing a simple (say 2 phase) spectrum with that of a very similar standard will give reasonable results.
If the impurities are smaller than about 4 microns the area analysed will be too small for an EDS on SEM and the smaller 'envelope' analysed in TEM 'wins'. But TEM analysis could never be 'quantitative' - e.g. +/- 1% in a hundred.
In TEM, beam diameter (plus a little) yields most X-rays, but the small analysis envelope is little help if the impurities are not uniform throughout the thickness of the specimen as well.
Few companies make TEM EDS/WDS standards because of their limited applications. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} -----------------------------------------------------------------------. } } We need to analise Cu, Ag, Fe, etc. as impurities in very small samples } of gold alloys. The best technique that we could find is thin foild EDS } using an analytical Hitachi 8100 TEM with a Ge detector. To improve } accuracy, we would like to use elemental standards, but we did not find } a supplier of EDS/TEM standards up to now. I would appreciate to receive } your suggestions. } } Best regards } Rui Vilar } DeMAT/IST } -- } ####################################### } Rui Vilar } Departamento de Engenharia de Materiais } Instituto Superior Ticnico } Av. Rovisco Pais, 1096 Lisboa Codex } Portugal } Tel.: -351-1-8418121 } Fax: -351-1-8418121 or -351-1-8418120 } Email: pcrvilar-at-alfa.ist.utl.pt } #######################################
Our facility is undergoing ISO certification. My EM lab is small, consisting of one SEM with BSE and EDS. I would appreciate any suggestions/warnings, etc. from your experiences with ISO certification.
You can use convergent beam diffraction to measure thickness. In principle, this relies on the measurement of the spacings of thickness fringes which are visible in the CBED disks. It is very well described in the book "Transmission Electron Microscopy", volume II, "Diffraction" by D. B. Williams and C. B. Carter (Plenum, 1996). For the development of the technique they quote works by P. M. Kelly et al, phys. stat. sol. A31 (1975) p. 771, and S. M. Allen, Phil. Mag. A43 (1981) p. 325.
This is probably the best bet, and may be fairly straightforward for gold foils.
Wharton Sinkler
On Wed, 8 Oct 1997, Nuno Braz wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are currently observing gold foils in TEM, along with EDS analysis. } However, in order to perform absortion corrections we need to know the } thickness of the sample. } Question: how can we determine sample thickness from TEM observations? } (We have a double tilt analytical holder). } } Email: pcrvilar-at-alfa.ist.utl.pt } Tel: 351 1 8418124 } Fax: 351 1 8418120 }
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Department of Materials Science and Engineering Northwestern University 2225 North Campus Drive Evanston, IL 60208-3108 tel: (847) 491-7809 fax: (847) 491-7820 email: sinkler-at-apollo.numis.nwu.edu
Now that the subject of holey-C grids has been raised, are there any vendors willing to guarantee contamination-free films? Most of the grids I've purchased from vendors will contain some silicon contamination, along with variable Na, Ca, K and Cl. Since I normally investigate silicates, Si contamination is very problematic. I assume the Si comes from diffusion pump oil. Have any of the vendors with "TEM quality control" checked for the cleanliness of their grids?
Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets Johns Hopkins University Baltimore, Maryland 21218 USA Phone: (410) 516-8342 Fax: (410) 516-7933 e-mail: klivi-at-jhu.edu
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Hank,
It just so happens... See: Material Residues on Stone Tool Edges: Is Optical Microscopy Missing an Opportunity. Microscope Vol 45:3 89-93 (1997). Just out. On first page is topic heading BLOOD TRACES. Let me know if you need a fax.
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Fellow micrsocpist, Has anyone had experience identifying blood residue on obsidian stone tools using sem. These tools are between 800 to 1000 years old. Does anyone know of any references that bases identification of blood on morphology or elemental analysis or has any hints for features that may be apparent?
Hank Adams Electron Microscopy Lab New Mexico State University, :"Home of the worst football team in the country" and proud of it! --IMA.Boundary.048223678 Content-Type: text/plain; charset=US-ASCII; name="RFC822 message headers" Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part Content-Disposition: inline; filename="RFC822 message headers"
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There are several methods you can use for this. Depending on the nature of your sample , some of these might be more " practical " than other methods.
1) You might be able to see a contamination spot on the sample. If so, then if you tilt the sample a known amount you can get a value of the thickness by measuring the distance between the contamination spots at the top and bottom of the foil.
2) You might be able to use convergent beam analysis. (see for example D.B. Williams Practical Analytical Electron Microscopy in Materials Science).
3) You can estimate the foil thickness from the number of thickness fringes seen using two beam conditions ( see for example Edington's book " Practical Electron Microscopy in Materials Science" ).
4) You can also use trace methods (see Hirsch et. al).
Jordi Marti
We are currently observing gold foils in TEM, along with EDS analysis. However, in order to perform absortion corrections we need to know the thickness of the sample. Question: how can we determine sample thickness from TEM observations? (We have a double tilt analytical holder).
} Mounting medium can add tens of } microns (or hundreds, if you're having a bad day!) to the effective } thickness of } the coverslip. Along with variability in thickness of coverslips (--even } those } that are nominally 0.17 mm--), that's the reason for the addition of } correction } collars onto lenses. } One way around that problem is to mount sections directly on the coverslip surface and then use the mounting medium to attach the coverslip the slide. With this approach, there is no chance of having to much mounting medium between the coverslip and specimen. But as Barbara Foster pointed out in another posting, we find great variation in actual coverslip thickness between manufacturers and within a box. We have also found supposedly high quality glass slides which varied significantly in thickness along the length of an individual slide. this can also lead to excessive mounting medium between the coverslip and slide if the coverslip spans a low point in the center of the slide. An inexpensive micrometer is the only way you can be sure you are buying a good product.
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)
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. Steve Chapman, That was so well put that I printed it into our operating manual. It still leaves me with the questions, "Is 150-200 hours all I can expect from our JEOL 5800LV filaments?, and should we expect less filament life if we run at low vacuum more often?" I have been told "no" to the second question, but I believe that may be wrong. As far as the first question goes, I have been working with probes so long that 200 hours just seems very low (even keeping in mind that SEM filaments work far harder). Does cooling the filament before venting the chamber extend filament life?? Filaments only cost ~$80 each, but we are soft funded, so saving a couple hundred over a year actually helps us alot. Thanx Christopher
} } "Filament Life" } The life of a filament in an electron probe has been mentioned during the } discussions of SEM filament life, they cannot be compared! } } A filament in a probe is being used to generate x-rays not to produce a } high resolution image, the cap design and component geometry is different } from a high resolution SEM. Those of you who use your SEM for EDX work } will know that with the correct geometry you do not need much beam current } to generate a good x-ray spectra; people use lower emission currents or } much smaller spot sizes to compensate for the SEM overcurrent. } } "Filament Base" } Filament bases indicate the environment in which that filament is being } forced to operate. A good vacuum and economy filament position (TEM style) } even in an SEM will give a base lightly coated with tungsten, which is } powder blue in colour. Drive the filament harder and the base will gather } even more tungsten and become much darker. If the vacuum environment is of } a poor quality the filament base will become yellow to orange in colour, } this IS an indication of a poor environment which, in spite of what the } vacuum gauges say, indicates a leak or an inefficient vacuum system. } } "Virtual Leaks" } Be aware that the practice of situating the diffusion pump or turbo pump } immediately below the specimen chamber does mean that filament life is } related to how dirty the specimen is. A gassy specimen will reduce the } pumping efficiency of the gun and thus filament life. A bad vacuum in the } gun produces discharge and thus instability but most of all you will smell } the problem when you open the gun; it has an oily-ozone smell! } } Steve Chapman } Senior Consultant } Protrain }
We have had good success in many cases using the Bremmstrahlung shape to determine an absorption correction. See "EMAG '87 - Anlytical Electron Microscopy", ed. G. W. Lorimer, Institute of Metals, London (1988) p. 7, or "Analytical Electron Microscopy 1987" ed. D. C. Joy, San Francisco Press, p. 225 for more information. In the work we reported there we used software we wrote ourselves. We now use "Desktop Spectrum Analyser" from the folks at NIH. I don't know if any of the commercial analyser software packages support this algorithm.
We have never, so far as I remember, used this method in gold, but I don't see why there should be any problem.
This method avaoids having to determine the sample thickness (notoriously difficult!) or having to make any assumptions about the sample geometry.
I'd be glad to provide more information if you e-mail me directly.
Tony Garratt-Reed
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
I am up against a brick wall in the calibration of a rotating compensator. For our ISO program, we must have all measurement devices calibrated by a traceable (NIST) standard if possible. Are there any ideas on how to calibrate a rotating compensator for birefringence measurements.
Jim Harper Amoco Fabrics and Fibers jeharper-at-amoco.com
The easy (or easiest) and accurate method to measure the thickness of the specimen with a known structure like gold is Two-beam CBED. From the rocking curves you see from the CBED pattern you can measure the thickness. If you use a Philips CM microscope, there is a freeware to measure the thickness from the CBED pattern. Although I never use that software by myself, I believe it is quite straigh forward. Or you can do a much more accurat thickness refinement by using J.M.Zou's refinement program or other similar program. Check out J.C.H.Spence's book.
Yifan
-- ********************************************************************** * 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 * **********************************************************************
Sorry, but we had a typo in my previous e-mail. Part A should have read Prepare a "saturated" solution not a "standard" solution. Hope this is clear. If not, please e-mail or call 802-878-6711 or fax at 802-878-8074.
Sounds like an infiltration problem. What is your procedure? Are your blocks too big? Is your infiltration time too short? Is your dehydrating agent wet? Are your components well mixed before use? Do you let the first 3 components mix well before adding the catalyst? Have you at any time let the surface of the sample dry so that further infiltration doesn't happen properly?
On Tue, 7 Oct 1997, Majid Ghoddusi wrote:
} Date: Tue, 7 Oct 1997 09:21:55 +1000 (GMT+1000) } From: Majid Ghoddusi {vp092327-at-student.uq.edu.au} } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM: Problem with Spurr's resin } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear All } } We are doing TEM on cultured Koala lymphocytes. We are having an on going } problem with Spurr's resin (we use medium Spurr's). In some cases resin } doesn't get polymerised and stays soft even after 3-4 days in sixty degree } oven. The interesting point is that it doesn't happen with every sample. } Even in a series of different samples which are processed at the same } time and embeded in the same batch of resin, some remain soft while the } others are quite Ok. } } We heard that some components of culture media may interfere with resin } polymerisation so we have been careful to wash the cultured cells properly } but it did not eliminate the problem. Any advice on how to tackle the } problem is highly appreciated. I would also be thankful if you suggest a } way to revive those samples which are embeded in soft resin. } } Regards } } M. Ghoddusi } } } Majid Ghoddusi } Division of Veterinary Pathobiology } The University of Queensalnd } QLD 4072 } Australia } } Tel: (07) 3365 2569 } Fax: (07) 3365 1355 }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Dear Nuno: Another way you can use to measure thickness, if you have the Gatan EL/P system, is EELS. By measuring the intensity of Zero loss and the first plasma peak, you can figure out the thickness. This method is described on Egerton's "EELS in Electron Microscopy" book. Good luck!
On Wed, 8 Oct 1997, Nuno Braz wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are currently observing gold foils in TEM, along with EDS analysis. } However, in order to perform absortion corrections we need to know the } thickness of the sample. } Question: how can we determine sample thickness from TEM observations? } (We have a double tilt analytical holder). } } Email: pcrvilar-at-alfa.ist.utl.pt } Tel: 351 1 8418124 } Fax: 351 1 8418120 }
} . . . } } "Filament Life" } The life of a filament in an electron probe has been mentioned during the } discussions of SEM filament life, they cannot be compared! } } A filament in a probe is being used to generate x-rays not to produce a } high resolution image, the cap design and component geometry is different } from a high resolution SEM. Those of you who use your SEM for EDX work } will know that with the correct geometry you do not need much beam current } to generate a good x-ray spectra; people use lower emission currents or } much smaller spot sizes to compensate for the SEM overcurrent. } } . . .
I agree that microprobes are designed quite differently than SEMs ... but I am not too sure to what extent this can be said of their respective guns. While I can well imagine a probe gun having design aspects for stability (... a cross-over less susceptable to mechanical or thermal variation ...), resultant beam currents for x-rays (edx or wdx) vs. imaging are a result of condenser lens settings. That is, both of my instruments (probe and SEM) use very similar emission currents (i.e., 60 to 80 microAmps) ... measured as the load on the HV power supply. While tungsten filament saturations for either type of gun do involve filament tip design and wehnelt geometries, I am willing to bet saturation (self-biasing as a result of "driving the filament heat") occurs within 25 degrees of eachother, and that there probably is as great a disparity between SEM manufacturers than SEM vs. probes. Of course, you should feel free to clarify what you mean by choosing lower "emission" currents and ""SEM overcurrent" ...
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/
Spaulding, Robert F wrote: } } I am looking for a source for the book, Quantitative Electron-Probe } Microanalysis by V. D. Scott and G. Love. My local bookstore is having } no luck. } } Thanks, } Robert F. Spaulding } Corning Incorporated } Characterization Science & Services } Microscopy & Microanalysis Dept. } SP-FR-01-8 } Corning, New York 14831 } } 607-974-3732 } fax 607-974-3385 } Internet: SpauldinRF-at-corning.com
Publisher is Ellis Horwood (New York, London, Toronto), ISBN 0-13-104050-2 and address is:
Ellis Horwood Limited A division of Simon & Schuster International Group Campus 400, Maylands Avenue Hemel Hempstead Hertfordshire, HP2 7EZ UK
Henrik -- Henrik Kaker SEM-EDS Laboratory Metal Ravne d.o.o. Koroska c. 14 2390 Ravne Slovenia Tel: +386-602-21-131 Fax: +386-602-20-436 SEM-EDS Lab http://www2.arnes.si/guest/sgszmera1/index.html MVD Database http://www.kaker.com/mvd/vendors.html Kaker.Com http://www.kaker.com
Although my primary responsibility is running our EM facility, I have served as an R&D team representative in obtaining ISO 9001 certification, and I have also been trained in auditing. Hopefully, I can give you some appropriate insight.
In order to provide you with appropriate information, I would need to know the nature of your organization and how your lab fits into it. But in general, I would say that your greatest challenge will be documentation. If you have an equipment log for your SEM and EDS (as you should), you have completed one aspect already. But in addition:
1.) You will probably need to generate some appropriate standard operating procedures (SOP's) for your equipment, including calibration and maintenance procedures. You should pay particular attention to how you schedule these, so that you will ensure compliance. Also, do not forget your standards; they are also subject to recalibration schedules.
2.) Sample traceability may be an issue for you as well. Be sure to have procedures describing how you handle and track them.
3.) If you have subcontractors who perform certain calibrations for you (e.g. picoammeters and stuff) you may need to have a document explaining how you assess them.
4.) Get some calibration stickers - you will need to tag any equipment that is subject to calibration. Equipment not requiring calibration may need reference only tags.
5.) You may need to conduct an equipment and software validation that includes installation, operation and processing (IQ, OQ, PQ) depending upon how the equipment is used. This can be quite extensive and time-consuming.
THE IMPORTANT THING TO REMEMBER WITH DOCUMENTATION IS THAT YOU DO WHAT YOU SAY YOU WILL DO. DO NOT GENERATE "ALL INCLUSIVE" DOCUMENTS THAT INCLUDE STEPS THAT YOU DO NOT NORMALLY DO - UNLESS YOU NEED TO CHANGE AND YOU PLAN TO STICK TO IT.
There are many other considerations that are too extensive to go into here, but if you still need information or would like to discuss this more, feel free to contact me direct.
Regards,
Bob Citron Chiron Vision Claremont, CA USA Bob_Citron-at-cc.chiron.com
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Our facility is undergoing ISO certification. My EM lab is small, consisting of one SEM with BSE and EDS. I would appreciate any suggestions/warnings, etc. from your experiences with ISO certification.
Majid: It is known that Spurr's resin has some incompatibilities for example I have seen problems using Kellenberger en bloc uranyl acetate staining with Spurr's resin. If this is the case it seems to affect some areas more than others in the block and is not due to mixing or infiltration. With proper usage Spurr's can infiltrate plant cells and moon rocks so it shouldn't be that hard to infiltrate cells in a monolayer. bob
Kenneth JT Livi wrote: =================================================== Now that the subject of holey-C grids has been raised, are there any vendors willing to guarantee contamination-free films? Most of the grids I've purchased from vendors will contain some silicon contamination, along with variable Na, Ca, K and Cl. Since I normally investigate silicates, Si contamination is very problematic. I assume the Si comes from diffusion pump oil. Have any of the vendors with "TEM quality control" checked for the cleanliness of their grids? =================================================== It is a correct statement that it is easy for grids to get contaminated as indicated. There is a reality check relating to cost, that is, we ourselves do not routinely check with EDS, each batch for the presence or absense of Si. But we do check periodically and will when specially requested, do a special batch check by EDS. This higher level of control represents slightly higher costs, that being the reason it is not done for every batch . If good housekeeping and cleanliness conditions are maintained, one can make "Si-free grids". In any case, we can produce what we ourselves determine (and believe) to be "Si-free" filmed grids as determined by our own EDS measurements on our in-house JEOL 100CX TEM. However, I am concerned about the "willing to guarantee" requirement. If you are in agreement that a "zero" amount of anything does not exist, then what detection limits are we talking about?
We have found at least a few of our customers probably have detection limits lower than what we are able to achieve when we do the control work. Clearly a small spot size XPS might find all kinds of things that would otherwise be "guaranteed to be free of" when done by EDS in a TEM.
There is another element to this equation: Some users "find" Si in our spectroscopically pure carbon mounts. We are convinced there is no silicon in our carbon mounts, at least if present at all, the level would be less than 1 ppm as determined by emission spectroscopy. Yet we are told by customers that Si is sometimes "detected". However, others have told us that the culprit is the Si in the Si (Li) detector being used. We know that at least some of the EDS systems (e..g like the older ones we have in house) seem to have some problem trying to model the "notch" for the background subtraction algorithms. So we have been operating on the theory that at least some of the so-called Si being detected in filmed grids is an artifact of the system being used, yet on the other hand, I am not saying it is always an artifact. After all, one CAN indeed end up with Si on filmed grids! The common practice of using silicone fluids in a vacuum evaporator is one already mentioned potential source.
The other comment would be that if one wants Si-free films, then to be on the safe side, at least we have for some time, used Santovac 5 in the diffusion pumps of the vacuum evaporators.
I hope that explains why we believe our holey carbon (filmed) grids are indeed "silicon-free". I would also expect that any other producer, taking the same precautions, should end up with grids that would be comparable.
I would welcome any further suggestions on this topic, either on-line or off -line.
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 ############################ ================================================== ------- FORWARD, End of original message -------
Although I work in medical research EM I am currently attending, in my spare time and for my own personal interest, a course in Horticulture. Part of that course involves the study of photosynthesis. Our instructor on the course does not have an EM photograph of a chloroplast which he can use as a "handout" for the class. I have no botanical EM material at all.
I was wondering if there is a kind soul on the List who would like to donate a scanned copy of a chlorplast. I can promise that it would be used for educational purposes only and not for any commercial purpose. Due acknowledgment would be printed on the image.
Ideally what I would like would be a JPEG, GIF or TIFF file which could be printed out on a decent laser printer and then the instructor could run off enough copies for the students on the course (about 50).
Failing that, could anybody with the relevant knowledge point me to a botany related website where I could download a public domain image of a chloroplast.
Quantitative Electron-Probe Microanalysis (Ellis Horwood Series in Physics and Its Applications) ~ Ships in 2-3 days Love G., et al / Paperback / Published 1995 Their Price: $77.00
Regards, Neal
Spaulding, Robert F wrote: } } I am looking for a source for the book, Quantitative Electron-Probe } Microanalysis by V. D. Scott and G. Love. My local bookstore is having } no luck. } } Thanks, } Robert F. Spaulding } Corning Incorporated } Characterization Science & Services } Microscopy & Microanalysis Dept. } SP-FR-01-8 } Corning, New York 14831
I am looking for a commercial lab in the New England area that I could send some samples out to be embedded, sectioned and TEM prints taken. If I could get the names of the labs and people to contact, I would appreciate it greatly. Please contact me off of the listserve so as not to infringe on those who are not interested. I will gladly supply more details about the samples to those who contact me.
David Bell Millipore Corporation phone: (781) 533-2108 fax: (781) 533-3143 e-mail: David_Bell-at-Millipore.com
You may visit my website at {http://www.ualberta.ca/~mingchen/pcell.htm} for the TEM image of a plant cell which has some chloroplasts in it. Also you should be able to print it out on your printer.
Best wishes,
Ming
On Thu, 9 Oct 1997, Stephen Griffiths wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi } } Although I work in medical research EM I am currently attending, in my } spare time and for my own personal interest, a course in Horticulture. Part } of that course involves the study of photosynthesis. Our instructor on the } course does not have an EM photograph of a chloroplast which he can use as } a "handout" for the class. I have no botanical EM material at all. } } I was wondering if there is a kind soul on the List who would like to } donate a scanned copy of a chlorplast. I can promise that it would be used } for educational purposes only and not for any commercial purpose. Due } acknowledgment would be printed on the image. } } Ideally what I would like would be a JPEG, GIF or TIFF file which could be } printed out on a decent laser printer and then the instructor could run off } enough copies for the students on the course (about 50). } } Failing that, could anybody with the relevant knowledge point me to a } botany related website where I could download a public domain image of a } chloroplast. } } With thanks in advance } } Regards } Stephen Griffiths. } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} } Stephen Griffiths } Visual Science Department } Institute of Ophthalmology } Bath Street, London. EC1V 9EL } e-mail:- s.griffiths-at-ucl.ac.uk (work address) } or stephen.griffiths-at-dial.pipex.com (home address) } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} }
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Kenneth JT Livi wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Now that the subject of holey-C grids has been raised, are there any } vendors willing to guarantee contamination-free films? Most of the grids } I've purchased from vendors will contain some silicon contamination, along } with variable Na, Ca, K and Cl. Since I normally investigate silicates, Si } contamination is very problematic. I assume the Si comes from diffusion } pump oil. Have any of the vendors with "TEM quality control" checked for } the cleanliness of their grids? } } Ciao for now, } Ken } } Kenneth JT Livi } Department of Earth and Planetary Sciences } 34th and Charles Streets } Johns Hopkins University } Baltimore, Maryland 21218 USA } Phone: (410) 516-8342 } Fax: (410) 516-7933 } e-mail: klivi-at-jhu.edu
Dear Kenneth Livi,
Kenneth JT Livi Wrote -
During the production of carbon film grids there are three (3) possible silicon contamination sources: 1) Residue in the evaporator 2) Evaporator Oil 3) Carbon Source
When we produce "silicon free" film we take the following precautions: 1) We thoroughly clean the evaporator prior to each evaporation cycle.
2) On all our evaporators silicon free oil is used.
3) We use pure carbon not graphite as a source. The impurity level does not exceed 2ppm
Since there is always a potential for silicon in the carbon the term "silicon free" is subjective but based on the above we feel we produce film with at most 1 ppm Si but it is impossible for LADD to guarantee the total absence of Si. :)
John Arnott PH: 802/878-6711 LADD RESEARCH FAX: 802/878-8074 13 Dorset Lane Williston, VT 05495
Dear all, I greatly should appreciate informations on experiences with existing/ or on dealers/companies of LASER PROJECTION SYSTEMS (connectable to PC/Video and TV Cams, Remote system). We know a company (ASK-System) in Europe which deals with LIGHT PROJECTION SYSTEMS for demonstration of slides, PC-data, etc. in teaching and else applications. Has anybody suggestions, informations (company/-ies, approx. price, combinations with periphery, necessary components) for us ? Such a Laser Projection System should work for projection screen dimensions of at the maximum approx. 2 x 3 m or little less, if available. Any suggestions and comments are welcome. Best wishes for the day sincerely yours Wolfgang MUSS Dept. Pathology LKA, EM-Lab, Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria/Europe Phone: ++43++662+4482-4720 Ext Fax: ++43++662+4482-882 Ext. e-mail: W.Muss-at-lkasbg.gv.at.
Does any one have any image processing and analysis scripts/macros/programs they are willing to share regarding the processing and analysis of cross section neurons. to include axon thickness, sheath size as well as to classify and count cross section samples based on size.
I would be willing to share one program I have (written for Kontron KS400).
Now that the subject of holey-C grids has been raised, are there any vendors willing to guarantee contamination-free films? Most of the grids I've purchased from vendors will contain some silicon contamination, along with variable Na, Ca, K and Cl. Since I normally investigate silicates, Si contamination is very problematic. I assume the Si comes from diffusion pump oil. Have any of the vendors with "TEM quality control" checked for the cleanliness of their grids?
Janet Woodward asked about laboratory experiences with ISO issues. I am laboratory director of Structure Probe, Inc., which has been accredited since 1980 in the subdiscipline of "microscopy" by the American Association for Laboratory Accreditation. We are currently accredited to the standard of ISO Guide 25, which is the international document which specifically applies to laboratories. A direct comparison with ISO 9000-series documents shows that the requirements are roughly the same, plus Guide 25 requires a demonstration of competence, which is not required by ISO 9000. While our experience has been evolutionary, and Janet is about to jump into the deep end of the pool without having had swimming lessons, some of the things which we have learned over the years might help.
1. Read the Directions: Unfortunately, there are a number of very specific requirements, and it is important that you know what they are. It is one thing to be involved in a discussion about whether what you do does or does not meet a particular requirement--it is another to be "blindsided" by a requirement of which you were not aware.
2. Documentation: The biggest problem for us in the evolution of our accreditation experience is the need for explicit procedure documents which cover every aspect of our technical operations. Writing procedures for the assembly of widgets from boxes of parts can be a challenge--writing procedures for doing contract microscopy in an independent laboratory environment, where the nature of the work is totally driven by the requirements of clients, can be a nightmare. We have written documents that are very heavy on such issues as how to document each and every micrograph so that it can be unequivocally traced to a specific sample and management review of each and every incoming project. At the same time, the documents allow a great deal of professional judgement for the individual analyst, provided that the procedures actually used are documented in the report.
3. Calibration: This is the "culture shock" area. You will be expected to support your results with a calibration trail, and that calibration will be expected to be traceable, where possible, to a national standards body. I have found it helpful to use the word "calibration" but think about "verification". We have a program in which each column instrument is run through a set of standard evaluations every month, and the results are recorded as micrographs and spectra in log books. As a result, we can go back and show you the performance history (magnification, resolution, etc.) of the instrument and, if push comes to shove, we can answer the critical question--"How do you know that, on October 5, 1895, the magnification of this micrograph was in fact 3,000X?"
4. Traceable Reference Samples: The actual core of the calibration program is the repetition and documentation, but the problem that seems to be causing the most pain is the traceability of the reference samples used. This is a moving target. What is going on is that there is no such thing, as this is written on October 9, 1997, as an accredited calibration laboratory for standards for microanalysis. There are materials available from the National Institute for Standards and Technology (NIST) in the U.S. and from comparable bodies in other countries, but they do not cover many of the areas which a good calibration program will include. Various suppliers, including SPI Supplies (tm), offer reference materials suitable for calibration programs, but these are not necessarily fully traceable--if someone says that a reference material is traceable, ask what the "uncertainty budget" is (get used to that term, it covers the fact that every time you extend a measurement from the ultimate reference, you become less and less certain what the "actual" value is).
I am happy to discuss any of these issues in detail--off line is probably better. I am also happy to conduct dialog with your assessor--some of these problems are more easily resolved by some who can speak "quality".
Disclaimer: I am employed by Structure Probe, Inc., parent company of SPI Supplies. I am also a member of the Accreditation Council of the American Association for Laboratory Accreditation.
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
I am trying to find the mailing addresses, email addresses, phone or fax for the following electron microscopists. I want to reproduce figures from their books in our web course and the publisher has informed me that the copyright has reverted back to the author. Unfortunately, the publisher (Elsevier) no longer has addresses for these authors. If you can help me, email me at gsosinsky-at-ucsd.edu.
1) Alan W. Agar 2) Ronald H. Alderson 3 Dawn Chescoe 4) J.H. Martin Willison 5) Arthur J. Rowe
I am looking for advice from those experienced with using Bouin's fixative.
A users of our lab is about to leave on a cruise and needs to preserve plankton at sea. In the past he has used Bouin's fixative, but now, due to the picric acid component, the hassle factor has gone way up.
He is looking for either a pre-made Bouin's so he does not have to deal with the picric acid issue, or a substitute fixative that would serve his needs.
If you can help, send the information to me and I will pass it along to him.
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
The Materials Research Science and Engineering Center (MRSEC) of the University of Maryland is looking for a post-doctoral fellow to perform research on ferroelectric and ferromagnetic oxide films using TEM. The MRSEC has very strong experience in the synthesis and characterization of these materials. We also have experitise in device fabrication. The position is available imediately with a starting salary of $32-35K depending on experience.
Experience on high resolution TEM is highly desirable. Interested candidates should submit their resume and a list of three references to Dr. Lourdes Salamanca-Riba either by regular mail or e-mail at either of the following addresses.
Materials and Nuclear Engineering Department University of Maryland College Park, MD 20742-2115
Thank you for your helpful responses to my question:
} What is the best commercial source of consistently } high-quality holey grids?
Ian Montgomery recommeds Agar Aids in England. He has been using the same grid for almost 30 years! That's my kind of quality production. British engineering is second to none. Ian, is that grid made of copper or kryptonite? ;-)
Larry Allard recommends grids produced by Structure Probe and Pella.
Brian Demczyk recommends SPI grids.
Markus F. Meyenhofer wrote "It's the fool behind the tool!!!". Thank you, Markus, for a touch of humor.
Representatives of Scott Scientific, Ted Pella, SPI Supplies, and Ladd Research wrote to praise their own products.
Does anybody have (good or bad) experience with any of the following or other fast freezing devices: Reichert KF80 Leica CPC Gentleman Jim (constructed by Alan Boyne)
Has anybody made a comparison?
Philip
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
I'm sending this message on behalf of the Department of Biology at Eastern Connecticut State University. Our microbiologist had to undergo emergency surgery and is not expected to return to his teaching duties for another 5 to 8 weeks. We have been able to cover most of his classes, but right now his introductory microbiology class has no instructor. The lecture meets M, W, and F from 9 to 10:00 AM and the lab meets on Thursday from 2 to 5:00 PM. We are in desperate need of a temporary replacement. If anyone is interested or knows of someone, please let me know. Of course, the individual would have to live within reasonable driving distance of the University (Willimantic, Connecticut). If these times don't fit your schedule, but you are still interested, let me know. We might be able to rearrange the time.
If we cannot find a replacement, we are open to suggestions. Perhaps someone knows of a course on tape or even an ongoing internet course.
Please respond immediately via Email. I will forward your message to our department chair.
Thanks,
Martin A. Levin Department of Biology Eastern Connecticut State University Willimantic, CT 06226 Phone: (860)465-4324 FAX: (860)465-5213
We would like control our microscope Philips 420/STEM by connecting it to a PC, Mac or Silicon Graphics. Can anybody give advice to make it effectively? Thanks. Stanislav Hucek Lab. of electron microscopy Institute of Parasitology ASCR Branisovska 31 370 05 Ceske Budejovice Czech Republic
O.k., resin wizards, I have a user looking a truely clear, i.e. water clear, resin for emmbedding some small objects (1-2 cm x 2 mm neoperene O-rings) for gifts as paper weights. Therefore sectionablity or LM / EM is not a factor but durability and hardness are - they would like "nice and hard" resin.
I realize that this is not a strickly scientific quiry - and I apologize to anyone offended by any triviality - but why not ask the experts, eh? After who among us hasn't emmbedded a cockroach, etc. and kept it on their desk?
Commercial vendors should feel free to respond.
Thank you.
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 the SILVER Scottish Microscopy Symposium will take place at the above = =0Avenue and the Organising Committee have arranged a Scientific Programme = which =0Awe hope will appeal to as many microscopists as possible. Here is = the finalised =0Aprogramme.
Low temperature strategies for immunocytochemistry: choice or compromise - = =0AJeremy Skepper, Cambridge
Combining stereology and for immunogold labelling in quantitative =0Aimmuno= electron microscopy of membranes - John Lucocq, Dundee
Confocal Microscopy with high light budget - Tony Wilson, Oxford, England
Green Fluorescent Protein (GFP) as a tag for plant virus movement studies -= Alison =0ARoberts, Dundee
Environmental Scanning Electron Microscopy - Dirk van der Vall, Eindhoven,= =0AHolland
Staining resin sections - why infiltration is (or should be) incomplete - R= ichard =0AHorobin, Sheffield
Efficient and unbiased 3D measurements in microscopy - Stereology - Vyvyan = =0AHoward, Liverpool, England
Minimal preparation procedures for SEM of botanical specimens - Stephan Hel= fer, =0AEdinburgh
Registration will be =A320. For more information either contact me or you c= an visit =0Aour web site for the latest information and also details of las= t years meeting. Web site- http://www.abdn.ac.uk/~nhi691/smg97.htm=20 (This will be updated next week)
Kevin Mackenzie Tillydrone E.M. Unit University of Aberdeen Tillydrone Avenue Aberdeen AB9 2NT
This the SILVER Scottish Microscopy Symposium will take place at the above = =0Avenue and the Organising Committee have arranged a Scientific Programme = which =0Awe hope will appeal to as many microscopists as possible. Here is = the finalised =0Aprogramme.
Low temperature strategies for immunocytochemistry: choice or compromise - = =0AJeremy Skepper, Cambridge
Combining stereology and for immunogold labelling in quantitative =0Aimmuno= electron microscopy of membranes - John Lucocq, Dundee
Confocal Microscopy with high light budget - Tony Wilson, Oxford, England
Green Fluorescent Protein (GFP) as a tag for plant virus movement studies -= Alison =0ARoberts, Dundee
Environmental Scanning Electron Microscopy - Dirk van der Vall, Eindhoven,= =0AHolland
Staining resin sections - why infiltration is (or should be) incomplete - R= ichard =0AHorobin, Sheffield
Efficient and unbiased 3D measurements in microscopy - Stereology - Vyvyan = =0AHoward, Liverpool, England
Minimal preparation procedures for SEM of botanical specimens - Stephan Hel= fer, =0AEdinburgh
Registration will be =A320. For more information either contact me or you c= an visit =0Aour web site for the latest information and also details of las= t years meeting. Web site- http://www.abdn.ac.uk/~nhi691/smg97.htm=20 (This will be updated next week)
Kevin Mackenzie Tillydrone E.M. Unit University of Aberdeen Tillydrone Avenue Aberdeen AB9 2NT
The Department of Materials at Oxford University will shortly be setting up an Advanced Analytical HREM Facility, funded by a major grant from EPSRC. This will be based on a JEOL 3000F (300kv FEG/TEM) with a comprehensive range of analytical techniques, including EDX, GIF holography, etc. It is anticipated that the Facility will be set up early in 1998 to extend the present extensive range (12) of TEM and SEM instruments and support facilities. We are planning to appoint a Senior Research Officer who will be responsible for the day-to-day running of the Facility, and who will develop a successful Outside Users' programme. This will be a key position, funded for three years in the first instance. The successful candidate will be someone with expertise in the above techniques and some relevant post-doctoral experiance. Salary will be in the range 15159-22785 pounds sterling depending on age and experience.
For further details, please contact directly: Dr. John Hutchison, Department of Materials, University of Oxford, ParksRoad, Oxford. OX1 3PH. UK. phone: +44 1865 273705 e-mail: john.hutchison-at-materials.ox.ac.uk
I have been searching for an electronic overlay font (preferably a Microsoft compatible "True Type" variety rather than an Adobe "Post Script") to facilitate labeling of gray-scale images. Is anyone aware of a source for such work saver?
In the pre-digital days, we were getting good results with the paste-on overlay letters that are still available from microscopy supply houses (e.g., I was partial to the Microscopist's Collection from SPI). These were the little black letters that were printed over slightly larger white outlines. Because of this contrasting outline, they were easily readable irrespective of the brightness of the image below.
Currently, we are barely getting by in labeling of electronic images by adjusting the font color to either white or black, according to what the brightness of the underlying image dictates. More often than not, this is ineffective in EM images with highly modulated brightness. On occasion, I have resorted to manually overlaying a black text over a white one in a bold version of the same font; this is tedious and does not work in all but the briefest annotations because the kerning of the two font styles does not compensate for the differences in the widths of the letters.
The simplistic solution of setting the overlay text box background color to non-transparent is not satisfactory because it often ends up obscuring the feature of interest.
Does anyone possess a suitably outlined font, or has come up with a creative solution to this vexing problem?
valdemar-at-fast.net Valdemar Furdanowicz Homer Research Labs Bethlehem Steel Co. Bethlehem, PA
} We would like control our microscope Philips 420/STEM by connecting } it to a PC, Mac or Silicon Graphics. Can anybody give advice to make } it effectively? } Thanks.
I don't know specifically about the 420/STEM, but we're using the 4pi SE-II board in a Power Mac to control our Phillips STEM.
Imageing plug-ins work with NIH-Image or Photoshop. EDS plug-ins work with (NIST) DTSA or FLAME.
We're at work on a library to call 4pi driver routines from XlispStat ( a graphical statistics package ), Python (and interpreted object-oriented language), Java and NIH-Image. ( We've been using prototypes of these long enough to finally figure out how to redo them right! ;-)
See: http://www.4pi.com
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| University of Virginia Health Sciences Center |--- ---| P.O. Box 10011 Charlottesville, VA 22906-0011 |--- All power corrupts and obsolete power corrupts obsoletely." - Ted Nelson
The "Analytical Methods" section of the following article includes operating conditions for electron microprobe analysis of Na-bearing zeolites. Analcime is not specifically addressed, but the described method should work for this mineral, too.
Ross, M., Flohr, M.J.K., and Ross, D., 1992, Crystalline solution series and order-disorder within the natrolite mineral group: American Mineralogist, v. 77, p. 685-703.
Back in the 1950s, the standard embedding medium for biological EM was a mixture of methyl and n-butyl methacrylates (~1:9), catalyzed with 2% Luperco CDB, and polymerized at 60oC (or by UV). That material (otherwise known as "plexiglass") was very clear and hard, and would satisfy your criteria. In fact, about 1958 I embedded a ~3.5" cockroach in methacrylate. I had encountered it in the hallway outside our lab (it looked like an approaching Volkswagen "beetle"). It was an escapee from another lab that was doing research on the nervous system of such critters.
