Where do your cells grow? I had problems with osmium fixation of cell cultures when they grew on plastic membranes, and a nice lady from this list server suggested that I should reduce the osmium tetraoxide with potassium ferrocyanide because the osmium reacts with the plastic polymer.
We now grow the cells on "Aclar film" and get good membrane contrast, with or without reducing the osmium tetraoxide. This film is supposed to be chemically insensitive and can withstand dehydration and embedding. The film has to be pre-treated with poly L lysine to prevent the cells from floating away, though.
yours
Gerd Leitinger
} } Question: I have recently been having problems getting good staining of the membranes of cultured cells with osmium. The cells were fixed with 2% glutaraldehyde and post fixed in 1% osmium tetroxide. I have tried 0.1M phosphate buffer, pH 7.4 and 0.1M cacodylate buffer pH 7.3. When using these the membranes were not visible at all. I also tried osmium with potassium ferrocyanide which worked well with phosphate buffer but left a black precipitate. With cacodylate buffer and potassium ferrocyanide the membrane definition was a little better but still not adequate. If anyone has any suggestions as to how to improve the membrane staining, they would be greatly appreciated. Thank you. } } --------------------------------------------------------------------------- }
Dr. Gerd Leitinger
Institut für Zellbiologie, Histologie und Embryologie Medizinische Universität Graz Harrachgasse 21 A-8010 Graz Austria
I have a question that takes up the recent permeabilisation issue:
Is it possible to permeabilise membranes with methanol instead of ethanol for pre-embedding immunoEM, and what effects does the methanol have on the ultrastructure of membranes?
I have some experience with pre-embedding immunocytochemistry of vibratome slices of nervous tissue. For this, I usually pre-treat the cells with up to 50% ethanol for a few minutes, and then add 0.05% saponin to all the antibody solutions, and usually get quite good staining in the outer regions of the slices.
I now wanted to do a similar procedure on cultivated cells and have found that the antibodies do not penetrate into the cells at all! For light microscopic immunocytochemistry, I have found that the antibodies only penetrate if these cells are pre-treated with 100% methanol. For immunoEM, I only tested my "usual" protocol of 10, 30, and 50% ethanol for a few minutes each. Does anybody know whether it is worth trying methanol for immunoEM, and perhaps applying the methanol for a longer time, perhaps 30 minutes?
thank you
Gerd Leitinger
Dr. Gerd Leitinger
Institut für Zellbiologie, Histologie und Embryologie Medizinische Universität Graz Harrachgasse 21 A-8010 Graz Austria
Ethanol and methanol will destroy all the membranes you have in the sample. You will see only some blobs of different densities instead of nicely preserved cells. For pre-embedding EM on cultured cells people use saponin 0.1% or digitonin (do not remember %), even Triton X-100 is too harsh.
Gerd Leitinger:
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Try adding 1% tannic acid to your glut and possibly the OsO4, Mallinckrodt cat. #1764 seems to work best -- I believe because it's a monomer, instead of various polymers. This helps with the membrane preservation. Is this for TEM or SEM?
Phil
} Email: rtemkin-at-mtsinai.on.ca } Name: Robert Temkin } } Organization: Hospital for Sick Children Research Institute, Toronto, Canada } } Title-Subject: [Microscopy] [Filtered] osmium fixation of cultured cells } } Question: I have recently been having problems getting good staining } of the membranes of cultured cells with osmium. The cells were fixed } with 2% glutaraldehyde and post fixed in 1% osmium tetroxide. I have } tried 0.1M phosphate buffer, pH 7.4 and 0.1M cacodylate buffer pH } 7.3. When using these the membranes were not visible at all. I also } tried osmium with potassium ferrocyanide which worked well with } phosphate buffer but left a black precipitate. With cacodylate } buffer and potassium ferrocyanide the membrane definition was a } little better but still not adequate. If anyone has any suggestions } as to how to improve the membrane staining, they would be greatly } appreciated. Thank you. } } ---------------------------------------------------------------------------
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 10:10:03 2005
I am posting this as a favour to Dr. Helen Reid at West Chester University:
The university is looking for a chemical microscopist for the Fall 2005 semester. If you are looking, or know anyone who is looking, for this type of position, please contact Helen directly at:
_____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 11:24:53 2005
Greetings, } It is really something to see one's name in print, especially } for being the person to go to for information on a } particular technique! Thanks Paul. } } With tissue cultured cells I have found it critical to fix them as } soon as possible after they come out of the incubator. } DO NOT WASH them! Simply decant the growth } medium then immediately and gently flood the cells with } the fixative. } } The fixative that works well on many different types of cultured } cells as well as pelleted material and small tissue pieces is as follows: } 1% Glutaraldehyde+1% Osmium tetroxide+0.05 M Phosphate Buffer } -at-6.2 pH on ice and kept dark. Usually 45 minutes is fine. } } Yes, this fixative will start to oxidize. To avoid this have all } the components } on ice; mix in the glutaraldehyde immediately before the fixative is needed } and allow sufficient volume to fix both the cells and the small amount of } protein that is left on the cells after decanting the medium. } } Dr. Tilney INSISTS that the Electron Microscopy Sciences } 8% Glutaraldehyde in the 10 ml vials be used } at all times for his actin studies and that it be kept in } a scintillation vial and discarded one week after opening } the vial. The only reason for using the EMS product is that } we always get excellent results and he refuses to even try } another company. } } With the phosphate buffer one needs to wash well with cold } (best available) water at least 3 times over 20 - 30 min. time } AND remember to rinse the entire vessel (top too) } at least once before adding cold 1% UA in water } overnight in the refrigerator to avoid the dreaded uranyl-phosphate } crystals.There is no light in our refrigerator so I do not need to worry } if it really goes out when the door is closed. } } If the cells are to be fixed in the flask/petri dish for face-on sections, } an ethanol dehydration is used and Ladd's LX-112 as an epon } substitute. These do not melt the plastic. } All other cases or cells grown in "Pernanox" dishes } are acetone dehydrated and any epon substitute can be used. } } Contact me off-line if more information is needed. } If there are several similar questions I'll post them. } } If this fixation does not show what you wish with the membranes } try using an objective apperature that is a size smaller than is } usually used in the TEM. } } Pat Connelly psconnel-at-sas.upenn.edu } Dept. of Biology, University of Pennsylvania } Philadelphia, PA 19104-6018
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 12:12:10 2005
I would be interested in hearing from the microscopy community what 3rd party software is available for looking at datasets obtained from spectrum imaging. As many of you know, spectrum images are 3d datasets which contain an EDX (or EELS) spectrum at each pixel from a STEM image. We have a Tecnai F20 FEG with STEM/EDX/EELS and want to probe deeper into extracting useful information from these datacubes; our software can generate basic element maps but we want to purchase software that is more flexible and comprehensive and perhaps interactive too. For instance, we would like to be able to highlight a subset of pixels in an image or element map and see these points appear in a ternary diagram or other type of plot. We can generate a spreadsheet of quantified elements or element ratios for each pixel in a STEM image and can export the spreadsheet. Thanks for your input.
Dave Joswiak Univ. of Washington Dept. of Astronomy Seattle, WA
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 13:23:25 2005
I am forwarding this on behalf of Mr. Goodblatt.. He would like to have an informal session at M&M in Honolulu regarding items he has mentioned. Please reply to him directly if you are interested.
} ------- Forwarded message follows ------- } From: Avrum Goodblatt {goodblat-at-mail.med.upenn.edu} } To: gwe-at-ufl.edu } Subject: [Microscopy] hawaii conference and LIMS } Copies to: luellen-at-mail.med.upenn.edu, jonni S. Moore } {moorej-at-mail.med.upenn.edu} , } qcyu-at-mail.med.upenn.edu } Date sent: Thu, 26 May 2005 12:01:48 -0400 } } } Managing data today has become more challenging due to the very large } datasets } being produced. It is not uncommon for a facility to turn out a full DVD } of data in one } day. Providing online access and backup becomes more complicated, especially } when one also has to protect instrumentation from viruses and other attacks. } } In addition, usage and billing records should be tied into the LIMS but } frequently } these are two completely different systems. With compliance much more on } the radar } today, and funds coming from a wider variety of sources, better } record-keeping is } essential } } I am IT coordinator for a group of 9 resource facilities called } PathBioResource } (including BioMedical Imaging) in Pathology and Laboratory Medicine at the } University of Pennsylvania. I would like to share with others what we are } doing and } learn from others ways to approach these issues. We might also find ways } to share } some tool development together in some open source fashion, or educate } vendors as } to what is needed. } } I will be in Honolulu at the conference from Sunday evening through Wednesday } evening. } } Avrum Goodblatt } goodblat-at-mail.med.upenn.edu } 215 573 0675 } } } } ------- End of forwarded message --------- } Avrum Goodblatt goodblat-at-mail.med.upenn.edu } BMCRC 215 573 0675
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 13:48:41 2005
I hope you are already familiar with the LISPIX package available through NIST (http://www.nist.gov/lispix/doc/contents.htm). It is free and should allow you to play with your data as you please. I have not used it myself so I cannot comment on it.
The other question will be the data format. I don't know that vendors afford export into a generic LISPIX format in a similar way as EDS data can be exported to MSA format. Hopefully such portability will be forthcoming. You can take the matter up with your EDS or EELS vendor.
Warren
At 12:11 PM 06/01/05, you wrote:
} I would be interested in hearing from the microscopy community what 3rd } party software is available for looking at datasets obtained from spectrum } imaging. As many of you know, spectrum images are 3d datasets which } contain an EDX (or EELS) spectrum at each pixel from a STEM image. We } have a Tecnai F20 FEG with STEM/EDX/EELS and want to probe deeper into } extracting useful information from these datacubes; our software can } generate basic element maps but we want to purchase software that is more } flexible and comprehensive and perhaps interactive too. For instance, we } would like to be able to highlight a subset of pixels in an image or } element map and see these points appear in a ternary diagram or other type } of plot. We can generate a spreadsheet of quantified elements or element } ratios for each pixel in a STEM image and can export the spreadsheet. } Thanks for your input. } } Dave Joswiak } Univ. of Washington } Dept. of Astronomy } Seattle, WA
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Inter/Micro 2005 Preliminary Program July 11-15, 2005 Talbott Hotel & McCrone Research Institute Chicago, Illinois
McCrone Research Institute (McRI) cordially invites you to join us at Inter/Micro 2005, an internationally recognized professional meeting dedicated to applied microscopy, held each year in downtown Chicago.
The Inter/Micro 2005 conference will span 5 days. The first 3 days of the conference will consist of a symposium, to be held at the Talbott Hotel, featuring daily 15-20 minute paper presentations, a Monday evening session with Brian J. Ford, Exhibitors (Tuesday and Wednesday), and McRI’s joint meeting with the State Microscopical Society of Illinois including a Banquet, Awards Ceremony, and Auction (Wednesday Evening). The symposium will be followed by a special 2-day workshop (July 14th and 15th), to be held at the laboratories and classrooms of McCrone Research Institute, entitled "Introduction to Techniques of Forensic Soil Comparison" conducted by Skip Palenik. Exhibitors and sponsor’s for Inter/Micro 2005 include: Leica Microsystems, Educational Products, McCrone Research Institute, Bruker Optics, Lomo America, State Microscopical Society of Illinois, Olympus America, and Tienta Sciences.
To exhibit or sponsor (deadline July 1, 2005) please visit: http://mcri.org/ExhibitorINFOandCONTRACT.pdf
The papers presented at Inter/Micro 2005 will be published in the International Journal on microscopy, The MICROSCOPE. For more information, call 312-842-7100 visit www.mcri.org or contact intermicro-at-mcri.org.
Bucher, Edith – Laboratorio Biologico - APPA Bolzano, Applied Aerobiology and Melissopalynology in South Tyrol (Northern Italy) - Realization of a Pollen Atlas
Chun-I Lee, James – University of Strathclyde, Diatom Distribution in Danshuei River
Laughlin, Gary – McRI, Indoor Air Quality: Microscopy of House Dust Particles
Speir, Jacqueline – McRI, Can Dispersion Aid in Amphibole Differentiation?
Chatfield, Eric – Chatfield Technical Consulting, TBA
Havics, Tony – QEPI, Asbestos: To Be a Fiber or Not To Be
12:00 p.m. – 2:00 p.m. Lunch
Ford, Brian J. – Gonville & Caius College, University of Cambridge, The Discovery of Giardia
Alden, Harry – Smithsonian Center for Materials Research and Education, Hair of the Dog, or How Blanket Fibers Can Get Your Goat
Welsh, Frank– Welsh Color and Conservation, Inc., Historic Paint Colors of Spanish St. Augustine
Hills, Linda – CTL Group, From Rocks to Bridges: An Introduction to Cement and Concrete Microstructure
Hagni, Ann – CTL Group, Failure Analyses of Construction Materials Using Optical Microscopy
Rantanen, Walter – Integrated Paper Services, Inc., Matching Matches (Part 2)
5:00 p.m. – 6:00 p.m. Cocktail Hour with Exhibitors: 2nd floor, Victoria Room
Wednesday, July 13 Chemical and Forensic Microscopy
Crowe, John – FDA Forensic Chemistry Center, The Characterization of Microchemical Test Resultant Crystalline Formations using PLM, FT-IR and Raman Spectroscopy
Bowen, Andrew – Stoney Forensic, Inc., Recent Contributions of Chemical Microscopy to the Analysis of Unknowns
Rothhaar, Katia – Tienta Sciences, Use of Drop Coated Deposition Raman for Detection of Explosives
Hollifield, Jeff – Micro Analytical, Flow Chart for Rapid Identification of Inorganic Compounds Using PLM
Ford, Brian – Gonville & Caius College, University of Cambridge, World's Worst Microscopy
Palenik, Skip – Microtrace, TBA
12:00 p.m. – 2:00 p.m. Lunch
Reffner, John – Smith’s Detection, TBA
Boltin, Randy – MVA Scientific Consultants, A Class in Forensic Microscopy
Lawrence, Gene – San Diego County Sheriff's Crime Lab, Marine Coatings - Components and Analysis
Grant, Ricky – 41st CST (WMD) US ARMY, Active National Guard, Weapons Of Mass Destruction Terrorism Experts and Forensic Microscopy
Hopen, Thomas – ATF Forensic Science Laboratory, PLM Characterization of Inorganic Constituents in Automobile Body Fillers
Hietpas, Jack – Microtrace, Microscopy of Explosives
Diaczuk, Peter – John Jay College of Criminal Justice, Determination of Entry vs. Exit Bullet Holes in Garments using Light Microscopy
6:00 p.m. SMSI Banquet and Auction, Victoria Room Dr. Osamu Shimomura, SMSI 2005 Émile Chamot Award Recipient
Regards, Christine
Christine Weaver Inter/Micro Coordinator McCrone Research Institute 2820 S. Michigan Avenue Chicago IL 60616
We would like to do some em immunolocalizations on pig intestinal tissue to detect virus infected cells. Can anyone direct me to some literature? Which would be the best way to start pre- or post-embedding labeling?
The antibodies, we will be using, work well in confocal microscopy on whole tissue with Triton permeabilization, or on paraffin sections
Thank you.
Tea Meulia
-- *************************************** Tea Meulia, PhD Director Molecular and Cellular Imaging Center Ohio State University/OARDC 1680 Madison Ave. Wooster OH 44691
tel.: 330-263-3836 or -3828 fax: 330-202-3563
http://www.oardc.ohio-state.edu/mcic
*****************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 15:17:01 2005
Hello everyone I have just had a request for the use of a somatoscope developed by Gaston Naessens as they want colour at 30,000 times.