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.umich.edu/~akc/
------------------------------------
On Fri, 10 Oct 1997 edelmare-at-casmail.muohio.edu wrote:
} O.k., resin wizards, I have a user looking a truely clear, i.e. } water clear, resin for emmbedding some small objects (1-2 cm x 2 mm } neoperene O-rings) for gifts as paper weights. Therefore } sectionablity or LM / EM is not a factor but durability and hardness } are - they would like "nice and hard" resin. } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu
I am looking to buy or borrow a used power supply for a 100 W Hg arc lamp (specifically the Osram 100 W/2, although I believe most DC arc lamp supplies will work). Please respond to
umbach-at-msc.cornell.edu
or reach me by phone at 607-255-7426
Thanks!
-- Dr. Kit Umbach Dept. of Materials Science and Engineering 360 Bard Hall Cornell University Ithaca, NY 14853
we want to open the .img images of our XL20 and need some program for this purpose. Every hint appreciated.
Thanks in advance.
Andreas Loewe
______________________________________________________________ Andreas Loewe Tel: +49-228-734-180 University of Bonn Fax: +49-228-734-205 Institute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
} I am looking for advice from those experienced with using Bouin's fixative. } } A users of our lab is about to leave on a cruise and needs to preserve } plankton at sea. In the past he has used Bouin's fixative, but now, due to } the picric acid component, the hassle factor has gone way up. } } He is looking for either a pre-made Bouin's so he does not have to deal } with the picric acid issue, or a substitute fixative that would serve his } needs. } } If you can help, send the information to me and I will pass it along to him. } } Thanks. } } Jonathan Krupp
Substitute: I've used Trump's, Karnovsky's and plain old 2% glut or 10% formalin in 0.1 micron filtered seawater for plankton. Do you guys have a copy of the UNESCO book on Collection and Preservation of Zooplankton? (This includes ciliates, etc.) Try John Pearse, he might have one.
One solution is to import your image files into Microsoft Word (other word processors may have similar functions) and use the WordArt tool to overlay your picture with the desired text. This tool will allow you to outline letters using most true type fonts and allows you to control pitch, text color, outline color, and outline thickness. This method is fairly quick and painless. One hint is that when you first start adding text it may disappear behind the image. To bring the text to the front click on the image with the second mouse button, click on "order" and select "send behind text". This will place the image behind the text so that the text remains visible.
You can then either save the image in Word format or use a screen capture program to save the image in other formats. If using a screen capture program (such as LView) you will be limited to the resolution of your screen minus window borders so this solution works only for smaller images (aprox. 970x570 pixels on a 1024x768 monitor). There may be a way to export larger images from Word but as I am a novice to using Word I haven't found one yet.
Dan Moore
At 12:17 PM 10/10/97 -0400, you wrote: } Dear Colleagues: } } I have been searching for an electronic overlay font (preferably a } Microsoft compatible "True Type" variety rather than an Adobe "Post } Script") to facilitate labeling of gray-scale images. Is anyone aware of a } source for such work saver? } [snip] } } Does anyone possess a suitably outlined font, or has come up with a } creative solution to this vexing problem? } } valdemar-at-fast.net } Valdemar Furdanowicz } Homer Research Labs } Bethlehem Steel Co. } Bethlehem, PA } }
I should have added that using Word you can rotate your labels and add arrows, circles, boxes, etc. to your images. You can even add cartoon balloons and let your subjects speak to you.
Vlademar, There is a software package called "Designer" it is sold by Micrografx. It is very powerful and will do what you need. The cost is about $600. It is worth while to check it out.
Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
valdemar wrote:
} I have been searching for an electronic overlay font (preferably a } Microsoft compatible "True Type" variety rather than an Adobe "Post } Script") to facilitate labeling of gray-scale images. Is anyone aware of a } source for such work saver? } } In the pre-digital days, we were getting good results with the paste-on } overlay letters that are still available from microscopy supply houses } (e.g., I was partial to the Microscopist's Collection from SPI). These } were the little black letters that were printed over slightly larger white } outlines. Because of this contrasting outline, they were easily readable } irrespective of the brightness of the image below. } } Currently, we are barely getting by in labeling of electronic images by } adjusting the font color to either white or black, according to what the } brightness of the underlying image dictates. More often than not, this is } ineffective in EM images with highly modulated brightness. On occasion, I } have resorted to manually overlaying a black text over a white one in a } bold version of the same font; this is tedious and does not work in all } but the briefest annotations because the kerning of the two font styles } does not compensate for the differences in the widths of the letters. } } The simplistic solution of setting the overlay text box background color to } non-transparent is not satisfactory because it often ends up obscuring the } feature of interest. } } Does anyone possess a suitably outlined font, or has come up with a } creative solution to this vexing problem?
This is the recipe that I use in Photoshop 4.0 to put Black on White scale markers, text, and symbols onto micrographs. The results look just like the rub-on transfers that I used to use.
1. create a layer (Photoshop 4.0 does this automatically with text tool) You must use Photoshop image mode for the layer option.
2. in that layer in the font and font size that you want, type the text. add a black line at an appropriate length and width and any other text, symbols, arrows, etc. that you want to put on the micrograph. By using the layer, you preserve the original micrograph in the background layer. you can use the info window to draw lines to particular lengths. If other layers are created when new text is added, merge those layers. Don't merge them with the background layer!
3. Select all (ctrl-A in the PC) The marquee will be around the whole layer.
4. You have to move the selected region up then down with an arrow key. (This is done in the PC with the Ctrl-shift-arrow key in the PC) what this does is to select all of the objects in the layer individually. A Marguee should be around each object.
5. Select the foreground color as white. (You are going to write a white border around each Marquee.)
6. Go to Edit-Stroke and select the width of the white line you want (Width) and select the Outside option. for 300 dpi images at about 4" x 5", I suggest a font size of about 14 (Arial) with a Width of about 3-4 pixels for the stroke width. This will write a white border 4 pixels wide around all of the selected black features.
7. Deselect (ctrl-D)
8. If you want to save this as image in another format such as TIF or BMP, then you have to Merge the layers and save the image in that mode.
Note: you should have anti-aliasing selected for all this.
This technique works very well for me. You can make them look like real Rub-ons with another technique and offsetting the white a little, but that is another story.
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."
Philip, I used a Gentleman Jim Plunge Freezer a million years ago and as I recall our tissue had freeze damage.
For a review try: Ryan, KP (1992) Cryofixation of tissues for electron microscopy: A review of plunge cooling methods. Scanning Electron Microsc., 6:715-743. I got that reference from Scott Russell's article (1997) Spray-Freezing Freeze Substitution of Cell Suspensions for Improved Preservation of Ultrastructure. Microscopy Research and Technique 38:315-328. He lists several references for freezing device reviews.
best regards, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Dear all,=20 (according to a suggestion of Scott WHITTAKER I place the posting to the = server once more) if there anybody still is out there interested in getting written = informations on "How do do it", product data sheets (in English or = German, if you like) of the silicone rubber used, the adress of the = Company (in the USA and Germany) and a sample mold out of my fabrication = (all free of charge)=20 PLEASE SEND an E-MAIL NOW (to get it from my desk) to:
W.Muss-at-lkasbg.gv.at (note: the "l" right to "-at-" is a small "L").
Thank you for your interest.=20 Best regards, have a nice and a calm weekend, W.MUSS
} I have been searching for an electronic overlay font (preferably a Microsoft } compatible "True Type" variety rather than an Adobe "Post Script") to } facilitate labeling of gray-scale images. Is anyone aware of a source for } such work saver? } } In the pre-digital days, we were getting good results with the paste-on } overlay letters that are still available from microscopy supply houses (e.g., } I was partial to the Microscopist's Collection from SPI). These were the } little black letters that were printed over slightly larger white outlines. } Because of this contrasting outline, they were easily readable irrespective } of the brightness of the image below. } (snip) } Does anyone possess a suitably outlined font, or has come up with a creative } solution to this vexing problem?
We use Adobe Photoshop. In version 4.0, be sure to select the outlined text tool so that the letters are surrounded by marquees when you are done typing. Type your text on the image. Before you de-select the text, go to the pallette region of the toolbar and exchange foreground (usually black) and background (usually white) colors. Then go to the "Edit" menu and select "Stroke" chose a 2-3 pixel stroke width "outside" the letter. This makes an outlined text in any font on your system.
-- Best Regards, John Minter
Eastman Kodak Company Phone: (716) 722-3407 Analytical Technology Division FAX: (716) 477-3029 Room 2142G Bldg 49 Kodak Park Site email: minter-at-kodak.com Rochester, NY 14562-3712 calendar: via PROFS
Seemed like PhotoStyler used to have an option for "drop shadowing". I would assume that PhotoShop also would have that eature. you could chose the amount and direction of shift of the shadow. I think the colors were also cusomizable.
However, I don't have acopy of either very accessible just now. For as much as it comes in handy, I can't imagine such a feature being dropped. } } Dear Colleagues: } } I have been searching for an electronic overlay font (preferably a } Microsoft compatible "True Type" variety rather than an Adobe "Post } Script") to facilitate labeling of gray-scale images. Is anyone aware of a } source for such work saver? } } In the pre-digital days, we were getting good results with the paste-on } overlay letters that are still available from microscopy supply houses } (e.g., I was partial to the Microscopist's Collection from SPI). These } were the little black letters that were printed over slightly larger white } outlines. Because of this contrasting outline, they were easily readable } irrespective of the brightness of the image below. } } Currently, we are barely getting by in labeling of electronic images by } adjusting the font color to either white or black, according to what the } brightness of the underlying image dictates. More often than not, this is } ineffective in EM images with highly modulated brightness. On occasion, I } have resorted to manually overlaying a black text over a white one in a } bold version of the same font; this is tedious and does not work in all } but the briefest annotations because the kerning of the two font styles } does not compensate for the differences in the widths of the letters. } } The simplistic solution of setting the overlay text box background color to } non-transparent is not satisfactory because it often ends up obscuring the } feature of interest. } } Does anyone possess a suitably outlined font, or has come up with a } creative solution to this vexing problem? } } valdemar-at-fast.net } Valdemar Furdanowicz ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
I am posting this for the principal investigator listed below. Please send all request and questions to the person at the address listed. Thank you
EXPERIENCED ELECTRON MICROSCOPIST
To participate in highly interactive and motivated lab of molecular biologists; analysis of gene knockout mice with emphasis on the cytoskeleton. Ph.D. is desirable; expertise in immuno EM is exxential. Long-term position. Submit r=E9sum=E9. and list of three references to:
Dr. Elaine Fuchs Howard Hughes Medical Institute The University of Chicago 5841 Sout Maryland Avenue MC1028 Room N314 Chicago, IL 60637 =46ax: 773-702-0141
Mei Lie Wong Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 email wong-at-msg.ucsf.edu
} One solution is to import your image files into } Microsoft Word (other word processors may have } similar functions) and use the WordArt tool
Why not import the image file into PhotoShop or Corel where there is a huge variety of text/drawing tools and little constraint on the size/resolution of the image file? You can do composites, layouts, cutouts, masks, etc. quite handily.
If you do, you will pellet your gold. As a matter of fact, this is one way to clean up your old reagents of protein that has come off.
On Thu, 2 Oct 1997, Pat Hales wrote:
} Date: Thu, 02 Oct 1997 09:34:31 -0700 } From: Pat Hales {hales-at-medcor.mcgill.ca} } To: microscopy-at-sparc5.microscopy.com } Subject: Gold Conjugated Antibodies } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } For years we have "airfuged" our fluorescent antibodies at 95 000 rpm before } use to eliminate any aggregates which may have formed during storage. Does } anyone know if this can (or should) be done with gold conjugated antibodies? } } TIA, } } Pat Hales } McGill University } Dept. of Anatomy & Cell Biology } hales-at-hippo.medcor.mcgill.ca }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Dear Richard, The nicest clear resin I have seen is one the geologists use called: transoptic. It is a hot-press resin that comes up hard and clear and in good contact with the material. I don't know any details of where to get it. You wrote: } O.k., resin wizards, I have a user looking a truely clear, i.e. } water clear, resin for emmbedding some small objects (1-2 cm x 2 mm } neoperene O-rings) for gifts as paper weights. Therefore } sectionablity or LM / EM is not a factor but durability and hardness } are - they would like "nice and hard" resin. } } I realize that this is not a strickly scientific quiry - and I } apologize to anyone offended by any triviality - but why not ask the } experts, eh? After who among us hasn't emmbedded a cockroach, etc. } and kept it on their desk? } } Commercial vendors should feel free to respond. } } Thank you. } } } 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
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I am a graduate student in the MBA program at Rockhurst College in Kansas City, Missouri.
For a class I am taking this semester, I am required to conduct some marketing research. This research will determine the public's attitudes toward a variety of consumer products.
Your email address was randomly chosen by me to be invited to participate in this program. If you would like to cooperate, you will be asked to answer via email a series of questionnaires. The surveys can always be answered by using the "RESPOND" feature of your email software and filling in the blanks on forms that will be sent to you.
Each survey should take only a couple of minutes to complete. There will be a small non-monetary gift awarded by email to those who complete all the surveys on time.
To participate in this project you must: - Be at least 18 years old. - Be a citizen of the United States. - Have an email address that ends with .gov, .com, .org, .net, or .edu.
If you would like to help me with this project, please send an EMPTY email to jdeshon-at-sky.net with only the words "YES SURVEY" (without the quotes) in the SUBJECT line. I will email you further instructions.
If you don't want to participate, kindly ignore this message. If I hear nothing from you, I will place your email address on my suppression file and you will never be bothered by me again.
Whether you participate or not, I hope you have a wonderful day.
I'm wondering whether anyone has remote TEM capabilities working via Internet connections, similar to remote SEM facilities that I've seen and heard about. I was told that remote TEM was not currently available. I would like to know whether remote TEM is useful and/or practical.
Casey Lu Assistant Professor of Biology Humboldt State University
I'm planning on submitting a grant proposal to NSF this fall to obtain matching funding for a TEM at Humboldt State University, located in far northern California. One of the questions that I must answer is "Why teach undergraduates TEM?" TEM is obviously a major tool used by cell biologists, and biologists in general must regularly interpret TEM-based information, but do they really benefit from learning the techniques? I would argue that they do greatly benefit and point out that students of TEM learn how to design and carry out rather involved biological investigations. They must be careful, persistent, and use reasoning to determine whether real changes have occurred in tissues/cells under investigation. I'm wondering if there are other benefits for students who learn TEM, such as possessing a valuable skill for jobs in the drug industry etc.
I would greatly appreciate any thoughts on the value of learning TEM as an undergraduate biology major.
Casey Lu Assistant Professor of Biology Humboldt State University
Casey: check out the URL: http://tpm.amc.anl.gov/MMC.
Also, details about Remote Microscopy at the High Temperature Materials Lab at ORNL are given at http://www.ms.ornl.gov/htmlhome/mauc/MAGRem.html.
Larry PS who told you remote TEM was not currently available?
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
We start to work in the field of image analysis in application to the light microscopy of steels, aluminium, titanium and copper alloys. Our problems are standard: grain size measurement, distribution of grain size, area of phases %, and so on. We try to assemble the simple system on the base of PC (486DX100). Now we looking for the software for this problems, but financial situation in Ukraine is rather bad. We have the preliminary information about the existing of free ware and shareware programs for quantitative metallography and image analysis for IBM PC compatible computers. The not expensive commercial programs in this field are interested for us too. We need for Your advise: where we can obtain or buy this programs ? The most convenient way for us is to transfer the files via E-mail. Ordinary and express mail services (for example UPS or DHL) are accessible for us too. We are very grateful for Your help.
With best regards Vladimir Pashinsky
Ukraine (fSU), Donetsk State Technical University, Department of Material Science
We start to work in the field of image analysis in application to the light microscopy of steels, aluminium, titanium and copper alloys. Our problems are standard: grain size measurement, distribution of grain size, area of phases %, and so on. We try to assemble the simple system on the base of PC (486DX100). Now we looking for the software for this problems, but financial situation in Ukraine is rather bad. We have the preliminary information about the existing of free ware and shareware programs for quantitative metallography and image analysis for IBM PC compatible computers. The not expensive commercial programs in this field are interested for us too. We need for Your advise: where we can obtain or buy this programs ? The most convenient way for us is to transfer the files via E-mail. Ordinary and express mail services (for example UPS or DHL) are accessible for us too. We are very grateful for Your help.
With best regards Vladimir Pashinsky
Ukraine (fSU), Donetsk State Technical University, Department of Material Science
Software such as Image Pro and Materials Pro are available through us, Evex analytical or through your Image Pro Reseller in the Ukraine.
For more information contact Medica Cybernetics at www.mediacy.com
Cheers Peter Tarquinio Evex Analytical
----------
Dear colleagues,
We start to work in the field of image analysis in application to the light microscopy of steels, aluminium, titanium and copper alloys. Our problems are standard: grain size measurement, distribution of grain size, area of phases %, and so on. We try to assemble the simple system on the base of PC (486DX100). Now we looking for the software for this problems, but financial situation in Ukraine is rather bad. We have the preliminary information about the existing of free ware and shareware programs for quantitative metallography and image analysis for IBM PC compatible computers. The not expensive commercial programs in this field are interested for us too. We need for Your advise: where we can obtain or buy this programs ? The most convenient way for us is to transfer the files via E-mail. Ordinary and express mail services (for example UPS or DHL) are accessible for us too. We are very grateful for Your help.
With best regards Vladimir Pashinsky
Ukraine (fSU), Donetsk State Technical University, Department of Material Science
Casey Lu asked about remote control of TEM/SEM over the internet
In addition to the reference that Larry Allard gave to the DoE2000 Materials Microcharacterization Collaboratory (http://tpm.amc.anl.gov/mmc) which has links and connections to the TPM programs at ANL, LBNL, NIST, ORNL, & the University of Illinois. You should also get a copy of the proceedings of Microscopy & Microanalysis '96 meeting in Minneapolis. The proceedings were published by the society through the Journal of the Microscopy Society of America (http://www.msa.microscopy.com).
At that meeting there was an entire day long session on TelePresence Microscopy in Education and Research. This symposium covered nearly all aspects of TelePresence from TEM to SEM. You can obtain copies of most all of the abstracts via the MSA site (http://www.msa.microscopy.com) in Adobe Acrobat PDF format, to find these you must go to "Past and Future MSA meetings" on the MSA site and then use the "View an On-line Copy of the 1996 Meeting Abstracts" link. Specify the search phrase "TelePresence Microscopy". There are 13 papers listed there, 6 of which deal with TEM.
A follow-up symposium will also be held at the Microscopy & Microanalysis 98 meeting in Atlanta. The meeting is scheduled for July 12-16th so mark your Calendar if your interested.
Thanks to everyone who responded to my query about marking fluorescent TEM screens. Most related that it can be done, carefully, with a soft lead pencil. I tried it, it worked. Contact me off-line if you'd like more info.
Owen
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
Has anyone performed image processing and analysis on nerve? Following is the question posed to me, I am clueless on what I would be measuring/doing regarding nerve analysis.
I do not have limited IA and EM experience with nerves would anyone recommend the best methods for fixation, and the types/concentration of fixatives used.
Input from Experts on neurology would be appreciated.
I suspect that there are probably reports of quantitative approaches to assessing peripheral nerves?? Does anyone have any suggestions?
Project:
I will be working with a diabetic drug, I believe designed to be an insulin "secretagog" (don't quote me on that) which in a previous mouse study caused a peripheral neuropathy morphologically similar to the classic spontaneous ageing neuropathy of mice. The hypothesis is that drug related changes in blood glucose contributed to the pathogenesis and thus it is a pharmacologic effect.
What questions should I be asking Gregory.Argentieri-at-pharma.novartis.com
---------- Von: Deutschlaender, Norbert, Path. An: Gregory.Argentieri-at-sandoz.com Betreff: AW: Image Processing and Analysis on Nerve Datum: Dienstag, 7. Oktober 1997 12:04
Dear Gregory, be careful not to mix up "diabetic neuropathy" (with hyperglycemia) and hypoglycemic damage. Isn't the drug you mention a compound producing hypoglycemia (followed by mainly cerebral alteration, i.e. in cortex and hippocampus; rarely in anterior horn neurons of spinal cord followed by axonal degeneration of peripheral nerves)? This is the first question you have to ask. Before having a clean qualitative neuropathologic diagnosis you better do not go into any morphometry of nerves (fiber density, fiber diameter, "roundness" of profiles, myelin sheet thickness etc.). At least in rats and humans the target for hypoglycemic damage is the central nervous system. Immersion of nerves with fixative (for instance Karnovsky's) turns mostly out to be adequate to perfusion, if you inhibit postmortal contraction (in situ fixation of nerve segment fixed to a wooden stick by 2 filament-knots before dissection, or at least fixation of segment attached to piece of firm paper).
Think further of doing fiber teasing, and using more distal nerve segments (tibial, sural nerve) instead of sciatic, because you may have a distally pronounced axonopathy in this model.
Norbert.
Dr. med. vet. Norbert Deutschlaender; Hoechst Marion Roussel Frankfurt/Germany. ---------- Von: Gregory.Argentieri-at-sandoz.com An: Gregory.Argentieri-at-sandoz.com; microscopy-at-Sparc5.Microscopy.Com Betreff: Image Processing and Analysis on Nerve Datum: Montag, 6. Oktober 1997 18:11
Greg, for your information, Rob Vogt at Erim (Environmental Research Institute of Michigan) has done extensive work in nerve resconstruction using Visilog. I believe he works on AXXON fibers or something similar.
If you want his phone or email, let me know. I can also fax you a paper that he has written on the subject if you are intereted.
Luc Noesis Vision Inc.
At 01:07 PM 10/12/97 -0500, Gregory.Argentieri-at-sandoz.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
---------------------------------------------------------------------------- -------- Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com http://www.noesisvision.com 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada ---------------------------------------------------------------------------- --------
We paste up our digital images onto pages using CorelDraw! CorelDraw! enables you to make an outline around your letters in a contrasting colour. So we can have black letters with a white minimal outline or vice-versa. It works with any typeface. And of course you can make the text any colour or shade of gray.
Mel Dickson Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
To convert XL.img files to tiff one can use either:
maketiff.exe (some Philips made program) DeBabelizer by Equilibrium opens .img files as well Graphicconverter by Lemkesoft (not sure)
Thanks again.
Andreas Loewe
______________________________________________________________ Andreas Loewe Tel: +49-228-734-180 University of Bonn Fax: +49-228-734-205 Institute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
} Dear colleagues, } } We start to work in the field of image analysis in application } to the light microscopy of steels, aluminium, titanium and } copper alloys. Our problems are standard: grain size } measurement, distribution of grain size, area of phases %, } and so on. We try to assemble the simple system on the } base of PC (486DX100). Now we looking for the software for } this problems, but financial situation in Ukraine is rather bad. } We have the preliminary information about the existing of free } ware and shareware programs for quantitative metallography } and image analysis for IBM PC compatible computers. The not } expensive commercial programs in this field are interested for } us too.
Dear Vlad,
there is quite a selection of shareware programs for the PC though I'm not sure how well they will perform on a 486. You might have to invest in a Pentium.
Try the following download sites:
Image Tool: http://ddsdx.uthscsa.edu/dig/itdesc.html ftp://maxrad6.uthscsa.edu ImageTool Mirror Sites Belgium - ftp://pa.cc.kuleuven.ac.be/pub/ImageTool Japan - ftp://gold.fish.kagoshima-u.ac.jp/pub/Win/science/ImageTool UK - ftp://micros.hensa.ac.uk/mirrors/uthscsa USA - ftp://wuarchive.wustl.edu/packages/graphics/image-tool
Osiris: http://www.expasy.ch/www/UIN/html1/projects/osiris/osiris.html OSIRIS software can be obtained free of charge from: Digital Imaging Unit University Hospital of Geneva 24 Micheli du Crest 1211 Geneva 14 - Switzerland Fax: (+41 22) 372 61 98 email : osiris-at-dim.hcuge.ch A special developper license is available for the full source code. Please contact us for more information.
Mage: ftp://suna.biochem.duke.edu/pub/PCprograms/
ScionImage: http://www.scioncorp.com ftp://scioncorp.com/ (the Macintosh program NIH image ported to the PC, but it seems to require a Pentium. Maybe You can get an older version for the 486.)
If You need any help, don't hesitate to ask.
Philip
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
POSTDOC POSITION IN COMPUTATIONAL SOLID STATE SCIENCES
At Department of Physics, Norwegian University of Science and Technology (NTNU), Trondheim, Norway there is a vacant postdoc position for a two years period. The postdoc will work in the Electron microscopy group, NTNU and the starting date is December 1st, other dates can be discussed. The position is financed by the Norwegian Research Council under a joint program between Professor Ragnvald Høier NTNU; Professor Johan Taftø, University of Oslo; Dr. Bjørn Anderson, SINTEF Materials Technology, Oslo and Dr. Jon Samset, IFE, Oslo.
Applicants should have a background in solid state theory and preferentially experience with computer modelling. The postdoc will be involved in material modelling problems closely connected to ongoing experimental activity ranging from early stages of nucleation and growth of new phases in aluminium alloys to bonding and ductility in intermetallics and EELS interpretation. Further, in addition to general material characterisation the group at NTNU is in particular strongly involved in development and application of quantitative convergent beam electron diffraction.
The computing facilities are good (CRAYT3E and UNIX workstations).
Applications should be sent to Professor Ragnvald Høier, Department of Physics, Norwegian University of Science and Technology, N-7034 Trondheim, Norway as soon as possible and before Nov. 10th. Phone +47-73-593588 Fax +47-73- 597710 e-mail ragnvald.hoier-at-phys.ntnu.no
More information can be obtained from: Dr. Knut Marthinsen, SINTEF Materials Technology, N-7034 Trondheim, Norway e-mail: knut.marthinsen-at-matek.sintef.no or
Dr. Randi Holmestad, Department of Physics, Norwegian University of Science and Technology, N-7034 Trondheim, Norway e-mail: randih-at-phys.ntnu.no
I am forwarding a question to the list from a colleague who is not a member of the list. Please respond to his address as requested. Thank you.
Hello, I am seeking information on the use of Cationized Ferratin in experiments to study extracellular structures on cellulolytic bacteria (or any others for that matter). I believe that the nature of the stain is such that it can cause proteins to aggregate on the surface of bacteria and cause inaccurate readings. If you have any information either in support or disproving this idea please let me know. I would love to discuss my research, as we are trying to get a publication ready. I am not a member of this list so please send your responces directly to me at the address below.
Here is an updated and slightly amended version of the EM technician job which I mailed to the list recently.
Informal enquiries to me
Chris
UNIVERSITY OF MANCHESTER SCHOOL OF BIOLOGICAL SCIENCES
RESEARCH TECHNICIAN, GRADE C
(ELECTRON MICROSCOPY)
Applications are invited for the position of electron microscope technician within the School of Biological Sciences Electron Microscope, Graphics and Photography Unit. The post will involve working closely within a team of technicians in the Unit; providing assistance with electron microscopy operation, sample preparation, image processing, image archiving, data interpretation, networking and computer software management and maintenance. Applicants should have working experience in electron microscopy and preferably a working knowledge of computers. The salary for this post is the grade C scale stlg10,735 - stlg12,037.
Applicants should be qualified to a minimum ONC/2A - level standard and preferably hold an HNC/BSc in a biological sciences subject, physics, computation or mathematics.
Application forms and further particulars are available from Mr A. Nicholas, School of Biological Sciences, University of Manchester, 2.205 Stopford Building, Oxford Road, Manchester, M13 9PT, UK. email anichola-at-fs1.scg.man.ac.uk
The closing date for applications in October 31, 1997.
The University of Manchester is an equal opportunities employer
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
I am physicist and so my SEM-EDX application field is inorganic material. I need to know whether a very small dried sample is blood. I have not any device to biological sample preparation. I have obtained the following EDX standardless semicuantitative analysis:
Unknow sample My own blood sample
C = 50.9 C = 47.4 O = 39.3 O = 44.6 Na= 0.5 Na= 1.2 Si= 0.1 Si { 0.1 (presence) P = 0.4 P = 0.2 S = 1.8 S = 1.4 Cl= 2.8 Cl= 2.7 K = 3.2 K = 1.6 Fe= 0.6 Fe= 0.4 Cu { 0.1 (presence) Cu { 0.1 (presence)
Are sufficient these data to conclude that the unknow sample is blood or furthermore human blood?. If not, what may I to do?.
Colleagues: Does anyone know of a good basic reference (textbook, tutorial, etc.) for fluorescence in situ hybridization (FISH)? Looks like I'm going into the FISH business, and I need to learn the basic techniques. Any help most humbly appreciated.
TIA-- Bev Maleeff SmithKline Beecham Pharmaceuticals Toxicology-US, UE 0462 709 Swedeland Road King of Prussia, PA 19406 610/270-7987 610/270-7202 fax Beverly_E_Maleeff-at-sbphrd.com
} I am physicist and so my SEM-EDX application field is inorganic } material. I need to know whether a very small dried sample is blood. I } have not any device to biological sample preparation. I have obtained } the following EDX standardless semicuantitative analysis: } } Unknow sample My own blood sample } } C = 50.9 C = 47.4 } O = 39.3 O = 44.6 } Na= 0.5 Na= 1.2 } Si= 0.1 Si { 0.1 (presence) } P = 0.4 P = 0.2 } S = 1.8 S = 1.4 } Cl= 2.8 Cl= 2.7 } K = 3.2 K = 1.6 } Fe= 0.6 Fe= 0.4 } Cu { 0.1 (presence) Cu { 0.1 (presence) } } Are sufficient these data to conclude that the unknow sample is blood or } furthermore human blood?. If not, what may I to do?. } } - Thank you in advance -
What is the sample? In what form did you get it? Are there square or cubical structures that are probably salt crystals (from a buffer solution)?
Have you examined the sample at low kV? {5kV, 1 or less better. You should still be able to see indications of (likely badly distorted) biconcave red cells (RBCs), and more likely platelets. Depending on how long the sample set before it dried, the platelets will be like tiny discs (smaller than the RBCs--say half the size or less, depending on how much the RBCs shrank) if it dried quickly. If the sample dried slowly, so the platelets were in fluid for a half hour or so, they will spread out and thin, with an irregular margin. They'll be larger in size, but very thin. 5kV is likely to be too high a voltage, likely you'll go right through them and not see the platelets. 1kV or less is needed. Examine your own blood sample first, starting with a fresh one, to find these structures, then go to the unknown. I'm not giving sizes, as there are too many variables for that to be meaningful, other than "smaller than RBCs", sizes of which in the SEM depends on how prepared and how dried (everything optimum, they're round about 8 microns in the SEM).
Any chance of getting the sample onto a formvar coated TEM grid, then putting that grid over a hole in the SEM stub? You'll have a better chance of seeing things if you're not imaging the substrate through the sample.
I am seeking information(micrographs, etc.) on the low-cycle-fatigue and high-cycle-fatigue fractures of ferritic malleable iron. The usual handbooks are of very little help. Thank you for any help.
My favorite program for creating and manipulating text is also CorelDraw. However, I just bought a package of plugins for use with Photoshop or Corel PhotoPaint. The package from Xaos (http://www.xaostools.com/products/index.html) cost $129 and included: TypeCaster, Paint Alchemy and Terrazo. The TypeCaster plugin will enable you to create highly visible text on your micrographs--it may be difficult to restrain your creative impulses. The other plugins may be useful for creating backgrounds for slides, web page images and as simple relief from the humdrum of scientific data.
I have no connection with Xaos financial, social or otherwise--wish I did! 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
Electron Probe filaments are just the same as are being used in the typic= al SEM. The difference is the position of the filament. If you place the filament a long way from the cap you need less heat because the bias fiel= d is more effective, the result is less evaporation, less emission current=
and a longer filament life. You do not need many electrons to generate enough x-rays for analysis compared with normal SEM imaging!
Saturation is saturation, it should be on the plateau of the graph. =
However the plateau may be moved higher or lower in the heat range depending upon the position of the filament and the amount of bias being used. I am afraid all those who have replied seem to be writing off electron gun importance, this is totally wrong, probe current is basicall= y number of electrons, get more from the gun and the current goes up. The gun sets the quality of an SEM image, set the gun up incorrectly and the rest of the system cannot compensate, you just run out of electrons!
Driving the filament hard means pushing it forward and increasing the bia= s field to constrain the beam but aiming in a Japanese instrument for 100 t= o 120uA emission current. This filament pushed forward increases the numbe= r of electrons being emitted from the cap, but this means more heat is required to reach saturation as more heat is lost to the cathode cap and because the bias field effect is weakened. More heat shortens the filame= nt life through evaporation. Increasing the bias constrains the electrons funnelling them together to try to achieve a small source. High performance requires a small high electron dense source which is improved=
further by using the correct anode-cathode distance of 1mm for every 2kV.=
Put very simply put a 50um source gives a 50A microscope, the condenser system giving about a 10,000X reduction. Reduce the source size but keep=
up the number of electrons it contains and you improve the instruments performance, 40um source gives a 40A microscope, it is possible to get mo= re than you paid for; the cost is filament life!
I can't vote positively for your results, although I am working in physical science as well.
In the range of my knowledge, a large error could be induced in the standardless EDS analysis for the light elements such as C and O, except you got a super-advanced detector and software to do the job. The results could be used to prove that the major contents of your sample are C and O, or possibly with H and N which you have not counted into the results or which are undetectable by the EDS.
The other reason I would not support your suggestion of the "human blood" is that the measurements of the minor contents such as Si, P, Na and Cu were less than 0.5%. They should be considered as the experimental error, rather than the evidence. The present of a few percent of S, Cl and K is also quite common in many organic samples, at least I met several times.
After all, I think that you should have more sufficient evidence to support your argument. The results of EDS could be trusted in many cases, but it failed to convince me this time.
Regards,
Charlie
Jose Luis Encinas wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi, } } I am physicist and so my SEM-EDX application field is inorganic } material. I need to know whether a very small dried sample is blood. I } have not any device to biological sample preparation. I have obtained } the following EDX standardless semicuantitative analysis: } } Unknow sample My own blood sample } } C = 50.9 C = 47.4 } O = 39.3 O = 44.6 } Na= 0.5 Na= 1.2 } Si= 0.1 Si { 0.1 (presence) } P = 0.4 P = 0.2 } S = 1.8 S = 1.4 } Cl= 2.8 Cl= 2.7 } K = 3.2 K = 1.6 } Fe= 0.6 Fe= 0.4 } Cu { 0.1 (presence) Cu { 0.1 (presence) } } Are sufficient these data to conclude that the unknow sample is blood or } furthermore human blood?. If not, what may I to do?. } } - Thank you in advance -
Can anyone tell me of a supplier of thin sections of minerals and rocks for educational purposes? I am told that Carolina Biological is a possibility, and have requested their catalog, which is long in coming. Are there additional suppliers?
I'm not looking for labs that make thin sections from rocks and minerals that the customer supplies.
W. Riedel Scripps Institution of Oceanography UCSD La Jolla, CA 92093-0220
San Francisco Microscopical Society and California Association of Criminalists Trace Evidence Study Group
Joint Dinner Meeting Announcement October 30, 1997 Berkeley, California
Speaker: Brian J Ford Topic: Antony van Leeuwenhoek and the Single Lens Microscope
We are delighted to announce a very special evening featuring the noted English scientist and author, Brian J. Ford. Have you ever wondered how 17th and 18th century microscopists could prepare images of insects, cells, bacteria, and all manner of microscopic organisms and structures without SEMs, DIC, PCM, confocal microscopes, scanning tunneling microscopes and all those other marvelous inventions of the 20th century? How can a single lens microscope reveal structures that are even now difficult to study with sophisticated research microscopes? How indeed! Come hear some amazing answers to these questions.
Our program this month features a presentation on Antony van Leeuwenhoek (1632-1723) and single lens microscopy. Professor Ford is the author of countless books and papers and has done extensive research on the historical development of microscopy, including work with original, 17th century Leeuwenhoek instruments and samples from the archives of the Royal Microscopal Society in London. Many American microscopists know Professor Ford through his annual presentations at Inter/Micro in Chicago.
For further information on this meeting please see a special web page prepared for this special occasion: The URL is http://ourworld.compuserve.com/homepages/steve_shaffer/announce.htm - check it out NOW!
-- Submitted by Stephen A. Shaffer MicroDataware sshaffer-at-microdataware.com (business) steve_shaffer-at-compuserve.com (personal)
Dear list, there were a lot of questions conserning image processing software - that is one more. Does anyone can point as to a free-, share- or commercial software for crystal images processing (excluding CRISP - we know it well)? What we need other than different filters is to measure lattice distorsions and spots shifts on the large area.
The CI number for Methylene blue is 52015. The CI number for Azure B (sometimes called Azure I) is 52010. There is no CI number for Azure II since it is a 1:1 mixture of the the two stains above. Could you simply mix 52015 and 52010? I have not done this, simply because the Azure II worked fine, and I don't have time to fix things which already work reliably. I cannot think of any reason why mixing the two stains rather than buying the azure II would alter the situation. We buy our methylene blue and our Azure II from Sigma. I do not know if the percentage of active stain varies per gram between batches. This is something one needs to be aware of given the vast interactions between LM stains and embedding epoxies. I also used to buy very nice stains from Fisher (but someone stole our Fisher Catalog)! PS. I do not have any commercial interest in either of the companies mentioned. The price of these stains can vary enormously so it pays to check several sources. Just make sure that the company states the CI number of the product they are selling. Please note that the stain improves with age. Make a lot and let it sit in the dark at rt. Also please note that if the stained section is not rinsed with alcohol (75% - destained, and then with 100%), the result will not be as brilliant as it should be. Bye, Hildy
It is out of my field also, but I understand there is a chemical which, when applied to blood will cause it to floresce under UV light.... Perhaps someone else cam be more specific.
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Hi,
I am physicist and so my SEM-EDX application field is inorganic material. I need to know whether a very small dried sample is blood. I have not any device to biological sample preparation. I have obtained the following EDX standardless semicuantitative analysis:
Unknow sample My own blood sample
C = 50.9 C = 47.4 O = 39.3 O = 44.6 Na= 0.5 Na= 1.2 Si= 0.1 Si { 0.1 (presence) P = 0.4 P = 0.2 S = 1.8 S = 1.4 Cl= 2.8 Cl= 2.7 K = 3.2 K = 1.6 Fe= 0.6 Fe= 0.4 Cu { 0.1 (presence) Cu { 0.1 (presence)
Are sufficient these data to conclude that the unknow sample is blood or furthermore human blood?. If not, what may I to do?.
I want to thank everyone that responded to my request for TEM labs. We are now in the process of wading through the myriad labs that were recommended. Hopefully, we will make a decision on which lab we will use within the next week. When we do, I'll be sure to contact each of you individually to let you know of our decision. Once again, thanks a bunch :-)
David Bell Millipore Corporation 80 Ashby Road Mailstop B2C Bedford, MA 01730
The largest perhaps is Ward's Scientific. They have many different sets of these. Pricy though.
Ward's Natural Science Establishment, Inc. 5100 West Henrietta Road Rochester, NK 14692-9012 1-800-962-2660 They may have a web page by now?
} Can anyone tell me of a supplier of thin sections of minerals and } rocks for educational purposes? I am told that Carolina Biological } is a possibility, and have requested their catalog, which is long } in coming. Are there additional suppliers? } } I'm not looking for labs that make thin sections from rocks and } minerals that the customer supplies. } } W. Riedel } Scripps Institution of Oceanography } UCSD } La Jolla, CA 92093-0220 ================================================ Michael L. Boucher Sr. mboucher-at-isd.net 13345 Foliage Avenue Apple Valley, MN 55124-5603 Ph 612-432-8836 ================================================
A few days ago, in response to an inquiry from Casey Lu about remote TEM operation, I posted the following:
Casey: } check out the URL: http://tpm.amc.anl.gov/MMC. } } Also, details about Remote Microscopy at the High Temperature } Materials Lab } at ORNL are given at } http://www.ms.ornl.gov/htmlhome/mauc/MAGRem.html.
It turns out that apparently the web page for the ORNL site does not display fully when addressed using Microsoft Internet Explorer. Netscape seems to work just fine. Also, earlier web pages up to htmlhome work OK on IE. We are checking to see what the problem might be. I will post when something is resolved, in case anyone else has encountered this problem.
Larry
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Electron microscopy specialist needed in spring 1998. Prepare ultra thin sections, and photogragh them using JEOL 1200EX. Darkroom, Adobe Illustrator/Photoshop.
Salary commensurate with experience.
Mail or fax resume and two letters of recommendation to:
Dr. Peter Sterling 123 Anatomy/Chemistry BLDG Department of Neuroscience University of Pennsylvania Philadelphia, PA 19104-6058
We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured by EMSCOPE) that is having problems attaining a low enough temperature prior to flushing with carbon dioxide. The manual suggests a temperature below 10C - we can rarely get to below 12C and frequently only get to 14C. Ambient temperature at the moment is only around 20C. Has anyone any suggestions? As usual we have no specifications for the electronic circuit nor a circuit diagram.
Thanks
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
Are we talking about Richardson's stain? (I don't have the reference to hand, but I could find it if anyone wants.) This is a 1:1 mixture of 1% methylene blue in 1% borax with 1% Azure II in water. It is my routine stain for thick sections and gives brilliant metachromatic results, even without an alcohol rinse.
Lesley Weston
On Tue, 14 Oct 1997, HILDEGARD CROWLEY wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Hi, } } The CI number for Methylene blue is 52015. } The CI number for Azure B (sometimes called Azure I) is 52010. } There is no CI number for Azure II since it is a 1:1 mixture of the the } two stains above. } Could you simply mix 52015 and 52010? I have not done this, simply } because the Azure II worked fine, and I don't have time to fix things } which already work reliably. I cannot think of any reason why mixing the two } stains rather than buying the azure II would alter the situation. } We buy our methylene blue and our Azure II from Sigma. I do not know if } the percentage of active stain varies per gram between batches. This is } something one } needs to be aware of given the vast interactions between LM stains and } embedding epoxies. I also used to buy very nice stains from Fisher (but } someone stole our Fisher Catalog)! PS. I do not have any commercial } interest in either of the companies mentioned. The price of these stains } can vary enormously so it pays to check several sources. Just make sure } that the company states the CI number of the product they are selling. } Please note that the stain improves with age. Make a lot and let it sit } in the dark at rt. Also please note that if the stained section is not } rinsed with alcohol (75% - destained, and then with 100%), the result } will not be as brilliant as it should be. } Bye, } Hildy }
Andrey L. Chuvilin wrote: -------------------------------------------------------------------. } } Dear list, } there were a lot of questions conserning image processing software - that } is one more. Does anyone can point as to a free-, share- or commercial } software for crystal images processing (excluding CRISP - we know it } well)? } What we need other than different filters is to measure lattice } distorsions and spots shifts on the large area.
Dear Andrey,
I know of 3 shareware programs that will measure and correct lattice distortions:
ICE: http://ncmi.bioch.bcm.tmc.edu/ftp.html http://condor.bcm.tmc.edu/3DEM/download.html ftp://ncmi.bioch.bcm.tmc.edu/pub/ICE/ which is a menu driven C-version of the Fortran-classic
MRC: http://www.EMBL-Heidelberg.DE/ExternalInfo/fuller/em_mrc_overview.html to obtain mail to: rac1-at-mrc-lmb.cam.ac.uk
You could also try EM (also a classic): to obtain mail to: hegerl-at-vms.biochem.mpg.de
A good source of information is the special edition of the Journal of Structural Biology, Vol. 116, No. 1, January/Februray 1996. It reviews most of the software generally used in the field.
Yours,
Philip
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
Dear all, in our lab the question of carcinogenicity of propylenoxide arose. Where can get hard data about it, and which alternative can be recommended in Epon-embedding. Thank you all, Norbert
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
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. The meeting is hosted by Jeffrey Horn of the Toxicology Research Laboratories at Lilly.
The program will begin at 8:30 a.m. and run until about 4:30 p.m. Lunch will be provided courtesy of Abbott Laboratories. In addition to podium presentations, there will be a tour of the Toxicology Facility at Eli Lilly.
The program includes a variety of topics - the proverbial "something for everyone" interested in microscopy and imaging. On the agenda are presentations about digital imaging, diagnostic electron microscopy, forensic electron microscopy, in situ labeling methods, and use of electron microscopy in the selection and development of pharmaceutical therapeutic candidates.
For more details about the program and meeting location, contact me by telephone at (847) 935-01014 or via E-mail at jane.a.fagerland-at-abbott.com.
Lilly is a secured facility, and pre-registration is requested to expedite entry and to allow us to plan for enough food for lunch. Please pre-register by contacting me at the phone or E-mail address above (E-mail is preferred).
Any supplier should be able to get you a MSDS (Material Safety Data Sheet) for it; ours came from Polyscience, 400 Valley Rd, Warrington, PA 18976, 215 343-6484. However WE DO NOT USE IT AT ALL AND HAVE NOT FOR OVER 15 YEARS. You can go straight from absolute ethanol into Eopn substitutes and Spurr as long as it is DRY. We use drying beads (molecular sieves) in all our 100% EtOH bottles. We keep only about 200-300 ml in a bottle, and when empty, we bake the beads. If there is silt, let it settle out and pipet off from the top. There has been a discussion of drying beads on this net recently.
On Wed, 15 Oct 1997, Deutschlaender, Norbert, Path. wrote:
} Date: Wed, 15 Oct 1997 10:07:00 +0200 } From: Deutschlaender, Norbert, Path. {DEUTSCHLAE-at-MSMPFEI.Hoechst.com} } To: "microscopy-at-sparc5.microscopy.c" {microscopy-at-sparc5.microscopy.com} } Subject: propylenoxide } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear all, } in our lab the question of carcinogenicity of propylenoxide arose. Where } can get hard data about it, and which alternative can be recommended in } Epon-embedding. } Thank you all, } Norbert }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
After years of always going from 100% ethanol to 100% propylene oxide before 1:1 epon:propylene oxide, I now go directly from 100% ethanol to 1:1 ethanol:epon and haven't seen any difference. Some resins (epon-araldite????) may require a propylene oxide intermediate but it doesn't seem to be required for epon only. } Dear all, } in our lab the question of carcinogenicity of propylenoxide arose. Where } can get hard data about it, and which alternative can be recommended in } Epon-embedding. } Thank you all, } Norbert
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)
Dear Microscopists, We are using chicken embrio fibroblasts (CEF) for Golgi traffic study. We need specific Golgi staining from one side (cis- or trans-). We have tried to stain Golgi with several lectines - Helix Pomatia (HP) , Wheat Germ Agglutinin (WGA), Limax Flavus Agglutinin (LFA), but without any success. We used preembedding technices with lectines conjugated with horseradish peroxidase. HP simply did not stain CEF Golgi. WGA and LFA stained practically only endosomal compartment and rarely trans-Golgi network, but not trans-cisternes of Golgi. All these lectines work good in other cell types as NRK or RBL. I would like to ask does anybody know any specific lectin staining protocol for these cells. Any suggestions about lectin staining of cis- or trans- Golgi cisternes would be greatly appreciated.
At 03:59 PM 10/10/97 +0200, Philip Koeck wrote: } Does anybody have (good or bad) experience with any of the following } or other fast freezing devices: } Reichert KF80 } Leica CPC } Gentleman Jim (constructed by Alan Boyne) } Has anybody made a comparison?
I have had a long history of involvement with these devices, and believe that they are all good instruments which produce comparable results. That being said, I have sold the Gentleman Jim for more than 15 years, first with Ted Pella and now with Energy Beam Sciences, so my prejudices are obvious- the Gentleman Jim is much, much less expensive than the other instruments mentioned.
Best regards, steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Donald P Robertson wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Could anyone suggest names of 3rd party vendors of LaB6 filaments } for an Hitachi H9000NAR. Thanks in advance for any advice. } Donald Robertson
Ladd Research supplies LaB6 filaments for Hitachi scopes under the catalog # 63070. If you wish a quote just contact us.
John Arnott Ladd Research ladres-at-worldnet.att.net tel 1-800-451-3406 fax 1-802-878-8074
I'm looking for a digital camera for an optical microscope that I can control from a program, such as a Visual Basic program, for setting up an automated imaging system. Anyone out there doing this type of thing?
} We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } Dear Ian, Peltier units can go bad. If you can measure the current they draw now and compare that with what they are supposed to draw, you can ascertain whether the unit is working as specified. Generally, the units are in the form of an array of individual devices. I don't re- member if these are series or parallel, but if one or more has shorted (series) or failed open (parallel), the unit will draw more/less current than it should. Since you have neither specs nor diagrams, this info may not be useful, but perhaps someone else can let you know what cur- rent a properly-working device draws. Good luck. Yours, Bill Tivol
I have also avoided propylene oxide for years for safety reasons. (Although I was warned of its flammability/volatility).
Alternatives: Usually I work with Spurr's resin (OK also carcinogenic but more easy to contain) and when dehydrating in acetone no transition solvent is needed. For ethanol, on the other hand, a transition solvent can be very useful in my experience with samples from pig skin to cell cultures. What to use? I have converted to tert-butyl glycidyl ether (t-BGE), available from Aldrich Chemical, Milwaukee, WI USA (cat. #25,171-2). This was recommended in a 1995 Biomaterials Workshop lecture by Hendrik K. Koerten in San Francisco, CA, sponsored by the Society for Biomaterials. It has been particularly useful to me in that it does not dissolve polymer materials/implants as readily as propylene oxide. Another name for this chemical is Butyl-2,3-epoxypropylether.
I have also used t-BGE with Araldite and EmBed 812 (an Epon-clone from Electron Microscopy Sciences), and it worked well.
Another good, or better, choice with the eponates is HPMA (hydroxypropyl methacrylate) which also is more kind to materials (cell culture plates) than propylene oxide.
I believe both HPMA and t-BGE are less flammable/carcinogenic than propylene oxide.
Karen
deutschlae-at-msmpfei.hoechst.com wrote: } } Dear all, } in our lab the question of carcinogenicity of propylenoxide arose. Where } can get hard data about it, and which alternative can be recommended in } Epon-embedding. } Thank you all, } Norbert
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
Title: Biological Science Technician (Plants) Lab: USDA-ARS, Salinas, CA
The USDA-ARS is seeking a biological science technician (plants) (GS-0404-6/7/8/9) for the Crop Improvement and Protection Research Unit in Salinas, CA. The incumbent will share responsibilities in electron microscopy of plant virus infections for ultrastructural characteristics. The incumbent will also assist in research involving molecular, serological, and biological studies of several plant viruses infecting sugarbeet and vegetables. Candidate must have a knowledge of electron microscopy, plant virology, and knowledge of microbiological techniques. Must be a U.S. citizen. Bachelors degree is desirable. Salary is commensurate with experience ($22,805-$40,300 per annum). For information regarding research program contact Gail C. Wisler or James E. Duffus (408)755-2835. For information regarding application procedures/forms contact Elsa Chavez at (408)755-2800. Applications must be postmarked by Nov 3, 1997. The USDA is an equal opportunity employer.
Gail C. Wisler USDA-ARS 1636 E. Alisal St. Salinas, CA 93905 (408)755-2835 FAX (408)755-2814 e-mail: gwisler-at- ASRR.ARSUSDA.gov
John R Reffner wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I'm looking for a digital camera for an optical microscope that I can control } from a program, such as a Visual Basic program, for setting up an automated } imaging system. Anyone out there doing this type of thing?Hi, John,
I believe most of the digital cameras have this capability. In the past, we have had some experience with those from Kodak, Leaf, and Pixera. Also, Polaroid just announced a new digital camera. I would also check to see what Optronics and Dage-MTI are carrying. RE: outlets - call your local system integrator or the manufacturer's directly. (The Kodak Division you need is the one in New Haven; the Polaroid Group is under Jim Landrigan). If you have trouble finding a local source, please email me for manufacturers' contacts.
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 ********** Americas First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
Title: Biological Science Technician (Plants) Lab: USDA-ARS, Salinas, CA
The USDA-ARS is seeking a biological science technician (plants) (GS-0404-6/7/8/9) for the Crop Improvement and Protection Research Unit in Salinas, CA. The incumbent will share responsibilities in electron microscopy of plant virus infections for ultrastructural characteristics. The incumbent will also assist in research involving molecular, serological, and biological studies of several plant viruses infecting sugarbeet and vegetables. Candidate must have a knowledge of electron microscopy, plant virology, and knowledge of microbiological techniques. Must be a U.S. citizen. Bachelors degree is desirable. Salary is commensurate with experience ($22,805-$40,300 per annum). For information regarding research program contact Gail C. Wisler or James E. Duffus (408)755-2835. For information regarding application procedures/forms contact Elsa Chavez at (408)755-2800. Applications must be postmarked by Nov 3, 1997. The USDA is an equal opportunity employer.
Gail C. Wisler USDA-ARS 1636 E. Alisal St. Salinas, CA 93905 (408)755-2835 FAX (408)755-2814 e-mail: gwisler-at- ASRR.ARSUSDA.gov
We have attempted to immuno-label one micron sections cut from LR White with primary antibody followed by TRITC or FITC conjugated secondaries. The sections are dried onto glass slides at 60C (the same temperature at which the blocks were cured), the immunolabel procedure performed, and a cover glass mounted with aquamount (UV clear). These experiments were completely unsuccessful in the fluorescent microscope, even though the same immunolabeling protocol works well in the EM with the antibodies and immuno-gold secondaries applied to the surface of ultrathin sections. I realize there are better ways to label tissue with antibody in preparation for fluorescence microscopy, but we are surprised by the negative result in these experiments. Does anyone have an explanation for the failure? Am I missing something simple? Is it because there is no penetration of primary and secondary into the tissue sections, therefore not enough bound TRITC or FITC to detect? It would be nice if this technique worked so that we could easily test the ability of our primaries to bind specifically to LR White embedded tissue.
Many thanks,
Doug Keene Laboratory for Ultrastructural Research Portland Shrine Research Unit ---------------------- Doug Keene DRK-at-shcc.org
OOPS! I share my husband's mailbox and erased the new address for Alwyn Eades. Where did he go? To avoid multiple responses, please limit reply to those who reside in Chicago, Ill. or immediate vic.
} I heard about a new x-ray microscopy technique that can image cellular } processes in real-time. Have you heard about this? } } } Bob Clark } } Hi ! Interesting idea, but it doesn=B4st seem very likely that it would be possible. According to a simple calculation we made, imaging of a cell would kill it within a few milliseconds, using x-rays. If someone has other information it would be interesting to share it !
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
A Research Fellowship is available from 1 January 1998 for a fixed period until 30 June 1999. The apointee will work on an electron microscope study of high-purity iron alloys to determine the mechanism of formation of coarse side-plate structures from grain boundary polygonal ferrite, and the effect of alloying elements on this transition, which would provide a means, via suppression of this reaction, to produce interlocking acicular ferrite microstructures from intragranular nucleation.
Applicants should have, or be submitting for, a PhD in Metallurgy, Materials Science or a related discipline. Experience in electron microscopy would be advantageous.
Salary will be on the scale for Research Staff Grade 1A within the range stlg15,159 - stlg18,494 p.a. according to qualifications and relevant experience.
Application forms and further particulars may be obtained from Professor D V Edmonds, Department of Materials, University of Leeds, Leeds, LS2 9JT, tel +44 0113 233 2341/8, fax +44 0113 233 3284, email d.v.edmonds-at-leeds.ac.uk
Closing date for applications 7 November 1997.
The University of Leeds promotes an Equal Opportunities Policy. _____________________________ 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.
Dear all, 10/16/97 unfortunately I don=B4t know at which point I=B4m entering = considerations and discussions (due to several failings of our Server). Following the discussion as far as I got the postings to my e-mail-box = (original question by Norbert Deutschlaender, Re Malcolm Haswell, Re = Sara E.Miller, Re Thomas E.Phillips , Re Karen Zaruba) for me there are = several points worth to consider: i) propyleneoxide (PO): why, when, which substitutes possible I was using PO for specimen preparation during my studies in Zoology = about 20 years ago. I was frustrated because of the odious smelling of = PO-vapors and its solving reactivity to safety wear like gloves and = plastic labware. When I got then for the first time a MSDS (material = Safety Data Sheet) on PO and searched for the chemical nature of the = substance I got aware of its properties as a carcinogen, I knew from the = beginning that it is a highly flammable solvent and there were severe = cautions in handling the substance....The main reason(s) for using this = solvent PO in my opinion was/were the high rate of volatilization at = room temperature (20 degr.C), which is, according to MSDS about 588 hPa = (at 33 degr.C: 980 hPa, compare data for Aceton and Acetonitrile, see = below): so in fact that "intermedium" (which as such should aid in the = miscibility of the hydrophilic alcoholic solutions with the hydrophobic = nature of the embedding resin, sometimes also said to be a = "clearing[??]agent") after several changes evaporated as a pure = hydrophobic solvent out from the resin. To produce "no shock treatment" = on the tissues which eventually could lead to severe ultrastructural = changes in morphological terms, that exchange between hydrophilic and = hydrophobic phases usually was/is done by including several changes of = the tissues=20 in relations of EtOH:PO e.g. 3:1, 1:1, 1:3, then pure PO e.g. 3 changes, then pure PO:Epon(-oxide) e.g. 3:1, 1:1, 1:3 then pure resin e.g. several changes to get rid of traces of the = solvent PO.
ii) It turned out many years ago that at least EPON (as a hydrophobic, = non water-miscible substance) only up to 96% would be hydrophobic. It = was said to have a hydrophilic phase at/up to a concentration of 4%, = namely "aquon"-phase. This allowed for an embedding only via 100%, = absolute water-free EtOH, provided several "protected" (against = watervapor-uptake) changes in pure such EtOH as well as several changes = in mixtures of abs.abs. EtOH / Epon were performed. In addition the = "infiltrating" time of each such step had to be prolonged (at least = doubled). I remember the days when the "new" hydrophilic resins like LR White or = the Lowicryls were not "born" yet and people tried to overcome = interactions of organic solvents with tissue antigens at the beginning = of ultrastructural localization of antigens by Antibodies.
iii) I don=B4t have experience with t-BGE (as mentioned by Karen = Zaruba). Karen mentions also the possibility to dehydrate by Acetone = rather than Ethanol. Yes, you don=B4t need an intermedium like PO, = provided the several changes of tissues as mentioned in i). As a = reference (maybe out of many possible/present references) I can quote: = BULLOCK et al (Eds, 1989): Techniques in diagnostic Pathology, Vol. 1, = p. 1 ff (Academic Press, ISBN: 0-12-681911-4) where it is stated that = "Acetone (is) slightly superior than Ethanol as dehydrating agent for = better preservation of Immunoreactivity/Antigenicity in processing Bone = Marrow Aspirates in Plastic". Acetone has a lower rate of volatilization = at room temperature (20 degr.C), which is, according to MSDS about only = 233 hPa (at 33 degr.C: no data available, compare data for Acetonitrile, = see below and PO, see above).
For several reasons (mainly for safety reasons) I use now for a long = time (} 10 years) a substitute for PO, namely ACETONITRILE (syn.: = Methylcyanide) for dehydration, or as an Intermedium, respectively. When = I saw the 2 page publication (CAVE: text p 38: : ACETONITRILE: = Substitute for Propylene Oxide in Tissue Processing for Transmission = Electron Microscopy ; CAVE: Illustrations erroneously are printed on = page 41 instead page 39) by:=20 TARNOWSKI Betty I., & SCHONBAUM Greg R. in the Proceedings of EMSA, = 42nd Ann. Meeting ( 1984) / Detroit I learned a lot of new aspects.
I do not know wether there is an earlier paper on that subject or an = other author(s) to be named for publishing first that substitution = method (please apologize).
Product info: ACETONITRILE: 2-cyanopropane-2-ol, (C 2 H 3 N), MW: 41.05 = g/mol, 1 liter =3D 0.78 kg; melting point: -46 degr.C., boiling point: 81 = degr.C, flash point: 5 degr.C, 100% miscible with water, free from = halogens; e.g. from SIGMA Order# A 3396 (Disclaimer: I have no interest in = SIGMA=B4s..... nor am I affiliated with Sigma....)
Acetonitrile (AN) has -compared to Acetone and PO- the lowest rate of = volatilization at room temperature (20 degr.C), which is, according to = MSDS about only 97 hPa (at 33 degr.C: no data available, compare data = for Acetone, and PO, see above). It is indeed a toxic substance = (methyl-cyanide): due to the reduced vapor-pressure you will have a = tremendous decrease in the possibility of inhaling the toxic vapors when = working with it. Additionally, AN is not classified as carcinogen, so if = you work carefully in a well-ventilated area, you wouldn=B4t need a = filter-mask or even a fume-cupboard (in practical work). At least you = should not eat or drink the solvent! The substance is fully (100%) miscible with water, so, in fact, you = could dehydrate and intermediate, infiltrate by/with only one solvent.=20 I haven=B4t tried a complete dehydration with increasing concentrations = of only Acetonitrile but I am convinced such would work (provided you = can prevent tissues from "rehydration" by hygroscopic saturation of the = solvent via water-vapors or high humidity).=20 As the substance at all has to be classified as toxic, flammable and = vapors to be a health-risk, I=B4m not using it as the only dehydrating = agent.
After I checked that substitution possibility in comparing control = specimen processings (i.e. PO vs. AN) and got aware of that no = alterations in ultrastructural morphology could be detected between PO = and AN-specimens, from 1985 on I have completely changed to the = following schedule for (automatically) processing my diagnostic tissue = samples (standardized protocol, approx. 1500-1600 blocks/year): increasing concentrations of alcohols (as usual), - 4 x EtOH absolutely waterfree (maybe there will be rehydration of = about=20 0.5-1%, when opening bottle and handling: this is no problem at all) (times: each 5/10/10/10 mins) - 3 x pure ACETONITRILE (each 5, 10, 15 mins) - infiltration by mixture=20 EPOXIDE RESIN : ACETONITRILE =3D 1:1 (at least 45 mins) (GLYCIDETHER 100,=20 formerly SERVA/now: BIOPRODUCTS) then - 3 x pure EPOXIDE RESIN (at least until tissue blocks have sunk down = to the bottom of the "infiltrating mold" or infiltrating glass).=20 Eventually I recommend the use of a bulb lamp (e.g. 60 W) to be = positioned approx. 10 cm above the infiltrating mold, at least of the = first pure resin step to aid in evaporation of Acetonitrile remnants in = the tissue/resin mixture. In doing that way, up to now no problems in infiltration- and = polymerization quality occured (except when wet tissue blocks exceeded = dimensions L/W/H of 4x5x1.5 mm, where one should increase duration of = the processing steps).
The suggestion of Sara Miller (or her method) to use molecular sieves = (drying beads) in all of their 100% EtOH bottles I have to mention that = I got serious problems with a following damage of the cutting edge of = our diamond knives when sectioning the blocks (due to silt produced from = the molecular sieve beads, which were infiltrating the tissues and = therefore embedded too). But: maybe this happened due to pouring of 100% = EtOH from the bottle rather than pipetting off from the top of the = solution.
At the end: answering the question of Malcolm Haswell "opinion about which is nastier: Spurr=B4s resin with alcohol or Epon with propylene oxide" I have to "confess": if you have to choose between "devil" and "satanas" you should try to reach the "purgatory" that means: Try to reduce toxicity, handling and healthy hazards, = maintain safety implications, be aware of "the devil and satanas".
Best regards to all Wolfgang MUSS EM-Lab of Dept. Pathol. LKA A-5020 SALZBURG, AUSTRIA/Europe
Hi, 10/16/97 unfortunately I am not aware of a commercial source of Uranyl formate, = which I think has been used as negative staining agent in former years. As an indication I found a fax notice from 1984 that a company in = Germany, namely PAESEL Chemicals (I wonder if they are still alive) = which offered 5 g of that substance at a price of US$ 53.-. Now I have had a look in my files and a phone call: the company is
PAESEL GmbH&Co=20 Moselstrasse 2 B D-63 452 HANAU Federal Republic of Germany phone: ++49++6181-18-700 Fax: ++49++6181-18-7070=20
They told me to have listed Uranyl-formate in quantities of 5 gs, no = price-information could be given at the moment.
Note added: If you still have to produce your uranyl-formate by yourself, you should = have a look on 2 original publications I found, dealing with the = fabrication of Uranylformate:=20
LEBERMANN R (1965): Use of Uranylformate as a Negative Stain, J. Molecul. Biol. 13, p.606 ff
and an alternative preparation method:
BRADLEY D.E. (1965) in J. Gen. Microbiology 38, p. 395 ff
Best wishes and luck to you
Wolfgang MUSS Dept. Pathology LKA, EM-Lab A-5020 SALZBURG, Austria/Europe.
Hi, 10/16/97 unfortunately I am not aware of a commercial source of Uranyl formate, = which I think has been used as negative staining agent in former years. As an indication I found a fax notice from 1984 that a company in = Germany, namely PAESEL Chemicals (I wonder if they are still alive) = which offered 5 g of that substance at a price of US$ 53.-. Now I have had a look in my files and a phone call: the company is
PAESEL GmbH&Co=20 Moselstrasse 2 B D-63 452 HANAU Federal Republic of Germany phone: ++49++6181-18-700 Fax: ++49++6181-18-7070=20
They told me to have listed Uranyl-formate=20
ADDED NEW: Order# 3 36 234
in quantities of 5 gs, no price-information could be given at the = moment.
Note added: If you still have to produce your uranyl-formate by yourself, you should = have a look on 2 original publications I found, dealing with the = fabrication of Uranylformate:=20
LEBERMANN R (1965): Use of Uranylformate as a Negative Stain, J. Molecul. Biol. 13, p.606 ff
and an alternative preparation method:
BRADLEY D.E. (1965) in J. Gen. Microbiology 38, p. 395 ff
Best wishes and luck to you
Wolfgang MUSS Dept. Pathology LKA, EM-Lab A-5020 SALZBURG, Austria/Europe.
Greetings, Doug Keene wrote: } } We have attempted to immuno-label one micron sections cut } from LR White with primary antibody followed by TRITC or } FITC conjugated secondaries. ... } These experiments were completely unsuccessful in } the fluorescent microscope, even though the same } immunolabeling protocol works well in the EM with the } antibodies and immuno-gold secondaries applied to } the surface of ultrathin sections. ....
Keep in mind that the propoprtion of surface area to volume is much higher for a 60 nm ultra section then for a 1 um semithin section. For LR White, if the ab can make it to antigens as deep as 15 nm (I am just guessing at the depth here), then it will find the antigen in half the ultrathin section volume but only in 3% of the semithin section. You may be able to improve your signal at the light level with one of the antigen retrival methods that are around. I have never done these myself, but I have heard that they will work. Alternatively, you can embed your samples in butyl-methyl methacrylate, which is extractable after sectioning and thus gives great access for your ab throughout the section volume. Of course, this resin is not so great in the em (although useable) and so this path would mean that you would have different preps for light and em work (although I believe that LR white is a methacrylate based resin). Hope this helps, Tobias Baskin
Propylene oxide can be omitted from an embedding protocol.
However! it is important to realize that 1)alcohol interferes with polymerization 2)acetone also interferes with polymerization 3)a mixture of PO and 812 is much less viscous than a mixture of 812 and acetone and alcohol 3) PO is actually a monoexpoxide and if remnants are left in the tissue it will become incorporated into the block. The critical issue in many infiltration steps is the viscosity of the monomers. Skin may require PO, liver can do nicely without it.
We have abondoned PO a long time ago. We use acetone instead. However, if the blocks are particularly large or difficult then we go to PO temporarily to insure adequate infiltration. We make absolutely sure that no acetone or alcohol is left in the mixture. This means changing 812 more often, and changing to clean vials after the tissue has been in the first undiluted 812 for one hour.
Dear list We are progressing nicely with digital imaging and printing from our microscopes. We have several networked printers that people are using. I now need to be able to charge accordingly for the printouts that people do. I am using WIN95 networking and NT4.0 server (and Novell 3.12 but want to stay away from that for network printing if I can). Is anyone using server software which will record which printer was used, who used it and how many printouts they did? NTserver event manager is sort of working but seems always to say 0 pages were printed. If anyone has gone down this route I would appreciate some comments.
Raining again in Manchester!
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
I routinly test 1 micron LR White sections on glass before going to the trouble of Immuno EM. I usually use peroxidase ABC which seems to enhance the signal enough to be visualized. However, the fluorescent signal is very weak unless you start stacking antibodies which can create more backround. So you need a very good optimized scope to be able to see the signal and your faint signal may be obscured by the backround signal of the aquamount which has higher fluorescence than some others. Prolong from Molecular Probes is very quiet and enhances Texas Red. Or my favorite for quiet and quick is: 70% Glycerol, 25% .5M Tris pH 9, 5% n-Propylgallate, heat to mix then pH to 7.4 and store in the fridge. For use: 1 drop, swirl, drain most of it off then coverslip. There should be barely enough to cover the coverslip.
Good Luck Doug
Robert Underwood Morphology Core Dermatology U of Wash. Seattle, WA On Thu, 16 Oct 1997, Doug Keene wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Fellow Microscopists: } } We have attempted to immuno-label one micron sections cut } from LR White with primary antibody followed by TRITC or } FITC conjugated secondaries. The sections are } dried onto glass slides at 60C (the same temperature at } which the blocks were cured), the immunolabel procedure } performed, and a cover glass mounted with aquamount (UV } clear). These experiments were completely unsuccessful in } the fluorescent microscope, even though the same } immunolabeling protocol works well in the EM with the } antibodies and immuno-gold secondaries applied to } the surface of ultrathin sections. I realize there are } better ways to label tissue with antibody in preparation } for fluorescence microscopy, but we are surprised by the } negative result in these experiments. Does anyone have an } explanation for the failure? Am I missing something } simple? Is it because there is no penetration of primary } and secondary into the tissue sections, therefore not } enough bound TRITC or FITC to detect? It would be nice if } this technique worked so that we could easily test the } ability of our primaries to bind specifically to LR White } embedded tissue. } } Many thanks, } } Doug Keene } Laboratory for Ultrastructural Research } Portland Shrine Research Unit } ---------------------- } Doug Keene } DRK-at-shcc.org } } }
Our lab is in the process preparing swatches of 50/50 ploycotton fabric contaminated with bacteria for SEM. The swatches will be processed using the fairly standard method, for this lab, of aldehyde, osmium, and ethanol.
HAS ANYONE HAD POSITIVE OR NEGATIVE RESULTS USING HMDS RATHER THAN CPD ON TEXTILES/CLOTH?
At 10:39 AM 10/16/97 GMT+1200, Ritchie Sims wrote: } Does anyone know of a commercially-available embedding resin kit } using methyl and/or butyl acrylate and/or methacrylate? } Preferably email and/or website contact.
There is a methyl methacrylate embedding kit manufactured by Heraeus Kulzer in Germany and sold under the name "Technovit 9100." Unfortunately, the activator, Percodox 16, an organic peroxide manufactured by Akzo Chemical, is classified as an explosive. This effectively prevents the kits from being shipped by air. Kulzer is working on a replacement protocol using a different activator but, in the meantime, the kits are not available.
Disclaimer: Energy Beam Sciences sells the Technovit GMA and (we hope) MMA kits in the United States.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Chris: Windows NT 4.0 should be able to to maintain a log for printers. We use NT 4.0 Workstation to record the user, the printer used, the name of the document printed, and the number of pages. We charge for prints from our TEK Phaser 440. This is how to set it up to log. 1. Start} settings} printers 2. double click the name of the printer you want to audit 3. Select Printer from the menu} properties} security} auditing 4. Select Add, select the name of the group you want to audit (I just select "Everyone"), select Add, then select OK, then select OK in the Properties page. 5. In the Printers page, select File} Server properties} Advanced} Log Spooler information events, then select OK. 6. Then restart the computer. 7. In the Event viewer in Administrative Tools it will record all print events in the System Event log.
I have found NT to be a great operating system once you figure out what you want to do. Hope this will help. Stanley L. Flegler, Assistant Director Center for Electron Optics Michigan State University US flegler-at-pilot.msu.edu
The UCF/Cirent Materials Characterization Facility has an immediate opening for an ion probe engineer with expertise in the operation and maintenance of SIMS equipment. Expertise in the operation and maintenance of an RBS accelerator is also a plus. The MCF at the University of Central Florida in Orlando is a 5400 sq. ft. facility which includes 2 FEG SEM's (Hitachi, JEOL), 2 TEM's (Philips 400T, JEOL 2000FX), a PHI Auger, 3 SIMS (PHI, Riber, Cameca IMS 3f), a JEOL 733 microprobe and an 1.7MeV RBS accelerator. The engineer will be responsible for the daily operation and maintenance of SIMS equipment. It is also expected that the engineer will work closely with MCF faculty, staff, students and industrial affiliates. UCF is close to Cirent Semiconductor (Lucent Technologies), Lockheed Martin, NASA Kennedy Space Center, Westinghouse, Universal Studios, Walt Disney World, Harris Corp., Pratt and Whitney, and others. Please send a resume and a list of references to: Dr. L.A. Giannuzzi, Director, UCF/Cirent Materials Characterization Facility, Mechanical Materials & Aerospace Engineering, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 or send an email Word attachment to: lag-at-pegasus.cc.ucf.edu. UCF is an equal opportunity affirmative action employer.
************************************************************************* 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 ----------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain *************************************************************************
We have an LKB 2208 Multiplate wax heater for mounting plastic boats onto glass knives. It seems to be running a few degrees cool as the wax solidifies too soon upon contacting the knife with the boat. Does anyone know of a way to increase the temperature?
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu
Deutschlaender, Norbert, Path. wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Dear all, } in our lab the question of carcinogenicity of propylenoxide arose. Where } can get hard data about it, and which alternative can be recommended in } Epon-embedding. } Thank you all, } Norbert
Norbet,
Propylene oxide is an NTP anticipated carcinogen and is an IARC category 2B (probable cacinogen. For further information contact National Toxicology Program Office at 919-541-0530 or the WHO Publications Center at 518-436-9686. Acetone and ethanol are viable alternatives to Propylene oxide. Ethanol should be used cautiously because it may inhibit epoxy polymerization.
Hope this helps,
Charles Duvic Chief Chemist Ladd Research tel 1-800-451-3406 fax 1-802-878-8074
Chris: I forgot one part of step 4. Step 4 should read: Select Add, select the name of the group you want to audit (I just select "Everyone"), select Add, then check Print Success, Failure, etc. as needed in the Printer Auditing page, then select OK, then select OK in the Properties page. Sorry for the confusion. Stanley L. Flegler, Assistant Director Center for Electron Optics Michigan State University US flegler-at-pilot.msu.edu
We have a couple you could use, but I'd need more info from you before I could recommend one; such as: when you say you are looking for digital, do you mean the image output, or simply a digital control?, color or monochrome?, what level of support of the camera control do you need?, etc.
Please feel free to contact me using the information below.
****************************** Jim Haley Applications Engineer I-CUBE 2411 Crofton Lane, Suite 14A Crofton, MD 21114 voice: (301) 858-0505 fax: (301) 858-0615 web site: http://www.i-cubeinc.com e-mail: haley-at-i-cubeinc.com ******************************
John R Reffner wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I'm looking for a digital camera for an optical microscope that I can control } from a program, such as a Visual Basic program, for setting up an automated } imaging system. Anyone out there doing this type of thing?
I read an article on X-ray microscopy in the Jan/Feb. 1996 issue =
of Biophotonics International (by Laurin Publishing), pg. 58. =
The article highlights work by researchers (Dr. Cathie Magowan) =
at the Ernest Orlando Lawrence Berkeley National Laboratory, =
studying interactions between malarial parasites and host cells. =
They studied living [I think] parasites in red blood cells over =
48 hours, in aqueous medium. The stated resolution of the system =
was 60 nm in X-ray mode; the microscope could be switched between =
visible light and X-ray modes on the same sample.
If anyone knows more about this I'd be interested out of =
curiosity. Being a biologist I don't really want all the gory =
details; I just want to know practicle applications, =
availability, cost, etc. of the instrumentation.
Thanks, Karen
bengt-at-mail.coxsys.se wrote: } =
} } =
} } I heard about a new x-ray microscopy technique that can image cellular=
} } processes in real-time. Have you heard about this? } } } } } } Bob Clark } } } } Hi ! } Interesting idea, but it doesn=B4st seem very likely that it would be } possible. According to a simple calculation we made, imaging of a cell } would kill it within a few milliseconds, using x-rays. If someone has o= ther } information it would be interesting to share it ! } =
} 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
-- =
Karen Zaruba =
kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
I'd like to de-embed some LX112 embedded plant leaf tissue so that I could examine it in the SEM but, never having done this before, I'm looking for advice, tips, references, protocols etc. Also, is it possible to de-embed thin sections already stained (uranyl acetate, lead citrate) and examined in the TEM? (Assuming I could get them off the grid....)