I was wondering if someone was pulling my leg but when I did a google search I got http://www.sumeria.net/tech/naessens.html and I am still wondering if someone has a huge hoax going. Has anyone got experience with this type of microscope?
Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada (2003-2005) University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 16:22:14 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rtemkin-at-mtsinai.on.ca) from http://www.microscopy.org/MicroscopyListserver/MLFormMail.html on Wednesday, June 1, 2005 at 08:06:18 ---------------------------------------------------------------------------
Email: rtemkin-at-mtsinai.on.ca Name: Robert Temkin
Organization: Hospital for Sick Children Research Institute, Toronto, Canada
Title-Subject: [Microscopy] [Filtered] osmium fixation of cultured cells
Question: Thanks for the responses but I guess I did leave out some details. I'm processing monolayers grown on glass coverslips. I do use en bloc staining with uranyl acetate. I'm using 2% aqueous UA for 45-60 minutes at room temperature. I use similar times for the glutaraldehyde and osmium fixations. I dehydrate with ethanol and embed in Epon. I separate the coverslip from the plastic with liquid nitrogen. The bizarre thing is that this protocol has worked well for the past few years but recently has given poor results. I'll try some of the suggestions but I'm wondering if the en bloc UA needs to be done overnight since I'm dealing with monolayers.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kevin.selkregg-at-us.vesuvius.com) from http://www.microscopy.org/MicroscopyListserver/MLFormMail.html on Wednesday, June 1, 2005 at 13:30:05 ---------------------------------------------------------------------------
Email: kevin.selkregg-at-us.vesuvius.com Name: Kevin Selkregg
Question: Even though this question does not appear microscopy related; the thermal expansion in this situation definitely affects the microstructure of the parent material I have been studying.
Anyone have any thermal expansion coefficient data for Sodium Aluminate. I am looking specifically for the coefficient values for NaAlO2 (1 Na2O : 1 Al2O3).
NIST-MAS-AMAS Workshop on Variable Pressure and Environmental Scanning Electron Microscopy (VPSEM-ESEM) for Imaging and Microanalysis: Roadmap 3 {http://www.nist.gov/esem}
Place: Lecture Room A and Poster Hall National Institute of Standards and Technology, 100 Bureau Drive, Gaithersburg, Maryland 20899 Dates: November 2 4, 2005
Co-organizers: NIST Surface and Microanalysis Science Division (837) NIST Precision Engineering Division (821) Microbeam Analysis Society (MAS) {http://www.microbeamanalysis.org/} Australian Microbeam Analysis Society (AMAS) {http://www.microscopy.org.au/}
From November 2 to 4, 2005, NIST will co-host a workshop on Variable Pressure Scanning Electron Microscopy (VPSEM) and Environmental SEM (ESEM) in conjunction with the Microbeam Analysis Society (MAS) and the Australian Microbeam Analysis Society (AMAS). Building on the highly successful AMAS Roadmap 1 and 2 workshops on VPSEM-ESEM organized by Brendan Griffin, this VPSEM-ESEM workshop, which constitutes Roadmap 3, will provide an opportunity for technical presentations (platform and poster), demonstrations, and discussions on leading topics in microscopy and microanalysis in VPSEM-ESEM. Special attention will be given to new breakthrough areas, such as combined VPSEM-ESEM-FIB (focused ion beam) instrumentation. Discussions will attempt to identify those critical issues where research is needed for further progress in VPSEM-ESEM. The program as it has been currently developed is attached below. We are seeking interested participants, proposed titles for presentations, as well as suggestions for additional topics and invited keynote speakers to include. In keeping with past NIST-MAS workshops, there will be no cost for attendance, but pre-registration is required so that the available lecture room space is not overwhelmed and NIST security requirements can be satisfied.
NIST-MAS-AMAS VPSEM-ESEM Workshop Tentative Program Wednesday, Nov 2, 2005
AM: Electron and ion interactions in the gas environment Organizer: Brad Thiel (SUNY-Albany) Topics: fundamentals, charge control, radiation damage
PM: Contrast mechanisms in VPSEM-ESEM Organizer: David Joy (Univ. Tennessee) and Brendan Griffin (U. Western Australia)
Funny you should mention fixation with a mixture of glutaraldehyde and paraforrmaldehyde. I just came out of the scope where I looked at some samples of drosophila egg chambers fixed with such a mixture, followed by overnight incubation in uranyl acetate (0.5%). Membrane contrast was unbelievable, but also the preservation of cytoplasmic elements such as ribosomes, actin filaments and clathrin coats. Why don't we use that technique for all our EM preps?!
Marc
On Wednesday, June 1, 2005, at 12:06 PM, Pat Connelly wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Greetings, } } It is really something to see one's name in print, especially } } for being the person to go to for information on a } } particular technique! Thanks Paul. } } } } With tissue cultured cells I have found it critical to fix them as } } soon as possible after they come out of the incubator. } } DO NOT WASH them! Simply decant the growth } } medium then immediately and gently flood the cells with } } the fixative. } } } } The fixative that works well on many different types of cultured } } cells as well as pelleted material and small tissue pieces is as } } follows: } } 1% Glutaraldehyde+1% Osmium tetroxide+0.05 M Phosphate Buffer } } -at-6.2 pH on ice and kept dark. Usually 45 minutes is fine. } } } } Yes, this fixative will start to oxidize. To avoid this have all the } } components } } on ice; mix in the glutaraldehyde immediately before the fixative is } } needed } } and allow sufficient volume to fix both the cells and the small } } amount of } } protein that is left on the cells after decanting the medium. } } } } Dr. Tilney INSISTS that the Electron Microscopy Sciences } } 8% Glutaraldehyde in the 10 ml vials be used } } at all times for his actin studies and that it be kept in } } a scintillation vial and discarded one week after opening } } the vial. The only reason for using the EMS product is that } } we always get excellent results and he refuses to even try } } another company. } } } } With the phosphate buffer one needs to wash well with cold } } (best available) water at least 3 times over 20 - 30 min. time } } AND remember to rinse the entire vessel (top too) } } at least once before adding cold 1% UA in water } } overnight in the refrigerator to avoid the dreaded uranyl-phosphate } } crystals.There is no light in our refrigerator so I do not need to } } worry } } if it really goes out when the door is closed. } } } } If the cells are to be fixed in the flask/petri dish for face-on } } sections, } } an ethanol dehydration is used and Ladd's LX-112 as an epon } } substitute. These do not melt the plastic. } } All other cases or cells grown in "Pernanox" dishes } } are acetone dehydrated and any epon substitute can be used. } } } } Contact me off-line if more information is needed. } } If there are several similar questions I'll post them. } } } } If this fixation does not show what you wish with the membranes } } try using an objective apperature that is a size smaller than is } } usually used in the TEM. } } } } Pat Connelly psconnel-at-sas.upenn.edu } } Dept. of Biology, University of Pennsylvania } } Philadelphia, PA 19104-6018 } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 16:53:58 2005
gee elaine, my first reaction was that this is an interesting question, and that i'm sure that speculation as to this new instrumentation will be running rife over your question. after checking the webpage you referred to, i would even say the speculation will be royal....
i couldn't help that - please, don't ask me to apologize.... the devil made me do it, ok....
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-9354 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 17:11:51 2005
Dear Robert. You have gotten some good suggestions so far. I might add a few things: You should try the reduced osmium- potassium ferricyanide with cacodylate buffer instead of phosphate and you can get rid of the ppt problem. Leave your cells in serum until you are ready to fix and keep them warm until ready to fix(right out of incubator). Sick cells will never look good no matter what you do. I embed cultured hippocampal neurons in Chang embedding molds then dissolve away the whole glass coverslip after embedding. Dissolve the coverslip with hydrofluoric acid(under hood)- that way you have the whole coverslip and lots of cells. If the coverslips are coated with matrigel they are harder to work with the nitrogen method.
For the thin sections, I stain in 5% aqueous UA a for 20 minutes or longer. Follow with a shorter(1-2 mins) in fresh lead stain. I do not do a prolonged UA stain en bloc-I microwave everything. Use a lower kV or smaller aperture to image cells. I would be happy to send you a detailed protocol. Good luck, JoAnn
Question: Thanks for the responses but I guess I did leave out some details. I'm processing monolayers grown on glass coverslips. I do use en bloc staining with uranyl acetate. I'm using 2% aqueous UA for 45-60 minutes at room temperature. I use similar times for the glutaraldehyde and osmium fixations. I dehydrate with ethanol and embed in Epon. I separate the coverslip from the plastic with liquid nitrogen. The bizarre thing is that this protocol has worked well for the past few years but recently has given poor results. I'll try some of the suggestions but I'm wondering if the en bloc UA needs to be done overnight since I'm dealing with monolayers.
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 17:19:37 2005
Is reduced osmium prepared with ferrocyanide or ferricyanide? I know which one I use.
Regards,
Paul Webster.
On 6/1/05 3:11 PM, "JoAnn Buchanan" {redhair-at-stanford.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Dear Robert. You have gotten some good suggestions so far. I might add } a few things: } You should try the reduced osmium- potassium ferricyanide with cacodylate } buffer instead of phosphate and you can get rid of the ppt problem. } Leave your cells in serum until you are ready to fix and keep them warm } until ready to fix(right out of incubator). Sick cells will never look good } no matter what you do. } I embed cultured hippocampal neurons in Chang embedding molds then dissolve } away the whole glass coverslip after embedding. Dissolve the coverslip with } hydrofluoric acid(under hood)- that way you have the whole coverslip and } lots of cells. If the coverslips are coated with matrigel they are harder } to work with the nitrogen method. } } For the thin sections, I stain in 5% aqueous UA a for 20 minutes or longer. } Follow with a shorter(1-2 mins) in fresh lead stain. I do not do a } prolonged UA stain en bloc-I microwave everything. } Use a lower kV or smaller aperture to image cells. } I would be happy to send you a detailed protocol. } Good luck, JoAnn } } Question: Thanks for the responses but I guess I did leave out some } details. I'm processing monolayers grown on glass coverslips. I do use en } bloc staining with uranyl acetate. I'm using 2% aqueous UA for 45-60 } minutes at room temperature. I use similar times for the glutaraldehyde and } osmium fixations. I dehydrate with ethanol and embed in Epon. I separate } the coverslip from the plastic with liquid nitrogen. The bizarre thing is } that this protocol has worked well for the past few years but recently has } given poor results. I'll try some of the suggestions but I'm wondering if } the en bloc UA needs to be done overnight since I'm dealing with monolayers. } } } } Department of Molecular and Cellular Physiology } Stanford University School of Medicine } Stanford, CA 94305 } 650-723-5856 } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 17:27:10 2005
Odd that there aren't any images associated with the websites lauding Naessens, don't you think? At least, not on any of the first page of Google hits I came up with.
Tamara
On Wed, 1 Jun 2005, Elaine Humphrey wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello everyone } I have just had a request for the use of a somatoscope developed by Gaston } Naessens as they want colour at 30,000 times. } } I was wondering if someone was pulling my leg but when I did a google search } I got } http://www.sumeria.net/tech/naessens.html } and I am still wondering if someone has a huge hoax going. Has anyone got } experience with this type of microscope? } } Elaine } } } -- } Dr. Elaine Humphrey } Director, BioImaging Facility } President, Microscopy Society of Canada (2003-2005) } University of British Columbia } 6270 University Blvd, mail-stop Botany } Vancouver, BC } CANADA, V6T 1Z4 } Phone: 604-822-3354 } FAX: 604-822-6089 } e-mail: ech-at-interchange.ubc.ca } website: www.emlab.ubc.ca } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 17:32:28 2005
Elaine, To save another exile from speculation ( Royal or otherwise) I look forward to a report on the demo when you orgainse it. :)
Chris
Christopher J Gilpin Ph.D. Assistant Professor Director, Molecular and Cellular Imaging Facility K1.A04 UT Southwestern Medical Center at Dallas 5323 Harry Hines Boulevard Dallas, TX 75390-9039 Phone +1 214 648 2827 Fax +1 214 648 6408
-----Original Message----- } From: paul r hazelton [mailto:paul_hazelton-at-umanitoba.ca] Sent: Wednesday, June 01, 2005 4:53 PM To: Elaine Humphrey Cc: microscopy-at-msa.microscopy.com
gee elaine, my first reaction was that this is an interesting question, and that i'm sure that speculation as to this new instrumentation will be running rife over your question. after checking the webpage you referred to, i would even say the speculation will be royal....
i couldn't help that - please, don't ask me to apologize.... the devil made me do it, ok....
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-9354 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 1 21:20:15 2005
Hmm, the only new optics I've heard about recently, also from Canada, is to do with the liquid crystal thin lenses developed by Galstian and Presnyakov
Vladimir V. Presnyakov, Karen E. Asatryan, and Tigran Galstian (2002) Polymer-stabilized liquid crystal for tunable microlens applications. Optics Express 10: 865 - 870
Vladimir V. Presnyakov and Tigran V. Galstian (2005) Electrically tunable polymer stabilized liquid-crystal lens. J. Appl. Phys. 97: 103101
} From: Tamara Howard {thoward-at-unm.edu} } Date: Wed, 1 Jun 2005 16:26:28 -0600 (MDT) } To: Elaine Humphrey {ech-at-interchange.ubc.ca} } Cc: microscopy-at-msa.microscopy.com } Subject: [Microscopy] Re: gaston naessens and somatid biology } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Odd that there aren't any images associated with the websites lauding } Naessens, don't you think? At least, not on any of the first page of } Google hits I came up with. } } Tamara } } On Wed, 1 Jun 2005, Elaine Humphrey wrote: } } } } } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------------- } } -- } } } } Hello everyone } } I have just had a request for the use of a somatoscope developed by Gaston } } Naessens as they want colour at 30,000 times. } } } } I was wondering if someone was pulling my leg but when I did a google search } } I got } } http://www.sumeria.net/tech/naessens.html } } and I am still wondering if someone has a huge hoax going. Has anyone got } } experience with this type of microscope? } } } } Elaine } } } } } } -- } } Dr. Elaine Humphrey } } Director, BioImaging Facility } } President, Microscopy Society of Canada (2003-2005) } } University of British Columbia } } 6270 University Blvd, mail-stop Botany } } Vancouver, BC } } CANADA, V6T 1Z4 } } Phone: 604-822-3354 } } FAX: 604-822-6089 } } e-mail: ech-at-interchange.ubc.ca } } website: www.emlab.ubc.ca } } } } } } |--------------------------------------------------| } Tamara Howard } Department of Cell Biology and Physiology } University of New Mexico - Health Sciences Center } Albuquerque, NM 87131 } thoward-at-unm.edu } |--------------------------------------------------| }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 2 04:14:47 2005
A total load of crap (forgive my bluntness). Anyone wishing to argue the point is welcome to do so directly with me, off the listserver (don't reply to all, just me).