} Our lab is in the process preparing swatches of 50/50 ploycotton } fabric contaminated with bacteria for SEM. The swatches will be } processed using the fairly standard method, for this lab, of aldehyde, } osmium, and ethanol. } } HAS ANYONE HAD POSITIVE OR NEGATIVE RESULTS USING HMDS RATHER THAN } CPD ON TEXTILES/CLOTH? } } Thanks, Lloyd.
I haven't tried HMDS on 50/50 polycotton, but I have used it with good success on cotton fibers and polycarbonate membranes, as well as other compounds. I would expect it to work fine, but it might be better if you specified the polymer in the blend. (Have you contacted the vendor of the HMDS?)
An excellent source for Image Analysis/ Imaging techniques for any topic is Dr. John Russ's book "The Image Processing Handbook" which is available from CRC press.
David Bell Millipore Corporation Mailstop B2C 80 Ashby Road Bedford, MA 01730
} I am trying to to find a reference for the Epon-Spurr resin recipe that is } used for microwave fixation. } } I have listed here that the resin can be poylmerized at 70 degree C for } 24 hours but I ned a reference that says I can do that.. } } Any suggestions or does anyone have the reference } } Thanks } } Eric } Fred Hutchinson Cancer Research Center } }
I have tried labelling 1 micron sections with lectins conjugated with TRITC and they worked exceptionally well.... that is the lectin was labelled with the TRITC tag..
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Tomorrow evening there will be a meeting in Chicago of the State Microscopical Society of IL (SMSI). The speaker is Riccardo Levi-Setti, director of the Enrico Fermi Institute at the Univ. of Chicago and he will speak about his work on trilobites. If you are interested in details, let me know for I will be going.
==================================================================== FALL MEETING OF THE APPALACHIAN REGIONAL MICROSCOPY SOCIETY (AReMS) October 23-24, 1997
Host: Clinch Valley College of the University of Virginia, Wise, VA
Registration: Oct. 23, Noon until 5:00 pm, Holiday Inn, Norton, VA (For additional information and pre-registration, see the bottom of this notice.)
Thursday, October 23, 1997, WORKSHOPS:
Workshop 1. 2:00 - 3:00 "HTML and Web Design" Led by Dr. Herb Brown, Dir. Instructional Technology, CVC. Will cover the basics of web page creation, using the Netscape Navigator Gold editor. Hypertext Markup Language (HTML) will be explained and interpreted. Participants will create their own basic web page with text, graphics, and hypertext links. Good web page design techniques will be fostered. Participants will only need basic computer skills.
Workshop 2. 2:00 - 3:00 "E-mail and networked communications" Led by Mr. Alex Edwards, Assoc. Professor of Computer Science, CVC.
Workshop 3. 2:00 - 3:00 "An Introduction to the SEM" Led by Dr. Stan Kunigelis, Assoc. Professor of Zoology, CVC Intended for students and other novices.
Workshop 4. 3:00 - 4:30 "Digital Imagery: A How-To Approach for Importing, Exporting, Managing, and Everything in Between". Led by Rick McGill, Eastman Chemical Company A. How to design your own image management database system using Microsoft Access. B. How to make your importing/exporting life easier, using some inexpensive commercial software packages.
SOCIAL HOUR: 6:00 - 7:00 (Open bar, Alumni Hall, CVC Campus)
BANQUET: 7:00 - 9:00, Alumni Hall, CVC campus
GASTRONOMIC DELIGHTS Cream of Artichoke Soup Peppered Duck with Chutney glaze, served over wild rice Tenderloin Medalions of Beef, with Burgundy sauce Orange-Ginger baby Carrots Twice-Baked Potatoes Almond Cream custard, with Raspberries White or Red Wine Coffee/Tea
SPEAKER: Dr. Loren W. Knapp, University of South Carolina "Science Education: Coming of Age in the 21st Century"
Friday, October 24, 1997 Room 220 New Classroom Building, Clinch Valley College
8:00 - 8:30 Registration and Coffee 8:00 - 8:30
8:30 - 8:50 Mr. B.J. Craven, Lorillard Research. "Ultrasonic leak detection for the microscopist"
8:50 - 9:15 Dr. Fred E. Hossler, Dept. of Anatomy, School of Medicine, ETSU "Intrinsic lymph nodes in the wall of the urinary bladder - structure and blood supply"
9:15 - 9:35 Mr. Eric Bond, University of Tennessee "The uses of Electron Microscopy in polymer morphology characterizations"
9:35 - 10:00 Dr. Bob Price, School of Medicine, Univ. of South Carolina "The effect of angiotension II on early embryonic heart development"
10:00 - 10:30 BREAK: VISIT THE EXHIBITS
10:30 - 11:00 AReMS Business Meeting
11:00 - 11:25 Dr. Loren W. Knapp, Dept. of Biology, Univ. of South Carolina "Spiculogenesis in Octocorals--Hard tissues: Hard Science"
11:25 - 11:50 Mr. Dave Calvert, Eastman Chemical Company "Of misconceptions and serendipity: Our microscopy and image analysis successes"
11:50 - 12:15 Mr. Mike Kiser, Clinch Valley College "An SEM analysis of development in the tapeworm Hymenolepis dimunata"
12:15 - LUNCH, CVC cafeteria. ---------------------------------------------------------------------- For additional information, pre-registration, and directions, contact Dr. J. Rex Baird, Dept. of Natural Science, Clinch Valley College, Wise, VA 24293 FAX: 540-328-0247 E-mail: jrb-at-clinch.edu Phone: 540-328-0201 AReMS Web site: http://www.clinch.edu/~jrb/arems.html Costs: Registration - Member: $10.00 Non-member: $15.00 Exhibitor: $75.00 Thursday Banquet: $25.00 Friday Lunch: $ 4.00 Annual Dues - Individual: $ 5.00 Corporate: $15.00
Housing: The following motels have reserved a block of rooms at the listed rates. Please mention AReMS when calling. They are side by side in Norton, approximately four miles from Wise.
Holiday Inn (540-679-7000) $54.50 (dbl) Should call before Oct. 15. Super 8 Motel (540-679-0893) $38.59 (1-4 persons) ==================================================================
The FBI Microanalysis Laboratory is seeking development of an application to archive spectra generated by SEM/EDXA, in order to manage large numbers of spectra used for analysis and reference purposes. The application would function somewhat similarly to databases presently used for other spectroscopies, such as FTIR. Database utilities currently provided by EDXA manufacturers consist simply of 1 - naming a spectrum, and 2 - retrieval of spectra by either recalling a specific spectrum by name, or comparing a spectrum to an entire directory of spectra by either a "matching" or quantitative comparison. In order to provide flexible search management, we would like an application that would additionally provide:
1. Attachment of text to spectra. This text would permit descriptors (key words) for each spectrum, which would be searchable with usual "AND/OR" operators, thereby permitting retrieval of spectral clusters based on similar criteria such as material, batch, use, source, date, etc. This spectral database would then function as a true relational database. Other functions, such as sorting, would also be possible. 2. Automatic display of spectra in "nested" fashion (overlayed, but vertically offset slightly), permitting display and critical comparison of numerous spectra simultaneously. 3. Attachment of images to spectra.
I am seeking advice from those who might have experience and/or interest in archiving spectra for the purpose of writing a RFP. If you have interest in the development of a database such as this, please contact me directly for more details.
Thank you. Dennis.
Dennis C. Ward voice: 202-324-2982 FBI fax: 202-324-4018 Microanalysis Laboratory e-mail: DCWard-at-juno.com
} } I heard about a new x-ray microscopy technique that can image cellular } } processes in real-time. Have you heard about this? } } } } } } Bob Clark
Perhaps you are thinking about biospectroscopy, in which you can follow cell processes by their effects on molecular signatures observed by confocal raman spectroscopy. For example, you can follow the movement of a DNA-binding organic molecule (I forget what it was) in 3D and real time as it crosses the membrane, traverses the cell and enters the nucleus. Needs considerable computer power and the 3D effects are calculated after the experiment.
Rosemary White Department of Ecology and Evolutionary Biology Monash University, Melbourne, Victoria 3168, Australia phone 61-3-9905 5670 fax 61-3-9905 5613 email r.g.white-at-sci.monash.edu.au
kszaruba-at-MMM.COM wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I read an article on X-ray microscopy in the Jan/Feb. 1996 issue } of Biophotonics International (by Laurin Publishing), pg. 58. } The article highlights work by researchers (Dr. Cathie Magowan) } at the Ernest Orlando Lawrence Berkeley National Laboratory, } studying interactions between malarial parasites and host cells. } They studied living [I think] parasites in red blood cells over } 48 hours, in aqueous medium. The stated resolution of the system } was 60 nm in X-ray mode; the microscope could be switched between } visible light and X-ray modes on the same sample. } } If anyone knows more about this I'd be interested out of } curiosity. Being a biologist I don't really want all the gory } details; I just want to know practicle applications, } availability, cost, etc. of the instrumentation. } } Thanks, } Karen } } bengt-at-mail.coxsys.se wrote: } } } } } } } } I heard about a new x-ray microscopy technique that can image cellular } } } processes in real-time. Have you heard about this? } } } } } } } } } Bob Clark } } } } } } Hi ! } } Interesting idea, but it doesnīst seem very likely that it would be } } possible. According to a simple calculation we made, imaging of a cell } } would kill it within a few milliseconds, using x-rays. If someone has other } } information it would be interesting to share it ! } } } } 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 } } -- } Karen Zaruba } kszaruba-at-mmm.com } 3M Company, 3M Center Bldg. 270-1S-01 } St. Paul, MN 55144 } "The opinions stated above are my own, not necessarily 3M's"Just a quick tag-along to Karen's notes... Having been involved in both applications support and microscope design, I would like to know all the gorey details.
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 ********** Americas First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
We have seen your message. We at Electron Microscopy Sciences have both a Methyl Methacrylate alone and a Methyl/Butyl Methacrylate kits available. You may see it at our Website where our complete 400 page catalog is up and working at www.emsdiasum or in our hard copy catalog. Please let me know if I may be of further assistance to you.
Sincerely,
Stacie Kirsch Electron Microscopy Sciences http://www.emsdiasum.com
} .... } } The FBI Microanalysis Laboratory is seeking development of an } application to archive spectra generated by SEM/EDXA, in order to manage } large numbers of spectra used for analysis and reference purposes. The } application would function somewhat similarly to databases presently used } for other spectroscopies, such as FTIR. ...
One problem, with respect to other spectroscopies, is the multitude of different SEM/EDX configurations ... for example, take-off angles and detector windows. Operators' choices for instrumental parameters (... e.g., keV ...) also run a gamut. Towards the end of having a "trustworthy" database, you might want to call all of us in for a conference (... any excuse to party ...).
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
I read warning that " ImagePC beta 1a" , a port of NIH Image to Windows 95 which requires installation of Microsoft's "DirectX, " driver causes computer's display to stop working correctly. I will appreciate feedback or suggestions from those using Image PC.
as many of you will be aware, it can be very difficult to accurately determine diffraction ring diameters in selected area diffraction patterns recorded from multicrystalline TEM specimens. Especially when rings are very spotty.
What I'd like to do is digitize the SAD pattern, locate the exact centre and intergrate pixel intensities through 180 degrees. This will result in a one dimensional diffraction pattern with pairs of spots either side of the undiffracted spot, which should be much simpler to measure.
I would like to know if anyone out there has a computer program to do this type of SAD pattern manipulation. I propose to use Photoshop and a flat bed scanner on a Macintosh to digitize the SAD patterns which would be saved as TIFF or some other suitable format. So I suppose the ideal solution to my problem would be a Photoshop plugin. I also use NIH Image so a plugin for this program would also be suitable.
I would really appreciate any help with a plugin, stand alone program (Mac or PC) or comments on how I should go about writing my own solution. I look forward to your replies,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
John Luft originally suggested 1-2 epoxy propane or propylene oxide as an intermediate solvent for epoxy infiltration because he reasoned that its chemical structure would allow it to be incorporated into the epoxy resin.
But I question if this could ever happen. Consider the boiling points of the three solvents which are being compared:
ethanol....... 78.3 Deg. C acetone....... 56 deg. C. epoxy propane 34.3 Deg. C.
Most epoxies are cured at 70 Deg or above. At this temperature acetone but especially epoxy propane will very rapidly evaporate from the mixture. But alcohol will not. Years back I mixed epoxy resin with 10 percent of each solvent and put the three samples into the curing oven at 70 deg. C. The acetone an epoxy propane mixtures cured perfectly normally. But the alcohol sample stayed sticky. I think it just doesn't evaporate.
Bottom line: since then my lab has always used acetone as a less hazardous alternative to epoxy propane. They both reduce the viscosity of epoxy resin about the same amount. The historically minded could check my 1968 paper-- Viscosity changes in Araldite during polymerization---- Laboratory Practice 17, pp 707-708.
Mel Dickson Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
} I read warning that " ImagePC beta 1a" , a port of NIH Image to Windows 95 } which requires installation of Microsoft's "DirectX, " driver causes } computer's display to stop working correctly. I will appreciate feedback or } suggestions from those using Image PC.
It's tricky. It took 2 installations but it worked fine on my Hitachi lap-top. OTOH, after a bit of work, I opted for ImageTools and removed ImagePC.
I had contact with Chris Cathcart about a DIC system for a Zeiss microscope , but unfortunately he gave me a wrong e-mail address so I can't send him my reply.Knows anybody his correct e-mail.
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Reply to: RE} Reference for use of Epon-spurr recipe
Dear Eric, You may want to look up Giammara,B.1993, Scanning 15:82-87 or "The = Microwave Tool Book", Chapter 10, Login and Dvorak, Beth Israel Hospital = Dept. of Pathology, Boston, MA. In addition, Gary Login has published = extensively regarding microwaves and microwave embedding. Linda Chicoine Center for Cell Imaging Yale University New Haven, CT 203-785-3646 http://info.med.yale.edu/cellimg
--------------------------------------
I am trying to to find a reference for the Epon-Spurr resin recipe that = is used for microwave fixation.
I have listed here that the resin can be poylmerized at 70 degree C for 24 hours but I ned a reference that says I can do that..
Any suggestions or does anyone have the reference
Thanks
Eric Fred Hutchinson Cancer Research Center
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Folks: Over the years the subject of immersion oils has come up several times, with the general agreement that Cargille oils are the best. The question is which one! A,B, NVH, FF, and DF Our application is almost entirely fluorescence microscopy. That would be types A, FF, and DF... but which is best and can that selection be used on an inverted scope?
I am sure that someone has done a comparative analysis, if so could you forward the conclusions Thanks
-- Simon C. Watkins Ph.D. Associate Professor Director CBI University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 Fax:412-648-2004 URL:http://sbic6.sbic.pitt.edu
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Folks: Over the years the subject of immersion oils has come up several times, with the general agreement that Cargille oils are the best. The question is which one! A,B, NVH, FF, and DF Our application is almost entirely fluorescence microscopy. That would be types A, FF, and DF... but which is best and can that selection be used on an inverted scope?
I am sure that someone has done a comparative analysis, if so could you forward the conclusions Thanks
-- Simon C. Watkins Ph.D. Associate Professor Director CBI University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 Fax:412-648-2004 URL:http://sbic6.sbic.pitt.edu
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10.30-11.10 Low voltage cryoSEM for x-ray microanalysis Patrick Echlin, Cambridge
11.10-11.50 Variable pressure SEM: an enhancement of cryomicroscopy. Roger Angold, RHM
11.50-12.20 CryoEM and research in cosmetics Philippe Hall=E9got, L=92Or=E9al Recherche
12.20-1.00 Trades Exhibition 1.00-1.45 LUNCH 1.45-2.00 Annual General Meeting
2.00-2.15 Freeze-drying of thin sections for x-ray microanalysis Alice Warley, UMDS, St. Thomas=92 Hospital, London
2.15-2.30 Freeze-substitution strategies for the retention of mineral for= EDX=20 and correlated immunogold labelling of Lowicryl HM20 sections. Jeremy Skepper, Multi Imaging Centre, Cambridge
2.30-2.45 Advances in cryotechniques for analytical microscopy of plant= tissue John Forsdyke, Oxford Microscopy Consultancy
2.45-3.00 Structure of a chaperonin ATP-ase mutant by cryoTEM and 3-D reconstruction. Jose Jimenej, Birkbeck College
3.00-3.30 TEA
3.30-4.00 Cryogenic light microscopy in the development of long term cryopreservation techniques for fungi. David Smith, International Mycological Institute, Surrey
4.00-4.30 Quantitative x-ray mapping of ion-transporting and=20 metal-sequestering epithelia of invertebrate tissues after hyperbaric freezing Carol Winters, University of Wales, Cardiff
4.30 Chairman=92s concluding remarks.
=A325 Registration includes coffee, tea, lunch and Trade Exhibition/Posters welcome
FURTHER INFORMATION FROM : =09
Secretary, CMG Georgina Godwin =09 International Mycological Institute=09 Bakeham Lane, Surrey web site : http://www.gla.ac.uk/Acad/IBLS/II/cryomg.htm=09 TW20 9TY
Tel: 01784470111 x 556 email: G.Godwin-at-Cabi.Org FAX: 01784 470909 Dr Laurence Tetley IBLS EM Centre Joseph Black Building University of Glasgow Glasgow G12 8QQ
email l.tetley-at-bio.gla.ac.uk =09 tel. 0141 330 4431 fax 0141 330 3516 =09 I & I Divisional web pages http://www.gla ac.uk/Acad/IBLS/II/ EM facility web pages http://www.gla.ac.uk/Acad/IBLS/II/em/mcb-em.htm
WinJade from MDI Inc. (XRD-at-AOL.com) is a program for x-ray diffraction analysis that has a software option for doing just that. I've helped them make a few modifications on how they extract the image from the digitized pattern. They now have 4 different ways of doing it. I have submitted an abstract to the International Conference on Metallurgical Coatings an Thin Films-98 being held in San Diego in April on the use of this program with multiphase samples with partial ring patterns. I also plan to publish the paper. If you correlate XRD data with electron diffraction data, this program is great. You can overlay JCPDS or NIST Crystal Database files directly on the image. You can reduce it to a one dimensional pattern that can be overlayed with the XRD patterns. this program has made phase identification from SAD patterns much simpler for me. They have a demo program. I don't know if it has the electron diffraction module in it. You should direct your questions to Quentin Johnson. -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."
as many of you will be aware, it can be very difficult to accurately determine diffraction ring diameters in selected area diffraction patterns recorded from multicrystalline TEM specimens. Especially when rings are very spotty.
What I'd like to do is digitize the SAD pattern, locate the exact centre and intergrate pixel intensities through 180 degrees. This will result in a one dimensional diffraction pattern with pairs of spots either side of the undiffracted spot, which should be much simpler to measure.
I would like to know if anyone out there has a computer program to do this type of SAD pattern manipulation. I propose to use Photoshop and a flat bed scanner on a Macintosh to digitize the SAD patterns which would be saved as TIFF or some other suitable format. So I suppose the ideal solution to my problem would be a Photoshop plugin. I also use NIH Image so a plugin for this program would also be suitable.
I would really appreciate any help with a plugin, stand alone program (Mac or PC) or comments on how I should go about writing my own solution. I look forward to your replies,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
Many thanks to the numerous responders to the propylene oxide-alternative. I hope that I did not break off an avalanche of fear because of the carcinogenicity problem. I am aware, that many labs in this country have been using the compound for many years, including my EM-lab. But without wishing to depriciate the potential danger of p.o., I believe that according to the experimental data in rats and mice (literature up to 1996), the compound seems to be a rather weak carcinogen, acting either after repeated local subcutaneous injections (rare sarcomas in rats) or after life-long inhalation of rather high doses ( very low incidence of nasal hemangiocarcinomas in mice). Thus, since p.o. is classified now as a possible carcinogen in humans, we should of course avoid it completely or reduce it to exceptional cases, but nobody should be anxious about exposition in the past. Norbert
What you're describing is similar to a procedure we use called EDIP - electron diffraction image processing. Basically, what we do is scan a TEM negative, digitize it, identify the center of the pattern, then "draw" a series of concentric rings going out form this central spot, increasing by one pixel at a time. We then take the entire integrated intensity inside each of these rings. By subtracting the integrated intensity successively from the next largest ring, i.e., subtract the integrated intensity of ring "Y" from the integrated intensity in ring "Z", then "X" from "Y". etc., one ends up with a "graph" of the diffraction pattern, containing all of the information. This is an extremely useful technique for identifying small precipitates, different phases, etc., particularly if there is a known element or compound included in the pattern which can be used to calibrate the graph. The full procedure is given below - John Philips wrote it up for internal use, so it may be a bit "site specific", but it should give you the useful software names and procedures. NOTE 1: The most recent version of Image Pro Plus contains a circular line profile feature, that does all this automatically. NOTE 2: John Russ of "The Image Processing Handbook" fame has expended a lot of effort on this very problem but from another angle, cf: "Application of the Hough Transform to Electron Diffraction Patterns", Journal of Computer- Assisted Microscopy, Vol. 1, No. 1, pp. 3-37 (1989). I heard that he is producing some software for Photoshop using Hough transforms due out in a month(?) that performs this procedure. Aanyway, good luck! It's a great technique. Please contact me off the listserver if you need more info.
This note describes a procedure for analyzing spots or ring electron diffraction patterns with an image analysis program and a personal computer.
Software: Image analysis program from Image-Pro plus for the PC from Media Cybernetics (www.mediacy.com). Plotting program from Origin Ver 4.0 from Microcal Software Inc.
Hardware: IBM compatible Pentium, Windows NT or equilavent; HP scanner with negative attachment.
Purpose: The analysis is used to do data extraction from electron diffraction patterns that is equivalent to a line scan plot of intensities vs distance (Rd) from the centre of the central spot on the diffraction pattern to some arbitrary user selected point outside the spots or rings near the edge of the photo.
Procedure: The negative to process is converted to a grey level tiff image with a flatbed scanner. A program written in the Auto_Pro macro language of Image-Pro finds the x,y coordinates of the central spot and calculates successive histograms of concentric circular areas of interest starting at the centre of the central spot and increasing by one pixel radius outward to some user-selected point.
Each of these values which are the sums of all the grey levels within these circular areas of interest are appended to a file for later processing with the Origin plotting program. A background image is created from the original using the background extraction feature of the Image-Pro program and a background' file is created using identical xy coordinates and circular histogram parameters. Processing: Subtracting the preceeding value from each of these histogram sums produces a table where each value is equilavent to adding up all the pixel values that lie on the original concentric circle that bounded the histogram. Plotting these values vs their row number is a line scan of intensities from the centre of the central spot to the selected position.(ie: intensity vs Rd). Similar processing of the background image gives a background line profile. Specific instructions: Scan the EDP negative as grey level with the highest resolution available( currently 200dpi on the HP scanner and fine black and white photo mode.). Save as a tiff file. Load the file into Image Pro Invert the file (only to get numbers for a graph that has the background(black) as zero and spots (white) giveing increasing numbers. Make sure you apply the inversion map to the image. select an AOI that includes the spots and rejects edges and various features that are not part of the pattern. duplicate/crop this area and minimize the original image for clarity of the display. create a background image from the duplicated image using the background extraction feature . run the macro diff_main' and follow the instruction on the screen. This macro will find the centre of the image and allow you to select an outer ring where the analysis will stop. This center and outer ring radius will be exactly the same on the foreground' image and the background' image. This is necessary to simplify later processing of the data. The files generated currently have to be edited because the first reading is extraneous and the current version of Image Pro adds two newline characters between readings when you append data to the file. To remove these use for example, Wordperfect and search for three newlines and replace them with one newline. This is essential for Origin or Excel to import the data as a continuous column of numbers. The file as saved has five items in a row and the last one is of interest to us since it is the sum of the grey values contained in the concentric circular histograms that the macro creates. The first four columns can be deleted if desired. The first readings are not accurate representations of the histogram because the size of the circle is only one pixel and it increments by one pixel radius for successive histograms. The first few readings are significant in that their existence defines the distance from the centre of the central spot to where ever the outer ring was chosen. The point of this exercise is to subtract the background image data from the foreground image data and to end up with a plot of Rd vs intensity and try to coorelate the intensities and peak positions with orientation on an unknown sample.
We do a lot of immunofluorescence and after upgrading our scope and looking for very low signals of insitus we under went a study of optimizing mounting media and immersion oils in order to get better signal to noise ratio. I tested every major brand of immersion oil several times and from old and fresh batches. The Cargilles were always highest in autofluorescence and the lowest was Nikon oil. It has always been a mystery why so many people like Cargille, we have several bottles that we gave up using years ago due to it obscuring our signals.
Bob Morphology Core
On Fri, 17 Oct 1997, Simon C. Watkins wrote:
} Folks: } Over the years the subject of immersion oils has come up several times, } with the general agreement that Cargille oils are the best. The } question is which one! A,B, NVH, FF, and DF } Our application is almost entirely fluorescence microscopy. That would } be types A, FF, and DF... but which is best and can that selection be } used on an inverted scope? } } I am sure that someone has done a comparative analysis, if so could you } forward the conclusions } Thanks } } } -- } Simon C. Watkins Ph.D. } Associate Professor } Director CBI } University of Pittsburgh } Pittsburgh PA 15261 } tel:412-648-3051 } Fax:412-648-2004 } URL:http://sbic6.sbic.pitt.edu } }
I've been contacted by a rep from Ted Pella with a new microwave item that allows complete TEM specimen preparation in just 3 hours (live tissue to grid in scope)! Is anyone using this technology regularly, especially for preparing plant tissues? If so, how is it working out for you?
An article in the 27 Sept. '97 issue of New Scientist, pp. 24-28, describes a x-ray microscope being developed at the Rutherford Appleton Laboratory in Oxfordshire, headed by Jie Zhang. The microscope is under development - uses a very bright X-ray laser pulse with a very narrow spectrum, between 2.2 and 4.4 nanometers. Perhaps that laboratory has a web site with more details. Haven't checked; just came across the article in the library.
On Thu, 16 Oct 1997, Bengt Stocklassa wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } =20 } =20 } } I heard about a new x-ray microscopy technique that can image cellular } } processes in real-time. Have you heard about this? } } } } } } =09=09=09Bob Clark } } } } Hi ! } Interesting idea, but it doesn=B4st seem very likely that it would be } possible. According to a simple calculation we made, imaging of a cell } would kill it within a few milliseconds, using x-rays. If someone has oth= er } information it would be interesting to share it ! } =20 } bengt } Bengt Stocklassa , Managing Director } Cox Analytical Systems AB=09| Phone: +46 31 7725300 } House of Innovations, CTH=09| Fax: +46 31 7725600 } 412 96 - Gothenburg, SWEDEN=09| E-mail: bengt-at-xco.se } =20 } =20
Dear Casey, I saw your message. There are many researches here in the United States that have moved over to Microwave technology. It started in Europe and South Africa and now is slowly moving over. The technology is grand and yes even for plant tissue it allows you to do all of the procedures associated with specimen prep for microscopy in the microwave and all in 3 hours or less. This includes dehydration, fixation, staining, polymerizing and evn immuno and decalcification work. We have done alot of work with the major movers who have published on the subject of microwave technolgy, including Gary Login from Harvard, and Dr. Boon who has published the Microwave Cookbook for Microscopists. It truly is a technology worth looking into. There are quite a few laboratory microwave oven manufacturers here in the states and we happen to be one of them. You may see what we have to offer at www.emsdiasum.com or just request a complete technical brochure from us. Please do not hesitate to contact us for further information or if we can give you any technical assistance. Sincerely,
Stacie Kirsch Electron Microscopy Sciences 215-646-1566
Dear colleagues, 8th of October 1997 I posted the following question to the server:
} } Dear all, I greatly should appreciate informations on experiences with existing/ = or on dealers/companies of LASER PROJECTION SYSTEMS (connectable to PC/Video and TV Cams, Remote system). We know a company (ASK-System) in Europe which deals with LIGHT = PROJECTION SYSTEMS for demonstration of slides, PC-data, etc. in teaching and else applications. Has anybody suggestions, informations (company/-ies, approx. price, combinations with periphery, necessary components) for us ? Such a Laser Projection System should work for projection screen = dimensions of at the maximum approx. 2 x 3 m or little less, if available. Any suggestions and comments are welcome. Best wishes for the day sincerely yours Wolfgang MUSS Dept. Pathology LKA, EM-Lab, Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria/Europe Phone: ++43++662+4482-4720 Ext Fax: ++43++662+4482-882 Ext. e-mail: W.Muss-at-lkasbg.gv.at. { {
Unfortunately up to now no suggestion or any answer was received. Since at the time of first posting our server had a lot of troubles and = problems it may be possible that the message was not transmitted in full = or at least garbled. I post our question once more again, asking anybody (also = selling/dealing companies) for suggestions, solutions. Thanking you in advance, have a nice sunday/weekend, Wolfgang MUSS Department of Pathology, 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) e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to -at- is a small "L")
We are planning to purchase a microscope to be used with Polaroid DMC-2000 camera for image analysis of immunohistochemical data. We have found an information on the web about new Nikon Eclipse E-600 and E-800 series microscopes which seem to suit our purposes. But we cannot locate any Nikon dealers in Russian that can provide us with the information on prices and purchase. We can make more complex steps towards purchase through one of the USA companies but can anyone tell what are at least the price ranges for these series of microscopes to know whether they'll fit into our budget?
Thanks in advance.
Konstantin Anokhin
----------------------- neuro-at-redline.ru
Laboratory of Molecular Neurobiology Institute of Physiology Russian Academy of Medical Sciences
Another correspondent claimed that Cargille' is subject to autofluorescense. True, if you are using the wrong type. I have no idea were Nikon is sourcing immersion oil, but you can be sure that several microscope manufacturers are "branding" Cargille's. Here is a copy from our online re immersion oils:
"The refractive index using incandescent light is: Type A 1.5482; type B 1.5468 and type NVH 1.5439. Request additional information if required:
Type B is the most used of these immersion oils. Types A & B have viscosities of 150 and 1250cSt respectively. For a greater gap between cover glass and objective, (or condenser and slide) type B is more desirable. Type A is less sticky and easier to clean up. For horizontal, inverted and inclined instruments and projection equipment, Type NVH with high viscosity of 21,000cST should be used. These three types are classed as low fluorescence, however, for serious fluorescence work types FF and DF - see below - are required.
Type DF combines very low fluorescence and ideal optical properties. It is recommended where specimen fluorescence is good and highest resolution is required. Type FF does not fluoresce but its optical characteristics are not quite perfect."
For fluorescent work the choice seems to be: DF when specimen have fairly strong fluorescence and ideal optical properties are required. If fluorescence is weak type FF should be used which has zero autofluorescence but is optically no quite perfect. 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, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} Date: Friday, 17 October 1997 22:35 } } Folks: } Over the years the subject of immersion oils has come up several times, } with the general agreement that Cargille oils are the best. The } question is which one! A,B, NVH, FF, and DF } Our application is almost entirely fluorescence microscopy. That would } be types A, FF, and DF... but which is best and can that selection be } used on an inverted scope? } } I am sure that someone has done a comparative analysis, if so could you } forward the conclusions } Thanks } } } -- } Simon C. Watkins Ph.D. } Associate Professor } Director CBI } University of Pittsburgh } Pittsburgh PA 15261 } tel:412-648-3051 } Fax:412-648-2004 } URL:http://sbic6.sbic.pitt.edu
My name is Myriam Aguirre and I am student of PHD of physics in Argentina. My e-mail is maguirre-at-citefa.edu.ar . I would like to ask you some questions about transmission microscopy. I am studying the structure of compound Mercury Cadmiun Telluride in composition x=0.2 (Hg0.8Cd0.2Te) by TEM. I have had some problems with Hg evaporation. I could not see the structure of the compound because the high temperature which is produced by the electron beam, evaporates the sample. I have tried with the sample cooled with liquid N but the beam goes on eating the sample. Are there solutions to this problem? Thank you in advance. Myriam Aguirre -------------------------------------------------------------------------
I had the same problem with two other Multiples. Pull the heat element out and surround it with some heat sink compound then reinsert it. That should be enough to raise the temp short of buying a new element. Any hardware or electronics store will carry it.
Fred Hayes The Dow Chemical Company Analytical Sciences, Microscopy 1897 bldg, E78 Midland, MI 48667 517-638-2203
} ---------- } From: wise-at-vaxa.cis.uwosh.edu[SMTP:wise-at-vaxa.cis.uwosh.edu] } Sent: Thursday, October 16, 1997 11:31 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: LKB Multiplate } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} as many of you will be aware, it can be very difficult to accurately } determine diffraction ring diameters in selected area diffraction patterns } recorded from multicrystalline TEM specimens. Especially when rings are } very spotty. } } What I'd like to do is digitize the SAD pattern, locate the exact centre } and intergrate pixel intensities through 180 degrees. This will result in a } one dimensional diffraction pattern with pairs of spots either side of the } undiffracted spot, which should be much simpler to measure. } This can readily be done using two of the operations in the SPIDER image-processing program. The operation which determines the center coor- dinates and ring radii works by marking from 3 to 20 points on the ring and least-squares fitting the center & radius to those points (for more than 3 selected). Once the center has been found, another operation can be used to calculate the rotational average--which is equivalent to integrating through 360 deg. If you need 180 deg specifically, SPIDER can take half the image, prepare a mirror (or rotated) image and produce a new image which would give you (2 times) the integrated value. This can be done for each half.
} I would like to know if anyone out there has a computer program to do this } type of SAD pattern manipulation. I propose to use Photoshop and a flat } bed scanner on a Macintosh to digitize the SAD patterns which would be } saved as TIFF or some other suitable format.
Several formats (including TIFF) can be converted to SPIDER format.
} So I suppose the ideal } solution to my problem would be a Photoshop plugin. I also use NIH Image } so a plugin for this program would also be suitable. } } I would really appreciate any help with a plugin, stand alone program (Mac } or PC) or comments on how I should go about writing my own solution. I } look forward to your replies,
I have written a stand-alone version (in addition to that incor- porated in SPIDER), and I'll be happy to send you the FORTRAN code. I think you can, then, rewrite it for Mac or PC. Yours, Bill Tivol
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Post-Doctoral Appointment in Electron Microscopy
A post-doctoral appointment for a person skilled in electron microscopy techniques is open in the Materials and Engineering Sciences Center at Sandia National Laboratories-California. The appointee will apply transmission electron microscopy to the characterization of metallurgical and ceramic materials. The candidate must have received a PhD in Materials Science, Chemistry, or Physics. Experience with diffraction contrast methods, HRTEM, and analytical electron microscopy, including EDS and EELS, is strongly desired. Experience using other microscopy methods, including SEM, is also desirable.
Send resume, with names of references, publication list, statements of research expertise, and copies of transcripts to:
Sandia National Laboratories c/o Anna Isham, MS 9111, HR Dept. -CA0041 P.O. Box 969 Livermore, CA 94551-0969
U.S. Citizenship is normally required. Sandia National Laboratories is an Equal Opportunity Employer/Affirmative Action Employer.
For the fourth time a subscribed to the microscopy discussionlist and three times I unsubscribed , because my main interest is LM , I do not work for my research on marine plankton with EM. Would it not possible to make 2 sublists one for LM and one for EM ? Now members who's main work is with LM are sometimes flooded with EM discussions and vice versa. Please give your opinion.
Dear Myriam, } } I would like to ask you some questions about transmission microscopy. I am } studying the structure of compound Mercury Cadmiun Telluride in composition } x=0.2 (Hg0.8Cd0.2Te) by TEM. I have had some problems with Hg evaporation. } I could not see the structure of the compound because the high temperature } which is produced by the electron beam, evaporates the sample. I have tried } with the sample cooled with liquid N but the beam goes on eating the } sample. } Are there solutions to this problem? Thank you in advance.
I have used high voltage (1.2 MV) and low dose imaging to ameliorate this problem. Since the interaction cross-sections with matter fall as the energy of the electron beam increases, the higher the energy (up to ~1 MV), the lower the amount of heat deposited by the beam. This is counter-intu- itive, but none-the-less true. Also, the lower the dose rate, the smaller the ultimate temperature rise. Our scope is equipped with a very sensitive intensified CCD, which allows rapid scanning of the specimen to locate the area of interest, and rapid focussing at low dose rates. Since this camera sacrifices some reso- lution to achieve high sensitivity, we record images on film, and we use LoDose or MRF32 x-ray film to get about one order of magnitude more sensi- tivity than from 4489 or SO163. I cannot tell you that this will be enough for you to be able to get images from your material, but it could be worth doing--at LN2 temperature, of course. Good luck. Yours, Bill Tivol
Greetings, Pieter Houpt wrote: } Dear microscopist, } } For the fourth time a subscribed to the microscopy discussionlist and } three times I unsubscribed , because my main interest is LM , I do not } work for my research on marine plankton with EM. } Would it not possible to make 2 sublists one for LM and one for EM ? } Now members who's main work is with LM are sometimes flooded with EM } discussions and vice versa. The answer here is for folks to make good use of the subject line. The clearer the subject line, the easier it is for us to pick the items to peruse. None of us has the same set of interests. I might read posts on TEM but not ion probe. Someone else may read anything in the life science but nothing in materials science. And so it goes. I think one list with good subject lines will serve the community better than a lot of separate lists. Just my few sou, Tobias Baskin
For my $0.02....I think many of us are involved in both LM and EM, or are interested in both. As long as people stick to using descriptive subject lines, there isn't really a problem sorting through messages to find the ones of direct interest. But isn't it all interesting, anyway? I read (or try to read) the materials stuff...miles/km over my poor head most of the me, but still interesting. Or am I just a hopeless geek?
So - I'd prefer not to see the list split along those lines. Although I could just subscribe to both, if it comes to that; and just deal with the cross-postings.
PLease excuse typing/grammar oddities - I'm midway through a techniques course and losing my marbles.
Tamara CSHL
On Mon, 20 Oct 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 microscopist, } } For the fourth time a subscribed to the microscopy discussionlist and } three times I unsubscribed , because my main interest is LM , I do not } work for my research on marine plankton with EM. } Would it not possible to make 2 sublists one for LM and one for EM ? } Now members who's main work is with LM are sometimes flooded with EM } discussions and vice versa. } Please give your opinion. } } Pieter Houpt FRMS } } The Hague The Netherlands. }
There is an immediate opening for an experienced SEM operator in our East Coast independent analytical research laboratory. Duties are highly varied and challenging, mostly materials science; the major responsibility is operation of a JEOL JSM-840 with EDS. Frequent client contact. Non-smokers only; drug screen required. Please send resume with salary history in confidence to ablackwood-at-2spi.com
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
I am looking for brief comments on laser printers for doing images from our SEM and CCD cameras.
We have an Apple 4/600PS printer now. It's OK, but slow. I would like to replace it with something like an Apple 12/640PS or HP LaserJet 5M.
Any advice regarding which would be a good choice for our lab (mostly Mac, but printer would be on ethernet) and how much additional memory would be good to get for doing grayscale images of several MBs?
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
I am looking for brief comments on laser printers for doing images from our SEM and CCD cameras.
We have an Apple 4/600PS printer now. It's OK, but slow. I would like to replace it with something like an Apple 12/640PS or HP LaserJet 5M.
Any advice regarding which would be a good choice for our lab (mostly Mac, but printer would be on ethernet) and how much additional memory would be good to get for doing grayscale images of several MBs?
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
My primary interest is in electron microprobery, nevertheless I feel that I get something out of the biolological EM and even LM material.
After all, most people do put in a "subject" field, and the "delete" button is, on my computer at least, very close to hand.
Please leave things as they are.
Ritchie
} For the fourth time a subscribed to the microscopy discussionlist and } three times I unsubscribed , because my main interest is LM , I do not } work for my research on marine plankton with EM. } Would it not possible to make 2 sublists one for LM and one for EM ? } Now members who's main work is with LM are sometimes flooded with EM } discussions and vice versa. } Please give your opinion.
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Dear Electron Beamers, Imagers, Probers and etc.'ers. Another great opportunity to touch base with fellow microscopists etc. in the Ottawa area.
WHEN: Wednesday, October 29, 1997, 3:00 p.m. to 8:30 p.m.
WHERE: Same place: RCMP Sgt.s Mess, HQ BLDG., 1200 Vanier Pkwy (at Hwy #417) Directions for the Sgts. Mess and parking available at entrance gate.
FORMAT: Same as before, very informal; any examples of your work in any format would be appreciated for conversation pieces. (Poster board stands will be available). Micrograph Competition "MOST INTERESTING MICROGRAPH"
NOTE: Sorry but no equipment demos: Computer demos can be accommodated to some extent but access to power is limited.
FOOD (free): Snacks, Coffee, Beverages - Served at 4:00 .p.m. Pizza and Subs - Served at 5:30 p.m. For those in need there will also be a cash bar available between 4:00 p.m. and 8:00 p.m.
COST: Its Free=A6 Thanks to some of our local vendors of equipment and supplies. (Who will also be there as colleagues in our field)
I WILL ATTEND (FAX BACK)
NAME PHONE FAX =09 =09 Dave Ballantyne tel:613- 998-6047 fax: 613-952-7325 e-mail: david.ballantyne-at-rcmp-grc.gc.ca Jeff Fraser tel613-: 993-8570 fax:613- 993-8566 e-mail: jeff.fraser-at-nrc.ca
I also would prefer a LM list. Perhaps it is too much work pro bono for the sysop. If list posters would use the prefixes suggested (LM, PLM, SEM, TEM), then it would be much easier to filter out the postings. To enforce this, the sysop would have to have a way to electronically bounce nonconforming postings. Perhaps Nestor would comment.
Cross fertilization is in my opinion essential to science. We all learn from others and I have benefited from the occasional tidbit of information from other fields. It would be a mistake to seperate "microscopists" just based upon a percieved differentiation in the illumination source, imaging modality, or field (Life Science / Physical Science).
I realize that there are a few people that have particuliar interests and they would prefer a seperate list, but it is not my intent to segregate the community but bring it together. With these comment in mind I will respectfully decline the request to create yet another list.
Modern Email programs such as Eudora, have the ability to filter messages based upon key words. To be honest with the volume of mail I receive on a daily basis it is the only way I can keep up. If everyone uses the subject line as it is intended and the delete key then it is a simple job to skip a whole range of messages.
Just for those that have forgotten here is a exerpt from the instructions on Subject lines....
As a courtesy to the readers of this list please indicate in the subject line of your message a reasonable descriptive title of your comment/inquiry. Also preface your description by the conventional abbreviation of the type of microscopy you are interested such as LM, IRM, XRM, TEM, SEM, AFM, STM, uProbe...... For example, if you are interested in optical microscopy and have a question about staining then a Title/Subject line for your message might be
Subject: LM - Need help on Staining Cells
or if you are interested in TEM analysis of dislocations then
Subject: TEM - Dislocation Loop Analysis and so forth.
Just a note about Apple printers. They have a technology called photograde which widens the gray scale and makes photos look better. For details, see their web page at:
http://imaging.apple.com/printers/pr-tech.html
We have that technology on a LaserWriter Pro 630 and I think it prints at 300 dpi as well as a LaserWriter Pro 810 at 800 dpi (but in less time). I do not know if other manufacturers have something like this.
The printer is not especially fast, and I don't know what you can do to really speed one up, except that you might as well put as much RAM in them as will fit.
Disclaimer: I do not sell Apple printers, I just like the ones we have!
Cheers,
John Vetrano js_vetrano-at-pnl.gov _______________________________________________________________________________
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Hi:
I am looking for brief comments on laser printers for doing images from our SEM and CCD cameras.
We have an Apple 4/600PS printer now. It's OK, but slow. I would like to replace it with something like an Apple 12/640PS or HP LaserJet 5M.
Any advice regarding which would be a good choice for our lab (mostly Mac, but printer would be on ethernet) and how much additional memory would be good to get for doing grayscale images of several MBs?
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
} Cross fertilization is in my opinion essential to science. } We all learn from others and I have benefited from the } occasional tidbit of information from other fields. It } would be a mistake to seperate "microscopists" just } based upon a percieved differentiation in the illumination } source, imaging modality, or field (Life Science / Physical Science). } } I realize that there are a few people that have particuliar } interests and they would prefer a seperate list, but it } is not my intent to segregate the community but bring it } together. With these comment in mind I will respectfully } decline the request to create yet another list.
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Dear Jon, I have been using an HP Laserjet 4P for about four years, now, to print images from my SEM and Quartz PCI. This printer (600 dpi) runs hard all day long and I usually need a new cartridge every three or four weeks from printing everyone's images in 8 X 10 format (instant blow-up). For reliability the HP's can't be beat. It has never given me the slightest problem. I have an additional 4MB of memory, on top of the 2MB it comes with. If you get a 1200 dpi printer, double that. This does an 8 X 10 print in about 1.5 minutes. It is the size of the printout that has the most effect on the speed. You wrote: } Hi: } } I am looking for brief comments on laser printers for doing images from our } SEM and CCD cameras. } } We have an Apple 4/600PS printer now. It's OK, but slow. I would like to } replace it with something like an Apple 12/640PS or HP LaserJet 5M. } } Any advice regarding which would be a good choice for our lab (mostly Mac, } but printer would be on ethernet) and how much additional memory would be } good to get for doing grayscale images of several MBs? } } Thanks. } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } Regards, Mary
snip=8A NOTE 1: The most recent version of Image Pro Plus contains a circular line profile feature, that does all this automatically. NOTE 2: John Russ of "The Image Processing Handbook" fame has expended a lot of effort on this very problem but from another angle, cf: "Application of the Hough Transform to Electron Diffraction Patterns", Journal of Computer- Assisted Microscopy, Vol. 1, No. 1, pp. 3-37 (1989). I heard that he is producing some software for Photoshop using Hough transforms due out in a month(?) that performs this procedure. Aanyway, good luck! It's a great technique. Please contact me off the listserver if you need more info.
This note describes a procedure for analyzing spots or ring electron diffraction patterns with an image analysis program and a personal computer.
Software: Image analysis program from Image-Pro plus for the PC from Media Cybernetics (www.mediacy.com). Plotting program from Origin Ver 4.0 from Microcal Software Inc.
Hardware: IBM compatible Pentium, Windows NT or equilavent; HP scanner with negative attachment.
Purpose: The analysis is used to do data extraction from electron diffraction patterns that is equivalent to a line scan plot of intensities vs distance (Rd) from the centre of the central spot on the diffraction pattern to some arbitrary user selected point outside the spots or rings near the edge of the photo.
Procedure: The negative to process is converted to a grey level tiff image with a flatbed scanner. A program written in the Auto_Pro macro language of Image-Pro finds the x,y coordinates of the central spot and calculates successive histograms of concentric circular areas of interest starting at the centre of the central spot and increasing by one pixel radius outward to some user-selected point.
Each of these values which are the sums of all the grey levels within these circular areas of interest are appended to a file for later processing with the Origin plotting program. A background image is created from the original using the background extraction feature of the Image-Pro program and a background' file is created using identical xy coordinates and circular histogram parameters. Processing: Subtracting the preceeding value from each of these histogram sums produces a table where each value is equilavent to adding up all the pixel values that lie on the original concentric circle that bounded the histogram. Plotting these values vs their row number is a line scan of intensities from the centre of the central spot to the selected position.(ie: intensity vs Rd). Similar processing of the background image gives a background line profile. Specific instructions: Scan the EDP negative as grey level with the highest resolution available( currently 200dpi on the HP scanner and fine black and white photo mode.). Save as a tiff file. Load the file into Image Pro Invert the file (only to get numbers for a graph that has the background(black) as zero and spots (white) giveing increasing numbers. Make sure you apply the inversion map to the image. select an AOI that includes the spots and rejects edges and various features that are not part of the pattern. duplicate/crop this area and minimize the original image for clarity of the display. create a background image from the duplicated image using the background extraction feature . run the macro diff_main' and follow the instruction on the screen. This macro will find the centre of the image and allow you to select an outer ring where the analysis will stop. This center and outer ring radius will be exactly the same on the foreground' image and the background' image. This is necessary to simplify later processing of the data. The files generated currently have to be edited because the first reading is extraneous and the current version of Image Pro adds two newline characters between readings when you append data to the file. To remove these use for example, Wordperfect and search for three newlines and replace them with one newline. This is essential for Origin or Excel to import the data as a continuous column of numbers. The file as saved has five items in a row and the last one is of interest to us since it is the sum of the grey values contained in the concentric circular histograms that the macro creates. The first four columns can be deleted if desired. The first readings are not accurate representations of the histogram because the size of the circle is only one pixel and it increments by one pixel radius for successive histograms. The first few readings are significant in that their existence defines the distance from the centre of the central spot to where ever the outer ring was chosen. The point of this exercise is to subtract the background image data from the foreground image data and to end up with a plot of Rd vs intensity and try to coorelate the intensities and peak positions with orientation on an unknown sample.
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-cime.uhd.epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
Hi, despite or better, just because being a greenhorn in using the = Listserver for informations of any +/- EM-related kind I do not vote = for separating the list into several specialized fields of interests. = Nestor Zaluzec=B4s suggestions in my opinion are a great deal.=20 I am learning, learning and learning and am interested to see also other = problems in neighboured areas of my main topic TEM/diagnosis/pathology = (by the way it is too much related and connected with LM due to the = necessity of correlative microscopy for diagnostic purposes).=20 I fully agree to the opinions of Tamara, Tobias, Ritchie and Nestor and = partially to corwinl-at-pt.cyanamid.com in respect to "subject lines" : if = requests would posted using systematic (?) Prefixes, it would help much. = Is there anybody who might have a suggestion how to adhere to "rules" = which must be designed very simple but obligatory??? Therefore: as Ritchie said: "No No No Please leave things as they are."
Best regards, have a nice day Wolfgang MUSS A-5020 SALZBURG, Austria/Europe
Mark Blackford asked for a soft to perform intensity integration along diffraction rings.
If you have access to a copy of Digital Micrograph from Gatan, you can easily add a plug-in named "rotational projection" (or some equivalent name) available on Gatan home site http://www.gatan.com/. It was initially designed to integrate rings in the Fourier transform of images. Thus if you enter your diffraction pattern, let say in TIFF (ie real numbers), you should first tell to the program to change the data into complex numbers and then run the averanging on the rings. If you know how to use the programming language of Gatan, you can also probably modify the data type required in the plug-in (it will then need less RAM to run). Don't forget that diffraction "rings" may deviate from perfect circles by some percent if you don't check and correct the astigmatism in diffraction mode!
Best regards
Philippe Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-cime.uhd.epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
Check the review by Kirz et al., Soft X-ray microscopes and their biological applications,1995, Q. Rev. Biophys., 28:33-130. Also, Chris Jacobsen and members of his group at SUNYSB had excellent presentations on the topic, including new developments, at MSA in Cleveland.
Regards, Michel **************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
Dear all, Does anybody know where I could obtain a sample of natural yttrialite (a thorium bearing yttrium silicate). I would like some to compare with some yttrium disilicate (the synthetic form of yttrialite) that we have made in our lab.
Thanks
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ 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. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
We have a variety of laser printers but my favorite is a Lexmark Optra L(?). It is on a heterogeneous (Mac/PC/UNIX) network and can handle large images without a problem. The printer easily handled a 24MB tiff image embedded into a frame in a Microsoft Word document although it took seven minutes to print. However, I typically print 2-3 640x480 8-bit images on a singe page, along with text, arrows, graphics, etc. and by the time I walk the 50 steps to the printer, the page is done. I'm a happy customer.
Also, check out the latest PC Magazine (Nov. 4) which is devoted almost entirely to printers.
------------------------------------------------ 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 --
I, too, am in the market for a new printer and have been extremely impressed with the HP and Epson stylus photo printer. It's so new on the market I had trouble finding someone with on to test. Try going through Epson or HP.
I was asked by a chemist to look at carbon nano tubes, as I'm a biologist I have no idea how to prepare the stuff. May I just put some powder on a grid and look at it or are there some special procedures ? Any commments , ideas and references are welcome.
TIA
Marc
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
Well... I believe that anyone is free to set up a separate listserv f=F6r Light Microscopy without nessecarily having to split the current microscopy listserv...
People are then free to decide what list/lists to subscribe... :)
} } We have a variety of laser printers but my favorite is a Lexmark Optra } L(?). It is on a heterogeneous (Mac/PC/UNIX) network and can handle } large images without a problem.
We also have the Lexmark (Optra R) and use pc/mac/unix with success. The printer is 1200 dpi but the best thing is the grey level capability. Try experimenting with gloss and semi-gloss papers for different appearance.
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
Due to an information of Dalene Josling (thank you very much!) that my = posting was not read-able because of more characters than 80 /line I = post the message once more to the Server (hopefully I was able to get = rid of the wider format, I counted 72 characters/line now):
Hi, despite or better, just because being a greenhorn in using the = Listserver for informations of any +/- EM-related kind I do not vote for = separating the list into several specialized fields of interests. Nestor = Zaluzec=B4s suggestions in my opinion are a great deal.=20 I am learning, learning and learning and am interested to see also other = problems in adjacent areas of my main topic TEM / diagnosis / pathology = (by the way this is too much related and connected with LM due to the = necessity of correlative microscopy for diagnostic purposes).=20 I fully agree to the opinions of Tamara, Tobias, Ritchie and Nestor and = partially to corwinl-at-pt.cyanamid.com in respect to "subject lines" : if = requests would be posted using systematic (?) prefixes, it would help = much (this also would be valid for the "Archives" -section). Is there = anybody who has a suggestion which, and how to adhere to "rules" which = must be designed very simple but obligatory???
Therefore: as Ritchie said: "No No No Please leave things as they are."
Best regards, have a nice day Wolfgang MUSS A-5020 SALZBURG, Austria/Europe
My interests and career have heavily involved both LM and EM, so their separation would mean that I would have to look at two lists instead of one. I hope that they can stay together, since I think they reinforce one another.
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.umich.edu/~akc/
Dear all, Please point me to some references related to quantitative measurement of lipid/phospholipid/membrane extraction due to conventional chemical processing of biological samples. Thank you in advance.
Yew Meng Heng E.M. Facility Health Sciences Centre McMaster University Hamilton, Ontario Canada
YES there is really such a thing - the Maryland Microscopical Society 25th semi-annual swap meet will be held on November 2, 1997 at the Holliday Inn in Calverton, MD (Exit 29B off Route 95, just North of the DC beltway) 10am - 4pm. 50 dealers expected - this is usually a good show with lots of used equipment. More info.?, contact J.F. Ptak, 202-337-0945.
I have been going for about 10 years, and this is a great place to pick up used equipment.
Instead of using wax for attaching the boats, use nail polish. You can get really cheap nail polish in black, blue, or green if you wish. I gave up on the LKB Multiplates years ago. The temperature is not right for properly drying the 1-micron sections, or staining with toluidene blue either. Joyce Craig Chicago State University
} I was asked by a chemist to look at carbon nano tubes, as I'm a biologist I } have no idea how to prepare the stuff. May I just put some powder on a grid } and look at it or are there some special procedures ? Any commments , ideas } and references are welcome.
We examine carbon nanotubes as follows:
1. suspend the specimen in a small volume of either acetone or methanol (trial and error will be needed to determine the exact dilution. so try different dilutions)
2. suspend the sample by swirling (some people sonicate for several minutes) and use a pipette to transfer a droplet onto a holey, filmed (Butvar or Formvar) and carbon coated grid (suggest purchasing from commercial source)
3. allow to air dry and examine in TEM looking for tubes suspended over the holes (best resolution)
The TEM should be set up for hi res, cooled traps over specimen area and a clean vacuum system is needed.
Good luck.
#################################################################### 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 agree that the Microscopy list should not be split. [Nestor needs *something* to keep him busy. :-)! ]
But there already is a list dealing mostly with light microscopy. It's the Histonet list. They do cover related topics as well, but most posts are about light microscopy, immunostaining, and tissues (no materials stuff). It's run by histotechnologists. Address:
{HistoNet-at-Pathology.swmed.edu} Subject: re: subscribe } } WHO SHOULD SUBSCRIBE? } Anyone interested in research or clinical applications of histology, } immunohistochemistry, in-situ hybridization pathology, and electron } microscopy may find Histonet informative and useful. Currently, there are } more than 850 subscribers from all over the world. Subscribers include } hospital employees from major urban centers and obscure remote locales, } university researchers, botanists and the employees of commercial } laboratories, government agencies, veterinary facilities and a wide } variety of commercial industrial ventures. } } WHO RUNS HISTONET? } The list is run by Linda R. Margraf, M.D. and Herb K. Hagler, Ph.D. using } hardware and software owned by the University of Texas Southwestern } Medical School, Department of Pathology in Dallas, Texas. If you have any } questions or problems with Histonet please contact Linda Margraf at } LMargraf-at-childmed.dallas.tx.us. } } HOW DOES THE LIST WORK? } This server, unlike many systems, uses ONLY ONE ADDRESS to send commands } to the computer and to post messages. The server will recognize commands } sent in the SUBJECT line of the message and only when they are spelled } exactly as listed below. Anything not identified as a command will be } circulated to EVERYONE on the list. } } The following is a list of commands the server recognizes: } } subscribe } Your address will be added to the list of subscribers. You will then be } able to send messages to this list that will be forwarded to all other } list subscribers. You will begin to receive all messages sent to the list } by other subscribers. } } subscribe digest } Your address will be added to the list of subscribers who receive a digest } instead of each forwarded message. A digest is a compilation of all the } messages received in a 24 hour period. It is sent to the digest } subscribers every night after midnight. Digest subscribers can post and } respond to messages the same as "real-time" subscribers.
} Well... I believe that anyone is free to set up a separate listserv f=F6= r } Light Microscopy without nessecarily having to split the current microscopy } listserv... } } People are then free to decide what list/lists to subscribe... :) } } / Martin
Hi, I had the same problem, so I can recommend you to make a suspention of this powder in alcohol, then put a drop on a grid with deposited thin film. Contrast will be poor, so you will need to use defocused image.
Vladimir Dusevich dusevich-at-ncsu.edu
Schmutz Marc wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi around the microscopy world, } } I was asked by a chemist to look at carbon nano tubes, as I'm a biologist I } have no idea how to prepare the stuff. May I just put some powder on a grid } and look at it or are there some special procedures ? Any commments , ideas } and references are welcome. } } TIA } } Marc } } ------------------------------ } SCHMUTZ Marc } IGBMC } 1 rue Laurent FRIES } BP 163 } F 67404 Illkirch Cedex } FRANCE } } Tel: +33 (0)388 653 330 direct } Fax: +33 (0)388 653 201 } email:schmutzm-at-lear.u-strasbg.fr } } ------------------------------
At 03:15 PM 10/21/97 +0900, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I used to view these regularly for a materials scientist. We simply put a small amount of the carbon sample in a solvent and used a sonicator to disperse it. Ten minutes should do a decent job. We then put drops of the suspension onto holey grids which were sitting on filter paper. When they were dry we put them in the TEM for viewing. Some of the nanotubes would sit on the edges of the holes in the film and could be viewed that way.
The nanotubes we viewed required very high magnification and very stable beam and vacuum conditions. Use liquid nitrogen on the pumps and on your decontaminator. Set up your instrument for high resolution conditions and let the electronics and gun stabilize for a while before viewing. Getting the best resolution was trickier by far than preparing the samples.
Good luck.
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Upon the advice of John MacKenzie, I began training our facility users to open their images in Photoshop, set the halftoning screening to 150 lpi, and using our HP LaserJet 4M+ to make working prints of scanning electron micrographs and, with less success, transmission electron micrographs. They are a vast improvement over the default settings.
However, four weeks ago I bought an Epson Stylus 800 ink jet color printer for home (list $449, cheaper at various outlets). It makes excellent working prints of SEMs and, when pushed to the limit (higest res, expensive paper), makes *nearly* publication quality prints. Fine, light, horizontal lines do show in areas of solid color, although they are less noticeable on a colleague's printer than on my own. They would be great for reviewers' copies and general-purpose applications. The colors (including black and greys) are snappier than on a dye-sublimation printer. (The dye-sub printer I have used shows the horizontal lines, as well, but they are more obscured by the slight blurriness of the process.)
Special ink-jet papers run from a few cents a sheet to about $.50 for the really good stuff. Replacement color and black ink cartridges are about $20 to $27, depending on source, and can be used up pretty quickly on images.
We will be buying this printer for the EM lab within the next week, primarily for good working prints and posters (it does banners, as well). It's a great buy for less than the cost of a dye-sub printer, but I don't expect it to do it all! I still use the darkroom...
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 don't know if you have a special place in your heart for LaserJet printers specifically, but if you don't, we are getting excellent results with the Epson Stylus Color 800 Ink Jet printer which prints up to 1440 dpi/8 bit image depth. I typically print four 512x512, 8 bit images on a page at 720dpi and it takes about 2 minutes. We are using it on a Novell network with no extra memory. There is special paper for printing -at- 720dpi or above which costs about $12 US for 50 sheets and there is also a special glossy paper which really makes the images look photographic, which sells for about $25 US for 15 sheets. If you have your heart set on a LaserJet printer to use plain paper, I also have used the HP 5P (now no longer sold as the 5P but the 6P has the same engine) which gives true 8 bit gray level shading and excellent pictures. I, personally could not see enlarging the images larger than 2 per page (3.5"x3.5"), but both of these printers have done an excellent job of eliminating the "P" word from the lab. The Epson, being color, has the added advantage of doing a great job with color presentation graphics and transparencies. The Epson sells for about $400 US.
David Bell Millipore Corporation 80 Ashby Road Bedford, MA 01730 (617)533-2108
As usual, I have no vested interest in either of the above mentioned companies, nor do I have anything against Polaroid! (Film may never die!)
Jonathan Krupp wrote:
jmkrupp-at-cats.ucsc.edu on 10/20/97 02:41:30 PM
To: Microscopy-at-Sparc5.Microscopy.Com cc: (bcc: David Bell)
Hi:
I am looking for brief comments on laser printers for doing images from our SEM and CCD cameras.
We have an Apple 4/600PS printer now. It's OK, but slow. I would like to replace it with something like an Apple 12/640PS or HP LaserJet 5M.
Any advice regarding which would be a good choice for our lab (mostly Mac, but printer would be on ethernet) and how much additional memory would be good to get for doing grayscale images of several MBs?
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Ritchie Sims wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } No No No } } My primary interest is in electron microprobery, nevertheless I feel } that I get something out of the biolological EM and even LM material. } } After all, most people do put in a "subject" field, and the "delete" } button is, on my computer at least, very close to hand. } } Please leave things as they are. } } Ritchie } } } For the fourth time a subscribed to the microscopy discussionlist and } } three times I unsubscribed , because my main interest is LM , I do not } } work for my research on marine plankton with EM. } } Would it not possible to make 2 sublists one for LM and one for EM ? } } Now members who's main work is with LM are sometimes flooded with EM } } discussions and vice versa. } } Please give your opinion. } } Ritchie Sims phone: 64 9 3737599 ext 7713 } Department of Geology fax: 64 9 3737435 } University of Auckland } Private Bag 92019 } Auckland } New Zealand
I agree with the concensus about keeping LM and EM together. We have been fighting a battle for more balanced approaches to integrated microscopy and having the two intertwined makes a lot of sense. My original training was with the Royal Microscopical Society, which integrates all types of microscopy. It was the general feeling then, and my continued feeling, now, that each of us has a lot to learn from rubbing elbows with neighboring technologies.
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 ********** Americas First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
We just make a dispersion of the powder ( say in methanol) and put a drop on a holey grid and we let it dry for a few minutes. We examine the nano tubes at about 100K to 300K. If you use regular carbon coated grids ( rather than holey grids) the image of the nano tubes will not be as clear because the carbon background will interfere.
I was asked by a chemist to look at carbon nano tubes, as I'm a biologist I have no idea how to prepare the stuff. May I just put some powder on a grid and look at it or are there some special procedures ? Any commments , ideas and references are welcome.
TIA
Marc
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
Question for anyone using a Lexmark Optra printer (specifically Optra Rn+) have you run into "spotting" problems with your prints? We've been having problems with ver fine "spotting" or "speckling" - randomly distributed black dots ranging from ~20 um to 300um (no I didn't actually measure them) all over the page. If so have you managed the a cure? I've clean everything I can think of serval times, without success. I've contacted Lexmark, but they weren't all that helpful and suggested sending it in for servicing. I haven't even gone through one cartridge yet!
Any suggetsions?
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
For those of you who print on CMYK printers like the Epson photo, how do you deal with the out-of-gamut colors like pure red or green or blue? It looks like my HP1600 (or is it the program I'm using, Photoshop) may be substituting a single shade of red for all out-of-gamut intensities of red in the original, which makes the print look saturated, lifeless, less detail than I expect. Since I am printing confocal images that are all red & green (& look beautiful on the screen), it's an obvious problem.
Is there a good solution (other than buying an RGB printer)? Is there an easy _automatic_ way to compress the dynamic range to span the colors available from CMYK printers, rather than clipping off the top intensities? or another approach?
Thanks! Richard Richard_Thrift-at-DepoTech.com p.s. it has struck me again how different the images look depending on the monitor used. I optimized the color levels using one monitor and they now look poor on another. Can you recommend color management systems, & indicate price? Thanks!
I've heard a few mentions of the BioQuant system in the past. Now I am considering it myself, and would like to hear a little more detail from users.
What do you like most/least about the system? How easy is it to handle intensity comparisons of stained specimens (BF and Fluor)? What camera option did you choose and how well does it work with a variety of specimen types (macro copy stand, micro BF, fluor, moving subject, etc)?? Does it really do excellent gel densitometry? How about colony counting? 3D measurements? Can it turn VCR tapes (short) into digital movies?
I'd appreciate any comments! Thanks as always, Karen
P.S. Regarding the Message Subject requirements, what about general subjects like digital imaging, printers, etc?? LM vs. TEM, etc. doesn't really fit.
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
} We also have the Lexmark (Optra R) and use pc/mac/unix with success. } The printer is 1200 dpi but the best thing is the grey level } capability. Try experimenting with gloss and semi-gloss papers for } different appearance.
In a message dated 97-10-21 18:20:52 EDT, tina-at-pbrc.hawaii.edu, wrote:
{ { However, four weeks ago I bought an Epson Stylus 800 ink jet color printer for home (list $449, cheaper at various outlets). It makes excellent working prints of SEMs and, when pushed to the limit (higest res, expensive paper), makes *nearly* publication quality prints. Fine, light, horizontal lines do show in areas of solid color, although they are less noticeable on a colleague's printer than on my own. They would be great for reviewers' copies and general-purpose applications. The colors (including black and greys) are snappier than on a dye-sublimation printer. (The dye-sub printer I have used shows the horizontal lines, as well, but they are more obscured by the slight blurriness of the process.)
Special ink-jet papers run from a few cents a sheet to about $.50 for the really good stuff. Replacement color and black ink cartridges are about $20 to $27, depending on source, and can be used up pretty quickly on images. } }
Have you tried Kodak Inkjet Photographic Quality paper yet? It works out at about 60 cents a sheet and indeed produces "near photographic quality" results.
I have tried everything available and am very pleased with this product.
I use it with a Hewlett Packard 694C DeskJet printer but I believe that it is compatible with the Epson Stylus printers also.
} Due to an information of Dalene Josling (thank you very much!) } that my posting was not read-able because of more characters than 80 /line } I post the message once more to the Server (hopefully I was }
For those who use Pegasus mail, there is a option "Reader as you are reading a message with thw option "Wrap long lines" very useful.
} "No No No } Please leave things as they are."
Since I am working in multidissiplinary environment which includes SEM, TEM, CONF, LM, EDS and hopefully STEM, and PEELS at a later stage. We do have users from Life sciences and Physical sciences and a separated list is unthinkable for me.
My 5 cents worth (SA) ~ 1.3 cents worth (US). Bad exchange rate!
A couple of people e-mailed me to thank me for mentioning the trick of increasing the halftone screening resolution, so I will repeat it for you all.
In Photoshop, under File select Page Setup..., then click on Screen... *Uncheck* Use Printer's Default Screen, and enter 150 lines/inch for Frequency. Leave Angle at 45 degrees and Shape as diamond, unless you want to experiment.
To set this as the new default for Photoshop, Alt-click on the Save button (Windows) or Option-click (I think) on Save for Mac.
I have found this trick to work on numerous printers, not just HP LaserJets. It even made my Brother fax/copier/scanner/200dpi printer at home print reasonable SEMs! For some of the printers it was more necessary to adjust gamma levels or just to lighten up the images than on others, as more ink will be applied.
This tip originally came from John MacKenzie at MSA in Cleveland, and was worth the entire cost of traveling all the way from Hawaii!
I've got a few fun tricks coming up in the next Microscopy Today, and a more complete Photoshop tutorial in another month or so. I'd be glad to incorporate any other tips you all may have, so don't hesitate to e-mail.
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* ****************************************************************************
Regarding the suggestion that the List Server should be be sub-divided into LM andEM. No! No ! No! Microscopy is a broad canvas and we can learn from all aspects of the subject and fromm different application disciplines within microscopy.
Patrick Echlin Multi-Imaging Centre Cambridge University
On Mon, 20 Oct 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 microscopist, } } For the fourth time a subscribed to the microscopy discussionlist and } three times I unsubscribed , because my main interest is LM , I do not } work for my research on marine plankton with EM. } Would it not possible to make 2 sublists one for LM and one for EM ? } Now members who's main work is with LM are sometimes flooded with EM } discussions and vice versa. } Please give your opinion. } } Pieter Houpt FRMS } } The Hague The Netherlands. }
Marc, The method I used was to disperse the powder in 2-propanol. Sonication helps but is not necessary. Then a drop of the suspension is dried on a holey carbon support film. If you do not use a holey carbon, you won't be able to distinguish the tubules from the background.
Regards, Eric Lachowski
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
Does anybody have a spare Pirani gauge for an Edwards high vacuum carbon coating unit E12E. I would be eternally grateful to any list server member who did or new how I could get hold of one (relatively cheap). Thanks
The Microscopical Society of Southern California (MSSC) is proud to invite anyone interested to a lecture by Brian J Ford, on October 28 (Tuesday)
Topic: Antony van Leeuwenhoek and the Single Lens Microscope
Our program this month features a presentation on Antony van Leeuwenhoek (1632-1723) and single lens microscopy. Professor Ford is the author of countless books and papers and has done extensive research on the historical development of microscopy, including work with original, 17th century Leeuwenhoek instruments and samples from the archives of the Royal Microscopal Society in London. Many American microscopists know Professor Ford through his annual presentations at Inter/Micro in Chicago. The (MSSC) is Located at the Crossroads Schools in Santa Monica. Crossroads is located on Olympic Boulevard and 20th street. Anyone interested may contact Larry Albright the Program Chair at: 310-399-0865 Days or 310 471-0424 evenings.. It will be an exiting evening.......Larry
"The beauty and genius of a work of art may be reconceived, though its first material expression be destroyed, but when the last individual of a race of living things breathes no more another heaven and another earth must pass before such a one can be again." William Beebe albrite-at-Plasma-Art.com 419 Sunset Avenue Venice CA 90291 310-399-0865 310-392-9222 FAX
You are invited to check out my web site at: www.Plasma-Art.com
} The Microscopical Society of Southern California (MSSC) is proud to invite anyone } interested to a lecture by Brian J Ford, on October 28 (Tuesday) } } Topic: Antony van Leeuwenhoek and the Single Lens Microscope } } Our program this month features a presentation on Antony van Leeuwenhoek } (1632-1723) and single lens microscopy. Professor Ford is the author of } countless books and papers and has done extensive research on the } historical development of microscopy, including work with original, 17th } century Leeuwenhoek instruments and samples from the archives of the } Royal Microscopal Society in London. Many American microscopists know } Professor Ford through his annual presentations at Inter/Micro in } Chicago. } The (MSSC) is Located at the Crossroads Schools in Santa Monica. } Crossroads is located on Olympic Boulevard and 20th street. } Anyone interested may contact Larry Albright the Program Chair at: } 310-399-0865 Days } or 310 471-0424 evenings.. } It will be an exiting evening.......Larry } } "The beauty and genius of a work of art } may be reconceived, though its first material expression be destroyed, but when the last individual of a race of living things breathes no more another heaven and another earth must pass before such a one can be again." William Beebe } albrite-at-Plasma-Art.com } 419 Sunset Avenue } Venice CA 90291 } 310-399-0865 } 310-392-9222 FAX } } } You are invited to check out my web site at: www.Plasma-Art.com }
Jonathan Swift 1733
So, naturalists observe, a flea Larry Albright Hath smaller fleas that on him prey; albrite-at-Plasma-Art.com And these have smaller fleas to bite'em, 419 Sunset Avenue And so proceed ad infinitum. Venice CA 90291 Thus every poet, in his kind, 310-399-0865 Is bit by him that comes behind. 310-392-9222 FAX Web Page.www.Plasma-Art.com Jonathan Swift 1733
I used to use a pyramitome to trim my blocks for TEM, and since I have relocated to another facility, I don't have the luxury of using it anymore. I was wondering if they are still available and would appreciate more information on getting one. Please reply to me at my email address, rather than cluttering up the listserver.
Thanks in advance,
Susan
Susan Carbyn Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
Recently our resident molecular biologists have requested us to fix and process suspensions of ovarian granulosa cells so that they can perform in situ hybridisations on them. Because of the limitations of the experiment and the in situ methodology this locks us into a number of factors. 1. the cells must be in suspension (ie they have been mechanically dispersed) 2. Paraformaldehyde must be the fixative 3. the tissue must be embedded in paraffin
We have tried fixing in suspension in 4% paraformaldehyde in phosphate buffered saline, centrifuging to a pellet, resuspending in a small volume of 2% agar followed by 8 hours of dehydration (ethanol), clearing (xylene) and infiltration/embedding in paraffin. However there appears to be a lot of damage to the cytoplasm and the nucleii are very condensed.Any thoughts at all on preparing cell suspensions would be appreciated.
Regards Peter Smith AgResearch Wallaceville Upper Hutt New Zealand Smithp-at-agresearch.cri.nz
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I use all forms of microscopy and would be more than a bit annoyed if I had to subscribe to multple lists. In spite of its being somewhat cumbersome I prefer the listserver to remain unchanged.
my $0.02(US) worth
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/
Privilege does not absolve one of ecological responsibility.
Greetings, Peter Smith asked alternatives to agar embedding for "loose" samples. We have had good luck with a trick we picked up from the cryofixation folks. We make wire loops out of very fine copper wire (36 gauge) and coat the loop with Formvar. The diameter of the loop can be a few mm or up to 7mm (we have never tried larger). You then collect your cells on the Formvar. For example, if the cells are in a small volume of solution, go "fishing" with the loop. You can make the Formvar surface more sticky by pretreatment with polylysine. Now what you do is to place a second Formvar coat over the loop, thus trapping your sample between two Formvar films. You will probably need to be sure that you don't have too great a puddle of liquid on your loop for this step. Some fiddling will be needed. We cast small rectangles of Formvar on water, with the narrow side of the rectangle a bit wider than the loop diameter, and the long side of the rectangle a bit longer than twice the loop diameter. Then you can very quicky dunk your loop with the samples onto the Formvar rectangle. Line up your loop with the middle of the rectangle, so it makes two squares, and plunge at right angles to the water. The Formvar rectangle just snaps to the loop. Now your cells are trapped inside. But the FOrmvar is readily permeable to your fixative, to your dehydration agent and even to paraffin. At the end of the "day", you can excise the loop the with a razor. In fact, if you really get 36 gauge copper, you can just cut through it with a double-edged razor blade. I hope this helps. If any of the above was foggy, please reply. Tobias Baskin
} Recently our resident molecular biologists have requested us to fix and } process suspensions of ovarian granulosa cells so that they can perform } in situ hybridisations on them. Because of the limitations of the } experiment and the in situ methodology this locks us into a number of } factors. } 1. the cells must be in suspension (ie they have been mechanically } dispersed) } 2. Paraformaldehyde must be the fixative } 3. the tissue must be embedded in paraffin } } We have tried fixing in suspension in 4% paraformaldehyde in phosphate } buffered saline, centrifuging to a pellet, resuspending in a small } volume of 2% agar followed by 8 hours of dehydration (ethanol), } clearing (xylene) and infiltration/embedding in paraffin. However there } appears to be a lot of damage to the cytoplasm and the nucleii are very } condensed.Any thoughts at all on preparing cell suspensions would be } appreciated. } } Regards Peter Smith } AgResearch Wallaceville } Upper Hutt } New Zealand } Smithp-at-agresearch.cri.nz }
To those more chemically minded than myself (i.e. everyone),
I've seen many notices that waste 2% osmium tetroxide can be reduced to less hazardous form by mixing with twice volume of corn oil. I'd like to implement this practice, but I also use a lot of 0.5% ruthenium tetroxide. Can this also be reduced with oil? The only thing I've heard of for RuO4 is sodium bisulfite, which is quite toxic/harmful itself. It would be great to find a single practice that works for both Os and Ru.