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street Saint Charles, Illinois 60174 phone (630) 513-7093 fax (630) 513-7092 email mailto:ars-at-sem.com web www.sem.com
On Wednesday, June 01, 2005 3:16 PM, Elaine Humphrey [SMTP:ech-at-interchange.ubc.ca] wrote: } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Hello everyone } I have just had a request for the use of a somatoscope developed by } Gaston Naessens as they want colour at 30,000 times. } } I was wondering if someone was pulling my leg but when I did a google } search I got } http://www.sumeria.net/tech/naessens.html } and I am still wondering if someone has a huge hoax going. Has anyone } got experience with this type of microscope? } } Elaine } } } -- } Dr. Elaine Humphrey } Director, BioImaging Facility } President, Microscopy Society of Canada (2003-2005) } University of British Columbia } 6270 University Blvd, mail-stop Botany } Vancouver, BC } CANADA, V6T 1Z4 } Phone: 604-822-3354 } FAX: 604-822-6089 } e-mail: ech-at-interchange.ubc.ca } website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 2 10:56:12 2005
A co-worker asked me to assist him with the following problem. He has metal 3 m.m. diameter discs that are about 0.25 m.m. thick. They are radioactive due to being neutron irradiated. A fixture has been made which resembles a punch and die to mechanically deform the discs at 300 degrees centigrade by hydralic pressure applied to a central rod about 1 m.m. In diameter. The "viewing hole" below the disc will be 1.5 m.m. in diameter by 3 centimeters long or "deep." What is needed is a telescopic device which can both illuminate the surface of the disc and transmit an image of it to a viewing device or camera which is several inches away.
Bernie Kestel Material Science Division Argonne National Laboratory 9700 South Cass Avenue Argonne, Illinois, 60439
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 2 11:02:09 2005
It's a lovely idea - the cure for cancer and everything else - and if you read it fast enough it almost makes sense. And it comes complete with a conspiracy theory: "Unfortunately, Rife's success attracted the attention and wrath of the American Medical Association (AMA) and the powerful pharmaceutical companies - the organised opposition of the medical fields", which makes it even more authentic.
-- Lesley Weston
} From: Elaine Humphrey {ech-at-interchange.ubc.ca} } Date: Wed, 01 Jun 2005 13:16:14 -0700 } To: microscopy-at-msa.microscopy.com } Subject: [Microscopy] gaston naessens and somatid biology } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Hello everyone } I have just had a request for the use of a somatoscope developed by } Gaston Naessens as they want colour at 30,000 times. } } I was wondering if someone was pulling my leg but when I did a google } search I got } http://www.sumeria.net/tech/naessens.html } and I am still wondering if someone has a huge hoax going. Has anyone } got experience with this type of microscope? } } Elaine } } } -- } Dr. Elaine Humphrey } Director, BioImaging Facility } President, Microscopy Society of Canada (2003-2005) } University of British Columbia } 6270 University Blvd, mail-stop Botany } Vancouver, BC } CANADA, V6T 1Z4 } Phone: 604-822-3354 } FAX: 604-822-6089 } e-mail: ech-at-interchange.ubc.ca } website: www.emlab.ubc.ca }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 2 12:10:46 2005
All you likely need is a consumer digital camera that has a video output for a TV monitor and an small achromatic lens of the right focal length mounted in front, and most of its surface apertured out to shield stray light. Some Coolpix cameras are nice because the front lens surface stays fixed while zooming or focusing.
If you mount a piece of a clean cover slip at the right angle just outside the hole and then have a bright collimated light beam reflected into the hole by the tilted glass, there should be little scattering and your camera setup should get a good hole bottom picture through the cover glass.
I am sure there are also many more fancy and expensive ways to do the same job. -- Roger
On Jun 2, 2005, at 10:55 AM, Bernie Kestel wrote:
} } } ---------------------------------------------------------------------- } -------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } } A co-worker asked me to assist him with the following problem. } He has metal 3 m.m. diameter discs that are about 0.25 m.m. } thick. They } are radioactive due to being neutron irradiated. A fixture has } been made } which resembles a punch and die to mechanically deform the discs } at 300 } degrees centigrade by hydralic pressure applied to a central rod } about } 1 m.m. In diameter. The "viewing hole" below the disc will be } 1.5 m.m. } in diameter by 3 centimeters long or "deep." } What is needed is a telescopic device which can both } illuminate the } surface of the disc and transmit an image of it to a viewing } device or camera which is several inches away. } } Bernie Kestel } Material Science Division } Argonne National Laboratory } 9700 South Cass Avenue } Argonne, Illinois, 60439 } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 2 14:39:47 2005
SEM/TEM Semiconductor and Nanotechnology Sample Prep Workshop June 7, 2005 Austin, TX
Cerium Laboratories LLC. and Sagitta Inc. will be holding a FREE workshop to present applications of automated polishing to sample preparations from semiconductor and nanotechnology materials for SEM, TEM, and AFM based evaluations.
Two sessions will be held. The morning session will concentrate on SEM prep, while the afternoon session will be addressing TEM and SRP-AFM techniques. There will be short presentations and live demonstration of automated polishing followed by SEM or TEM imaging of these samples using equipment at CeriumLabs. A possibility exists for preparation of specimens brought by participants and imaging sessions on June 8 and 9. Please contact us to set them up.
For more information or to register, please contact: Jerzy Gazda - Jerzy.Gazda-at-ceriumlabs.com - 800-538-8450, Ext. 51453; or Efrat Raz - Efrat.Raz-at-sagitta.com - 408-249-9060.
Please RSVP
Jerzy
****************************************************** Jerzy Gazda, Ph.D. CeriumLaboratories L.L.C.
Supervising Engineer 5204 E. Ben White Blvd. - MS 512 Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 jerzy.gazda-at-ceriumlabs.com ******************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 2 20:12:34 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (brian.leonard-at-sunhealth.org) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, June 2, 2005 at 16:33:18 ---------------------------------------------------------------------------
Email: brian.leonard-at-sunhealth.org Name: Brian Leonard
Organization: Sun Health Research Institute
Title-Subject: [Microscopy] [Filtered] Human leukocyte embedment and thin sectioning
Question: Does anyone have a protocol for em processing of buffy coat from human whole blood? I plan on doing post-embedding immuno on thin sections of leukocytes. The ultrastructure of the cells processed with my current method is terrible. I fractionate EDTA-treated whole blood over a Ficoll gradient to get leukocytes, and then fix the buffy coat cells at 4 deg C in 2.5% glut/4% para in 0.1M PB (or pbs). After pelleting, I resuspend the cells in low-melting agarose (37-40 deg) to put the cells in a stable matrix for the remainder of the processing. After dissecting the gel matrix into ~1mm cubes, the remainder of the processing is standard em processing for tissue sections, with final embeddment in L.R. White. Any help is much appreciated. Thanks
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lcgould-at-med.cornell.edu) from http://microscopy.com/MLFormMail.html on Thursday, June 2, 2005 at 19:47:23 ---------------------------------------------------------------------------
Organization: Joan & Sanford I. Weill Medical College
Title-Subject: [Microscopy] [Filtered] viaWWW: osmium fixation of cultured cells
Question: Robert, For membrane preservation I use a fixation protocol given to me years ago by someone who studied photoreceptors (tons of membrane). Her recipes were as follows: Primary fix: 2.5% glut, 4% pfa and 0.2% picric acid in 0.1M cacodylate buffer, pH 7.3 Post fix: 1% osmium tetrox., 1.5% Pot. ferricyanide (aqueous)
I also en bloc stain with Ur Ac.
for the picric acid (my office of environmental health and safety just loves me) I keep the smallest jar available with the crystals fully covered with water. I use the resultant saturated solution in my primary fix (2 ml picric acid sol'n to 40 ml fix).
The primary fix is based on one published by Somogyi and Takagi in Neuroscience Vol 7 No 7 pp1779-1783, 1982
Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (h.cao-at-genesis.co.nz) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, June 2, 2005 at 22:12:27 ---------------------------------------------------------------------------
Email: h.cao-at-genesis.co.nz Name: Helen Cao
Organization: Genesis R & D Ltd
Education: Graduate College
Location: Auckland, New Zealand
Question: Dear all,
I am having some problems in examining fluorescence in skin tissues. I injected (intradermally ) a fluorescence marker into mouse skin, then fixed skin tissues in 4% PFA at 4C overnight, washed in PBS (3 times, 20min each time at 4C), 20% sucrose at 4C overnight. The skin tissues were then embedded in OCT and frozen over liquid nitrogen. No good sections could be obtained, the microanatomical structures in the sections were fragmented. I tried to freeze fresh skin (unfixed) and obtained very good morphology. Any suggestions on how to preserve the morphology and fluorescence in skin tissues? Can I freeze fresh skin tissues to get good sections, and then fix the tissues before examine the fluorescence? Or, can I fix skin tissues in 4% PFA, get paraffin sections and then examine fluorescence? Do these methods quench the fluorescence?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (adriana-at-cab.cnea.gov.ar) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, June 3, 2005 at 07:29:20 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: TEM EDS for Philips CM200 UT
Question: Dear microscopists,
We are interested in buying an EDS detector for a Philips CM200 TEM with an ultratwin lens. We would like to have user experience on the performance of the following EDS systems:
a) Noran System SIX model provided by Thermo Electron, or similar models, that have no shutter, are non-retractable but have automatic FET protection.
b) EDAX Genesis 2000 model, that is non-retractable and has no shutter protection (because of the ultratwin lens).
We are particularly concerned with the damage to the detector when the microscope operates at low magnification. We would like to know whether the detector can be permanently damaged if it receives a high dose of radiation, or if the only consequences are practical (wait until the system decays).
All comments will be appreciated.
Thank you for your help.
Adriana
---------------------------------------------------------- Adriana CondÛ CONICET Researcher Metals Physics Group Centro AtÛmico Bariloche 8400 San Carlos de Bariloche ARGENTINA e-mail: adriana-at-cab.cnea.gov.ar http://www.cab.cnea.gov.ar/cab/invbasica/metales fax: +54 2944 445299 TE: +54 2944 445290 ----------------------------------------------------------
I work in a Dermatology/Laser Research Lab and we cut frozen sections of skin all the time for fluorescent studies and confocal work.
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:h.cao-at-genesis.co.nz] Sent: Thursday, June 02, 2005 11:41 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (h.cao-at-genesis.co.nz) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, June 2, 2005 at 22:12:27 ---------------------------------------------------------------------------
Email: h.cao-at-genesis.co.nz Name: Helen Cao
Organization: Genesis R & D Ltd
Education: Graduate College
Location: Auckland, New Zealand
Question: Dear all,
I am having some problems in examining fluorescence in skin tissues. I injected (intradermally ) a fluorescence marker into mouse skin, then fixed skin tissues in 4% PFA at 4C overnight, washed in PBS (3 times, 20min each time at 4C), 20% sucrose at 4C overnight. The skin tissues were then embedded in OCT and frozen over liquid nitrogen. No good sections could be obtained, the microanatomical structures in the sections were fragmented. I tried to freeze fresh skin (unfixed) and obtained very good morphology. Any suggestions on how to preserve the morphology and fluorescence in skin tissues? Can I freeze fresh skin tissues to get good sections, and then fix the tissues before examine the fluorescence? Or, can I fix skin tissues in 4% PFA, get paraffin sections and then examine fluorescence? Do these methods quench the fluorescence?
I'm looking for some advice. I understand Adobe has issued Photoshop 8 and called it Photoshop CS2. I am currently using Photoshop 6 with Reindeer Graphics's Fovea Pro V3. Is anyone actually using Photoshop CS2 and is it better than Photoshop 7 and will my Povea Pro 3 work with it. I don't want to be too far behind the curve as technology changes, but I don't want to be the public debuger either!
Thanks!!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
This e-mail transmission, and any documents, files or previous e-mail messages attached to it may contain information that is confidential or legally privileged. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that you must not read this transmission and that any disclosure, copying, printing, distribution or use of any of the information contained in or attached to this transmission is STRICTLY PROHIBITED. If you have received this transmission in error, please immediately notify the sender by telephone or return e-mail and delete the original transmission and its attachments without reading or saving in any manner. Thank you.
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 3 10:43:44 2005
Brian: I have fixed leukocytes that were grown on coverslips but I think that the same technique may solve your problem. First: keep PBS away from any cells that you ever wish to do EM on. This makes a noticeable difference. LM /confocal investigators do not seem to worry about it. Second: try fixing the buffy coat at room temperature unless it is already cold - then cool it down if protocol requires it. Third: the fixative may work better if it was less concentrated. 1%glutaraldehyde seems to be the top concentration for immuno. studies but freshly prepared (para)formaldehyde is used in many labs by itself or in combination with glutaraldehyde. Others will be sure to remark on this point. You are welcome, Pat Connelly Dept. of Biology Univ. of Pennsylvania Philadelphia, PA 19104-6018 psconnel-at-sas.upenn.edu =================== Below is the result of your feedback form (NJZFM-ultra-55).
Email: brian.leonard-at-sunhealth.org Name: Brian Leonard Organization: Sun Health Research Institute Title-Subject: [Microscopy] Human leukocyte embedment and thin sectioning Question: Does anyone have a protocol for em processing of buffy coat from human whole blood? I plan on doing post-embedding immuno on thin sections of leukocytes. The ultrastructure of the cells processed with my current method is terrible. I fractionate EDTA-treated whole blood over a Ficoll gradient to get leukocytes, and then fix the buffy coat cells at 4 deg C in 2.5% glut/4% para in 0.1M PB (or pbs). After pelleting, I resuspend the cells in low-melting agarose 37-40 deg.) to put the cells in a stable matrix for the remainder of the processing.After dissecting the gel matrix into ~1mm cubes, the remainder of the processing is standard EM processing for tissue sections, with final embeddment in L.R. White. Any help is much appreciated. Thanks
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 3 10:52:07 2005
Photoshop 8 came out with CS1. I am using it. I use lots of complex scripts and the Photoshop 8 scripting was worth my money. It has lots of other great things, but most of them are of no use to microscopists (unless you want to forge your data :-) David
On Jun 3, 2005, at 8:30 AM, frank.karl-at-degussa.com wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } I'm looking for some advice. I understand Adobe has issued Photoshop } 8 and } called it Photoshop CS2. I am currently using Photoshop 6 with } Reindeer } Graphics's Fovea Pro V3. Is anyone actually using Photoshop CS2 and } is it } better than Photoshop 7 and will my Povea Pro 3 work with it. I don't } want to be too far behind the curve as technology changes, but I don't } want } to be the public debuger either! } } Thanks!! } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 } } } 330-668-2235 Ext. 238 } } } This e-mail transmission, and any documents, files or previous e-mail } messages attached to it may contain information that is confidential or } legally privileged. If you are not the intended recipient, or a person } responsible for delivering it to the intended recipient, you are hereby } notified that you must not read this transmission and that any } disclosure, } copying, printing, distribution or use of any of the information } contained } in or attached to this transmission is STRICTLY PROHIBITED. If you have } received this transmission in error, please immediately notify the } sender } by telephone or return e-mail and delete the original transmission and } its } attachments without reading or saving in any manner. Thank you. } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 3 10:54:58 2005
I'm using Adobe Creative Suite CS2 Premium to produce Microscopy Today. The package includes Photoshop CS2. The first thing I did after installing the suite was to install my Fovea Pro 3 filters, etc. Everything works perfectly. What you see in Microscopy Today is testament. The July issue, in the works, will be the first one done all in CS2. There are some very valuable improvements in PS 7 (CS) and more in PS 8 (CS2). Adobe is beginning to recognize the scientific market to our considerable benefit.
I have no interest in Adobe other than being a very satisfied user.
Ron Anderson, Editor Microscopy Today
frank.karl-at-degussa.com wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 3 12:23:26 2005
I'm using Photoshop CS, which is Photoshop 8 (the number shown when it first opens), with Fovea Pro 3 and Optipix 3 with no problems whatsoever. I don't know the numerical designation for Photoshop CS2 but will learn more about it tomorrow in a class.
Damian Neuberger, Ph.D.
We have a Philips EM420T on which one of the push-button function selector switches (The M button for image mode) is starting to go bad. (It sometimes does not engage smoothly and the OL current then changes.)
Replacement parts for these old instruments are depressingly expensive.
Does anyone have a decommissioned Philips TEM of the same vintage, and, if so, would you be willing to give us (or sell us at a reasonable price) spare parts like the aforementioned switch?