Also, does anyone know if buffer solutions (cacodylate or phosphate) interfere with OsO4 reduction by corn oil?
Thanks for your help, Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
may-be this question doesn't belong here, but I am totally stuck so I still ask it: I have recently taken some 120 Mb of digitized pictures with a CCD-camera equiped microscope using IPlab software on a MacIntosh. Does anybody know a windows program that can read IPlab's TIFF format (12 bit greyscale, stored in 2 bytes, I think) and convert it to any windows-recognized format? These data are important for me. Thanks in advance. Kees Jalink
--
Kees Jalink The Netherlands Cancer Institute, dept. of Cell Biology H1 Plesmanlaan 121 1066CX Amsterdam, the Netherlands 020-5121982 (tel) / 020-5121989 (fax) kees-at-NKI.NL (email) / 0297-320248 (tel at home)
Dear Karen, for over ten years now I (use my OsO4-solutions) and dispose of my used = up working solutions in a way which is very safe, but uncommon. I=B4ve = had posters on that at MSA-Meeting Boston 1992 (also included in the = Proceedings, 50th Ann. Meeting, p. 754/755: MUSS W.H.: "Pro=B4s for a = multiple/repeated use and safe disposal with recovery of OsO4 = solution(s) in routine EM") For about 1300-1600 specimen blocks to osmicate I need only 4-6 g = OsO4/year. I don=B4t use the cornoil-disposal method, because I think Os a valuable = metal which shouldn=B4t be wasted off the drain/ down the sink ("with a = plenty flush of water" !! as has been proposed by several people, which = in my opinion would not fit ecological considerations!) but be recovered = and recycled. I learned my lession at the 1992 Boston Meeting as well as within the = last 5 years: I tried people to convince that it is possible to reduce = the amount of OsO4 used for staining/postfixation of specimens AND = recover all of the Os from solutions as used (like buffers, like = mixtures etc.) as this is possible in Switzerland, where a regular Os = recovery and recycling has been established for all EM-Labs aided by the = SGOEEM=3DSwiss Society for EM since 1972!!=20 It is not easy to convince people about that though the process is very = easy, simple, effective in my experience and more or less costs are low. = I am doing it. I have, unfortunately, no experience in "deactivating" = RuO4, but I think, considering my scanty knowledge from chemistry = lessions and 17 years experience in disposal/recovery of OsO2/Os (+/- = pure), this should be no problem at all. If you want to get informed about the reasons, why and how I do it: = please don=B4t hesitate to contact me by e-mail.
Best wishes for a lucky day to you and all of the community
Wolfgang MUSS EM-Lab, Dept. Pathology, LKA A-5020 SALZBURG, Austria/Europe.
I fix cells in suspension on regular basis and use agar to encapsulate the cells before pelleting them. Fix cells in suspension, spin down gently to remove excess fix and resuspend in minumum fix. Make 2% agar and keep at 48 deg to avoid solidifying the agar. Mix equal vol of concentrated cell suspension and agar, solidify the suspension and then run up the agar piece as you normally would any tissue.... hope this helps. However if the tissue is losing some of the morphological integrity then you may need to change the fixative or may be add low concentrations of glut to it. Neelima Shah,UOP morphology core.
At 08:47 AM 10/23/97 +1300, Smith, Peter wrote: } } Recently our resident molecular biologists have requested us to fix and } process suspensions of ovarian granulosa cells so that they can perform } in situ hybridisations on them. Because of the limitations of the } experiment and the in situ methodology this locks us into a number of } factors. } 1. the cells must be in suspension (ie they have been mechanically } dispersed) } 2. Paraformaldehyde must be the fixative } 3. the tissue must be embedded in paraffin } } We have tried fixing in suspension in 4% paraformaldehyde in phosphate } buffered saline, centrifuging to a pellet, resuspending in=A0 a small } volume of 2% agar followed by=A0 8 hours of dehydration (ethanol), } clearing (xylene) and infiltration/embedding in paraffin. However there } appears to be a lot of damage to the cytoplasm and the nucleii are very } condensed.Any thoughts at all on preparing cell suspensions would be } appreciated. } } Regards =A0=A0=A0=A0 Peter Smith } =A0=A0=A0=A0=A0=A0 AgResearch Wallaceville } =A0=A0=A0=A0=A0=A0 Upper Hutt } =A0=A0=A0=A0=A0=A0=A0 New Zealand } =A0=A0=A0=A0=A0=A0 Smithp-at-agresearch.cri.nz=A0=A0=A0=A0=A0 } =A0=A0=A0=A0=A0 } } Attachment Converted: "c:\eudora\attach\LM cell suspensions"=20
Has anybody evaluated the Agfa T5000 or T8000 or the ScanMate 11000, 3000, F8 or F8 Plus for use in high resolution EM (2D crystals, helical arrangements or singe particles of proteins)? We're trying to decide which scanner to buy and we're specifically looking at geometric accuracy of the scan and modulation transfer functions. So far we have data on the Zeiss SCAI, the Scitex Eversmart Pro and an Eikonix scanner.
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
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I have had all of my nerve and muscle questions answered in; Myology by Andrew G. Engel and Betty Banker, Vol I and II, McGraw-Hill. I have no idea of the year of the latest edition or the price. My edition (1986) has 2106 pages. Kate Connolly
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Reply to: RE} molviol distributor?
Dear Reinhard:
We buy our supply of Mowiol from Calbiochem (LaJolla, CA 92039) = Tel.1-800-854-3417 The catalogue number is 475904 and 100g costs $44US. Hope this helps.
Linda Chicoine Center for Cell Imaging Yale University New Haven, CT 203-785-3646 http://info.med.yale.edu/cellimg
--------------------------------------
Hi,
I am looking for the fluorescent antifading mounting medium MOVIOL. Can someone out there tell me who the distributor is?
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What makes you think that sodium bisulfate is toxic? The Merc Index gives its LD50 (in rats) as 115 mg/Kg, and says that it is used as a preservative and bleach in food.
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
You might want to begin with Image Alchemy from Handmade Software (http://www.handmadesw.com/index.html) or Thumbs Plus from Cerious Software (http://www.cerious.com). While I don't know if they have direct translators for IPLab files, they can handle many types of conversion.
NIH Image may work also. I don't have the URL handy. Does anybody else have it?
If you can't translate the files directly, can you use IPLab to translate them into an intermediate, platform-independent format? TIFF? JPG? Photoshop?
------------------------------------------------ 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 --
Thanks a lot to all who gave me some nice tricks to observe carbon nanotubes. It helped me a lot and due to your nice indications I've got correct images at the first shot.
Marc
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
In a message dated 97-10-23 03:19:53 EDT, Kees Jalink wrote:
{ { Does anybody know a windows program that can read IPlab's TIFF format (12 bit greyscale, stored in 2 bytes, I think) and convert it to any windows-recognized format? } }
PaintShopPro, ver. 4.14, from JASC will read 6 different TIFF subformats. These are: uncompressed, Huffman compresssed, pack bits compressed, LZW compressed, Fax Group 3 compressed, and Fax Group 4 compressed. None of these are 12 bit grayscale. They range thru 1, 4, 8, and 24 bit/pixel color scales. Attach a single file to an e-mail to me and I will try to change the image to something else, such as a gif or a jpg, your choice. PSP supports 35 different formats.
Yours truly, Steve Stokowski Stone Products Consultants Concrete Petrographers 10 Clark Street, Suite A Ashland, Massachusetts, 01721 USA 508-881-6364 http://members.aol.com/CrushStone/index.htm
My safety officer has told me that treating the osmium as noted may bring charges of reprocessing hazardous waste, for which the University doesn't have a license. They insist I give them the Osmium as is for disposal.
} To those more chemically minded than myself (i.e. everyone),
} I've seen many notices that waste 2% osmium tetroxide can be } reduced to less hazardous form by mixing with twice volume of } corn oil. I'd like to implement this practice, but I also use a } lot of 0.5% ruthenium tetroxide. Can this also be reduced with } oil? The only thing I've heard of for RuO4 is sodium bisulfite, } } which is quite toxic/harmful itself. It would be great to find a } single practice that works for both Os and Ru.
} Also, does anyone know if buffer solutions (cacodylate or } phosphate) interfere with OsO4 reduction by corn oil?
} Thanks for your help, vKaren
} -- } Karen Zaruba vkszaruba-at-mmm.com } 3M Company, 3M Center Bldg. 270-1S-01 vSt. Paul, MN 55144 } "The opinions stated above are my own, not necessarily 3M's"
--------------------------------------------------------------------- COME TO OUR OPEN HOUSE "INNER SPACE/OUTER SPACE" IT IS FREE, AND EVERYONE IS WELCOME Saturday, November 15, 1997 4-8 pm SEE ELECTRON MICROSCOPES TELESCOPES AND IN OPERATION FOR MORE INFORMATION, SEE MY WEB PAGE OR CALL 594-6182 --------------------------------------------------------------------- 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
} windoff-at-mail.uni-mainz.de Reinhard, MOWIOL Cat.#475904, is distributed in the USA by CALBIOCHEM-NOVABIOCHEM Corp. of LaJolla, CA phone 800 628 8470
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Salut to the listmembers, } } may-be this question doesn't belong here, but I am totally stuck so I } still ask it: } I have recently taken some 120 Mb of digitized pictures with a } CCD-camera equiped microscope using IPlab software on a MacIntosh. Does } anybody know a windows program that can read IPlab's TIFF format (12 bit } greyscale, stored in 2 bytes, I think) and convert it to any } windows-recognized format? These data are important for me. } Thanks in advance. } Kees Jalink } } -- } } } Kees Jalink } The Netherlands Cancer Institute, dept. of Cell Biology H1 } Plesmanlaan 121 1066CX Amsterdam, the Netherlands } 020-5121982 (tel) / 020-5121989 (fax) } kees-at-NKI.NL (email) / 0297-320248 (tel at home) } You might try Wang Image (now a standard free imager that comes with windows 95, or is availiable as freeware I believe) We use it to convert JEOL tiffs to tiff formats that Lview/NIH can read. It seems to read many different tiff formats but only saves in one (which I don't know, but a tiff saved by wang image suddenly becomes readable by all of our other viewer programs). \ *shrug* hope it helps} Christopher Institute of Meteoritics Albuquerque, NM USA
An issue that won't go away? I think there are legitimate reasons for this.
Like the sys-op and others with similar views expressed here, I believe in cross-fertilization among the hosted disciplines, and at least skim each message routed through the MSA Server.
Nevertheless, I do appreciate the other view. All inclusiveness is nice, but we all have different depths of interest, time, and patience.
In addition, MSA Mail Reflector messages arrive jumbled with more pressing and time sensitive professional and personal mail. The messages are rarely labeled in the subject field with the suggested key-words, and none bear a prefix (e.g. "MSA:") identifying them as emanating from this forum. I wade through it all-at-once, unable to apportion time by priority &| interest. How many have discarded a critical bill or letter intermingled with the mountain of trash stuffed in the post box? Need that experience be re-enacted in the electronic medium?
A fix does exists in most e-mail browsers in the form of the recipient defined filters (electronic or visual); some e-mail might get redirected to the MSA\LM folder, some to MSA\EM folder, some to MSA\Bio folder, ..., some to MSA\MISC folder, ... - to be perused as time and interests allow, and the reminder to the TrashBin folder. Without those key words, though, it's tough to set up the filters adequately.
However, the sys-op's recent appeal for voluntary use of the key-words in the subject field (please excuse this, Nestor) is naive; after a brief excursion into compliance, most of us return by expediency to the minimum that will get by. As the MSA Mail Reflector is currently set up, there are no simple solutions to the problem.
Suggested changes to the MSA Mail Reflector to address these issues:
(1) Filter the incoming postings for approved key-words in the subject field (for inclusiveness, "MISC." among them) and bounce back non-compliant mail with appended reminder of the policy and a list of acceptable key-words to choose among. (A new key-word may be added to the approved list when the traffic on the issue warrants separating it from the "MISC".) The filtering should also effectively purge the forum of machine generated SPAM that may spill in here inadvertently.
(2) Automatically prefix the subject field with the "MSA:" key-word. This, above the current practice by the MSA Server of inserting the MSA logo into the body text, will aid the subscribers in determination of the source and in sorting of e-mail prior to wading into the content.
(3) Create and ENFORCE a policy for vendors to use "VENDOR" as perhaps the last key-word in the subject field. Such feature would permit the commercial members to maintain their valuable contributions without bothersome censorship, but with ample warning for the rest of us. I am tiring of tripping over the same outfit indulging here in thinly disguised attempts at brand recognition & promotion; a disclaimer at the end of the message is too little too late. There ought to be a recognized, regulated, and thus less insidious venue for this stuff. (If non-compliance continues unabated, a special filter at the server might be programmed to append the VENDOR key-word automatically, perhaps with addition of the vendor's name, for the recalcitrant few.)
I do appreciate that all of this would fall on Nestor, who can't be looking for more work at the moment. But, in my opinion, it would address the root causes for the recent gripes without inconveniencing those who like things just the way they are.
So much for my 50 cents worth.
Otherwise, a great service, and under no circumstances I would unsubscribe.
A few weeks ago I posted a querry to the listserver wondering why I was not getting good fluorescent labeling using otherwise successful primary antibodies applied to the surface of one micron thick LR White sections followed by secondary TRITC or FITC conjugates. I sincerely thank those that responded, and I am now sharing the responses:
I have tried labelling 1 micron sections with lectins conjugated with TRITC and they worked exceptionally well.... that is the lectin was labelled with the TRITC tag...
Eric Rosen
I suspect the problem lies in the different secondaries, maybe try the fluorescent-gold-secondaries that are commercially available to test fluorescence and gold label at the same time. The product is called fluoronanogold and is available from Nanoprobes Inc. You can tap into their website (http://www.nanoprobes.com) to obtain detailed protocols or contact them at 516-444-8815. They also put out a newsletter.
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
I routinly test 1 micron LR White sections on glass before going to the trouble of Immuno EM. I usually use peroxidase ABC which seems to enhance the signal enough to be visualized. However, the fluorescent signal is very weak unless you start stacking antibodies which can create more backround. So you need a very good optimized scope to be able to see the signal and your faint signal may be obscured by the backround signal of the aquamount which has higher fluorescence than some others. Prolong from M olecular Probes is very quiet and enhances Texas Red. Or my favorite for quiet and quick is: 70% Glycerol, 25% .5M Tris pH 9, 5% n-Propylgallate, heat to mix then pH to 7.4 and store in the fridge. For use: 1 drop, swirl, drain most of it off then cove rslip. There should be barely enough to cover the coverslip.
Good Luck Doug
Robert Underwood Morphology Core Dermatology U of Wash. Seattle, WA
We had the same disappointment a few years back when we went from great IHC labelling at the EM level to nothing with half micron LR sections. We got round the problem in two ways. One, we gold labelled the half micron LR sections then silver intensified and photographed with white light. Two, we went back to embedding in wax, cut 5 to 10 micron sections, de-waxed and got excellent FITC-IHC labelling. Although we were reticent to go to wax embedding (19th century technology) it rendered available such enormous areas of antigenic sites that the IHC was excellent plus we could use the sections for in situ work and the fluorescent apoptosis kits.
If you really want to go to dissolvable resins then check Frank's work in: Gubler, F. (1989). Immunoflourescence localization of microtubules in plant root tips embedded in Butyl-Methyl Methacrylate. Cell Biol International Reports, Vol. 13, No. 1, January, 1989.
Eric Hines Microscopy Centre CSIRO Entomology Canberra
Have you tried to do regular gold followed by silver enhancement or gold toning on the semi-thin LR White sections? Then you'd even be using the same secondaries on your tests. I have no ideas on the fluorescence working (or not), but could you please post any responses to the server?
A few weeks ago I posted a querry to the listserver wondering why I was not getting good fluorescent labeling using otherwise successful primary antibodies applied to the surface of one micron thick LR White sections followed by secondary TRITC or FITC conjugates. I sincerely thank those that responded, and I am now sharing the responses:
I have tried labelling 1 micron sections with lectins conjugated with TRITC and they worked exceptionally well.... that is the lectin was labelled with the TRITC tag...
Eric Rosen
I suspect the problem lies in the different secondaries, maybe try the fluorescent-gold-secondaries that are commercially available to test fluorescence and gold label at the same time. The product is called fluoronanogold and is available from Nanoprobes Inc. You can tap into their website (http://www.nanoprobes.com) to obtain detailed protocols or contact them at 516-444-8815. They also put out a newsletter.
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
I routinly test 1 micron LR White sections on glass before going to the trouble of Immuno EM. I usually use peroxidase ABC which seems to enhance the signal enough to be visualized. However, the fluorescent signal is very weak unless you start stacking antibodies which can create more backround. So you need a very good optimized scope to be able to see the signal and your faint signal may be obscured by the backround signal of the aquamount which has higher fluorescence than some others. Prolong from M olecular Probes is very quiet and enhances Texas Red. Or my favorite for quiet and quick is: 70% Glycerol, 25% .5M Tris pH 9, 5% n-Propylgallate, heat to mix then pH to 7.4 and store in the fridge. For use: 1 drop, swirl, drain most of it off then cove rslip. There should be barely enough to cover the coverslip.
Good Luck Doug
Robert Underwood Morphology Core Dermatology U of Wash. Seattle, WA
We had the same disappointment a few years back when we went from great IHC labelling at the EM level to nothing with half micron LR sections. We got round the problem in two ways. One, we gold labelled the half micron LR sections then silver intensified and photographed with white light. Two, we went back to embedding in wax, cut 5 to 10 micron sections, de-waxed and got excellent FITC-IHC labelling. Although we were reticent to go to wax embedding (19th century technology) it rendered available such enormous areas of antigenic sites that the IHC was excellent plus we could use the sections for in situ work and the fluorescent apoptosis kits.
If you really want to go to dissolvable resins then check Frank's work in: Gubler, F. (1989). Immunoflourescence localization of microtubules in plant root tips embedded in Butyl-Methyl Methacrylate. Cell Biol International Reports, Vol. 13, No. 1, January, 1989.
Eric Hines Microscopy Centre CSIRO Entomology Canberra
Have you tried to do regular gold followed by silver enhancement or gold toning on the semi-thin LR White sections? Then you'd even be using the same secondaries on your tests. I have no ideas on the fluorescence working (or not), but could you please post any responses to the server?
A "RFQ" for products which, to the best of our knowledge are similar to those offered by you ,was placed with us by one of our clients.
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} } Salut to the listmembers,
Where to get freeware Wang Image (as *.zip files ?), on which URL address ? J o z e f Stankovic
} You might try Wang Image (now a standard free imager that comes with } windows 95, or is availiable as freeware I believe) We use it to convert } JEOL tiffs to tiff formats that Lview/NIH can read. It seems to read } many different tiff formats but only saves in one (which I don't know, } but a tiff saved by wang image suddenly becomes readable by all of our } other viewer programs).
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Dear Steve Stokowski, I will try to send you single file (*.spe or *.lab) by e-mail and you`l to change the spectrum image to jpg or gif format, O.K. ? To send you file better by e-mail or FTP ?
Yours sincerely J o z e f Stankovic
} In a message dated 97-10-23 03:19:53 EDT, Kees Jalink wrote: } } { { Does anybody know a windows program that can read IPlab's TIFF format (12 } bit greyscale, stored in 2 bytes, I think) and convert it to any } windows-recognized format? } } } } PaintShopPro, ver. 4.14, from JASC will read 6 different TIFF subformats. } These are: uncompressed, Huffman compresssed, pack bits compressed, LZW } compressed, Fax Group 3 compressed, and Fax Group 4 compressed. None of } these are 12 bit grayscale. They range thru 1, 4, 8, and 24 bit/pixel color } scales. Attach a single file to an e-mail to me and I will try to change } the image to something else, such as a gif or a jpg, your choice. PSP } supports 35 different formats. } } Yours truly, } Steve Stokowski } Stone Products Consultants } Concrete Petrographers } 10 Clark Street, Suite A } Ashland, Massachusetts, 01721 USA } 508-881-6364 } http://members.aol.com/CrushStone/index.htm
Vachik Hacopian wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Charles Duvic of Ladd Research wrote: } } } Acetone and ethanol are viable alternatives to Propylene oxide. Ethanol } } should be used cautiously because it may inhibit epoxy polymerization. } } What is meant here by the word "cautiously"? Is it in reference to the } ethanol not being absolutely dry? } } Vachik Hacopian
Dr. Hacopian, Perhaps " cautiously" was too strong of word. I simply meant that care must be taken to ensure that all ethanol is removed from the speciman before any attempt at polymerization. Some investigaters have have noted that a "small" amount of ethanol in the speciman doesn't seem to affect polymerization of the block. Since "small" is subjective and not clearly defined I think it is best to wash the speciman with a resin mixture until all (or most) ETOH is removed.
To Eriko: unfortunately it is not possible to communicate with you by direct = e-mail (see below). Eriko, you did=B4nt send with your postal adress! Please forward it to me Thank you very much Wolfgang MUSS ------------------------------------------------------------
----- Transcript of session follows ----- mail: Cannot append to /var/mail/terao
----- Message header follows ----- Received: from aesculap.lkasbg.gv.at by fisher.sc.ucl.ac.be = (5.x/SMI-SVR4) id AA15831; Mon, 13 Oct 1997 09:16:53 +0200 Received: (from smap-at-localhost) by aesculap.lkasbg.gv.at (8.6.12/8.6.9) = id IAA27090 for {terao-at-bani.ucl.ac.be} ; Mon, 13 Oct 1997 08:11:18 +0100 Received: from unknown(10.1.101.6) by aesculap.lkasbg.gv.at via smap = (V1.3) id sma027088; Mon Oct 13 08:11:03 1997 Received: from c1pa008.lkasbg.gv.at (c1pa008.lkasbg.gv.at [10.1.42.8]) by hermes.lkasbg.gv.at (8.8.5/8.8.5) with SMTP id LAA06414 for {terao-at-bani.ucl.ac.be} ; Mon, 13 Oct 1997 11:09:34 +0200 Received: by c1pa008.lkasbg.gv.at with Microsoft Mail id {01BCD7B8.54DF35C0-at-c1pa008.lkasbg.gv.at} ; Mon, 13 Oct 1997 09:13:55 = +-200 Message-Id: {01BCD7B8.54DF35C0-at-c1pa008.lkasbg.gv.at}
Hello,
after several unsuccessful attempts to stain PE ultrathin cuts with the Kanig method I would like to ask if somebody has been successful and is willing to share his protocol with me. The specimen is already cut with an cryo-ultramicrotome and the cuts are lying on copper grids.
TIA,
Petra -------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu
We are exploring the use of ambient atomic force microscopy to study the surface of transition metal oxide. We would like to contact research groups who are involved in the simulation of water adsorption on transition metal oxide(v2o5,MoO3) Who are the contacts for this type of work?
} may-be this question doesn't belong here, but I am totally stuck so I } still ask it: } I have recently taken some 120 Mb of digitized pictures with a } CCD-camera equiped microscope using IPlab software on a MacIntosh. Does } anybody know a windows program that can read IPlab's TIFF format (12 bit } greyscale, stored in 2 bytes, I think) and convert it to any } windows-recognized format? These data are important for me. } Thanks in advance. } Kees Jalink } } -- } } } Kees Jalink } The Netherlands Cancer Institute, dept. of Cell Biology H1 } Plesmanlaan 121 1066CX Amsterdam, the Netherlands } 020-5121982 (tel) / 020-5121989 (fax) } kees-at-NKI.NL (email) / 0297-320248 (tel at home)
I'd do it the other way. Usa Graphic Converter, a widely available Mac shareware application, to do the convertion on the Mac. Graphic converter can read about 50 graphic formats and saves to about 40, including a range of PC formats. Not sure if it will read in your particular TIFF files, but if not, you could contact the author - he has added in a number of proprietary formats on request.
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
I use Spurr's resin for general e.m. histology and LR White (London Resin Company Limited) for most other things. They can both be mixed with absolute alcohol and so I can avoid the problems with propylene oxide.
I know that Spurr's is more carcinogenic than other epoxides but at least it isn't as volatile as propylene oxide which also has an annoying habit of melting many types of gloves. But I would be interested to hear anyone else's opinion about which is nastier: Spurr's resin with alcohol or epon with propylene oxide.
Malcolm Haswell E.M. Unit University of Sunderland
Disclaimer - these are my opinions and not necessarily those of my employer.
----------
Norbert
I use Spurr's resin for general e.m. histology and LR White (London Resin Company Limited) for most other things. They can both be mixed with absolute alcohol and so I can avoid the problems with propylene oxide.
I know that Spurr's is more carcinogenic than other epoxides but at least it isn't as volatile as propylene oxide which also has an annoying habit of melting many types of gloves. But I would be interested to hear anyone else's opinion about which is nastier: Spurr's resin with alcohol or epon with propylene oxide.
Malcolm Haswell E.M. Unit University of Sunderland
Disclaimer - these are my opinions and not necessarily those of my employer.
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Peggy Brannigan wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Hello all, } } I'd like to de-embed some LX112 embedded plant leaf tissue so that I could } examine it in the SEM but, never having done this before, I'm looking for } advice, tips, references, protocols etc. Also, is it possible to de-embed } thin sections already stained (uranyl acetate, lead citrate) and examined } in the TEM? (Assuming I could get them off the grid....) } } Thanks,
Peggy,
We at LADD sell LX-112 and one procedure we know of that may work for you is prepared as follows:
1. Prepare saturated solution of KOH in absolute ethanol. Let stand overnight. 2. Pour off supernatant fluid. This is the epoxy solvent. 3. Trim block of excess epoxy resin. 4. Let speciman soak in solvent until resin is removed. Time will vary depending on size of block. 5. Wash with several changes of absolute ethanol. 6. Prepare for SEM
NOTE This solution may do damage to your speciman. I suggest trying this procedure on only one or two of your less critical samples.
Good luck,
Dr. Charles Duvic Ladd Research tel 1-800-451-3406 fax 1-802-878-8074
Below is the result of your feedback form. It was submitted by (gusev-at-ipm.sci-nnov.ru) on Friday, October 24, 1997 at 07:05:18 ---------------------------------------------------------------------------
Email: gusev-at-ipm.sci-nnov.ru Name: Gusev S.A.
School: IPM RAS, Nyzhnii Novgorod University
Zip: 603600
Question: We have made periodical 2-D arrays of the magnetic single domain particles. The size of the each particle ~30 nm. We measured their cooperative properties and would like to see the state (the direction and the magnetude of magnetization) of the each particle. Who can help us to realize our dream by means of the electron microscopy?
I just wanted to thank you all for your help. I just replaced the Tonner cartridge in our Lexmark Optra, tidded up the printer, and the first test print out was free from spots, speckles, blemishes! We're greatly happy with our Lexmark once again.
Take home lesson - when printing any sort of images, even a lite quaitity mixed with mostly text, the 7K-pages and 15K-pages are not anywhere in the ball park (we started having problems -at-~2,500 pages and totally unacceptible at 3K pages with our 7k-page cartridge)
Thank you.
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
As I was asked to put a summary on the server so here it is. Thank's again to every body who answered me.
Marc
Just take a small amount of the Soot and put it in a vial and add Acetone (10ml) and then ultrasonicate the solution for 3-4 mins. Then you need to place a few drops on a carbon coated grid, let the solvent evaporate and put the grid in the scope. First use low mag to locate the material on the grid. It is best to use 100-200 kV to image the tubes. KMJ
I sssm to recall that the powder can be suspended in a solvent (benzene or toluene-I don't remember which). A single drop onto a carbon coated copper mesh grid is sufficient.
Hi, I had the same problem, so I can recommend you to make a suspention of this powder in alcohol, then put a drop on a grid with deposited thin film. Contrast will be poor, so you will need to use defocused image.
Marc, There are many reports of TEM on carbon nanotubes. However, you can sonicate the powder containing the nanotubes in ethanol or water, then place a small aliquot on the TEM grid and wick away the excess liquid. It's important to make sure the carbon actually gets suspended in the liquid you are sonicating with. I use a pipette to help mix the suspension quickly.
We examine carbon nanotubes as follows:
1. suspend the specimen in a small volume of either acetone or methanol (trial and error will be needed to determine the exact dilution. so try different dilutions)
2. suspend the sample by swirling (some people sonicate for several minutes) and use a pipette to transfer a droplet onto a holey, filmed (Butvar or Formvar) and carbon coated grid (suggest purchasing from commercial source)
3. allow to air dry and examine in TEM looking for tubes suspended over the holes (best resolution)
The TEM should be set up for hi res, cooled traps over specimen area and a clean vacuum system is needed.
Good luck.
I used to view these regularly for a materials scientist. We simply put a small amount of the carbon sample in a solvent and used a sonicator to disperse it. Ten minutes should do a decent job. We then put drops of the suspension onto holey grids which were sitting on filter paper. When they were dry we put them in the TEM for viewing. Some of the nanotubes would sit on the edges of the holes in the film and could be viewed that way.
The nanotubes we viewed required very high magnification and very stable beam and vacuum conditions. Use liquid nitrogen on the pumps and on your decontaminator. Set up your instrument for high resolution conditions and let the electronics and gun stabilize for a while before viewing. Getting the best resolution was trickier by far than preparing the samples.
We just make a dispersion of the powder ( say in methanol) and put a drop on a holey grid and we let it dry for a few minutes. We examine the nano tubes at about 100K to 300K. If you use regular carbon coated grids ( rather than holey grids) the image of the nano tubes will not be as clear because the carbon background will interfere.
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
} as many of you will be aware, it can be very difficult to accurately } determine diffraction ring diameters in selected area diffraction patterns } recorded from multicrystalline TEM specimens. Especially when rings are } very spotty.
This is true even if "signatures" between patterns are obviously different by eye, at least for large unit cells. Before the days of digitization, we developed a method for making sense of such patterns from mixed-mineral specimens (esp. interplanetary dust particles collected in the earth's stratosphere) which was published in Micron around 1980. Let me know if you would like the reference.
} What I'd like to do is digitize the SAD pattern, locate the exact centre } and intergrate pixel intensities through 180 degrees. This will result in a } one dimensional diffraction pattern with pairs of spots either side of the } undiffracted spot, which should be much simpler to measure.
One of our attempts in those early days involved "photographic" azimuthal averaging: putting film on a turntable whose center aligned with the projected pattern center, and then exposing the film for a couple revolutions. I don't recommend this at all!
} I would like to know if anyone out there has a computer program to do this } type of SAD pattern manipulation. I propose to use Photoshop and a flat } bed scanner on a Macintosh to digitize the SAD patterns which would be } saved as TIFF or some other suitable format. So I suppose the ideal } solution to my problem would be a Photoshop plugin. I also use NIH Image } so a plugin for this program would also be suitable.
The image processing language Semper has verbs for both locating the pattern center very precisely, and for azimuthal averaging while treating images not simply as bytes but as (real or complex) floating point numbers. If you get access to a Semper interpreter (my contact address for the company keeps getting lost), I'll be happy to share our macros with you.
} I would really appreciate any help with a plugin, stand alone program (Mac } or PC) or comments on how I should go about writing my own solution. I } look forward to your replies,
We've written a Visual Basic program for doing this, but I'm not sure it's sufficiently error-trapped for outside distribution. If Java programs could read local system data files on the request of a local user (can they?), I could probably whip together a program that everyone could use in a couple of hours.
In a message dated 97-10-24 02:38:53 EDT, you write:
{ { I will try to send you single file (*.spe or *.lab) by e-mail and you`l to change the spectrum image to jpg or gif format, O.K. ? To send you file better by e-mail or FTP ? } }
Dr. J o z e f Stankovic:
I do not know what to do with a .spe or .lab format. Sorry. Maybe somebody on the Listserver can help.
I have been a happy user of this system for about 5 years(just upgraded to the WIN 95 version). The answer to all of your question but one is yes it can do that.. I don't know about the vidio conversion though.. But than again I use Adobe for that. On Tue, 21 Oct 1997 kszaruba-at-MMM.COM wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Listers, } } I've heard a few mentions of the BioQuant system in the past. } Now I am considering it myself, and would like to hear a little } more detail from users. } } What do you like most/least about the system? How easy is it to } handle intensity comparisons of stained specimens (BF and Fluor)? } What camera option did you choose and how well does it work with } a variety of specimen types (macro copy stand, micro BF, fluor, } moving subject, etc)?? Does it really do excellent gel } densitometry? How about colony counting? 3D measurements? Can } it turn VCR tapes (short) into digital movies? } } I'd appreciate any comments! } Thanks as always, } Karen } } P.S. Regarding the Message Subject requirements, what about } general subjects like digital imaging, printers, etc?? LM vs. } TEM, etc. doesn't really fit. } } -- } Karen Zaruba } kszaruba-at-mmm.com } 3M Company, 3M Center Bldg. 270-1S-01 } St. Paul, MN 55144 } "The opinions stated above are my own, not necessarily 3M's" }
The file is about 2.0 MB and will expand to 2.2 MB after decompression.
At 08:10 AM 10/24/97 +0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The November meeting of the Mic. Soc. of Northeast Ohio will welcome S. Frank Platek, of the FDA - Forensic Chemistry Center (Cincinnati, OH) as our featured speaker on Thursday, November 6, 1997. Details are as follows:
Title: "Forensic Microscopy Related to Product Tampering and Counterfeiting" The evening's presentation will 'focus' on the Forensic Chemistry Center's multiple microscopy investigation of product tampering and counterfeiting of foods, beverages, pharmaceuticals, and medical devices. Specifically, the use of stereoscopic, polarizing and comparison light microscopy, as well as scanning electron microscopy, and energy dispersive x-ray analysis in State and Federal cases. Techniques for case analysis and some uncommon used of image analysis; backscattered electron imaging and x-ray mapping will be featured along with case histories.
Place: Case Western Reserve University (Medical School) Andrews Conference Center, room 6306
Time: 5:45 Reception/Social Hour 6:30 Presentation 8:00 Dinner at Club Isabella Menu includes: Soup or salad; choice of Grilled Salmon and Pasta Primavera; OR Herb Roasted Chicken Breast; OR Pasta Sauteed with Fresh Spinach. Dessert Tray and Coffee for $20.00 (make checks payable to MSNO).
Meal choices and reservations must be made by noon, Monday November 3, 1997 to Valerie Woodward, MSNO Secretary (216)447-5408 (voice) or woodward-at-brk.bfg.com (E-mail).
does anyone know where I can find a source for the laser that fit on the Gatan Duo mill, and are connected with the auto-terminator with a bnc connector? Any hints are welcome: maker, vendor, etc.
I would be grateful if you would bring the position below to the attention of potential candidates.
Many thanks,
Steve Pennycook
{fontfamily} {param} Times {/param} {bigger} POSTDOCTORAL POSITION IN MATERIALS PHYSICS
OAK RIDGE NATIONAL LABORATORY, SOLID STATE DIVISION
A postdoctoral position is available in the Electron Microscopy Group led by Dr. Stephen J. Pennycook at ORNL, in collaboration with Prof. Elizabeth Dickey of the University of Kentucky. Although the position will be joint between ORNL and the University of Kentucky, the post-doctoral researcher will be on permanent assignment at ORNL. The research programs to be pursued in this position include: (1) Structure and Chemistry of Novel Nanotube Materials and (2) Interface Structure and Bonding in High-Temperature Oxide Composites. In the nanotube project, materials with novel physical properties are being developed at the University of Kentucky through selectively doping and functionalizing single wall nanotube (SWNT) bundles. Critical to the project is a fundamental understanding of the dopant distribution in the SWNT bundles and the subsequent effect on structure and electronic properties. In the oxide composite program, we seek to understand the atomic structure and chemistry of interfaces in these high-temperature structural materials and to develop correlations between the atomic structure and mechanical behavior of the interfaces. Both projects will require complementary atomic-scale electron imaging and electron energy loss spectroscopy (EELS), so the successful candidate should have practical experience in both techniques.
The Solid State Division has some of the world's finest facilities for atomic scale imaging of materials: a VG Microscopes HB603 300 kV scanning transmission electron microscope with a 1.26=C5 probe size provides direct, Z-contrast imaging capabilities for interfaces in materials. A VG Microscopes HB501UX 100 kV microscope with a high sensitivity parallel EELS capability provides atomic resolution spectroscopy. The Electron Microscopy Group has two Silicon Graphics=20 workstations with the Molecular Simulations' Cerius 2 package incorporating the CASTEP pseudopotential code. The Division has a number of additional workstations, and access to extensive parallel computing capabilities, including the Intel Paragon XP/S 35 and XP/S 150 with 512 and 2048 processors respectively.
I noticed that you mentioned SPE and LAB files which I recognize as being from a Link system. We also have one. Those files will not convert readily since they are a proprietary format. But we have a couple of approaches that we use to embed Link spectra in a document.
First, the file may be pasted into a document with the following procedure. - Prepare the spectrum for printing and begin the print dialog. The print preview window will display. - From the EDIT pull down menu select COPY. - Switch to the application you wish to paste into. - From the EDIT pulldown menu select COPY SPECIAL and paste the spectrum in as a picture (not as text which copies in only the header information). - The spectrum will appear as a resizable graphic line drawing with spectrum and axis labels.
Second, a snapshot of the spectrum window may be processed. - Prepare the spectrum for printing, but stop short of starting the printing process. - Activate the zoomed spectrum or main spectrum window (your choice), and press Alt-Prtscrn. This will copy a bitmap of the active window just as it appears to the clipboard. - Switch to the application of choice such as Word or MS-Imager (I like Imager). Paste the clipboard contents, or use the File New From-Clipboard function in Imager. The new image may be cropped to eliminate the unwanted features.
This second procedure works with any Windows (3.x or 95) application window. Hope this help.
At 10:29 AM 10/25/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Some of the user's here are embedding plant tissue into LR White after doing an acetone dehydration (I know it's not the recommended dehydrating solution, so don't nag me on that = ) ). My question is: Is there a stain that is soluble in acetone that we could use in our last 100% acetone step that would stain the plant tissue so it is visible for embedding and sectioning. When we use ethanol for dehydrating we use a 0.1% safranin O and it turns the plants a lovely pink without affecting the immunostaining capacity. Is there a stain out there that would work in acetone & do the same thing?
If anybody out there knows of one....
I'm dyeing to hear about it.
Freeing the radicals (acetone, that is) in Berkeley,
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
Dear All: I am forwarding this request for a donation of an oil immersion microscope from my son, Rene Jordan, who is a volunteer working for the Peace Corps in the Philippines. The microscope is needed for the detection of Malaria. I hope this is not un unreasonable solicitation, I am not familiar at all with the value of such a microscope. Abount my son: He graduated with a degree in environmental science from UCI, Calif. and joined the Peace Corps in April. After his 3 months training in Manila he was sent to one of the 7000 islands for coastal resource management, mainly sea turtles and coral reefs. The small village he is in has no phone (how nice), no paved roads and every night at 9 the generator is turned off except Tuesdays were it runs till 10 for the church service. To get his mail he travels once a month for 6 hours to the next bigger city. If you have one of those microscopes standing around please let me know or you can contact him direct: Rene Jordan - PCV PCSDS 3/F Capitol Complex Puerto Princesa City, 5300 Palawan, Philippines Thank you very much, Peter Jordan
Hello, I use eosin B (Kodak cat# 117 7930) in 100% ethanol to stain the tissue before embedding in LRWhite for immunostaining. So far I have never had any problems with the immo-detection of various antigens . Eosin B can be dissolved in acetone as well.. I use at 1%..hope it helps Neelima Shah. Morphology core, UOP
} Boarders, } } =A0=A0=A0=A0=A0=A0=A0 Some of the user's here are embedding plant tissue= into LR White } after doing an acetone dehydration (I know it's not the recommended } dehydrating solution, so don't nag me on that =3D )=A0 ).=A0 My question= is:=A0 Is } there a stain that is soluble in acetone that we could use in our last 100% } acetone step that would stain the plant tissue so it is visible for } embedding and sectioning.=A0 When we use ethanol for dehydrating we use a } 0.1% safranin O and it turns the plants a lovely pink without affecting the } immunostaining capacity.=A0 Is there a stain out there that would work in } acetone & do the same thing? } } =A0=A0=A0=A0=A0=A0=A0 If anybody out there knows of one.... } } =A0=A0=A0=A0=A0=A0=A0 I'm dyeing to hear about it. } } } Freeing the radicals (acetone, that is) in Berkeley, } } } Paula=A0=A0 =3D ) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu=20
} Thanks a lot to all who gave me some nice tricks to observe carbon } nanotubes. It helped me a lot and due to your nice indications I've got } correct images at the first shot.
} Marc ---------------------------------------------response------------------------- ----------------- Marc:
If you have time and the inclination I would be interested to see a brief description of the technique you finally used.
I have a question which has undoubtedly been answered before...! We are going to buy a framegrabber and a camera. We use a Power Mac 8200 and want to replace the old greyscale framegrabber with a color one. The camera is used in order to capture images of plants under development, but also for capturing images of cells under a microscope. Should we go for digital cameras? What is the difference between an AV card and a framegrabber?
Can somebody help us in order to take the right decision?
Thanks for all the suggestions and help for tranferring IPlabs 12-bit grey-scale TIFF format (MacIntosh) to windows. I tested some 20 different commercial and freeware viewers, converters, etc. In my hands, the only one that worked without any problems is Thumbsplus (ver3.10) by Philip Crews (Cerious software; available as shareware). Just copy the files to a PC directory, rename them to *.tif, and Thumbsplus will read them, correctly noting that it is 16-bit greyscale (last 4 bits not used) and motorola byte order. Nice piece of software for 65$.
((Salut to the listmembers,
may-be this question doesn't belong here, but I am totally stuck so I still ask it: I have recently taken some 120 Mb of digitized pictures with a CCD-camera equiped microscope using IPlab software on a MacIntosh. Does anybody know a windows program that can read IPlab's TIFF format (12 bit
greyscale, stored in 2 bytes, I think) and convert it to any windows-recognized format? These data are important for me.))
Kees Jalink The Netherlands Cancer Institute, dept. of Cell Biology H1 Plesmanlaan 121 1066CX Amsterdam, the Netherlands 020-5121982 (tel) / 020-5121989 (fax) kees-at-NKI.NL (email) / 0297-320248 (tel at home)
Inside IPLab spectrum you can change the data types from 16 bit to 8 bit images. It is under the math menu, change data types. Once you get it to 8 bit, the images should be readable in any Mac or PC program that can open tiff files.
One other note, I use MacLinkPlus to transfer tiff images between PC and Mac and visa versa.
Hope this helps, Lou Ross
Senior Electron Microscope Specialist 101 Geological Sciences Building University of Missouri Columbia, MO 65211 (573) 882-4777, 882=5458 (fax) geosclmr-at-showme.missouri.edu www.missouri.edu/~geosclmr/ebaf.html
Hello Microscopy world! We have been asked to attempt to microscopically evaluate collagen films. The sheets are approximately 50% hydrated and are approximately 500 um thin. The client wishes to see if there are differences evident in the cross-linking that occurs as the film sheets are produced under various conditions (varying concentrations of constituents). It is necessary to evaluate them as close to their natural state as possible. Any suggestions? Thanks for accepting this challenge! Tracey Pepper Bessey Microscopy Facility Iowa State University
We would like to find a computer program to determine the misorientation that exists across a grain boundary from electron diffraction patterns. Specifically, if the electron diffraction patterns of adjacent grains were obtained, does a program exist which, by using the two ED patterns, would describe the grain boundary geometry by determining the grain boundary plane and rotation axis that exists between the two grains? We are interested in performing this analysis on non-cubic materials which possess a hexagonal or rhombohedral structure.
Thanks in advance.
Owen
============================= 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
Hello Microscopy World! We have been asked to microscopically evaluate thin sheets of collagen films. They are approximately 50% hydrated and about 500 um thin. Our client wishes to see the differences in the cross-linking of the collagen produced under various conditions. It is necessary to evaluate them under as natural conditions as possible. Any suggestions? Thanks! Tracey Pepper Bessey Microscopy Facility Iowa State University
Software to calculate misorientation from Kikuchi patterns is quite common, however typically homemade (I used to have one written in FLEXTRAN, running on Tracor Northern analyzers, but that is history). I enclose below a list of those microscopist which I know having written such programs recently. Stefan and Robert both have software which could already have build in your application to determine grain boundary indices (you need to determine the direction of the grain boundary with respect to the orientation of the unit cell of the grain of interest as well the inclination of the grain boundary in the foil, typically obtained by a tilt analysis). Stefan also has a routine build in which is very much useful for Burgers vector analysis. All programs are suited for on-line analysis and are PC based.
Stefan Zaefferer: stefan-at-isma.u-psud.fr Robert Schwarzer: robert.schwarzer-at-tu-clausthal.de Paul Baggethun: baggeth-at-sms.emse.fr
Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
} ---------- } From: Owen P. Mills[SMTP:opmills-at-mtu.edu] } Sent: Monday, October 27, 1997 10:19 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM - Electron diffraction software } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am interested in getting some feed back on Digital Capture packages from various vendors. I specificaly want to place system on a JEOL 35C SEM. I aminterested in knowing how well the various vendors perform specificaly SemiCaps. If anyone has any information such as pros and cons to any one particular system I would realy like to hear it. Please send emails to rgarcia-at-cs.wright.edu. Thanks.
Roberto Garcia Electron Microscopy Facility Manager Wright State University
Thank you to those who responded so far to my question on reduction of OsO4 and RuO4 to less hazardous states using corn oil.
So far no one has responded with experience with RuO4 and corn oil - any others lurking out there want to add 2 cents? I did get a very detailed reply from Wolfgang Muss on using ethanol to inactivate OsO4. Also I have never heard of the "reprocessing hazardous waste" issue that Steven Barlow brought up. Sounds like a wonderfully forward-thinking, environmentally responsible policy to me. [Must have been designed by the same folks that allow liability issues re:children to make it too costly for our neighborhood to have a park.] Does this reprocessing issue apply outside San Diego?
Answers to two questions: The corn oil procedure for OsO4 appears in a "Technical Data Sheet" from Electron Microscopy Sciences, with the following references given: (I haven't read them, just the data sheet)
Cooper, K. Neutralization of Osmium Tetroxide in case of accidental spillage and for disposal. Bulletin of The Microscopical Society of Canada. 1988. 8:24-28. (Also I think I saw similar article in Don Grimes' free publication, Microscopy Today).
Lunn, G.;Sansone, E.B. Osmium Tetroxide. Destruction of Hazardous Chemicals in the Laboratory; Program Resources, Inc. Frederick, MD; p211-213.
The other question was about the hazards of Sodium Bisulfite. I am just going by an MSDS I have for CAS # 7631-90-5 from Sigma-Aldrich. This states that sodium bisulfite is Toxic, Harmful by Inhalation and Contact, causes Severe Irritation, and is a Possible Sensitizer. Taking into account the fact that even bread pudding would probably show up as harmful by inhalation on an MSDS, still the designations of Toxic, Sensitizer and "Severe" Irritant are worth noting.
Still appreciate any further comments! Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
I am a post doc working in nanotechnology at Washington University in St. Louis. I have a general question regarding calibration standards for microscopes. We are researching possible uses of a collections of small monodispersed particles and it occurred to us that there might be a use for them in calibrating microscope systems.
Specifically, I am interested in scanning probe microscopy (AFM,STM,MFM), electron microscopy (SEM,TEM) and light microscopy (both optical and near-field optical scanning microscopy).
What sort of small particles are used (if any) to calibrate these instruments? We are interested in the composition and size.
If collections of small particles are not generally used, would there be an interest in the microscopy community for a collection of small particles (~100 nm to 10 microns) to be used as a calibration or any other sort of aid.
I would be very interested in your reply,
Thanks very much
-Stephen Irons
********************** Washington University Department of Physics CB1105, 1 Brookings Drive St. Louis, MO 63130
Iam an international student accepted in the biology department at New York University. I have received my Master's degree in microbiology. I would like to know more about your scholarship for graduates. Thank you
I am experiencing an excessive charging in my SEM. The tested samples are conductive, the specimen mount, holder and SEM stage are all well grounded, but charging persists, precluding succesful imaging. Secondary and backscattered electron images are equally affected. These samples do not exhibit charging when analyzed under the same conditions (AV) in another SEM. Many different samples were tested in these two SEMs and the results are consistent.
It appears that some instrumental factor is involved. I would appreciate the input on the subject. Thank you very much in advance.
Chris Terlecki Applied Analytical Sciences ph: 714-434-6894 email: aas-at-pacbell.net
Dear Karen, } } So far no one has responded with experience with RuO4 and corn oil - any } others lurking out there want to add 2 cents?
Just a guess, but since Ru is also a group VIII metal, and since Ruthenium red is used to stain the lipid components of membranes, an un- saturated oil should combine with Ru in much the same way as with Os.
} Taking } into account the fact that even bread pudding would probably show up } as harmful by inhalation on an MSDS, ...
As the recent discussion of the MSDS of H2O would indicate. Yours, Bill Tivol
I've been asked to find out what software people are using to simulate electron diffraction patterns, specifically selected area diffraction patterns. We would like to know the name of each software package, supplier, approximate price, computer platform required, etc. Your comments on accuracy and ease of use would be most welcome.
We would also like to hear from people who have had bad experiences with commercial software. But please contact me directly, not via the listserver. We don't want to upset anyone unnecessarily.
Thankyou,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
I am trying to view air samples captured on a teflon filter with a Cambridge SEM. Is there a standard procedure that I should follow?
Joe Wise Director, W. M. Keck Math/Science Institute Crossroads School for Arts and Sciences 1714 Twenty-First Street Santa Monica, CA 90404 (310) 829-7391
Stephen Irons wrote: ============================================== I have a general question regarding calibration standards for microscopes. We are researching possible uses of a collections of small monodispersed particles and it occurred to us that there might be a use for them in calibrating microscope systems. ============================================== You might want to consider the Mag*I*Cal TEM Calibration sample, and unlike polymeric calibrated microspheres, there is nothing that is "changed" by the electron beam. Full details about the Mag*I*Cal sample can be found on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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} Date: Mon, 27 Oct 1997 21:08:35 -0800 } To: jwise-at-kmsi.org (Joe Wise) } From: Mary Mager {mager-at-unixg.ubc.ca} } Subject: Re: air samples } } Dear Joe, } I have done exactly that on 60 different air filtered samples. My only comment would be to use Nucleopore-type filters (polycarbonate) if possible, because they are much smoother and show the particles up better than Teflon. The Teflon filters are quite rough and do not carbon coat well, so particles are harder to see and charging is a problem. Just cut out a small piece, attach to a stub, carbon coat and observe. } } } I am trying to view air samples captured on a teflon filter with a Cambridge } } SEM. Is there a standard procedure that I should follow? } } } } Joe Wise } } Director, W. M. Keck Math/Science Institute } } Crossroads School for Arts and Sciences } } 1714 Twenty-First Street } } Santa Monica, CA 90404 } } (310) 829-7391 } } } } e-mail: jwise-at-kmsi.org } } http://www.kmsi.org } } } Regards, } Mary }
Chris - and other microscopists - If indeed the specimen and stage are grounded, you should check that you are not using excessive beam current. Also, a large condenser spot or a huge final aperture can be the problem. Since BS too is affected grounding of the scintillator cannot be the problem. If a bare specimen mount does not charge, then the specimen itself is likely the problem. The other SEM may use just a little less beam current and a partially conducting specimen would then not charge up. If the problem is specimen related, one of the more common problems would be the umbrella effect; where the specimen is well coated on the upper surfaces, but these surfaces prevent a continuos good coating on the under sides. Visualise a stack of cannon balls or a mushroom: Coating from above would not be effective. Such specimens can be sputter coated by laying the pin type mount on the side, coating and then turning the specimen on the opposite side of the pine and giving it a second coating. At least this is a more interesting problem then fiddling with digital files. Oh, did you use that address "experts" deliberately? I understand the word means 'squirt under pressure'. 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, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au ----------
} Dear SEM experts, } } I am experiencing an excessive charging in my SEM. The tested samples } are conductive, the specimen mount, holder and SEM stage are all well } grounded, but charging persists, precluding succesful imaging. Secondary } and backscattered electron images are equally affected. These samples } do not exhibit charging when analyzed under the same conditions (AV) in } another SEM. Many different samples were tested in these two SEMs and } the results are consistent. } } It appears that some instrumental factor is involved. I would } appreciate the input on the subject. Thank you very much in advance. } } } Chris Terlecki } Applied Analytical Sciences } ph: 714-434-6894 } email: aas-at-pacbell.net
} I am experiencing an excessive charging in my SEM. The tested samples } are conductive, the specimen mount, holder and SEM stage are all well } grounded, but charging persists, precluding succesful imaging. Secondary } and backscattered electron images are equally affected. These samples } do not exhibit charging when analyzed under the same conditions (AV) in } another SEM. Many different samples were tested in these two SEMs and } the results are consistent. } } It appears that some instrumental factor is involved. I would } appreciate the input on the subject. Thank you very much in advance. } } } Chris Terlecki } Applied Analytical Sciences } ph: 714-434-6894 } email: aas-at-pacbell.net
My first reaction is that it isn't charging if you see the effect is BSE images. You first need to really determine if it is the specimen or SEM. Same effect with entirely different specimens? If you move the same specimen between the different SEMs, is it always only in the one SEM? Use scan rotation - does the effect stay with the specimen or move with the scan direction. I guess I'd suspect some fault in the scan generation if it is really to do with the SEM - electrical fault in scan generation, or fault in coils, or connection to coils?
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
We are interested in cryosections of plant material especially for elemental analysis with EDX. I would like to get informations about the necessary equipment (cryostat or ultracryomicrotomes) and experiences. Thanks in advance
Heike Buecking Dr. Heike Buecking University of Bremen UFT Plant Physiology and Plant Anatomy Leobener Str. D 28359 Bremen Germany TEL: +49-421-218-2954 or TEL: +49-421-218-7283 FAX: +49-421-218-3737 e-mail: heibueck-at-uft.uni-bremen.de FAX: +49-421-218-3737 e-mail: heibueck-at-uft.uni-bremen.de
I presume that for some reason you cannot coat the sample even though they are conductive. A wiff ie 5nm AuPd on the surface should overcome the charging. Also, turn down the wick on your microscope ie low kV, low beam current.
Patrick Echlin Cambridge
On Mon, 27 Oct 1997, Janusz Chris Terlecki wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear SEM experts, } } I am experiencing an excessive charging in my SEM. The tested samples } are conductive, the specimen mount, holder and SEM stage are all well } grounded, but charging persists, precluding succesful imaging. Secondary } and backscattered electron images are equally affected. These samples } do not exhibit charging when analyzed under the same conditions (AV) in } another SEM. Many different samples were tested in these two SEMs and } the results are consistent. } } It appears that some instrumental factor is involved. I would } appreciate the input on the subject. Thank you very much in advance. } } } Chris Terlecki } Applied Analytical Sciences } ph: 714-434-6894 } email: aas-at-pacbell.net }
It's not clear from your message what you are looking at. Are you really looking at air in which case use a helium cold stage and condense the stuff to a solid. If you are looking at stuff suspended in air see Chap 11 in "SEM & XRMA' Goldstein et al Plenum 1992.
Good luck.
Patrick Echlin Cambridge UKOn Mon, 27 Oct 1997, Joe Wise wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am trying to view air samples captured on a teflon filter with a Cambridge } SEM. Is there a standard procedure that I should follow? } } Joe Wise } Director, W. M. Keck Math/Science Institute } Crossroads School for Arts and Sciences } 1714 Twenty-First Street } Santa Monica, CA 90404 } (310) 829-7391 } } e-mail: jwise-at-kmsi.org } http://www.kmsi.org } }
I have a question regarding what polymer resin to use for embedding a sample for (TEM) observation. The sample is a powder, which is likely to be moisture and CO2 sensitive. I am thus concerned that a condensation polymer would induce changes in the sample. We have been grinding the sample to produce a suspension for TEM specimen preparation. However, we would like to use ion milling, as this will be more likely to preserve the microstructure.
Does anyone know of a good embedding polymer for a moisture and CO2 sensitive sample, which is also high-vacuum compatible?
Wharton
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Department of Materials Science and Engineering Northwestern University 2225 North Campus Drive Evanston, IL 60208-3108 tel: (847) 491-7809 fax: (847) 491-7820 email: sinkler-at-apollo.numis.nwu.edu
Look for a grounding wire that may be disconnected. If your instrument capable of measuring the absrobed current, see if that is operating normally. A disconnected wire to ground from the stage will cause any sample to charge. Check continuity between the stage and the microscope body. Move the stage around while checking; you may have an intermitten contact.
-Scott Walck ----------
I would like to observe specimens on TEM substrates before and after ashing.
Specifically what I am trying to do is place a specimen on a suitable TEM substrate. Image the specimen and then image the specimen after ashing the specimen on the TEM substrate.
So far I have not been successful. I have tried germanium substrates on nickel and copper grids substrates. I also have tried gold on gold. The substrates decompose in all cases. I am surprised that the gold on gold decomposed.
I would be appreciated of any information related to this effort.
Regards,
Larry Murphy Group Leader, Analytical Section Cabot Corporation
I went through the MSA pages on the internet but could not get any information regarding the scholarships for graduate students. Please give a phone number or address or E-mail address to whom I can contact. thank you
I would like to put forward the presence of cellulose (or other major algal/Phaeophyceae/ cell wall components) in biological samples using fluorescence microscopy. I have heard about the existence of specific fluorochromes binding to cellulose. Could anyone give me more information about this.
If none of the other suggestions cure the problem, check for column contamination. A small bit of insulating material at the final lens or in the column can deflect (often erratically) the incident beam.
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
I have always used Calcofluor white M2R to visualize cell walls. I'm pretty sure it is specific for cellulose (I did find it's specificity described in writing, problem is that I was the author) but you might want to check.
================= C. John Runions Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
} I have a question regarding what polymer resin to use for embedding a } sample for (TEM) observation. The sample is a powder, which is likely to } be moisture and CO2 sensitive. I am thus concerned that a condensation } polymer would induce changes in the sample. We have been grinding the } sample to produce a suspension for TEM specimen preparation. However, we } would like to use ion milling, as this will be more likely to preserve } the microstructure. } } Does anyone know of a good embedding polymer for a moisture and CO2 } sensitive sample, which is also high-vacuum compatible?
} Wharton -
You don't say what TYPE of powder you're dealing with, but I'm wondering why you "would like to use ion milling, as this will be more likely to preserve the microstructure". Have you considered ultramicrotomy? There are standard embedding protocols, but selection of the right one depends on the sample.
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/
I believe that you better use ultramicrotomy to prepare powder samples for TEM. Particularly your sample is sentitive to drying and CO2. First, you embed your powder into resin which leads to completely enclose your powders from the environment. Then use Ultramcrotomy to cut thin-sections. I believe you will finds some places to prepare this type of TEM samples.
Larry Murphy wrote: ================================================== I would like to observe specimens on TEM substrates before and after ashing.
Specifically what I am trying to do is place a specimen on a suitable TEM substrate. Image the specimen and then image the specimen after ashing the specimen on the TEM substrate.
So far I have not been successful. I have tried germanium substrates on nickel and copper grids substrates. I also have tried gold on gold. The substrates decompose in all cases. I am surprised that the gold on gold decomposed. ===================================================== A technique has been developed that takes advantage of a SiO2 filmed grid followed by oxygen plasma etching. We at SPI, have used this method since the mid-1970's (credit for discovering it, so far as I know, is Dr. John T. Stasny who was working for SPI at the time) and it is useful for both materials science as well as life science thin sectioned samples:
1] Thin section the sample using normal diamond knife techniques but pick up the sections on SiO2 coated grids. 2] Do your pre-ashing TEM examination; the SiO2 film structure will not appear greatly different from what you are used to seeing with carbon. 3] Then place the grid in an oxygen plasma etcher and using pure oxygen, back fill to as low as possible a vacuum, and then "etch" for 10-20 seconds which should be enough for removal of organics in a thin section. The reason for the maximum vacuum for the backfill so to have the highest possible partial pressure of oxygen, thereby resulting in the fastest possible etching time. The oxygen plasma of course will not etch the SiO2 film. Of course, one does need a good stable SiO2 film. 4] You should be able to put the grid back into the TEM and photograph the same identical area after etching.
Tell me how it comes out. The plasma etcher (isotropic) being used should be operated at not more than 100 watts, other wise too much heat is generated. If you have a leaky system, letting in nitrogen, not too much has to be let in to drastically reduce the etching rate. The SiO2 film is made by evaporation of SiO at 10 -5 torr or better. When you buy them commercially, they are much cheaper than the alternatives you said you (unsuccessfullly) tried previously.
Disclaimer: SPI Supplies has been producing stable SiO2 films for this particular application and also offer the SiO for anyone wanting to make their own support films. We also produce the Plasma Prep II Plasma Etcher so we would have an obvious interest in seeing more of this sort of work being done. See our website, given below, for more information.
Chuck
PS: In a few weeks we will put up on our website a nice life science example of this kind of etching on a SiO2 filmed grid.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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I'm translating a microscope manual from Japanese and stuck with description that goes like, "The image position (of the eyepiece?) is set at 10 mm from the abutting joint."
Can anyone decipher my lousy translation and tell me what it means and if my diction is right?
Thanks in advance.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
Dear all, we are trying to determine the orientation of rutile crystals (20 microns) in a jarosite matrix. When we tried beamrocking, the rutile disappeared, probably because the jarosite acted as a flux. Has anybody any experience of this problem or any ideas as to how we might overcome it?
TIA Eric
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE +44 1224 272934 e.lachowski-at-abdn.ac.uk
} Greetings! } } I have a question regarding what polymer resin to use for embedding a } sample for (TEM) observation. The sample is a powder, which is likely = to } be moisture and CO2 sensitive. I am thus concerned that a condensation } polymer would induce changes in the sample. We have been grinding the } sample to produce a suspension for TEM specimen preparation. However, = we } would like to use ion milling, as this will be more likely to preserve } the microstructure. } } Does anyone know of a good embedding polymer for a moisture and CO2 } sensitive sample, which is also high-vacuum compatible? } } Wharton Wharton,
It=D5s an interesting question you have. All of the conventional epoxy r= esins (O.K. At least the ones that I=D5ve used) are anhydride cross-linked . T= he polyester polymerization that I recall involves the formation of a ester = bridge between the two carboxyl groups of the anhydride and an epoxy group at ea= ch site. Thus, the reaction (been a long time since organic chem, mind you)= is a condensation rather than an addition (e.g. polyethylene ). Some water is exchanged at the business end of the anhydride during these reactions, bu= t there shouldn=D5t be much generated overall. (Just how long is beginning = to show, I think). Anyway, a low viscosity resin like VCD (Spurr=D5s resin) might = be the ticket. If you limit the amount of flexibilizer (DER 736, Quetol, etc.) = you can get these resins quite hard and the resulting sections, especially if= given a light carbon coating are relatively beam-stable.
Other things will cross-link epoxies, polyamines, for example but I=D5m y= et less aware of their polymerization reaction than for what the biologists around here use. The other commonly used class of resins are = the acrylics. Most of the ones in common use are proprietary formulations. = The old, conventional acrylics aren=D5t very beam stable.
Having embedded it, do you then plan ion milling? I=D5ve not done this w= ith an epoxy embedded powder sample but, epoxy glue lines in sandwitched (mostly= Si) tend to erode quickly, at my hands. Might be grim for a powder. If you t= ry the ultramicrotome, you=D5ll need something other than water to float your films on or do it dry (sounds li= ke you=D5d need a really hard resin for that to work at RT; we can=D5t do an= hydrous cryo sections in our lab, here in the cellar ;-). You might be able to g= et thin enough or nearly thin enough with a tripod polisher, but you=D5ll ne= ed to work out an anhydrous system for, at least, the final colloidal silica st= ep.
I think I=D5d try to work out a tripod system for this, $0.02. Good luck.
Cheers, John Heckman TEM Supervisor Center for Electron Optics Michigan State University
A number of people have asked for more details as to my previous query.
First some brief background. Cabot has developed a new reinforcing filler for elastomers, Carbon-Silica Dual Phase Fillers. Naturally we are working on characterizing several aspects of these new fillers.
I would like to assess the morphology of the silica phase by removing the carbon phase. Ashing the sample in bulk and viewing by TEM provides some useful information. I would like a higher level of information regarding the silica phase.
So what I have tried to do is image the dual-phase aggregates on a TEM substrate, then place the TEM substrate into a muffle furnace for 2 hours at 550C in air. The intention is to then view the ashed aggregates (now only silica). Unfortunately the substrates I have tried (germanium and gold) have decomposed. By decompose I mean the substrate is no longer on the grid or severely folded onto the grid bars. I am very surprised that this also happens with a gold substrate on a gold 400 mesh grid!!
The carbon in the dual phase filler begins to decompose about 500C. The dimension of the aggregates are in the range of 100nm give or take.
Regards,
Larry Murphy Group Leader, Analytical Section Cabot Corporation
I'm looking for the source of the quote: "There are lies, damn lies, and statistics."
No, this has nothing to do with interpretation of EDS spectra despite what the cynics may say!
Thanks in advance
Harry ------------------------------------------------ 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}
} I have a question regarding what polymer resin to use for embedding a } sample for (TEM) observation. The sample is a powder, which is likely to } be moisture and CO2 sensitive. I am thus concerned that a condensation } polymer would induce changes in the sample. We have been grinding the } sample to produce a suspension for TEM specimen preparation. However, we } would like to use ion milling, as this will be more likely to preserve } the microstructure. } } Does anyone know of a good embedding polymer for a moisture and CO2 } sensitive sample, which is also high-vacuum compatible? } Is it possible to examine the powder without embedding? I have looked at sediment particles which have been suspended in water and placed on a grid. These have been examined after the water has evaporated. Since water is not good for your particles, another liquid would have to be used, but the procedure should work equally well. It is also very quick to try. Yours, Bill Tivol
We are using our new Ted Pella 3450 microwave for processing potato tissue and have some questions: 1) Do the fumes created by polymerization of LR White stay in the equipment and so might hinder future use for immunolabeling? 2) What would be the best grids for Protein A-gold labeling (on LR White)? 3) What is the best temp. limit for alcohol dehydrations? Thanking you in advance for your help... Bonnie Compas
I am posting the following query for a client of mine. Please respond to MICROSCOPY as usual, or to me privately off-line. Thanks.
Gib Ahlstrand
----------------------------------------------------------------------------- I have some encapsulated flavor particles primarily coated with modified starch and sucrose and coated secondarily with fat. To look at how the flavor particles are coated with fat, confocal microscopy is used. I have stained fat with nile blue. I wonder what is a good stain for modified starch. Any other staining tips you can give for fat & starch using confocal microscopy will be appreciated.
I have been having particular difficulty with a batch of fish eggs I have tried to prepare for SEM. They were fixed in glut. followed by osmium and then dehydrated to 100% EtOH. I have then dried them from liquid CO2 in a CPD. A few eggs have survived the drying intact, the rest either exploding or collapsing.
I have extended the periods in EtOH, and soaking in the CO2 (with lots of flushing), and have slowly vented the gas to minimize any 'shock'. I have tried taking the eggs from 100% EtOH through a graded series of Freon before drying, but the eggs shrivel up by the time I get to 50% Freon. I also tried drying from Peldri, going in a graded series from 100% EtOH to 100% Peldri and letting them soak in the liquid Peldri overnight. By that stage, they had again collapsed.
I guess freeze-drying might be my next move, but thought I'd ask if others may have suggestions on how to get these specimens through the CPD stage. I have processed fish eggs before, but these seem to be quite robustly encapsulated.
Thank you.
Carolyn J. Emerson email: cemerson-at-plato.ucs.mun.ca
Biology Department Memorial University St. John's, NF A1B 3X9 Tel: (709) 737-7515 Fax: (709) 737-3018
I recall it was Mark Twain in "Life on the Mississippi". It was connected to a discussion as to how fast the mouth of the Mississippi was moving north and how soon it would be in Tennessee instead of Louisiana.
At 09:49 AM 10/29/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
According to my source, The New Dictionary of Thoughts by T. Edwards et al and published in 1959 (okay, I ain't that new either) by Standard Book Company, the quote is actually:
There are three kinds of lies - lies, damnable lies, and statistics. Commander Holloway H. Frost
I pesonally prefer:
Statistics are no substitue for judgement. Henry Clay
Shalom from Jerusalem, Azriel Gorski
On Wed, 29 Oct 1997, Crossman, Harold wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I'm looking for the source of the quote: "There are lies, damn lies, } and statistics." } } No, this has nothing to do with interpretation of EDS spectra despite } what the cynics may say! } } Thanks in advance } } Harry } ------------------------------------------------ } 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} }
I get to much mail. WHY do people not reply directly to inquiry posters'!
______________________________________________________ Anand Patel Harvard School of Public Health 94 Beacon Street #78 Cardiovascular Biology Laboratory Somerville, MA 02143 Building II, Room 127 677 Huntington Avenue tel & fax: (617) 354-5132 Boston, MA 02115 USA
} Hello, Does anyone know how to open an IP Lab 12 bit greyscale image file } captured from a Photometrics Sensys CCD camera, as a Raw file in } Photoshop? All you have to do is to convert the 12 bit image to 8 bit. You can do this with IPLab, go to Math and select "to byte as shown" and save your file. Now you'll be able to open your stuff in Photoshop. Let me know if you have any problem
--Ciprian Have fun and keep the sun on your back and a smile on your face. __________________________________________________________ Ciprian A. Almonte University of Pittsburgh Center for Biologic Imaging Pittsburgh, PA 15261
} } } Hello, Does anyone know how to open an IP Lab 12 bit greyscale image file } captured from a Photometrics Sensys CCD camera, as a Raw file in } Photoshop? } } Bob } Morphology core Lab } U of Washington } Hi Bob, It seems that IPLab must be gaining popularity, as this question has been posed a few times recently. The solution, according to Micheal Mort, Ph.D. (president of Scanalytics, Inc., the makers of IPLab software) is to go to the Math menu (once the image is contrast adjusted), and select the command "to byte as shown" then save as a tiff file. Dr. Mort has been most gracious and prompt with my questions regarding their software, and can be reached via email at mmort-at-iplab.com He also informed me that they have software IPLab/Windows at a discount price (since you already proably have IPLabs/Mac). I have no financial interest in any of the above mention firms, just someone like yourself, learning as I go......
Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
Hi Carolyn, You might try some of the mordant techniques using glutaraldehyde, tannic acid, guanidine HCl and osmium by: Gamliel, Scanning Elec. Microsc. 1985: IV; pp1649-1662, 1985. or Osmium, tannic acid, uranyl acetate as per: Shroeter, etal., J. Elect. Microsc. Techn. v1 pp219-225, 1984.
Judy Murphy published two nice reviews covering non-coating techniques which may also help to strengthen cells against collapse: Scanning Electron Microsc. 1978, vol II, pp 175-194 Scanning Electron Microsc. 1980 , vol I, pp 209-
Klaus Peters also published a paper titled "Improved handling of structural fragile cell-biological specimens by the exchange method." J. of Microscopy v 118, pp 429-441.
I could dig them out and FAX to you if you do not have access to these journals.
good luck
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/ Privilege does not absolve one of ecological responsibility.
I am trying to do the same thing, but on yougert. I am using fast green for protein and nile blue for fat, but what fluorochrome can be used in the 647nm wavelength of the Argon/Krypton laser to identify starch?
list
William R. McManus Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305 1-801-797-1920 billEMac-at-cc.usu.edu
On Wed, 29 Oct 1997 09:45:32 -0800 (PST) BECOMPAS-at-CSUPomona.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are using our new Ted Pella 3450 microwave for processing potato tissue } and have some questions:
Bonnie: you will probably get lots of different responses to these questions. Here are mine, based on at least some experience with the same equipment:
} 1) Do the fumes created by polymerization of LR White stay in the equipment } and so might hinder future use for immunolabeling?
Can you vent your 3450 to the outside? Certainly, this would be recommended. We have noticed that if samples of LR White are not covered during polymerization, the media does seem to sublimate, then recondenses and polymerizes on all surfaces (this is our experience during the polymerization of LR White in a nitrogen rich chamber heated to 60C). Using the microwave, you should consider submersing beem capsules, sealed with parafilm under the cap, in water. This will insure that LR white fumes do not enter the microwave during polymerization. Use the temperture limiting probe to limit the temperture of the water to 70 C and polymerize for 60 minutes, or set to 80C and polymerize for 45 minutes. A 500 ml beaker with recirculated water at 10 to 20 C should also be included in the microwave during polymerization.
} 2) What would be the best grids for Protein A-gold labeling (on LR White)?
We use formvar coated Nickel grids. We prefer slot grids, but use whatever you prefer. You may want to use uncoated nickel grids if you wish to label both sides for a double labeling protocol.
} 3) What is the best temp. limit for alcohol dehydrations?
A progressively lower temperture scheme is best. Start with 30% EtOH at 0C for 10 min, then lower to -10 C for another 10 min. Add 50% at -10C, then lower to -20C. All subsequent steps to 1:3 90% EtOH:LRW should be at -20C. We also leave the samples overnight in 100 % LRW at -20 C, then raise the temp to ambient for another hour or so prior to thermal polymerization. We might be to careful. You could work in the cold room (brrrrr!) to obtain -20C.
Feel free to phone if you would like to trade secrets. (503)- 221-3434.
} Thanking you in advance for your help... } Bonnie Compas
In regard to Lawrence Murphy's request, the utilization of a low energy plasma containing oxygen would work to selectively remove the carbonaceous material without effecting the silica. In this process, disassociated oxygen is created by the plasma. The oxygen radicals combine chemically with the carbon, converting it to CO and CO2. By utilizing a plasma type which possesses ion energies of less than 25 eV, the carbon is reduced without any alteration to the substrate.
Please feel free to contact me directly for more specific information relating to the process.
Kind regards,
Paul
Paul E. Fischione, President E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 Phone 412-325-5444 FAX 412-325-5443 e-mail paul.fischione-at-internetmci.com www.fischione.com ---------- } From: Lawrence_Murphy-at-cabot-corp.com } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Ashing Samples on TEM Substrates } Date: Wednesday, October 29, 1997 10:24 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } A number of people have asked for more details as to my previous query. } } } First some brief background. Cabot has developed a new reinforcing filler } for elastomers, Carbon-Silica Dual Phase Fillers. Naturally we are working } on characterizing several aspects of these new fillers. } } I would like to assess the morphology of the silica phase by removing the } carbon phase. Ashing the sample in bulk and viewing by TEM provides some } useful information. I would like a higher level of information regarding } the silica phase. } } So what I have tried to do is image the dual-phase aggregates on a TEM } substrate, then place the TEM substrate into a muffle furnace for 2 hours } at 550C in air. The intention is to then view the ashed aggregates (now } only silica). Unfortunately the substrates I have tried (germanium and } gold) have decomposed. By decompose I mean the substrate is no longer on } the grid or severely folded onto the grid bars. I am very surprised that } this also happens with a gold substrate on a gold 400 mesh grid!! } } The carbon in the dual phase filler begins to decompose about 500C. The } dimension of the aggregates are in the range of 100nm give or take. } } Regards, } } Larry Murphy } Group Leader, Analytical Section } Cabot Corporation } }
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I have troubles with a box of 10 year old Philips W SEM filaments. It is very difficult or almost impossible to obtain a good saturation or worse, increasing the filament current the beam current decrease. With new ones no problem. These filaments were forgotten for some year but were always stored in a clean place. My question is: is this an aging effect or the filaments are defective? Is there any possibilty of recovery ? (for instance cleaning with an acid bath ?) Thanks for help
------------------------------------------- Giorgio Gasparotto Dipartimento di Scienze della Terra e Geo-Ambientali Piazza di Porta S. Donato 1 40126 Bologna Italy Tel. 51 243.556 FAX 51 243.336 WWW: http://geode.geomin.unibo.it Internet e-mail gaspar-at-geomin.unibo.it -------------------------------------------
Anand wrote: } } I get to much mail. WHY do people not reply directly to inquiry posters'! } Very simple. Other people might be following the thread and waiting for the replies. Anything that might be of general interest should be posted to the whole list. It saves us sending inquiries about possible replies to the inquiry.
That is my opinion, anyway.
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
} We are using our new Ted Pella 3450 microwave for processing potato tissue } and have some questions: } 1) Do the fumes created by polymerization of LR White stay in the equipment } and so might hinder future use for immunolabeling?
This is a function of the instrument's ventilation system. If the vent fan is sufficiently powerful (the one in our microwaves is rated at 100 cubic feet per minute), there should be no problem with fumes.
{snip}
} 3) What is the best temp. limit for alcohol dehydrations?