Or can anyone suggest a less expensive alternative than buying a $1500 switch from FEI?
Thanks to anyone with any suggestions! _________________________________
Joseph Kulik Research Associate Materials Research Institute The Pennsylvania State University 194 MRI Bldg University Park, PA 16802
David is right. Photoshop CS2 is PS 9, not PS 8 as I said in my previous post. Check out Adobe site to see a list of new and improved features.
Ron Anderson
David Elliott wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Photoshop 8 came out with CS1. I am using it. I use lots of complex } scripts and the Photoshop 8 scripting was worth my money. } It has lots of other great things, but most of them are of no use to } microscopists (unless you want to forge your data :-) } David } } } On Jun 3, 2005, at 8:30 AM, frank.karl-at-degussa.com wrote: } } } } } } } ----------------------------------------------------------------------- } } ------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------- } } -------- } } } } I'm looking for some advice. I understand Adobe has issued } } Photoshop 8 and } } called it Photoshop CS2. I am currently using Photoshop 6 with } } Reindeer } } Graphics's Fovea Pro V3. Is anyone actually using Photoshop CS2 and } } is it } } better than Photoshop 7 and will my Povea Pro 3 work with it. I don't } } want to be too far behind the curve as technology changes, but I } } don't want } } to be the public debuger either! } } } } Thanks!! } } } } Frank Karl } } Degussa Corporation } } Akron Technical Center } } 3500 Embassy Parkway } } Suite 100 } } Akron, Ohio 44333 } } } } } } 330-668-2235 Ext. 238 } } } } } } This e-mail transmission, and any documents, files or previous e-mail } } messages attached to it may contain information that is confidential or } } legally privileged. If you are not the intended recipient, or a person } } responsible for delivering it to the intended recipient, you are hereby } } notified that you must not read this transmission and that any } } disclosure, } } copying, printing, distribution or use of any of the information } } contained } } in or attached to this transmission is STRICTLY PROHIBITED. If you have } } received this transmission in error, please immediately notify the } } sender } } by telephone or return e-mail and delete the original transmission } } and its } } attachments without reading or saving in any manner. Thank you. } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 3 17:03:34 2005
Could I ask those who do pre-embedding immunogold labeling routinely to contact me off-line? I would like to discuss with you a few issues regarding silver enhancement. Thank you in advance.
Hong Emory SOM EM
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 3 22:26:45 2005
In a message dated 6/3/05 11:14:03 AM, frank.karl-at-degussa.com writes:
} I'm looking for some advice. I understand Adobe has issued Photoshop 8 } and } called it Photoshop CS2. I am currently using Photoshop 6 with Reindeer } Graphics's Fovea Pro V3. Is anyone actually using Photoshop CS2 and is } it } better than Photoshop 7 and will my Povea Pro 3 work with it. I don't } want to be too far behind the curve as technology changes, but I don't } want } to be the public debuger either!
CS2 (aka Photoshop 9) has quite a few new capabilities, but whether you need them is hard to say. The handling of 16 bit images is quite a bit smoother and more complete than in PS7, and the layers capability is also much better. The Fovea Pro plugins work just fine with CS2, but you will have to re-record any actions you may have set up to automate Photoshop as they do not translate smoothly from PS9 to PS9.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Sat Jun 4 08:19:44 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dgrimm-at-tulane.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, June 3, 2005 at 09:50:36 ---------------------------------------------------------------------------
Email: dgrimm-at-tulane.edu Name: Deborah Grimm
Organization: Tulane University
Title-Subject: [Microscopy] [Filtered] Used Microscopes
Question:
AMRAY 1700 SEM, has not been under vacuum for over 1 year, but should be operational, as-is where-is, any reasonable offer accepted.
JEOL JSM 820 SEM, analog, probably a parts machine. Any reasonable offer accepted.
We need to remove these microscopes from our lab by June 17th. Please e-mail me if you have any questions.
The latest issue of Microscopy Today carries another of Jerry Sedgewick’s articles about Photoshop techniques. Like every one of his others, it contains much that is flat-out wrong or misleading, omits important information, and even the parts that will actually work are by far not the best, easiest, or most useful ways to accomplish the same purpose. I’ve tried in the past to send corrective information (which has never gotten any response from either Sedgewick or from Ron Anderson, who continues to accept these articles even though several people besides me have complained about them and pointed out the numerous errors). I suppose the need to fill up the magazine with unreviewed and blatantly incorrect stuff outweighs any other criteria, which is unfortunate because it reflects badly on other articles that are contributed by serious and respectable scientists and doesn't help the magazine achieve respectability.
Anyway, to the subject: The article concerns methods for combining stereo pair images to produce color anaglyphs. It is important to note some errors. First, the drivel about converting 16 bit to 8 bit images is incorrect. You can directly construct anaglyphs with either 8 or 16 bit images (or even a mixture, which is admittedly unlikely), since Photoshop’s latest two versions (8 and 9, also known as CS and CS2) handle 16 bit images just fine. And even if you do want to convert a 16 bit image to 8 bits, the procedure described in the article and shown in Figure 1 is wrong. That is the method (which was published some time ago in this same magazine by Brent Neal) applies to converting a 12 bit image to 16 bits, and has nothing whatever to do with the subject at hand. If for some reason you do want to convert a 16 bit image to 8 bits, just select Image-} Mode-} 8 bits per channel. Done.
But the main part of the article concerns putting together two images into the color channels of a composite anaglyph. There is no reason whatever to use the cumbersome procedure described, with extra black images and the merge channels routine and awkward steps to align things. Just do the following (this works with versions of Photoshop at least back to 6 - I haven’t tried older ones but it probably works with 5 as well): 1) Select-} All and Edit-} Copy the left eye image to place it on the clipboard 2) Change the mode of the right eye image from Grayscale to RGB Color. This will not change the appearance of the image on the screen, but if you look at the Channels palette you will see that there are now three (RGB) channels all containing the same data. 3) Click on the Red channel in the palette and paste in the image. The Sedgewick article omits the important fact that the convention for viewing stereo anaglyphs is to use the red channel for the left eye and either green or blue for the right, and that is the way glasses are constructed. It’s difficult to tell in the printed article, but it appears that the example shown in Figure 2 was put together backwards. 4) Click the eye symbol for RGB to display all of the channels. The original right eye image is now shown in both green and blue, which works with either the red/green or red/blue glasses and is brighter and easier to see than using just one of those two channels.
You may be finished now, and can crop the image if desired and save it. But if the alignment of the images is imperfect - either there is a vertical shift between the images, or one is slightly rotated, or you just want to change the horizontal displacement to alter the impression of depth - you can do it now without any of the “take it apart, fiddle around, and try again†advice in the Sedgewick article. The red channel is still selected from the steps above. You can shift it (this is usually done more controllably with the arrow keys than with the mouse - use the arrows to move the red channel a pixel at a time, or depress the shift key as well to move it ten pixels at a time), or even rotate it using the Edit-} Transform function. The key here is that you are viewing the full color anaglyph through your glasses, because you turned on the RGB visibility in step 4, and can therefore immediately see what your shifting of the image does visually, which gives precise control. All of the stuff in the Sedgewick article about locking or unlocking the layer, splitting and merging the channels, creating an additional image, and so on, is just unnecessary and confusing, and makes the task more difficult.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Sat Jun 4 10:03:24 2005
There is a demo version of CS2 that you can download and run. It has a lot of fun new stuff, a lot definitely NOT useful for scientific imaging. But two features for making figures that may be worth the ungrade are 1.) the ability to shrink and reenlarge images without throwing out the original pixels (I think this is from Illustrator) and 2.) a more sensible font selection tool.
} Is anyone actually using Photoshop CS2 and is } } it } } better than Photoshop 7 and will my Povea Pro 3 work with it. I don't } } want to be too far behind the curve as technology changes, but I don't } } want } } to be the public debuger either!
_________________________________________ Michael Cammer Analytical Imaging Facility and Dept. ASB Biophotonics Innovation Laboratory Albert Einstein College of Medicine 1300 Morris Park Avenue, Bronx, NY 10461 718-430-2890 Fax 718-430-8996 work: http://www.aecom.yu.edu/aif/ personal: http://coxcammer.com/
From MicroscopyL-request-at-ns.microscopy.com Sat Jun 4 16:31:31 2005
I'm trying to customize a control application for a BX Olympus motorized microscope but is short of a device driver kit for it (DDK). My problems are determining the exact communications protocol to connect the serial port to the controlling box.
Any one has the DDK or the protocol description?
Regards,
Berns B.
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 6 09:27:32 2005
Hoax/fraud/BS, pick one. Certainly not true.You might quote the requestor an outrageous price and see how they respond. :-)
Geoff
} On Wed, 1 Jun 2005, Elaine Humphrey wrote: } } } ------------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } } } Hello everyone } } I have just had a request for the use of a somatoscope developed by } } Gaston Naessens as they want colour at 30,000 times. } } } } I was wondering if someone was pulling my leg but when I did a google } } search I got } } http://www.sumeria.net/tech/naessens.html } } and I am still wondering if someone has a huge hoax going. Has anyone } } got experience with this type of microscope? } } } } Elaine } } } } } } -- } } Dr. Elaine Humphrey } } Director, BioImaging Facility } } President, Microscopy Society of Canada (2003-2005) } } University of British Columbia } } 6270 University Blvd, mail-stop Botany } } Vancouver, BC } } CANADA, V6T 1Z4 } } Phone: 604-822-3354 } } FAX: 604-822-6089 } } e-mail: ech-at-interchange.ubc.ca } } website: www.emlab.ubc.ca } } } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 6 10:31:19 2005
Hello there, First I wish to express my gratitude towards those list members who helped me out with the electrical schematics and manuals for the Cambridge S-120. Thanks alot!
Secondly, the samples seem to be charging/arching in a pulsating manner, even when the sample is thoroughly grounded. I know the column is somewhat dirty, do anyone know if this could be related to the trouble. If not do anyone have any ideas as what might be the cause.
Are you certain the sample is grounded? We have some coated samples that it seems we can never put enough gold on to eliminate charging. The samples are insulating and rough. I can only examine small aggregates of particles without encountering too much charging. I would try looking at a piece of well-grounded, clean metal to be sure there is no chance of it being the sample.
I went through that exercise with a dirty column a couple years ago. The service engineer strongly suspected the sample, but when I loaded a bare sample holder and still got drifting, he was finally convinced. He tore our column down ALL the way and found some contamination deep within. I think our scope was seven-years-old at the time. The location does not get dirty so fast that it requires cleaning at every preventative maintenance, but we will want to watch for problems and probably clean it at least every 4 or 5 years.
Warren
At 10:29 AM 06/06/05, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
The samples seem to be charging/arching in a pulsating manner, even when the sample is thoroughly grounded. I know the column is somewhat dirty, do anyone know if this could be related to the trouble. If not do anyone have any ideas as what might be the cause. Skage Hem {SHem-at-laurentian.ca} ============ Skage, Does your Cambridge S-120 have a spark plug or someplace where there is a gap like a spark plug? [Stop laughing!]
I have a pulsing when a fleck of carbon gets built up inside the gap of the spark plug in my Philips 200. It took weeks to find this answer many years ago and it has occured only once in the last 10 years. The cure is to run a piece of paper through the gap which is very small to remove the carbon without needing to remove the spark plug or readjust the gap space.
Pat Connelly psconnel-at-sas.upenn.edu The University of Pennsylvania Department of Biology Philadelphia, PA 19104-6018 215-898-7145
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 6 11:21:44 2005
I am considering buying Reindeer Graphics Focus Extender 1.0, and would like to hear feedback from actual users. A search of the Web turned up little except copies of the original press release. I would like to know about ease of use, how well the program handles various types of subjects, and how it stacks up (pun intended) against similar programs. On a related topic, does anybody know of a source for a computer driven Z-drive that doesn't cost a fortune?
Ralph Common Michigan State University Division of Human Pathology
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 6 11:25:11 2005
I tried two other sets of specimens and they all heated up. The ambient temperature is 70F. The heated temperature is 95F.
One specimen is a sputter coated (Pt) #1 cover slip stuck on a pin stub via Nisshin EMS carbon tab. The slip is offset slightly so I can ground it using Pella colloidal silver.
When I put this in the chamber, no beam, for about 10 minutes, I pull it out and it is hot.
Then I did three Al film on silicon pieces on stubs which were attached using Buehler wax but grounded using the Pella colloidal silver. This large holder also got up to 95F in chamber vacuum without any beam.
After some amount of time in air, the specimens seem to not heat up. What is going on?
Strange. But it messes up image collection due to large amount of drift.
gary g.
At 08:30 AM 5/31/2005, you wrote: } Just a wild guess, but would the sample have heated up the same if mounted } the same way and placed in a vacuum? I wonder if their might be something } galvanic going on with all the metals involved. Maybe the beam had nothing } to do with it. } } Warren } } At 04:49 PM 05/29/05, you wrote: } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 6 11:44:10 2005
Asylum Research is seeking a Technical Sales Executive to manage its UK sales operations. The candidate should have experience both in sales and technical applications for scanning probe/atomic force microscopy (SPM/AFM) or scientific instrumentation/metrology tools. Additional information on the position may be found at http:// www.AsylumResearch.com.
Terry Mehr Director of Marketing Asylum Research www.AsylumResearch.com
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 6 11:54:28 2005
We have scavenged the optics column from a defunct ISI WB6 electron microscope (tungsten filament) in order to build an electron interferometer. Does anybody out there have any technical specifications, engineering drawings, electrical schematics, etc. for this equipment? We already have the user manual and an adjustment manual.
Specifically, while we have been able to roughly focus and scan the beam using our own power supplies, it would be really nice to know, for example, coil currents that correspond to given focal lengths from a particular lens. Since we're doing everything "by hand", we would love to know where all the beam focal points are located. Dunno if that information is publicly available, but I thought I'd ask. Thanks for your time!
--Ben McMorran
-- Ben McMorran Research Assistant, Atom Optics Group Department of Physics University of Arizona 1118 E 4th St Tucson, AZ USA
ph. 520-621-2688
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 6 17:37:09 2005
"Introduction to the Techniques of Forensic Soil Comparison" Workshop Conducted by Skip Palenik Inter/Micro 2005 McCrone Research Institute July 14-15, 2005
This 2-day workshop, conducted by Skip Palenik, renowned expert in the field of forensic microscopy and microanalysis, introduces innovative techniques for the examination and interpretation of soil samples and soil comparison evidence. Using a combination of several techniques (including visual and microscopical methods, filtration, ultra-sonication, sieving, and chemical treatments) soil samples can be separated into useful fractions. You will learn and practice Skip's modified fractionation techniques to investigate the most distinguishing particles that might ordinarily be overlooked, present in low quantities, or hidden by clay and humic materials. This workshop is unique in that it emphasizes a microscopical approach on the level at which soil is most fundamentally distinct, i.e., on a particle-by-particle basis. Forensic scientists from around the world have verified that the use of these techniques on soil evidence has helped them to identify the original source of an unknown sample in addition to facilitating the process of comparison of known and unknown samples from almost any geographical location.
The workshop will be conducted in the laboratories and classrooms of McCrone Research Institute in Chicago, Illinois. The cost of the workshop is $200 which includes the use of equipment, teaching materials, lunch, snacks, and refreshments. Please register early http://mcri.org/IM_03homep.html, class size is limited.
Regards, Christine
Christine Weaver Inter/Micro Coordinator McCrone Research Institute 2820 S. Michigan Avenue Chicago IL 60616
Problem uncovered. Reason why is not totally clear.
The situation is that if the colloidal silver is applied and then the specimen is put in the SEM after about two to three hours, it gets hot. Leave the specimen in air or out for a day, put in SEM, no heat. OK. So, what seems to be happening is that the binders in the colloidal silver product outgasses rapidly in a vacuum and accelerates an otherwise slow and un-noticed heating when done in air over a long period of time.
So, the solution is to either vacuum dessicate the stub before putting in the SEM or wait over night before putting into the SEM.
Any other explanations?
gary g.
At 09:54 AM 6/6/2005, you wrote: } Well, it seems to me that you can't have a sample heating up that much } without the stage getting hot too. And you'd need a fair bit of } electrical current dissipating in resistive heat to cause it, must be } several hundred mA (depending how much material is getting hot). Seems } like you have a hot stage. Perhaps a dodgy motor on the stage drive, or } heat transmitted from a diff pump (can't see how though). Or friction, } does it depend how much you move the stage about? I guess the stage is } warm to the touch? } } When you say: } } 'After some amount of time in air, the specimens seem to not } heat up. What is going on?' } } Do you mean the specimen has been in air for a while, you put it back in, } and it doesn't get hot - or do you mean when the chamber is vented up to } air, the specimen cools down. (I guess the latter, cooling by convection } currents in the air then.) } } So I guess I would look for electrical current going into the stage (eg } pull the plug from the stage drive controller board, see if the effect } goes away), friction, or something connected to the stage that is warm.. } Interesting one though, I'd be intrigued to know what it turns out to be. } } Good luck } } Richard } } } ________________________________________ } Richard Beanland } Analytical Services } Bookham Inc } Caswell } Towcester } Northants } NN12 8EQ } United Kingdom } Tel. +44 1327 356362 } Fax. +44 1327 356775 } http://www.bookham.com } ________________________________________ } } } -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: 06 June 2005 17:24 } To: MSA listserver } Subject: [Microscopy] [MICROSCOPY] specimen heating up } } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 6 18:43:28 2005
For manuals, contact Nikon directly (email/phone) and they will send you a manual or a copy of one. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Sat, 4 Jun 2005, km602223-at-comcast.net wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Does anyone have an instruction manual for a Nikon Fluophot microscope? } Thank you, } Kathleen }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 6 20:02:51 2005
Who said that laws cannot be broken? Speeders do it all the time. Red light runners...etc. Let's give someone a ticket!
All I can say is that this is what I see. Try the experiment for yourself and see what you get. perhaps there is a variable in this situation that I am missing.
gary g.
At 06:14 PM 6/6/2005, you wrote: } Huh? You've uncovered a violation of the second law of thermodynamics } where the } transition liq } gas, or adsorbed specie } gas is exothermic in spite of the } increase in free energy???? } } John } } Gary Gaugler wrote: } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------- } } } } Hi all: } } } } Problem uncovered. Reason why is not totally clear. } } } } The situation is that if the colloidal silver is applied } } and then the specimen is put in the SEM after about two to } } three hours, it gets hot. Leave the specimen in air or } } out for a day, put in SEM, no heat. OK. So, what seems } } to be happening is that the binders in the colloidal silver } } product outgasses rapidly in a vacuum and accelerates an } } otherwise slow and un-noticed heating when done in air over } } a long period of time. } } } } So, the solution is to either vacuum dessicate the stub } } before putting in the SEM or wait over night before putting } } into the SEM. } } } } Any other explanations? } } } } gary g. } } } } At 09:54 AM 6/6/2005, you wrote: } } } Well, it seems to me that you can't have a sample heating up that much } } } without the stage getting hot too. And you'd need a fair bit of } } } electrical current dissipating in resistive heat to cause it, must be } } } several hundred mA (depending how much material is getting hot). Seems } } } like you have a hot stage. Perhaps a dodgy motor on the stage drive, or } } } heat transmitted from a diff pump (can't see how though). Or friction, } } } does it depend how much you move the stage about? I guess the stage is } } } warm to the touch? } } } } } } When you say: } } } } } } 'After some amount of time in air, the specimens seem to not } } } heat up. What is going on?' } } } } } } Do you mean the specimen has been in air for a while, you put it back in, } } } and it doesn't get hot - or do you mean when the chamber is vented up to } } } air, the specimen cools down. (I guess the latter, cooling by convection } } } currents in the air then.) } } } } } } So I guess I would look for electrical current going into the stage (eg } } } pull the plug from the stage drive controller board, see if the effect } } } goes away), friction, or something connected to the stage that is warm.. } } } Interesting one though, I'd be intrigued to know what it turns out to be. } } } } } } Good luck } } } } } } Richard } } } } } } } } } ________________________________________ } } } Richard Beanland } } } Analytical Services } } } Bookham Inc } } } Caswell } } } Towcester } } } Northants } } } NN12 8EQ } } } United Kingdom } } } Tel. +44 1327 356362 } } } Fax. +44 1327 356775 } } } http://www.bookham.com } } } ________________________________________ } } } } } } } } } -----Original Message----- } } } From: Gary Gaugler [mailto:gary-at-gaugler.com] } } } Sent: 06 June 2005 17:24 } } } To: MSA listserver } } } Subject: [Microscopy] [MICROSCOPY] specimen heating up } } } } } } } } } } } } } } } ----------------------------------------------------------------------- } ------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ----------------------------------------------------------------------- } -------- } } } } } } I tried two other sets of specimens and they all heated up. } } } The ambient temperature is 70F. The heated temperature is } } } 95F. } } } } } } One specimen is a sputter coated (Pt) #1 cover slip stuck on } } } a pin stub via Nisshin EMS carbon tab. The slip is offset } } } slightly so I can ground it using Pella colloidal silver. } } } } } } When I put this in the chamber, no beam, for about 10 minutes, } } } I pull it out and it is hot. } } } } } } Then I did three Al film on silicon pieces on stubs which were } attached using } } } Buehler wax but grounded using the Pella colloidal silver. } } } This large holder also got up to 95F in chamber vacuum without } } } any beam. } } } } } } After some amount of time in air, the specimens seem to not } } } heat up. What is going on? } } } } } } Strange. But it messes up image collection due to large } } } amount of drift. } } } } } } gary g. } } } } } } } } } } } } At 08:30 AM 5/31/2005, you wrote: } } } } Just a wild guess, but would the sample have heated up the same if } mounted } } } } the same way and placed in a vacuum? I wonder if their might be } something } } } } galvanic going on with all the metals involved. Maybe the beam had } nothing } } } } to do with it. } } } } } } } } Warren } } } } } } } } At 04:49 PM 05/29/05, you wrote: } } } } } } } } } } } } } -------------------------------------------------------------------- } ---- } } } ------ } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------------- } ---- } } } ------- } } } } } } } } } } Hi Listers: } } } } } } } } } } I've encountered a once only situation that I cannot } } } } } explain. Perhaps someone has some ideas about what } } } } } is going on. } } } } } } } } } } I put a specimen in the SEM for examination and when } } } } } I pulled it out via the load lock, the specimen and } } } } } holder were about 10 degrees hotter than ambient. } } } } } My experience has been that whatever they go in at, } } } } } they come out the same. This was simple SE imaging } } } } } at 10KV with 30u High Current in Zeiss Supra 55VP } } } } } in high vacuum mode. Specimen current was around 145pA. } } } } } } } } } } The specimen was a copper tube that had been plated } } } } } with silver on the outside and inside. The piece was } } } } } one half of the tube and about .5" long, attached to } } } } } a pin stub using colloidal silver. } } } } } } } } } } EDS showed presence of K, O, Ag, Cu, and C. There were } } } } } distinct areas of organic material that I take to be } } } } } cyanide from the plating process. But how could SEM } } } } } analysis of a specimen cause it to heat? } } } } } } } } } } Any ideas? } } } } } } } } } } gary g. } } } } } } } } } } } } ======================================================================= } } } This e-mail is intended for the person it is addressed to only. The } } } information contained in it may be confidential and/or protected by } } } law. If you are not the intended recipient of this message, you must } } } not make any use of this information, or copy or show it to any } } } person. Please contact us immediately to tell us that you have } } } received this e-mail, and return the original to us. Any use, } } } forwarding, printing or copying of this message is strictly prohibited. } } } No part of this message can be considered a request for goods or } } } services. } } } =======================================================================
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 6 21:21:21 2005
I tried three pieces of Si coated with colloidal conductive silver and began dessicating them in a Duniway chamber. This got down to about 50mT using a Varian dual vane mech pump.
After thirty minutes, the Zeiss holder is not hot. So, next is moving it into the E-6 Torr SEM chamber. More to follow.
gary g.
At 05:58 PM 6/6/2005, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 6 22:29:04 2005
Well, I think this problem is solved/isolated. The laws of thermo are preserved. The SEM is a Zeiss Supra 55VP. It has an Edwards XDS10 scroll pump and Varian TMP. It is a dry system. Specimens are introduced via a Fjeld load lock. All together, they are great. The current problem seems to be the stage rotate motor.
It turns out that depending on some variables, the Zeiss Supra 55VP stage rotate motor gets very hot. This translates to a hot specimen holder and hot specimens.
I see no way to avoid this other than to turn off R until a new specimen needs it. This is not optimum. But it seems to be the only solution. There were many start up problems with this unit and the stage was one of them. R was a big issue. I guess that it has not gone away.
Thus, colloidal silver is not the culprit/cause.
gary g.
At 06:31 PM 6/6/2005, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 6 23:09:30 2005
} To: DrJohnRuss-at-aol.com } Subject: [Microscopy] RE: correction for latest Microscopy Today
John
I have read with interest your recent Listserver posting of a few days ago. There is useful information in your text, specifically your methodology within Photoshop tools to construct stereo pairs. However, I note with concern that the first paragraph of your posting contains content which is in violation of the Microscopy Listserver rules.
This concern is not something to be taken lightly and I will quote from the Microscopy Listserver rules, which as subscribers EVERYONE receives a copy of (http://www.microscopy.com/Rules.html), and is a condition of their continued subscription. Specifically item #4 of those rules states:
---------------------------------------------------------------------- 4.) This forum is not to be used as a platform to accuse or defame any individual or organization in any negative manner. You may disagree with any comment posted and post a reply, but this server may not be used to spread misleading, derogatory or disparaging comments under any conditions. ----------------------------------------------------------------------
In my opinion, you have crossed the line with your initial comments. It is fine to ask questions and/or offer alternatives or comments about any posting on the Listserver as that is certainly one of the purposes of this forum. However, in your posting you are making comments to the Listserver about an article which has no relation to the Listserver, the text for this technique never appeared in the Listserver or its archives, nor has anyone asked a question concerning this technique on the Listserver. Using this forum to disparage anyone is against our rules, and both Microscopy-Today and Jerry Sedgewick are no different than anyone else. I have not stood by idly when comparable situations happened in the past, nor will I do so in this situation.
I reiterate, posting information on your technique to create stereo pairs using Photoshop is fine as well as offering it as an alternative to methods of others. It is also fine that you disagree with Sedgewick (I disagree with alot of people and have disagreed with you on various issues over the years). Since you feel so strongly about this particular article in MT, I would encourage you to write a contributed article and submit it to MT about the methodoloy you just outlined, however, using the Listserver is an inappropriate forum to voice complaints or views about MT and/or Sedgewick.
If you (or anyone for that matter) feel that you are being ignored or have problems with MSA or one of its subsidiaries, please feel free to contact me off-line to discuss the situation and please send me a copy of any documentation you have submitted to the parties in question. As a former Council member I will be willing to bring your concerns to the attention of the appropriate individuals within MSA, or to the Council as appropriate.
I insist that EVERYONE using the Microscopy Listserver follows proper channels when they have problems with other organizations or individuals (MSA related or not) and no one is permitted to use the Listserver as a forum to disparage any individual or organization.
Nestor Your Friendly Neighborhood SysOp & The Microscopy Listserver SysCop
------------------------------------------------------------------------------- } From: DrJohnRuss-at-aol.com } Date: Sat, 4 Jun 2005 09:19:58 EDT } Subject: [Microscopy] correction for latest Microscopy Today } To: microscopy-at-msa.microscopy.com
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From MicroscopyL-request-at-ns.microscopy.com Tue Jun 7 10:29:57 2005
Mass Spectrometry Engineer / Technician Job Opening at Schafer Corporation
Schafer Corporation is seeking an engineer / technician for its Mass Spectrometry Group at its Vallecitos Laboratory near Sunol, California. This position supports a multi-million dollar operation of thermal ionization and secondary ion mass spectrometers and support instruments, and interacts with a team of mass spectrometry scientists, engineers and technicians. The candidate will also be expected to directly participate in the analysis of samples and in the maintenance of auxiliary equipment. Schafer Vallecitos Laboratory performs materials characterization and related analytical services on commercial and government contracts. The activities of the group include chemical analysis of materials on a production basis, maintenance of several custom-built mass spectrometers, development of new or improved mass spectrometric techniques, and quality assurance of analytical data.
This position requires an individual who has proven technical abilities. The ideal candidate must have a strong background in science or engineering, or a related field of physical or chemical science, and experience operating mass spectrometers or other analytical equipment in a laboratory environment. The candidate must be able to operate independently with little supervision. Other desired qualifications include:
- Expertise in data evaluation and quality control. - Ability to write clear scientific and technical reports. - Bachelor's degree in physical science or engineering. - Minimum 4 years of laboratory experience.
Schafer offers a comprehensive array of benefits including dental, health and life insurance, 4 weeks of vacation annually, and a 401k program. US Citizenship required, as well as qualification for a DoD security clearance - An Affirmative Action, EEO employer promoting diversity in the workplace.
Respond to job code VAL200502 - at jobs2-at-schaferlabs.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 7 11:17:29 2005
Dear Skage, There are a few other possibilities that come to mind. First, you sample may be grounded to the stub, but have you traced the ground all the way? Check the sample holder, stage and the wire that goes to the feedthrough. My stage grounding goes through a BNC plug, in case you want to test the specimen current, and sometimes the plug needs cleaning because the oxide film builds up and stops the grounding current. Remember the current is picoamps, so it doesn't take much to stop it. I have had the grounding wire in the chamber broken because it got caught in the o-ring when the chamber was pumped down. The other thing to check is the scintillator. The aluminum coating on the scintillator can become worn off by micro-discharges, until the bare plastic button is showing, and this can build up a charge. A new scintillator button is a good idea, or a new evaporated layer of Al and check that the button is well grounded. Of course dirt in the column or stage can cause these problems, as well. ----- Original Message ----- } From: "Skage Hem" {SHem-at-laurentian.ca} To: {Microscopy-at-microscopy.com} Sent: Monday, June 06, 2005 8:29 AM
I am working with digital TEM images that need flat field correction and background subtraction to reduce fixed, uneven illumination and noise (lint & electronic noise), respectively. I have a program that came with the digital camera but it is cumbersome to use (at least, so far).
In PhotoShop (version 6 and CS1) I can flatten the field using a high pass filter but have been unable to use PS to subtract noise (original image minus empty field). In PS, I tried using the Image/Calculations feature as well as Image/Apply Image features. No combination gave the desired effects.
Am I asking too much of PhotoShop, or simply not understanding how to achieve this in PS? Maybe both, eh?
Any suggestions would be appreciated.
Thank you. -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 7 14:10:51 2005
We need to make 20-30µm sections of 200µm thick stainless steel wire coated with a polymer. The object is to be able to do some chemical analysis on the sections so as to visualize chemical distribution in the polymeric coating. The sections will be imaged using Ramon technology.