In my experience, the alcohols should be kept at or below 40 degrees C.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Thanks for the rapid and, as always, very informative responses to my question about collapsing/exploding fish eggs as they are dried for SEM viewing. Among the suggestions were:
1. Use OTOTO methods or other non-coating techniques to 'strengthen' the sample and reduce the possibility of shrinkage.
2. Puncture or cut the eggs in half to allow passage of fluids (not really feasible in my case, but no doubt useful for others).
3. Try a cryo-stage examination.
4. Try processing/drying from HMDS.
5. Do not allow ANY exposure to air during the handling of the eggs.
6. Keep the sample from being directlly exposed to the incoming CO2, ie invert the coverships in the dryer.
OTOTO and HMDS seem the next routes for me to go. THanks again to those who responded.
Carolyn J. Emerson email: cemerson-at-plato.ucs.mun.ca
Biology Department Memorial University St. John's, NF A1B 3X9 Tel: (709) 737-7515 Fax: (709) 737-3018
Thanks to everyone who replied. I'm going to conduct a seance to see if I can get Benjamin Disraeli, Mark Twain, William Shakespeare, the authors of the Bible, Commander Holloway H. Frost, and Winston Churchill in the same venue to argue the TRUE source. If possible, the debate will be moderated by Peter J. Stratham of Oxford Instruments.
------------------------------------------------ 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 --
Help! My nephew was given a E.Leitz-Wetzlar microscope and wants to trade it for a piece of art. He has no idea what it is worth and will probably come out on the short end of stick. Here is the info he has given me - model# 264460, dated July 1926. The microscope is brass and black ceramic, has 2 lens for top, testing oil emergence? and comes with black case that says, "The Max Worcher & Sons Co. Surgical Instruments, Cinn. Ohio. If anyone can help I would be grateful as I would hate to see him make a mistake. Thank You.
"Very simple. Other people might be following the thread and waiting for the replies. Anything that might be of general interest should be posted to the whole list. It saves us sending inquiries about possible replies to the inquiry."
I agree - I often read about problems or solutions I didn't know existed, but which are nonetheless relevant to my work. A listserver whose sole purpose is to initiate a series of private conversations seems like a waste of technology to me.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
The 24V power supply on my AMRAY 1000B SEM has recently blown out. Are there any kind souls out there with a spare power supply or know of a source where this one can be rebuilt?
Many thanks,
Dennis Shubitowski University of Michigan School of Dentistry dshubito-at-umich.edu
Developmental Biologist Department of Zoology Miami University Oxford, Ohio
We are seeking applicants for an Assistant Professor tenure-track position to begin in August, 1998. Ph.D. in zoology or biology and postdoctoral experience required. Individuals with expertise in any area of Animal Developmental Biology are invited to apply, but preference will be given to those who use electron microscopy as a major research tool. We expect this person to develop an independent research program in developmental biology that will enhance the department's research capabilities.
Teaching responsibilities include: (1) a sophomore level course in developmental biology; (2) participation in a team-taught introductory biology course; and (3) and advanced course in a specialty area. The successful applicant will be expected to seek external funding to support his/her research and to supervise and advise graduate and undergraduate students. Advancement will be based on teaching, research, and professional service, with primary emphasis on teaching and research.
Miami University is a state-assisted institution in SW Ohio. The department has excellent research facilities; the EM facility is well-equipped for SEM, TEM, cryopreservation, and confocal microscopy (see http://www.muohio.edu/~zoocswis for more details about the department and our facilities). The department has strong Ph.D. and M.S. programs, 32 faculty members, several postdoctoral researchers, and 50 graduate students on the Oxford campus.
Interested persons should submit a curriculum vitae, a statement of teaching philosophy, a description of current research and long-term research interests, and should arrange for three letters of recommendation and transcripts of graduate and undergraduate academic work to be sent to:
Dr. Douglas H. Taylor, Chair of Zoology, Miami University, Oxford, OH 45056.
Review of applications will begin on 1 December, 1997, and continue until the position is filled. Miami University offers equal opportunity in employment and education.
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
} Dear List, } } I am trying to do the same thing, but on yougert. I am using fast } green } for protein and nile blue for fat, but what fluorochrome can be used } in the 647nm wavelength of the Argon/Krypton laser to identify } starch? } } billEMac-at-cc.usu.edu
I'm not a confocalist (watch carefully as I insert my foot in my mouth) but starch will stain with PAS, and Schiff's is fluorescent... acriflavine-Schiff's (excitation 480, emission 550-600) will fluoresce yellowish orange, pararosaniline-Schiff's (excitation 560, emission 625) will glow red.
I realise that neither will work at 647nm, but do you have any other wavelengths available?
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department School of Medicine University of Auckland Private Bag 92019 Auckland New Zealand
Medical research laboratory has an opening for a full time position as Research Technician. Candidates should have a Bachelor's Degree and previous experience in many of the following techniques: tissue processing, microtomy, light and electron microscopy, photography and darkroom, immunohistochemistry, and in situ hybridization. Familiarity with computers is useful but not essential.
- Available Immediately -
Please send a resume and a list of 3 personal references (address & phone number) to: David H. Hall, Ph.D. Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
Send by mail [or by email to: hall-at-aecom.yu.edu]
AECOM is located in a residential section of the north Bronx, with good subway, bus and highway connections to Manhattan, Long Island, Westchester County, and New Jersey.
Anand wrote 10/30/97: } =20 } I get to much mail. WHY do people not reply directly to inquiry = posters'! }
Philip Koeck wrote 10/30/97: "Very simple. Other people might be following the thread and=20 waiting for the replies. Anything that might be of general interest should be posted to the whole list. It saves us sending inquiries about possible replies to the inquiry.
That is my opinion, anyway."
--=20 Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
The fact that a lot of postings/messages is sent via the MSA-Server is = obvious. Sometimes I also think about how to manage that mass of = informations. BUT: still I am reading them and if of interest I store = them according to a list of items created from Microsoft Exchange. The only problem I get with messages containing no "header" or = "concern", sometimes it is a RE, but it is sent like a "Q: question" so = I loose time in storing such informations for rewriting the = "concerns"-header. So my request to Users of the MSA-Server would be: Please adhere to the = regulations of the Server. Thank you very much.
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
Dear all, all silicone rubber mold "packages" as requested are mailed since = Tuesday, 28th of October, including product data sheets, instructions as = well as at least one self fabricated mold. 30 colleagues were interested in getting such information.
Within the last 3 weeks I fabricated approx. 45 molds which had to stand = 2 days for aeration on air (condensation process), then they were = tempered for at least 30 hours at 140 degr.C. Those embedding molds are tested and approved for use with epoxide-resin = (EPON 812 substitute Glycid-ether 100 from SERVA/BIOPRODUCTS) but not = with hydrophilic resins like LRWhite or Lowicryls.
I hope that you are satisfied with the informations you got.=20
For all those who were included in the first "mail" (~ 10/7-10/97): for U.S.A.residents: please call or contact the=20 WACKER SILICONE COMPANY at ADRIAN, Michigan (as indicated in my = text-info).
For residents in other countries than U.S.A.:
As I was informed by WACKER SILICONE GmbH Munich, all inquiries = concerning the ELASTOSIL R silicone masses, orders, info-requests etc. = should be addressed to:
DRAWIN-Vertriebs-GmbH c/o Mr. Helmut Klug or Mrs. Cornelia POHL=20 (they know about my information to you) Rudolf-Diesel-Strasse 15 D-85 521 OTTOBRUNN/RIEMERLING (which is situated near MUNICH) phone: ++49++ 89 - 6 08 69 - 0 FAX: ++49++ 89 - 6 08 69 - 250
It is a sub of WACKER SILICONES and unfortunately they (WACKER) didn=B4t = mention this address in their info-brochures.
If there is any questions more, please e-mail directly. If there is anybody of the colleagues who told me his interest but = didn=B4t get the package within the next week or so, please also e-mail = directly.=20
If there is anybody else who wishes to receive also that information = package, please request it also preferably by direct e-mail.=20
At the moment I=B4m out of glass-flasks/bottles for evacuating the = silicone rubber mass. Also I have to order new silicone mass, since all = of my own has gone. So please be patient.
I greatly should appreciate a short note when/that you got your = "package".
I thank you all for your interest and wish "good luck" for the first = self-fabricated mold meeting your special needs.
Best regards and have a nice day/night/weekend
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
} } } "Only the small minded will keep things in order the genius will overlook the chaos". { { { Today I am small minded.
Anand wrote 10/30/97: } =20 } I get to much mail. WHY do people not reply directly to inquiry = posters'! }
Philip Koeck wrote 10/30/97: "Very simple. Other people might be following the thread and=20 waiting for the replies. Anything that might be of general interest should be posted to the whole list. It saves us sending inquiries about possible replies to the inquiry.
That is my opinion, anyway."
--=20 Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
The fact that a lot of postings/messages is sent via the MSA-Server is = obvious. Sometimes I also think about how to manage that mass of = informations. BUT: still I am reading them and if of interest I store = them according to a list of items created from Microsoft Exchange. The only problem I get with messages containing no "header" or = "concern", sometimes it is a RE, but it is sent like a "Q: question" so = I loose time in storing such informations for rewriting the = "concerns"-header. So my request to Users of the MSA-Server would be: Please adhere to the = regulations of the Server. Thank you very much.
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
To AParent and microscopist: In the end that nephew will do has he pleases. There are some considerations which I have made: Art, presumably a painting, can be very appealing for some time, but after some years they seem dated or inappropriate. The truest general saying about art is "today's art is tomorrows trash" - and the exceptions (I venture less than 1%) prove that rule. I see art as decorative materials and not an investment.
That microscope, when properly displayed is an attractive statue. In twenty years time, chances are overwhelming that the microscope will have doubled in price and that the other piece of art will be very hard to sell at any price. Incidentally, in the history section of our links pages is an internal links to my two antique Leitz microscopes. I believe that either should be worth about US$2000 in the right market - sorry they are not for sale; they no longer make antique microscopes, just plenty of paintings. 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, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } From: AParent103-at-aol.com } } Help! My nephew was given a E.Leitz-Wetzlar microscope and wants to trade it } for a piece of art. He has no idea what it is worth and will probably come } out on the short end of stick. Here is the info he has given me - model# } 264460, dated July 1926. The microscope is brass and black ceramic, has 2 } lens for top, testing oil emergence? and comes with black case that says, } "The Max Worcher & Sons Co. Surgical Instruments, Cinn. Ohio. If anyone can } help I would be grateful as I would hate to see him make a mistake. Thank } You.
I spend most of each day preparing and analyzing samples using light microscopy and FT-IR microscopy. Fingerprints are more than a nuisance, so its frequent trips to the sink for a little soap and water. In the winter I find my fingertips become dry and cracked. Lotion is out during work hours. Cotton gloves shed linters. Finger cots provide some relief.
Anybody have other suggestions?
James "Fingers" Martin :) Williamstown Art Conservation Center
Stephen Edgar wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } } Dear List, } } } } I am trying to do the same thing, but on yougert. I am using fast } } green } } for protein and nile blue for fat, but what fluorochrome can be used } } in the 647nm wavelength of the Argon/Krypton laser to identify } } starch? } } } } billEMac-at-cc.usu.edu } } I'm not a confocalist (watch carefully as I insert my foot in my } mouth) but starch will stain with PAS, and Schiff's is fluorescent... } acriflavine-Schiff's (excitation 480, emission 550-600) will } fluoresce yellowish orange, pararosaniline-Schiff's (excitation 560, } emission 625) will glow red. } } I realise that neither will work at 647nm, but do you have any other } wavelengths available? } } Regards } } Stephen Edgar } } Electron Microscope Unit, Pathology Department } School of Medicine } University of Auckland } Private Bag 92019 } Auckland } New Zealand } } email address: s.edgar-at-auckland.ac.nz } Phone : +64-9-3737599 extn 6473 (GMT + 12h) } Fax : +64-9-3737459
Gentlemen,
An interesting alternative to trying to find a third fluorophore: Starch has a unique response to polarized light: it exhibits a maltese cross. Just as you might combine regular epi-fluorescence with, let's say, transmitted light phase contrast, why couldn't you combine fluorescence with polarized light? It would really only take adding one polarizer over the transmitted light source and a second, in crossed position, to the barrier filter ... or, if you have access internally to the confocal system, somewhere before the detector.
If you try this combination, please send me an image.
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 ********** Americas First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
I've got a question about plasma "etching" or "ashing" procedure to remove outer organic surfaces of polymer samples.
Another lab who did this for us in the past had a procedure where they did the etching in two steps: 1. oxygen plasma 2. argon plasma. (The other lab won't talk to us anymore, but that's another story...)
I can't figure out what what the argon plasma is supposed to do. The function of the oxygen plasma is clear: to oxidize the carbon and hydrogen, producing volatile substances. A mixed oxygen/argon plasma might make sense too, to control the ablation rate for instance. But the reason for doing pure oxygen and then pure argon eludes me.
Am I being dense? Can anybody give me a tip on this? (I'm a chemist, for goodness sake, but I still can't figure it out!)
We have found that the `pseudo-leather' gloves available from Agar Scientific (and I guess most other em accessory suppliers) are good for long term wear. The surface eventually starts to break up with wear but until then they are fine. They are the only gloves we will wear when handling HT components as all others we have tried leave something behind on the surface. They retail here at about 3.2 pounds sterling a pair.
Ron
Usual disclaimer - I am purely a satisfied customer. =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
} } after several unsuccessful attempts to stain PE ultrathin cuts with the } Kanig method I would like to ask if somebody has been successful and is } willing to share his protocol with me. } The specimen is already cut with an cryo-ultramicrotome and the cuts are } lying on copper grids. }
Here at Reading, many years ago, we used to do chlorosulphonation of PE by (more or less) the Kanig method. One always stained FIRST, then cut the section.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I am looking for documentation useful to my generating an operating procedure in a Good Laboratory Practice (GLP) environment for documenting transmission, archival, and handling of telepathology case sessions. Could anyone direct me to an area where I might review other protocols currenlty acceptable and in use Greg Argentieri Novartis Pharmaceuticals Corp 59 Rt 10 Bldg 406 Rm 121 East Hanover, NJ 07936 973-503-8617 Fax 973-503-6339 E-mail Gregory.Argentieri-at-pharm.novartis.com
One can get fairly small refrigerators that can be dedicated to osmium and glut. We double containerize the os solutions. First in a Teflon screw top bottle and then in a jar with a screw top and seal in the lid (intended to food preservation) We hope this minimizes the os vapor leackage. We store cacodylate in the regular chemical refrigerator. We also have a refrigerator reserved for biologicals such as enzymes and antibodies, away from any fixative vapors that might denature them. Os waste is stored in large screw top bottles which originally contained solvents. It is in the fume hood at room temp until it is collected by our safety dept. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } . At 11:03 AM 10/31/97 +1100, you wrote:
} Fellows Biological Users } } I am seeking information on the type of refrigerator and types of storage } currently used by biological departments using: } } osmium tetroxide + waste, and } } glutaradehyde } } sodium cacodylate } } } Thankyou in advance for your help } } } Barry } EM Unit } UNSW } } } ******************************************************* 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
You need to say if you are handling any chemicals that require protection, in which case you might try nitrile gloves, which are thin, strong, chemically resistant, and do not cause latex allergies. If you don't need gloves, you could try something moisturizing after washing. I use a product called Eucerin, a water-in-oil emulsion that's expensive, but a jar will last you a long time.
Leonard Corwin, Principal Research Chemist Fort Dodge Animal Health, Animal Health Research Analytical Princeton, NJ 08543-0400 corwinl-at-pt.cyanamid.com
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Here's a new thread. Hand care for microscopists.
I spend most of each day preparing and analyzing samples using light microscopy and FT-IR microscopy. Fingerprints are more than a nuisance, so its frequent trips to the sink for a little soap and water. In the winter I find my fingertips become dry and cracked. Lotion is out during work hours. Cotton gloves shed linters. Finger cots provide some relief.
Anybody have other suggestions?
James "Fingers" Martin :) Williamstown Art Conservation Center
} Here's a new thread. Hand care for microscopists. } } I spend most of each day preparing and analyzing samples using light } microscopy and FT-IR microscopy. Fingerprints are more than a nuisance, } so its frequent trips to the sink for a little soap and water. In the } winter I find my fingertips become dry and cracked. Lotion is out during } work hours. Cotton gloves shed linters. Finger cots provide some relief. } } Anybody have other suggestions? } } James "Fingers" Martin :) } Williamstown Art Conservation Center
I use Vaseline Intensive Care lotion, or any similar hand lotion. Small amount 2-3 times a day work better than a larger amount once a day.
After polishing my sample I immersed it in acetone to dissolve the cement and after it had separated from the glass support I examined it by OM . At that point I noticed that a thin solid film of the colloidal, non-crystallizing silica, had formed on the surface of the sample. Is there a safe way of removing this film ?? I've tried soaking the sample in the colloidal suspension and in water but it did no seem to help.
I suspect the film formed because I failed to wash the sample properly before putting it in the acetone but I hate the idea of starting all over again.
I need some help on taking picture using a Zeiss STEMI SR. Some how the image in the camera is not focused uniformly. The image (and therefor the picture) is best focused at the central spot, and becomes blur at the edges, most noticeably on one side. When I raise or lower the scope (or stage), the image kind of move laterally, not up and down. I could not figure out why, except suspecting the alignment of the scope may be off. Any suggestions and comments are welcome.
I use a normal refrigerator for storage of all chemicals. We are required to label all refrigerators at the facility. Those refrigerators for food storage are labeled "For Food Only". Those refrigerators which contain chemicals (any type of chemical) are labeled: "No Food or Beverage / BioHazard" or something to that effect. The labels are on the outside of the refrigerators and are fairly large and usually brightly colored.
Glutaraldehyde is stored in regular screw-cap bottles.
Osmium crystals are stored in the original shipping vials and packaging in the refrigerator until they are diluted for use.
Osmium crystals are solubilized accordingly: The whole vial containing crystals is dropped into water or buffer to solubilize. The solution is made up in a 25-30 ml glass-stoppered reagent bottle, this bottle is placed inside a second slightly larger glass screw-cap bottle. The second bottle contains cotton in the bottom to cushion the first bottle. The second + first bottles are then placed inside a third still larger, about 550ml capacity plastic Bel-Art Products jar. The Bel-Art jar is brown with a white, opaque screw cap which eliminates light. We also put cotton or paper towelling in the bottom of the Bel-Art jar to cushion the glass bottles. The outside of both glass bottles are wrapped with parafilm before placing in the next bottle. It's not necessary to wrap the outside of the Bel-Art jar with parafilm. The whole lot is left in the hood overnight to solubilize the osmium.
For storage: The series of nested bottles/jars is placed into a normal refrigerator.
Osmium is extremely volatile and you will find that the parafilm becomes black upon storage indicating leakage.
Following the above storage procedure, we've never had a problem with a blackened refrigerator but I've heard of that happening in other places.
At 11:03 AM 10/31/1997 +1100, Barry Searle wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Delilah F. Wood United States Department of Agriculture Western Regional Research Center 800 Buchanan Street Albany, CA 94710
We have ethe same problem since we handle clinical samples and are constantly washing with antiseptic soap.
Sleep in gloves with lots of Vaseline on your hands. In the day time, chapstick applied just to the edges of the cracks (not all over your hands) can help if cracks are not on the very tips. At least you can apply it at lunch, etc. You can glue cracks together with superglue (a hint from my surgeon friend). Eucerine is a good hand cream recommended by my pharmacist; you might also try bag balm (for cows utters, available at farm supply stores) for times when you aren't at work.
Sara Miller
On Thu, 30 Oct 1997, James Martin wrote:
} Date: Thu, 30 Oct 1997 20:40:20 -0500 (EST) } From: James Martin {James.S.Martin-at-williams.edu} } To: microscopy-at-sparc5.microscopy.com } Subject: hand care for microscopists } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Here's a new thread. Hand care for microscopists. } } I spend most of each day preparing and analyzing samples using light } microscopy and FT-IR microscopy. Fingerprints are more than a nuisance, } so its frequent trips to the sink for a little soap and water. In the } winter I find my fingertips become dry and cracked. Lotion is out during } work hours. Cotton gloves shed linters. Finger cots provide some relief. } } Anybody have other suggestions? } } James "Fingers" Martin :) } Williamstown Art Conservation Center }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Our GW Champerscope works great. Since we installed it, no one has hit = the polepiece yet with the ancillary detectors. We have had it attached = to the JEOL 35C and the Hitachi 520 and plan to put it on our Electroscan = depending on the geometry AND once it gets installed.
The standard disclaimer applies of no financial interest in the above.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 __________________________________________________________________________= _____
Hi:
I am thinking of getting an IR viewing system for our SEM chamber. Any advice or true to life adventures you can share?
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
------------------ RFC822 Header Follows ------------------ Received: by sjdccd.cc.ca.us with ADMIN;31 Oct 1997 08:54:03 -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, 30 Oct 1997 18:53:06 = -0800 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) id SAA23594 for dist-Microscopy; Thu, 30 Oct 1997 = 18:24:23 -0600 Received: from cats.ucsc.edu (rumpleteazer.UCSC.EDU [128.114.129.45]) by = Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id SAA23590 for = {microscopy-at-sparc5.microscopy.com} ; Thu, 30 Oct 1997 18:24:22 -0600 Received: from cats-po-1 (root-at-cats-po-1.UCSC.EDU [128.114.129.22]) by cats.ucsc.edu (8.8.5/8.8.4.cats-athena) with SMTP id QAA16740 for {microscopy-at-sparc5.microscopy.com} ; Thu, 30 Oct 1997 = 16:35:41 -0800 (PST) Received: from [128.114.25.225] by cats-po-1 (8.6.13/4.8) id QAA13257; = Thu, 30 Oct 1997 16:35:27 -0800 Message-Id: {199710310035.QAA13257-at-cats-po-1} Mime-Version: 1.0 Content-Type: text/plain; charset=3D"us-ascii"
{fontfamily} {param} New_Century_Schlbk {/param} TWO RESEARCH ASSOCIATE POSITIONS AVAILABLE
(1) Research Associate position in a project to determine the structure of crossbridges in contracting insect flight muscle. Fast freezing followed by freeze substitution enables us to trap force bearing myosin crossbridges with millisecond time resolution. Electron tomography is then used to produce 3-D images that preserve the variable structure of the crossbridges for subsequent analysis. Mechanical records on stiffness and tension are recorded up to the moment of freezing.=20 Parallel time resolved X-ray diffraction data of the diffrent contractile states is also available with which to compare the EM data with native muscle structure. Methods have been developed over the past year to (1) classify and group the variable crossbridge forms for subsequent averaging that improves the signal to noise ratio in the reconstructions and (2) adjust and refine the atomic structure of the myosin head in order to fit it into the in situ envelope from the 3-D reconstruction. The research offers a unique opportunity to learn electron tomography, correspondence analysis of 3-D motifs and atomic modeling within the envelope of a low resolution envelop and at the same time provide unique information on the structural changes that occur in situ in contracting muscle. The project involves primarily computing to obtain the 3-D images and computer modeling to interpret the structure.
Recent Publications:
Holger Schmitz, Mary C. Reedy, Michael K. Reedy, Richard T. Tregear, Hanspeter Winkler, Kenneth A. Taylor. Electron tomography of Insect =46light Muscle in Rigor and AMPPNP at 23=B0C. {italic} J. Mol. Biol. {/italic} 264, 279-301 (1996)
Holger Schmitz, Mary C. Reedy, Michael K. Reedy, Richard T. Tregear, Hanspeter Winkler, Kenneth A. Taylor. Tomographic 3-D Reconstruction of Insect Flight Muscle Partially Relaxed by AMPPNP and Ethylene Glycol. {italic} J. Cell Biol. {/italic} (in press, November, 1997)
(2) Research Associate position in a project to determine the structure of alpha-actinin by electron crystallography. Project is funded through January 2001. We have obtained crystals of several different isoforms of alpha-actinin using lipid monolayers. The crystals are large in extent and to date yeild structural information to at least 10 Angstroms resolution. =20
Applicants for both positions must have a PhD degree. Salary is negotiable based on relevent experience. =20
The successful applicant will become a member of the Structural Biology program at Florida State University that includes 3 protein X-ray crystallography groups, three macromolecular NMR groups, 2 EPR groups and 2 electron microscopy groups. Facilities for electron protein crystallography include the above mentioned CM300-FEG, a Philips CM120, 3 Gatan cryotransfer systems, a Gatan wide angle TV camera, a Perkin-Elmer PDS1010M densitometer, 2 surveying optical diffractometers. Inquiries and applications should be made to Dr. Kenneth Taylor, Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306-4380. Tel: (850) 644-3357. FAX (850) 561-1406. E-mail: taylor-at-bio.fsu.edu. Applicants should provide a CV and the names and addresses of 3 former mentors or knowledgeable individuals who can provided reference letters. =20
Has anyone figured out how to segregate from other mail the e-mail originating in this forum?
I use Internet Mail & News (v4.70) bundled with Microsoft IE 3.02. The "Inbox Assistant" in "Mail" menu is meant to assist in automatic segregation to sub-folders, but I have been unable to make it work with stuff from the MSA list server.
Any thoughts, suggestions, or recommendations for mail browsers better able to do the job?
More likely than not, the soap you use is the culprit. I find pure glycerine soap not drying nor chapping, even if used to excess. (In US, check your pharmacy for Nutrogena - UNSCENTED, or the same thing under the pharmacy name but at 1/2 price.)
Most "hand lotions" contain perfume in alcohol base and do more harm than good. At lunch, end of work, and night time, I work-in a good quantity of "A and D Ointment" - the variety WITHOUT ZnO. (Its customary use is to relieve diaper rash, and it can be found in baby-needs.)
By next morning, fingers are fine.
valdemar-at-fast.net No stock in either product.
---------- } From: James Martin {James.S.Martin-at-williams.edu} } To: microscopy-at-sparc5.microscopy.com } Subject: hand care for microscopists } Date: Thursday, October 30, 1997 8:40 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Here's a new thread. Hand care for microscopists. } } I spend most of each day preparing and analyzing samples using light } microscopy and FT-IR microscopy. Fingerprints are more than a nuisance, } so its frequent trips to the sink for a little soap and water. In the } winter I find my fingertips become dry and cracked. Lotion is out during } work hours. Cotton gloves shed linters. Finger cots provide some relief. } } Anybody have other suggestions? } } James "Fingers" Martin :) } Williamstown Art Conservation Center }
Cindy Bennett wrote: =================================================== I've got a question about plasma "etching" or "ashing" procedure to remove outer organic surfaces of polymer samples.
Another lab who did this for us in the past had a procedure where they did the etching in two steps: 1. oxygen plasma 2. argon plasma. (The other lab won't talk to us anymore, but that's another story...)
I can't figure out what what the argon plasma is supposed to do. The function of the oxygen plasma is clear: to oxidize the carbon and hydrogen, producing volatile substances. A mixed oxygen/argon plasma might make sense too, to control the ablation rate for instance. But the reason for doing pure oxygen and then pure argon eludes me.
Am I being dense? Can anybody give me a tip on this? (I'm a chemist, for goodness sake, but I still can't figure it out!) ================================================== Argon is used, in general, when one would want to remove a metal oxide and maybe some metals, cases where the aggressiveness of CF4 or other reactive fluorine based bases is not required. We are talking now about real "etching" and not just "plasma cleaning". I have heard of "formulas" where people have diluted oxygen with argon in order to moderate the etching of an organic surface. But I am unaware of any published literature that describes any benefit of first etching with pure oxygen and then etching with pure argon. Indeed, if the objective was to etch a polymer surface to reveal structure and morphology, to then expose it to argon would, at least to me, seem to be counter productive. The only exception I can recall was in the study of aluminum lithographic printing plates, the polymer emulsion layer was first etched off with oxygen and then the oxide layer was etched somewhat with argon to better "open" up the pore structure. The aluminum oxide was being "etched" slightly with the Ar.
I always hate to second guess another laboratory without hearing their rationale for doing something in some particular way. Here is the USA, an independent laboratory, especially one operating under ISO Guide 25 guidelines, and offering such a service would be expected to answer such a question in their report to the client. Since ISO Guide 25 had its origins in your corner of the world, I am surprised that laboratories there are not operating to the same standard?
Disclaimer: SPI manufactures a plasma etcher and would have an interest in seeing more people using this technique in microscopy. More information can be found on our website below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The stereo microscope is using only one of the optical paths to form the image on the film plane. Thus you will notice some side-to-side movement of the image as you focus the microscope. If your depth of field is limited and the subject is flat on the microscope stage (or base plate), only the center of the image will be in focus, the left and right edges being above or below the plane of focus. } -----------------------------------------------------------------------. } } Hi, there: } } I need some help on taking picture using a Zeiss STEMI SR. Some how the } image in the camera is not focused uniformly. The image (and therefor the } picture) is best focused at the central spot, and becomes blur at the } edges, most noticeably on one side. When I raise or lower the scope (or } stage), the image kind of move laterally, not up and down. I could not } figure out why, except suspecting the alignment of the scope may be off. } Any suggestions and comments are welcome. } } Jim
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
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
RE} EM: plasma etching procedure 10/31/97
I only reason I can think of for a final pure Ar plasma is to purge the chamber and pumping system of all pure oxygen prior to venting. I'm not aware of anyone else who does this, but it might be a safety concern for somebody.
********************************************************** Jake Schaper Product Analysis Lab Motorola, Inc. 1300 N. Alma School Rd. Chandler, Arizona 85224 Mail Drop CH240 Phone 602-814-4756 **********************************************************
--------------------------------------
Hello wise ones,
I've got a question about plasma "etching" or "ashing" procedure to remove outer organic surfaces of polymer samples.
Another lab who did this for us in the past had a procedure where they did the etching in two steps: 1. oxygen plasma 2. argon plasma. (The other lab won't talk to us anymore, but that's another story...)
I can't figure out what what the argon plasma is supposed to do. The function of the oxygen plasma is clear: to oxidize the carbon and hydrogen, producing volatile substances. A mixed oxygen/argon plasma might make sense too, to control the ablation rate for instance. But the reason for doing pure oxygen and then pure argon eludes me.
Am I being dense? Can anybody give me a tip on this? (I'm a chemist, for goodness sake, but I still can't figure it out!)
Message-Id: {3.0.2.16.19971031143030.425fbb24-at-inka.mssm.edu} X-Sender: massimo-at-inka.mssm.edu X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.2 (16)
At 01:20 AM 11/1/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I use Eudora as my email system, so I don't know if the following will work with Internet Mail & News.
In Eudora you can set filters which can look at any part of the message (header, sender, body, etc.) and take one of several actions (e.g. send to a specified mailbox). I have set up a mailbox "microscopy" and a filter which looks in the body of each incoming message for the prologue which is added by the Microscopy list server, in particular for the words "Microscopy ListServer". If these words are found, the message is automatically sent from the "In" mailbox to the "microscopy" mailbox. Your system should have an equivalent feature. If not, I suggest you take a look at Eudora (I have no financial interest in the company, I just like the product).
Sincerely,
Massimo Sassaroli _____________________________________________________ Massimo Sassaroli, D.Sc. Department of Physiology and Biophysics Box 1218 Mount Sinai School of Medicine One Gustave L. Levy Place New York, NY 10029-6574
A rapidly growin OEM of surface inspection equipment specializing in the development of state of the art micropscopy techniques is in need of a goal orientated leader to direct all production development and ensure that goals & objectives of plans are met within prescribed time frames and funding parameters. Requirements: BS & 10 years of related management experience. Diverse technical background, emphasis in optics and confocal microscopy. Experience: managing mutidiciplinary projects, intellectual property, team environment. Send resume to Kovex Corporation, 3711 Lexington Avenue N., Shoreview, MN. 55126 FAX: (612) 486-9785 or e-mail: kovex-at-spacesmar.net
} I am thinking of getting an IR viewing system for our SEM chamber. Any } advice or true to life adventures you can share?
We have a GW Chamberscope that uses infrared illumination attached to our Hitachi 2460N variable pressure SEM. Excellent system, extremely useful, no problems. I don't know how we got along without one. One suggestion: if you can, adjust the point of view to permit you to nearly look directly down on the sample (as is done with the Topcon) to better correlate to the SEM view.
#################################################################### 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 have a question about the importance of cover slip thickness. Namely, how important is it to use a cover slip thickness for which the objective is designed? For example, if I'm using a 40X, NA 1.3 oil immersion objective which has the number 0.17 on it (corrected for a 170 micron cover slip thickness), what would be the effect on image quality if I used a cover slip with, say, a 300 micron thickness?
Also, what is the definition of "working distance?" I understand it to be the distance between the top of the cover slip and the lens of the objective, but I want confirmation of this definition.
Thank you very much,
Brian Haab U.C. Berkeley bhaab-at-zinc.cchem.berkeley.edu
Dan Kremser dkremser-at-levee.wustl.edu =============================================================
JOINT FALL MEETING
MISSOURI-ILLINOIS-KANSAS MICROBEAM ANALYSIS SOCIETY & CENTRAL STATES MICROSCOPY SOCIETY
Washington University Conference Center, St. Louis, Missouri
November 14, 1997
PROGRAM:
Chair: Lou Ross University of Missouri-Columbia
8:30-9:00 Registration, Vendor Display Setup, and Welcome Coffee.
9:00-9:10 Welcome
9:10-9:30 Rebecca Morlando 3M Company, Columbia, Missouri. "SEM to Confocal for Measuring Vias in Flexible Circuits"
9:30-10:00 Jingyue (Jimmy) Liu Analytical Sciences Center, Monsanto Company,St. Louis, MO "Resolution and Surface Sensitivity in Low Voltage SEM"
10:00-10:20 Dan Kremser Department of Earth and Planetary Sciences, Washington University, St. Louis, Missouri. "Coupling Laser Ablation with a Sector Field ICP-MS: Calibration Enhancement through the use of Smithsonian Microprobe Standards"
10:20-10:40 Morning Break---Coffee We thank Ed Immer/Electron Microscopy Sciences for support for our refreshments this morning.
Chair: Don Parker Briem Engineering, St. Louis, Missouri
10:40-11:40 Greg Meeker, US Geological Survey-Denver MAS tour speaker who will present the Chuck Fiori Memorial Technology Presentation.
"Standards for X-ray Microanalysis: How We Get Them, How We Use Them & Why We Will Always Need Them"
11:50-1:00 LUNCH See menu and details below on this page.
Chair: Mike Veith Department of Biology, Washington University
1:00-1:20 William Randle Missouri State Highway Patrol, Jefferson City,MO. "A Microchemical Test for Alkylamines of Water Gel Explosives"
1:20-1:40 L. Shannon Holliday Renal Division, Washington University Medical School, St.Louis, Missouri. "Use of Light, Electron and Confocal Microscopy in the Study of Osteoclastic Bone Resorption"
1:40-2:00 Gayleen Cochran Plant Biology Department, Southern IllinoisUniversity-Carbondale. "Spermatogenesis in Equisetum; a Correlated TEM and Fluorescence Microscope Study"
2:00-2:20 Angel R. Maden Plant Biology Department, Southern IllinoisUniversity-Carbondale. "Comparative Ultrastructure of Lycopidiaceae Spermatozoids"
2:20-2:40 Afternoon Break---Refreshments
2:40-3:00 Cheryl A. Nickerson Department of Biology, WashingtonUniversity, St. Louis, Missouri. "Histological Effect of Salmonella Infection on Murine Tissue"
3:00-3:20 Robert Mark Friedman Analytical Sciences Center, Monsanto Company, St. Louis, Missouri. "Imaging and Analysis with Time-of-Flight Mass Spectrometry and Related Techniques" *
3:30- Business Meetings
*
LUNCH DETAILS
Lunch will be a Deli Buffet consisting of Shaved Ham, Turkey, Roast Beef, American and Swiss Cheeses, various sandwich toppings, assorted Breads, Fruit Salad and Pasta Salad. Drinks and Desserts are included. The cost is $5.00 per person. Local eateries are located a short walking distance from the Conference Center for those who would rather dine offsite.
We need confirmation of those who plan to eat the Deli Buffet, By 10:00 AM, Tuesday November 11. Please contact Dan Kremser by phone (314-935-5605) or email (dkremser-at-levee.wustl.edu). Please confirm even if you have done so on the earlier mailing request. -- *************************** 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
Another excellent product for hand repair is Extra Emollient Night Cream by Mary Kay. A jar costs $11 and lasts for a very long time. A physician friend of mine uses it and it has healed horrible skin cracking caused by latex allergy. Mary Kay consultants can be found either on the Web at www.marykay.com or in the white pages of the phone book.
} -----Original Message----- } From: Sara Miller [SMTP:saram-at-acpub.duke.edu] } Sent: Friday, October 31, 1997 11:22 AM } To: James Martin } Cc: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: hand care for microscopists } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } We have ethe same problem since we handle clinical samples and are } constantly washing with antiseptic soap. } } Sleep in gloves with lots of Vaseline on your hands. In the day time, } } chapstick applied just to the edges of the cracks (not all over your } hands) can help if cracks are not on the very tips. At least you can } apply it at lunch, etc. You can glue cracks together with superglue } (a } hint from my surgeon friend). Eucerine is a good hand cream } recommended } by my pharmacist; you might also try bag balm (for cows utters, } available } at farm supply stores) for times when you aren't at work. } } Sara Miller } } On Thu, 30 Oct 1997, James Martin wrote: } } } Date: Thu, 30 Oct 1997 20:40:20 -0500 (EST) } } From: James Martin {James.S.Martin-at-williams.edu} } } To: microscopy-at-sparc5.microscopy.com } } Subject: hand care for microscopists } } } } } ---------------------------------------------------------------------- } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } ---------------------------------------------------------------------- } -. } } } } } } Here's a new thread. Hand care for microscopists. } } } } I spend most of each day preparing and analyzing samples using light } } microscopy and FT-IR microscopy. Fingerprints are more than a } nuisance, } } so its frequent trips to the sink for a little soap and water. In } the } } winter I find my fingertips become dry and cracked. Lotion is out } during } } work hours. Cotton gloves shed linters. Finger cots provide some } relief. } } } } Anybody have other suggestions? } } } } James "Fingers" Martin :) } } Williamstown Art Conservation Center } } } } Sara E. Miller, Ph. D. } P. O. Box 3020 } Duke University Medical Center } Durham, NC 27710 } Ph: 919 684-3452 } FAX: 919 684-8735
Vlademar et al: I have written two rules which cover 90% of all MSA mail. Rule 1) When Event is "new Item" type is "mail" contents are "TO: "microscopy" Move to Folder MSA. Rule 2) same as above except contents are "CC: "microscopy" " Move to Folder MSA. I hope this helps. bob m