We need suggestions as to embedding resin, type of microtome and type of blades that may work for sections of this thickness and samples of this type.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 7 14:30:04 2005
There are three classic approaches to levelling background. Some are easier to do in Photoshop than others. 1) If you can measure a background (ie. collect an image with no sample, or a uniform sample present) then you can use layers modes to get the difference between the original and the background. Unfortunately, depending on how your camera works you may need to either subtract the background or divide by it (if the camera is log or linear, respectively). Photoshop doesn't offer a ratio layers mode (these are the same options as in the Calculations dialog), so you would have to use a plugin (e.g., the Reindeer Fovea Pro suite which can either subtract or divide. 2) If you can remove the features from the image (e.g. if they are skinny in one dimension) to leave a background representation, then you can use that. In Photoshop you can do a slightly crude but probably adequate job of a grey scale morpholological closing or opening using the Filter-} Custom-} Maximum or Minimum routine. For example if the features are dark on a bright background, you would first use Maximum to remove the features and then Minimum with the same radius to restore the scale of the background irregularities. This background is then removed as in step 1. 3) In many cases, the gradually varying background from nonuniform illumination can be modelled mathematically. In Fovea Pro, a cubic polynomial is fitted to either the bright or dark parts of the image to construct a function for removal. Sometimes you can accomplish the same thing with a large low-pass filter (e.g., a Gaussian smooth). This blurs out the features to the point where the image shows just the background, which you can then use as in step 1.
John Russ =======
In a message dated 6/7/05 3:03:50 PM, bozzola-at-siu.edu writes:
} I am working with digital TEM images that need flat field correction } and background subtraction to reduce fixed, uneven illumination and } noise (lint & electronic noise), respectively. I have a program that } came with the digital camera but it is cumbersome to use (at least, } so far). } } In PhotoShop (version 6 and CS1) I can flatten the field using a high } pass filter but have been unable to use PS to subtract noise } (original image minus empty field). In PS, I tried using the } Image/Calculations feature as well as Image/Apply Image features. No } combination gave the desired effects. } } Am I asking too much of PhotoShop, or simply not understanding how to } achieve this in PS? Maybe both, eh? } } Any suggestions would be appreciated.
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 7 14:44:34 2005
I suspect that slicing through stainless steel wire without disturbing its polymer coating on a microscopic level is going to be a huge challenge.
The hard wire would probably mess up knives and if you try to grind a flat on the end of the wire, the abrasive will get embedded in the interface.
I would suggest simply slicing/stripping off the coating and looking at that in cross section since its surface chemistry is not going to change with the removal of the wire in contact.
-- Roger
On Jun 7, 2005, at 2:10 PM, Debby Sherman wrote:
} } } ---------------------------------------------------------------------- } -------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } This is for the material Scientists among you, } } We need to make 20-30µm sections of 200µm thick stainless steel } wire coated } with a polymer. The object is to be able to do some chemical } analysis on } the sections so as to visualize chemical distribution in the polymeric } coating. The sections will be imaged using Ramon technology. } } We need suggestions as to embedding resin, type of microtome and } type of } blades that may work for sections of this thickness and samples of } this } type. } } Thanks, } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 7 15:59:40 2005
-----Original Message----- } From: Debby Sherman [mailto:dsherman-at-purdue.edu] Sent: Tuesday, June 07, 2005 12:10 PM To: message to: MSA list
This is for the material Scientists among you,
We need to make 20-30µm sections of 200µm thick stainless steel wire coated with a polymer. The object is to be able to do some chemical analysis on the sections so as to visualize chemical distribution in the polymeric coating. The sections will be imaged using Ramon technology.
We need suggestions as to embedding resin, type of microtome and type of blades that may work for sections of this thickness and samples of this type.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 7 16:09:47 2005
Here's a favorite Photoshop recipe for background leveling digital TEM images (until someone suggests a better one). It uses Gaussian Blur as mentioned by J. Russ.
1. Open image or paste into new PS window. 2. Duplicate layer (twice) 3. Apply Gaussian Blur (50 pixels or your choice) to top layer 4. Invert image 5. Overlay image (blending mode option from Layers window) 6. Merge down
Repeat leveling operation 2 to 4 times until artifacts appear or contrast washes out. Record as Action with different blur settings of choice.
For a more inclusive view of background leveling, see Russ tutorial at reindeergraphics.com/tutorial/chap2/defect06.html
Larry Thomas } ---------- } From: John J. Bozzola } Sent: Tuesday, June 7, 2005 12:01 PM } To: Microscopy-at-msa.microscopy.com } Subject: [Microscopy] high pass filter versus flat field correction } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I am working with digital TEM images that need flat field correction } and background subtraction to reduce fixed, uneven illumination and } noise (lint & electronic noise), respectively. I have a program that } came with the digital camera but it is cumbersome to use (at least, } so far). } } In PhotoShop (version 6 and CS1) I can flatten the field using a high } pass filter but have been unable to use PS to subtract noise } (original image minus empty field). In PS, I tried using the } Image/Calculations feature as well as Image/Apply Image features. No } combination gave the desired effects. } } Am I asking too much of PhotoShop, or simply not understanding how to } achieve this in PS? Maybe both, eh? } } Any suggestions would be appreciated. } } Thank you. } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/~image/ } ############################################################## } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 7 16:12:32 2005
Debbie, Second John's suggestion whole heartedly, you can lift out a small sample with no trouble at all, let me know if you need help, Bobby Hooghan,
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, June 07, 2005 3:59 PM To: Debby Sherman; message to: MSA listDebbie,
Debbie; FIB.
John Mardinly Intel
-----Original Message----- } From: Debby Sherman [mailto:dsherman-at-purdue.edu] Sent: Tuesday, June 07, 2005 12:10 PM To: message to: MSA list
This is for the material Scientists among you,
We need to make 20-30µm sections of 200µm thick stainless steel wire coated with a polymer. The object is to be able to do some chemical analysis on the sections so as to visualize chemical distribution in the polymeric coating. The sections will be imaged using Ramon technology.
We need suggestions as to embedding resin, type of microtome and type of blades that may work for sections of this thickness and samples of this type.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 7 18:35:54 2005
It's no lie that FIB would be the best solution to this problem, but what if you don't have a FIB?? It is actually easier to cut metals very thin i.e. 20-80 nm. At this thickness, they can be examined in the EFTEM, which will analyze and map the elemental composition at high resolution.
Bill McManus Mt Ogden Scientific Services -Providing full service electron microscopy for you 950 W Kershaw, Suite E Ogden UT 84401 toll-free: 877/311-6677 direct: 801/745-2583 cell: 435/757/2975 fax: 435/514-1781 conniemoss-at-relia.net www.mtogdensci.com
------------------------------- Mardinly, John said: } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Debbie; } FIB. } } John Mardinly } Intel } } } -----Original Message----- } } From: Debby Sherman [mailto:dsherman-at-purdue.edu] } Sent: Tuesday, June 07, 2005 12:10 PM } To: message to: MSA list } Subject: [Microscopy] Sectioning stainless steel wire } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } This is for the material Scientists among you, } } We need to make 20-30µm sections of 200µm thick stainless steel wire } coated } with a polymer. The object is to be able to do some chemical analysis on } the sections so as to visualize chemical distribution in the polymeric } coating. The sections will be imaged using Ramon technology. } } We need suggestions as to embedding resin, type of microtome and type of } blades that may work for sections of this thickness and samples of this } type. } } Thanks, } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy } } } } c } } }
======
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 7 21:05:12 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sdunning-at-uga.edu) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, June 7, 2005 at 10:57:01 ---------------------------------------------------------------------------
Email: sdunning-at-uga.edu Name: Sarah Dunning
Organization: University of Georgia
Education: Graduate College
Location: Athens, Georgia
Question: I read in the literature that AFM's achieve "atomic resolution." How does one go about calculating this mathematically? Is it a matter of the tip size, geometry, tip-sample sensitivity, etc., all folded into one equation?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (przybylowicz-at-tlabs.ac.za) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 6, 2005 at 12:44:43 ---------------------------------------------------------------------------
Email: przybylowicz-at-tlabs.ac.za Name: Wojciech Przybylowicz
Organization: Materials Research Group, iThemba LABS
Title-Subject: [Microscopy] [Filtered] Search for the second hand automatic refilling system
Question: Our laboratory is looking for the automatic refilling device (7019-11006) that operates with Reichert-Jung FC4 Ultracut Cryosystem. It is composed of the control module (6527-00001), dewar with solenoid valve (7019-31030) and heating element inside, and cooling chamber (7019-1150). If you know of any laboratory willing to offer it to us for free or for a reasonable price, please let me know.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kristinat-at-hotmail.com) from http://www.microscopy.com/MLFormMail.html on Tuesday, June 7, 2005 at 11:09:35 ---------------------------------------------------------------------------
Question: I am looking for a 620 or 625 nm UV filter for a 96-well plate reader. I will get the details of model number, etc. of the plate reader it would need to fit. I am doing Minimal Inhibitory Concentration susceptibility testing on Staphylococcus aureus with gossypol for my Master's thesis research project. The Professor in the Biology department that has a 96-well plate reader that I could use to read my results does not have a 625 nm UV filter that I need. His plate reader is 10 years old, so it is pretty old.
ImageJ http://rsb.info.nih.gov/ij/ is bit harder to use but will do perfectly remove a background image from form an image by taking a blank back ground image an using the Process} math} difference to remove the blank frame from the image. I leaves an image that need to be put on a flat back ground but I haven't got a round toit yet.
ImageJ is a free tool developed by NIH and run on almost all platforms. It has a large development community writing plug ins and background is mentioned in several pulgins.
Gordon Couger Stillwater, OK www.couger.com/gcouger
} In a message dated 6/7/05 3:03:50 PM, bozzola-at-siu.edu writes: } } } } I am working with digital TEM images that need flat field correction } } and background subtraction to reduce fixed, uneven illumination and } } noise (lint & electronic noise), respectively. I have a program that } } came with the digital camera but it is cumbersome to use (at least, } } so far). } } } } In PhotoShop (version 6 and CS1) I can flatten the field using a high } } pass filter but have been unable to use PS to subtract noise } } (original image minus empty field). In PS, I tried using the } } Image/Calculations feature as well as Image/Apply Image features. No } } combination gave the desired effects. } } } } Am I asking too much of PhotoShop, or simply not understanding how to } } achieve this in PS? Maybe both, eh? } } } } Any suggestions would be appreciated. } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 8 04:09:50 2005
I would have concerns about surface contamination from FIB which could affect the results, not to mention changes in the polymer itself from damage to the ion beam. This isn't too different from the optoelectronic packages we section on occasion, which can have everything from stainless steel through glass to adhesives. It's not clear to me why you need a thin section for Raman - can't you just pot the wire in an appropriate resin (we use Struers but I'm sure other suppliers are just as good) and grind/polish as for a standard metallographic section? When there are changes in material hardness it can be difficult to keep the surface flat, but a little experimentation for your application with different pads should get you there. This should give you a nice clean and flat surface for analysis, and a few polishing pads and a bit of resin is slightly cheaper than a FIB.
-----Original Message----- } From: Debby Sherman [mailto:dsherman-at-purdue.edu] Sent: 07 June 2005 20:10 To: message to: MSA list
This is for the material Scientists among you,
We need to make 20-30µm sections of 200µm thick stainless steel wire coated with a polymer. The object is to be able to do some chemical analysis on the sections so as to visualize chemical distribution in the polymeric coating. The sections will be imaged using Ramon technology.
We need suggestions as to embedding resin, type of microtome and type of blades that may work for sections of this thickness and samples of this type.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 8 06:26:11 2005
Is anyone aware of a method for accelerating oxidation while observing with ESEM?
tia :o) michael shaffer (709) 737-8346 (voice) (709) 737-2589 (FAX) {www.micro-investigations.com} {www.esd.mun.ca/epma/} Alexander Murray Bldg, Rm ER4063 Memorial University of Newfoundland St. John's, NL Canada, A1B 3X5
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 8 06:31:53 2005
HIROX-USA, INC. } } Hirox USA, distributor of Industrial Video-Microscope systems, is seekng a } Sales Engineer who has prior experience with the sale of capital equipment. } The job requirements include: Demonstration of the instruments, travel and } the ability to lift equipment up to 100LB. } The position is for the New Jersey area. This industry is growing } rapidly and our customers include many Fortune 500 companies. see } {http://www.hirox-usa.com} www.hirox-usa.com Email to: } {mailto:mail-at-hirox-usa.com} mail-at-hirox-usa.com or Fax resume to 201.342.7322
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 8 11:53:03 2005
I just got a request from someone in China if there are any labs available in China that can do SEM/EDS work. Can anyone provide contact information?
Stu Smalinskas, P.E. Metallurgist SKF North American Technical Center Plymouth, Michigan (734) 414-6862 stu.smalinskas-at-skf.com
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 8 13:34:07 2005
To add, this request was from a manufacturer of automotive mirrors. They need EDS work on the layers of the mirrored surface. I’ve been doing this work for their customer, but they want to have it done locally for themselves.
Stu
--- Changhui LEI {clei-at-uiuc.edu} wrote:
} Contact Shanghai Jiaotong U, Fudan U. } } They should have good TEM, SEM, lab. } } Or you can contact SMICS. They have SEM/EDS. But I } do not know if they like to check metals for you as } SMICS is a semiconductor manufacture. } } Ch } } ---- Original message ---- } } Date: Wed, 8 Jun 2005 09:52:18 -0700 (PDT) } } From: Kestutis Smalinskas {smalinskas-at-yahoo.com} } } Subject: [Microscopy] Any SEM/EDS labs in Shanghai, } China? } } } } } } I just got a request from someone in China if there } } are any labs available in China that can do SEM/EDS } } work. Can anyone provide contact information? } } } } Stu Smalinskas, P.E. } } Metallurgist } } SKF North American Technical Center } } Plymouth, Michigan } } (734) 414-6862 } } stu.smalinskas-at-skf.com
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 8 13:37:50 2005
I need a working/repairable scanning stage, ideally already set up for Leitz Orthoplan or Ergolux-compatible mounting dimensions. I've essentially abandoned trying to get my Ergolux's servo-driven stage working again due to the unavailability of parts and diagnostic info. The eventual goal is to drive the unit with an Oasis-4i to create an automated image capture system. If you have applicable stage hardware available for a reasonable price, please contact me. Thanks in advance.
-- Scott Griffith ISES-LLC 9745 Steeplechase Drive Franktown, CO 80116 303-660-2541 voice 303-660-2542 fax skod-at-ises-llc.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 8 15:06:32 2005
I'm trying to characterize two kinds of diffusion pump oils that I found in my "back room":
1. Balzers-Oil 71 für Diffusionspumpen. Ch 76-946
2. Trafo Olie, Dialac C #132250466801)
They both look like they have been around for some years. Does anyone know what kind of oils they are, eg. Silicone based or not?
My TEM and SEM have dedicated diff pump oils supplied by the manufacturer, but I'm wondering if either of the ones mentioned above would be OK to use in the diff pump of my Denton vacuum evaporator, which I use for routine carbon or gold-palladium coatings.
Thanks in advance!
Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-2785 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 8 16:16:27 2005
The second position (Trafo Olie, Dialac C) is a transformer oil- don't use it in vac. pumps. It is cheap - under $4 per gallon. If it is old and discolored- dispose of it. The first one (Balzers-Oil 71) must be OK for diff. pump use but:
a) is it in good condition (clean? clear? not oxydized/contaminated)?
b) you must completely clean traces of previously used oil from diff pump (these oils don't perform well if at all when mixed), unless you know for sure that diff pump was previously filled with the same type oil;
c) unless mistery oil is Santovac or Pentavac (very viscous at room temperature, like honey, clear, either colorless or has very light yellow color), it is cheap and IMO isn't worth the effort of trying it with uncertain results. But I would definitely try old Santovac, especially for a large diffusion pump, since it costs $1 and more per cc. Otherwise don't bother.
Vitaly Feihgold SIA 2773 Heath Lane Duluth, GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com
----- Original Message ----- } From: "Gib Ahlstrand" {ahlst007-at-umn.edu} To: "Microscopy Listserver" {Microscopy-at-MSA.Microscopy.com} Sent: Wednesday, June 08, 2005 4:05 PM
Hi Pat I agree with Debby. We have one too but I think your nearest cryo sem is University of New Brunswick, Frederickton. Elaine
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-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada (2003-2005) University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 8 17:05:13 2005
Vitaly right I would not put unknown quality diff oil into my pump: if it went wrong, you will spend days to clean pump up. Personally, I am using Santovac - it's serving my DV502A for 11 years without any problems: vacuum 5x10-7 torr after a few hours, no LN2. Sergey
At 01:58 PM 6/8/2005, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (acassell-at-mail.arc.nasa.gov) from http://www.microscopy.com/MLFormMail.html on Wednesday, June 8, 2005 at 19:04:33 ---------------------------------------------------------------------------
Email: acassell-at-mail.arc.nasa.gov Name: Alan Cassell
Organization: NASA Ames Research Center
Title-Subject: [Microscopy] [Filtered] MListserver: Gatan video capture card?
Question: We have a bottom mount Gatan fiber optic coupled camera on our JEOL2000FX TEM and we would like to use a video capture card to grab frames from the monitor for the camera. Has anyone tried regular commercial video cards? And if so, which ones work the best?
I've done it, and it certainly worked first time, no problems. I used old ISA cards, (well - they weren't old when I used them!!) I think an M-vision MV100 and a Synoptics card - possibly a Syntillate, but I'm not sure about that. Of course you would use newer ones, but I would be quite confident that any one with a standard TV input would work fine. It would all depend on the software that came with them - the "demo" software will allow you to capture and save images, but may not be as nice as you would like. You probably wouldn't want to get into writing your own software, and 3rd. party software can be expensive.
Tony.
At 09:14 PM 6/8/2005, you wrote:
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********************************************* Anthony J. Garratt-Reed, M.A., D.Phil MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 ********************************************* Phone: (617) 253-4622 Fax: (617) 258-6479 *********************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 9 07:16:40 2005
In an 1997 catalog from Pfeifer/Balzers I have the following indications :
Synthetique oil 71A :
vapor pressor 2.10-8 mbar at 20°C good chemical resistance Good heat and oxydation resistance Optimale pressure domaine 10E-3 to 5E-7 mbar Ultimate pressure : {6E-9 with LN2 trap {4E-7 with water trap {1E-6 with air trap (what is an air trap ????)
Recommended for metalurgie, electronic tube manufacturing, E-beam welding, sputtering, thermal evaprators, etc. Not for particules accelerators.
So no problem for your evaporator, but it seems to be a rough cheap oil, and not a Santovac quality one. A bit less good than DC 704.
If you send me your fax number, I can fax you the catalog sheet, but it is in french. It gives the specifications from 61A, 71A, DC704, AN175 and Santovac in regard.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi Listers, } } I'm trying to characterize two kinds of diffusion pump oils that I found in } my "back room": } } 1. Balzers-Oil 71 für Diffusionspumpen. Ch 76-946 } } } 2. Trafo Olie, Dialac C #132250466801) } } } They both look like they have been around for some years. Does anyone know } what kind of oils they are, eg. Silicone based or not? } } My TEM and SEM have dedicated diff pump oils supplied by the manufacturer, } but I'm wondering if either of the ones mentioned above would be OK to use } in the diff pump of my Denton vacuum evaporator, which I use for routine } carbon or gold-palladium coatings. } } Thanks in advance! } } Gib } -- } Gib Ahlstrand, Scientist } Electron Optical Facility, University of Minnesota, CBS Imaging Center, } 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 } (612)624-2785 FAX, ahlst007-at-tc.umn.edu } http://www.cbs.umn.edu/ic/ } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 10 08:28:26 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stacey.Andringa-at-uc.edu) from http://www.microscopy.com/MLFormMail.html on Friday, June 10, 2005 at 08:25:18 ---------------------------------------------------------------------------
Question: I have typeII cells that I want to label with HRP-gold and SPA-gold. Someone else has made the labels for me. I have a protocol to label the cells, but I want to make cytospin slides and check them to see if the cells are really labelled. I have a Vector kit that uses DAB (SK-4100). How can I block endogenous proteins? Thanks.
The Department of Molecular, Cellular, and Developmental Biology and the Neuroscience Research Institute are sponsoring an advance course on light microscopy. This 5-day workshop will be offered from September 5 through September 9, 2005 and will consist of lectures and laboratory exercises that will run from 9 am to approximately 5 pm each day. The seminar/workshop will be intensive, enabling participants to develop theoretical and hands-on expertise with light microscopes. Attendees will interact closely with the instructors while using modern research grade microscopes, cameras, and computers. The seminars and laboratories will cover basic optical theory and how it pertains to increasing contrast (signal to noise ratio) in biological samples. Fundamental techniques such as fluorescence, phase contrast, Nomarski Differential Interference Contrast, and darkfield imaging will be taught and attendees will use microscopes equipped with these optical enhancement accessories. In addition, the theory and practice of electronic image acquisition (analog and digital) will be presented and attendees will work with low-light cameras, digital image processing computers, and morphometric programs. There are five research grade microscopes, five electronic imaging cameras, two computer workstations, and one confocal microscope. With a maximum enrollment of 10 students, there will be ample opportunity to work with all of the microscopes and cameras. For those so interested, intensive hands-on instruction and guidance on the confocal microscope will be provided.
For a fuller description of the workshop please check the web address below. Enrollment forms can be completed online and this workshop provides an opportunity to have a working-vacation in Santa Barbara, California.
6/1/04, Tindall, Randy D. wrote: } ------------------------------------------------------------------------------ } Has as anybody noticed a second round of changes in this film? ... } } This leads me to think that there's a possibility that this film was } reformulated yet again at some point, since nothing obvious changed in } our procedures. Any similar experiences out there? Randy, I was justs cleaning out a lot of email messages from my file and re-read your much longer message. Did you ever find out if the 4489 was changed a second time?
I still have 3 boxes of 2004-06 which from my calculation should be "original film" but the other stock I have is 2005-06 which would fall into the time frame that your film batch might have been when you wrote the message. Does this make sense?
Storm just hit Bye- Pat
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 10 14:03:19 2005
I didn't see your original message so my reply is late. Yes, I have noticed a difference in the "new" 4489 film since it was changed. We have had to reset out TEM to accommodate for the much darker images that suddenly started to appear.
It seems like they are working at getting everyone to convert to digital in a very covert way.
Paul.
On 6/10/05 11:24 AM, "Pat Connelly" {psconnel-at-sas.upenn.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } 6/1/04, Tindall, Randy D. wrote: } } -----------------------------------------------------------------------------} } - } } Has as anybody noticed a second round of changes in this film? ... } } } } This leads me to think that there's a possibility that this film was } } reformulated yet again at some point, since nothing obvious changed in } } our procedures. Any similar experiences out there? } Randy, } I was justs cleaning out a lot of email messages from my file and } re-read your much longer message. } Did you ever find out if the 4489 was changed a second time? } } I still have 3 boxes of 2004-06 which from my calculation should be } "original film" } but the other stock I have is 2005-06 which would fall into the time } frame that your } film batch might have been when you wrote the message. Does this make sense? } } Storm just hit Bye- } Pat }
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 10 14:59:10 2005
That explains a lot! We've been having darker negatives later too, but I had assumed that it was either a change in temperature or developer dilution that had caused it.
Marc
On Friday, June 10, 2005, at 02:59 PM, Webster, Paul wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Dear Randy, } } I didn't see your original message so my reply is late. Yes, I have } noticed } a difference in the "new" 4489 film since it was changed. We have had } to } reset out TEM to accommodate for the much darker images that suddenly } started to appear. } } It seems like they are working at getting everyone to convert to } digital in } a very covert way. } } Paul. } } } On 6/10/05 11:24 AM, "Pat Connelly" {psconnel-at-sas.upenn.edu} wrote: } } } } } } } ---------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ----------------------------------------------------------------------- } -------} } - } } } } 6/1/04, Tindall, Randy D. wrote: } } } } ----------------------------------------------------------------------- } ------} } } - } } } Has as anybody noticed a second round of changes in this film? ... } } } } } } This leads me to think that there's a possibility that this film was } } } reformulated yet again at some point, since nothing obvious changed } } } in } } } our procedures. Any similar experiences out there? } } Randy, } } I was justs cleaning out a lot of email messages from my file and } } re-read your much longer message. } } Did you ever find out if the 4489 was changed a second time? } } } } I still have 3 boxes of 2004-06 which from my calculation should be } } "original film" } } but the other stock I have is 2005-06 which would fall into the time } } frame that your } } film batch might have been when you wrote the message. Does this make } } sense? } } } } Storm just hit Bye- } } Pat } } } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 10 15:37:13 2005
Thanks to the help from David Frey of Zeiss NY, the problem is software related. Versions of the SmartSEM prior to 5.00.08 had the problem. Field service here has 09. So this should fix the problem. If you have the heating problem, contact your service folks for a newer software version.
PS: disabling stage R does not remove drive from the motor. It just denies R commands.
gary g.
At 08:24 PM 6/6/2005, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Jun 10 16:22:33 2005
We've been having the same problem. Kodak has also changed the instructions on how to mix D-19 developer, though, and that may be compounding the problem. The old (paper envelopes) instructed adding the package to 3.8 liters of water, while the newer (plastic) envelopes have you start with 3 liters, dissolve the solution, then bring the final volume to 3.8 liters. Unfortunately, I threw out my last paper envelope a few weeks before mixing a batch with the stuff in the plastic envelope so I'm not sure whether the weight of the old (???) and new packages (551 grams) are the same or not.
Does anyone know how much powder is in a 1 gallon (3.8 liter) size paper envelope?
Heather Owen
Dr. Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee Lapham Hall, P.O. Box 413 Milwaukee, WI 53210 USA
Phone: (414)229-6816
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 10 16:27:45 2005
Oops. I had an empty Dektol envelope instead of an empty D-19 envelope. The actual weight of the plastic 3.8 liter size package is 607 grams, not 551 grams.
Friday afternoon brain slippage.
Heather Owen
Dr. Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee Lapham Hall, P.O. Box 413 Milwaukee, WI 53210 USA
Phone: (414)229-6816
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 10 16:50:20 2005
having some D-19 of older stock (just getting into the next order), the package size with the paper bag is 12gms less than for the new plastic bags. this difference could be just packaging. oh, yeah, that is for D-19, not dektol. our friday afternoon brain seizure has not yet hit up her in the frozen north....
as far as technique, the original procedure in my books - like 30+ years ago - was make it up in 3L at 37-50C, let cool, and then bring up to 3.8L volume. that is the way i've always done it wherever i have gone.
hope that trivia helps.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-9354 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 10 16:56:29 2005
Heather, I don't know about the paper bags as we don't have any of those, but we do have the cans that pre-date the paper bags and they have 595 grams with the instructions to add powder to 1 U.S. gallon (3.8 liters) at 100F (38C). Greg
Heather A Owen wrote:
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-- -- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
From MicroscopyL-request-at-ns.microscopy.com Sat Jun 11 22:14:04 2005
Stu Smalinskas wrote: ============================================== I just got a request from someone in China if there are any labs available in China that can do SEM/EDS work. Can anyone provide contact information? ============================================== See URL http://www.2spi.com/agntdist/china.html
The first firm listed is run by Dr. Susan Tai, formerly an executive with FEI Far East. She knows everyone who does this kind of work in China.
The third firm, KYKY, is a Chinese manufacturer of SEMs and at least at one time would run commercial samples for clients in their demo lab in Beijing. I have seen their newest model operate and it produces quite a good image. They are physically located next door to the large and very well equipped lab of the Academia Sinica (Chinese Academy of Sciences) so if they could not do what was needed, I am sure the lab next door could.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 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 ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Sat Jun 11 23:07:13 2005
Our electron microprobe laboratory has a chiller and sizable air conditioner, and thanks to Facilities Management at our university, there's little chance of getting them moved out of the lab. Since I cannot move the noise sources out of the lab, I've been thinking about installing acoustic panels to help with noise control. I've seen this done at a few other electron microscope labs, and the staff seemed happy with the results. I've been looking online at acoustic panels, and there are various materials, shapes, and designs out there. Has anyone else done this? What materials and shape did you use? Are you happy with the results? Any problems (shedding of fibers or dust, etc)? Is there anything you wish you'd done differently? Any suggestions or opinions are welcome.
Thanks in advance,
Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu/
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 12 11:08:16 2005
I personally would avoid the use of foam-based materials such as Sonex, as they degrade with time (~10years) and become a real mess to clean up and replace. We have begun using absorber/barrier blanket material, such as provided by Netwell Noise Control (and others), which does a great job and looks like it will last virtually forever. Get on Google and check it out. You can have special enclosures made to your own design, to isolate pumps and chillers, for example. Larry
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From MicroscopyL-request-at-ns.microscopy.com Mon Jun 13 00:43:33 2005
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From MicroscopyL-request-at-ns.microscopy.com Mon Jun 13 06:51:22 2005
Ellery, I haven't done this with our SEM lab, but I have used sound baffles in our Noble Gas lab. That lab has two mass specs with 7 rough pumps and 5 turbo pumps between them. Those pumps plus all of the electronic unit cooling fans add a fair amount of noise to the lab. I originally wanted sound baffling mounted to the walls, but our design and construction people would not allow it. Instead they recommended hanging 2' x 4' sound baffles from the suspended ceiling grid. They gave me a recommended layout which I tried to follow as best I could. The overall effect was excellent. The baffles we used are basically a tight weave of fiberglass enclosed in a poly sealed 'bag'. No dust collects on them as happens with wall mount egg crate material. They had two grommet wholes along one edge and they came with standard T-bar clips to hang them. I just used zip ties to connect the grommet to the hangers. Unfortunately, I don't know who makes the products as our design people did all the purchasing and had everything delivered to the lab. I hung them myself because I don't trust our facilities people working above our instruments.
Mike
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From MicroscopyL-request-at-ns.microscopy.com Mon Jun 13 09:37:14 2005
I have also run into some decision making regarding acoustic panels for a high end FESEM room. This is of major concern in a growing number of labs installing the newer very high end TEMs and SEMs. We are going to have a platform presentation about designing a high end facility in the Core Facility Management session at M&M2005. The session is from 1:30 to 3:30 on Wednesday.
I would really appreciate those who have information on acoustic noise remedies (as well as vibration and electrical interference remedies) to contact me. If you are going to be at M&M2005 I would invite you to participate in this session to share this information.
Another topic that will be introduced during the session, with additional discussion to follow the session (either at the Convention Center or at dinner Wednesday evening) is that of LIM (large Image Management) software and related issues (large data storage and backup, billing, authorization).
The discussion will be introduced by Avrum Goodblat and will revolve around two major issues:
a. LIMS software b. how to hook LIMS to the microscope. This is not always so easy to do - it depends both on the software and the nature of the utilities provided by the microscope manufacturer.
The Core Facility Management session, organized by the Focused Interest Group on Facility Operation, is designed to address topics of current interest. These threads often appear on the list serve a few months before the actual meeting and cannot be included in the meeting program. Since the session is geared to discussion of current topics of interest, we have a great deal of flexibility. Please take advantage of this and plan to bring information and participate fully in these discussions.
On 6/11/05 11:06 PM, "Ellery Frahm" {frah0010-at-tc.umn.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Microscopy folks, } } Our electron microprobe laboratory has a chiller and sizable air } conditioner, and thanks to Facilities Management at our university, } there's little chance of getting them moved out of the lab. Since I } cannot move the noise sources out of the lab, I've been thinking } about installing acoustic panels to help with noise control. I've } seen this done at a few other electron microscope labs, and the staff } seemed happy with the results. I've been looking online at acoustic } panels, and there are various materials, shapes, and designs out } there. Has anyone else done this? What materials and shape did you } use? Are you happy with the results? Any problems (shedding of } fibers or dust, etc)? Is there anything you wish you'd done } differently? Any suggestions or opinions are welcome. } } Thanks in advance, } } Ellery } } -------------------- } Ellery E. Frahm } Research Fellow & Manager } Electron Microprobe Laboratory } University of Minnesota - Twin Cities } Department of Geology & Geophysics } Lab Website: http://probelab.geo.umn.edu/ } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 13 12:15:52 2005
June 30, 2005 is the final deadline for submitting entries to the 31st Nikon International Small World Competition. The competition, open to all microscopists, was created in 1975 to recognize excellence in photography through the light microscope, and to applaud the efforts of those involved with photomicrography.
The competition is open to all types of light microscopy, and there are no restrictions to the type of specimens or techniques used (brightfield, darkfield, phase contrast, interference contrast, fluorescence, confocal, deconvolution, mixed techniques, etc.). Participants may obtain the rules, entry forms and enter their digital images directly at http://www.nikonsmallworld.com/. For those submitting entries in film format, the image and entry form should be sent to: Nikon Small World, 1300 Walt Whitman Road, Melville, NY 11747-3064. The first and second of twenty prizewinners will receive a selection of Nikon products and equipment worth $3,000 and $2,000 respectively.
Winners will be selected mid-summer. Each year, exhibits containing the winning entries are displayed at museums and science centers throughout North America. Many of these spectacular images have been featured on the covers of prestigious scientific publications and journals.
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 13 13:56:22 2005
Job Title: Histotechnologist III/EM UCLA Title: Histotechnologist III Job No.: H33530 Work Hours: Monday-Friday, 8:00am-5:00pm Work Location: CHS A3-240 Minimum Salary: $20.98 / $3651 Maximum Salary: $29.37 / $5110 Layoff Referral Deadline:
Job Duties: Provide administrative, technical, and maintenance support to the Electron Microscopy Department. Process, embed and cut EM blocks and tissue while maintaing excellent quality. Prepare: thin sections, staining, special processing, preparation of knives, reviewing and scanning of grids, and digital photography. Peforms independently a wide variety of technical and non-technical procedures. Perform processing of primary specimens, reagent preparation, operating, cleaning and maintaing work area and laboratory equipment, troubleshooting specimen/test order problems, filing, answering phones to respond to questions and participating in special studies. Oversee staff in their assigned area to see that all procedures meet the protocol stated. Tract laboratory supplies and places orders as necessary. Monitoring inventories of media, supplies and reagents used in diagnostic testing, quality control/quality assurance support, computer oepration (report printing) and data entry and retrieval. Enter data into the Tamtron system, maintain and print logs and labels. Perform daily recordkeeping.
Job Qualifications: Working knowledge of basic chemistry, biology, and anatomy with minimum of 5 years experience in Electron Microscopy. Skill in sectioning difficult tissues. Skill in performing all ranges of testing and procedures in Electron Microscopy. Ability to learn running variety of automated stainers and other equipments. Skill in Tamtron system or ability to learn very fast. Ability to maintain the work area clean and follow the safety procedure. Ability to speak clearly and distinctly using appropriate English vocabulary. Ability to communicate effectively with resident and faculty in regard to the different issues. Ability to communicate with mutual respect and consideration for diversity among individual. Ability to work as part of team and cover the missing area voluntarily. Able to maintain confidentiality. Willingness to participate in teleconferences and other continuing education program. Able to modify work schedule to meet the needs of the department. Ability to maintain the quality control. Ability to perform any other duties or tasks as requested by the sueprvisor.
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 13 16:26:37 2005
Dear Lists, A problem has come up with using the objective aperture in conjunction with the low dose kit. When the aperture is centered in the Exposure state, it is off-center in the Focus state by an amount that varies with the focus shift. The problem is the same using TEM mode or EFTEM mode. Logically, this should be related to image shift pivot points, but that alignment is right on, and a complete set of TEM-HM alignments did not solve the problem. Both our local service guy and an expert FEI guy have looked at the problem, but have not been able to solve it (although we thought they had on a couple of occasions). Has anyone else seen this, and, if so, what fixed it. TIA. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 13 20:11:25 2005
} I was of the understanding that } this problem was a fundamental one related to the image shifts and the } objective apertures, but if you find out otherwise, could you please } send an } email to either of the lists? } Dear Lists, According to David Mastronarde, the problem is inherent in the Tecnai, since the obj aper is not in the back focal plane, and the alignment procedures do not account for this. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 14 08:34:22 2005
Dear Group - what do you believe current usage for TEM or field emission TEM is right now for biological applications? Thanks in advance for your input Barbara
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 14 09:13:37 2005
It's funny. For the first 16 years I have been at Wellman, we had two full-time TEM people (myself included) who had non-stop projects to do. Then, about 3 years ago, (somewhat coinciding with the purchase of the confocal microscope),TEM work dried up; literally. Now, with only myself running it, TEM work is picking up again. I currently have 3 projects to scope. Our work is stricly biological (I am looking at bacteria, retinal lesions and tattooed skin). We are a large laser research center at MGH and we collaborate with a number of the departments. (I don't just do the TEM for the group; I train people on the confocal and am doing molecular biology as well).
We are still not up to the volume of work (TEM) that we did and probably will never reach that level of work again, but it is nice to get back to basic ultrastructure.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Barbara Maloney [mailto:maloneyb-at-fiu.edu] Sent: Tuesday, June 14, 2005 9:31 AM To: microscopy-at-msa.microscopy.com
Dear Group - what do you believe current usage for TEM or field emission TEM is right now for biological applications? Thanks in advance for your input Barbara
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 14 11:38:13 2005
} Dear Group - what do you believe current usage for TEM or field emission TEM } is right now for biological applications? } Thanks in advance for your input } Barbara {maloneyb-at-fiu.edu} ========================== Barbara, I am currently the only TEM Technologist in my biology department and since my PI is retiring in September it will also be the end of my employment here. Over the past several years we have had some collaborators who could not get research quality TEM done in their own departments or institutions. They realized that biochemistry, confocal microscopy, etc. showed that the experiments were working but could not give the ultrastructural confirmation that was ultimately needed for understanding and hence to get the publication of their data accepted. More than once I have heard the words, "That is exactly what I thought was happening but I could not prove it."
A grant request has been made for a new TEM for our department, the present scope being 39 years old. Ten investigators are making plans to use the scope (hoping that the grant is funded) so it seems that there is still a need for TEM [and I might add that I am attempting to make known to them that it may be wise to have an experienced technologist around!].
For these reasons, I state that TEM has not been abandoned only "put on a back burner" while other techniques have caught the glamor of the day. As more investigators see the importance of confirming their results on the ultrastructural level or re-discover the sense of wonder at seeing what has not been observed or reported previously, TEM will regain its place in biological applications.
Just my thoughts and feelings, Pat Connelly psconnel-at-sas.upenn.edu Dept. of Biology Univ. of Pennsylvania Philadelphia, PA
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 14 12:36:54 2005
Hi All, I was just asked about how to process mousse embryos (embryonic days 9.5, 10.5 and 11.5) for SEM specifically to look at the limb buds. I"ve done TEM of limb buds, but not SEM. Would anyone out there care to suggest a protocol (if it differs substantially from "standard" fixation-dehydration and CPD)? Thanks, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 14 14:25:21 2005
Hi Lee, When we processed mouse embryos for SEM ( in '92) we had excellent results using "standard" fixation, dehydration and CPD. Frank
At 02:19 PM 6/14/2005, Leona Cohen-Gould wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Jun 14 14:55:19 2005
The Materials Science Division at the Naval Research Laboratory in Washington, DC, seeks postdoctoral fellow candidates to carry out TEM and FIB-based investigations of nanostructured materials. Potential projects include analysis of natural and synthetic nanoparticles, e.g., 1 to 10 nm particles condensed in the upper atmosphere and/or laboratory grown by reverse micelle synthesis; FIB-based micromanipulation and analysis of cosmic dust samples (STARDUST Mission and dust from stars pre-dating the Sun), and in situ STM-TEM studies of nanowires. NRL has extensive facilities for microscopy studies, including newly installed JEOL 2200FS and FEI Nova 600 DB-FIB instruments. Experience in HAADF imaging, EDS and EELS spectrum imaging and/or FIB operation are desired. A Ph.D. in physics, chemistry, materials science, or geology is required. Qualified candidates will be expected to submit a proposal for the August 1, NRC fellowship deadline, but start dates prior to the October completion of the review process are possible. US citizens or permanent residents only. For more information, contact Rhonda Stroud (stroud-at-nrl.navy.mil).
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 14 15:33:00 2005
http://www3.interscience.wiley.com/cgi-bin/abstract/102524606/ABSTRACT Has the SEM embedding procedures
Grose R, Harris BS, Cooper L, Topilko P, Martin P. 2002. Immediate early genes krox-24 and krox-20 are rapidly up-regulated after wounding in the embryonic and adult mouse. Dev Dyn 223: 371-378. Links Has the exact fix
David
On Jun 14, 2005, at 11:19 AM, Leona Cohen-Gould wrote:
} } } ---------------------------------------------------------------------- } -------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } Hi All, } I was just asked about how to process mousse embryos (embryonic } days 9.5, 10.5 and 11.5) for SEM specifically to look at the limb } buds. } I"ve done TEM of limb buds, but not SEM. Would anyone out there } care to suggest a protocol (if it differs substantially from } "standard" fixation-dehydration and CPD)? } Thanks, } Lee } -- } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy & Histology Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175 } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 14 15:35:55 2005
On Jun 14, 2005, at 6:30 AM, Barbara Maloney wrote:
} what do you believe current usage for TEM or field emission TEM is } right now for biological applications? } Thanks in advance for your input
Dear Barbara, All structural biology benefits from TEM (although not all uses it). We are doing both tomography, for high-resolution 3-D imaging of biological specimens, and single-particle analysis, for very-high-resolution determination of the structures of protein complexes. Almost all our work in on frozen-hydrated unstained specimens, which is as close to the native state as can presently be achieved. So the bottom line is that (FE)TEM is the best technique for determining the high-resolution structure of (nearly) native biological specimens. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 14 15:39:49 2005
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Curtains? Works well in many cases and a lot cheaper than specialist panels. If they don't work, you won't have lost much; if they do, you'll probably have saved quite a bit. -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 15 11:39:16 2005
Hello Everybody, I have a user who wants to process fish eggs (salmonids) for LM. He embedded eggs in paraffin but infiltration was insufficient and samples could not be cut at all. Is there a better protocol to handling these kind of samples? We tried Spurr with dehydration time extended to 1h in each concentration of alcohol and resin infiltration overnight. Still did not work. The interior was not properly infiltrated and sample was shrunk. Thanks Dorota
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 15 15:07:20 2005
} On 6/14/05 3:35 PM, "Larry Stoter" {larry-at-cymru.freewire.co.uk} wrote: } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------} } - } } } } } } -----------------------------------------------------------------------------} } } - } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } ----------------------------------------------------------------------------- } } } -- } } } } } } Microscopy folks, } } } } } } Our electron microprobe laboratory has a chiller and sizable air } } } conditioner, and thanks to Facilities Management at our university, } } } there's little chance of getting them moved out of the lab. Since I } } } cannot move the noise sources out of the lab, I've been thinking } } } about installing acoustic panels to help with noise control. I've } } } seen this done at a few other electron microscope labs, and the } } } staff seemed happy with the results. I've been looking online at } } } acoustic panels, and there are various materials, shapes, and } } } designs out there. Has anyone else done this? What materials and } } } shape did you use? Are you happy with the results? Any problems } } } (shedding of fibers or dust, etc)? Is there anything you wish you'd } } } done differently? Any suggestions or opinions are welcome. } } } } } } Thanks in advance, } } } } } } Ellery } } } } } } -------------------- } } } Ellery E. Frahm } } } Research Fellow & Manager } } } Electron Microprobe Laboratory } } } University of Minnesota - Twin Cities } } } Department of Geology & Geophysics } } } Lab Website: http://probelab.geo.umn.edu/ } } } } Curtains? Works well in many cases and a lot cheaper than specialist } } panels. If they don't work, you won't have lost much; if they do, } } you'll probably have saved quite a bit.
} Larry, } Can you provide any specifics about curtains such as what to look for, } possible sources, etc. } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy }
Nothing special - in the JEOL UK Applications Lab we just went to the local department store and ordered up some curtains of the correct size and hung them against the wall, ceiling to floor. Went for a fairly heavy, non-synthetic material, as we thought synthetics might cause problems with static electricity. Even to my ears, the room acoustics were very noticeably deadened. They weren't exactly cheap, since we needed quite a lot and it was quite a heavy material. On the other hand, I'd bet it cost less than specialist acoustic tiles.
PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 15 20:08:38 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dgarrett-at-unt.edu) from http://www.microscopy.com/MLFormMail.html on Wednesday, June 15, 2005 at 10:06:28 ---------------------------------------------------------------------------
Email: dgarrett-at-unt.edu Name: Da vid Garrett
Organization: University of North Texas
Title-Subject: [Microscopy] [Filtered] SEM general JEOL 5800 CMD list
Question: Can anyone supply me with a list of the features that are controled with the CMD key. To change the load current press CMD type bias use focus and mag to change. Thats the only one I know. Thanks, David
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (chenkaic-at-msu.edu) from http://www.microscopy.com/MLFormMail.html on Wednesday, June 15, 2005 at 23:27:20 ---------------------------------------------------------------------------
Email: chenkaic-at-msu.edu Name: Kai-Chun
Organization: MSU
Title-Subject: [Microscopy] [Filtered] EPON and Prion
Question: Dear all,
My advisor and I want to visulize prion protein with EM. We want to use negative stainning. However, it is somewhat difficult because of the safety level of EM lab, so we are thinking about how to deactivate prion. I heard that EPON may deactivate prion, but I can not find the references to support that. I wonder if anyone has this kind of experiences to handle prion protein. Do you think EPON can deactivate prion protein?
i seem to have lost a very good response to the recent posting asking about prion infectivity - raised by ralph common. someone on the list responded with information from previous work of theirs, where they had checked to determine retention of catalytic infectivity of prions after osmication.
i seem to not have that particular response saved. could any one on the list who has the note, or the original respondant, please send me another copy - it was excellent, probably the best of the whole group of responses. i wish to have it in my notes for future reference.
thanks
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-9354 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 16 09:27:58 2005
} Question: Dear all, } } My advisor and I want to visulize prion protein with EM. } We want to use negative stainning. } However, it is somewhat difficult because of the safety level of EM lab, so we are thinking about how to deactivate prion. } I heard that EPON may deactivate prion, but I can not find the references to support that. } I wonder if anyone has this kind of experiences to handle prion protein. } Do you think EPON can deactivate prion protein? } } IMHO, the main danger when you work with prion protein infected tissue is the possible inhalation or ingestion of the contaminated material. Therefore, if you will cover the grid with some strong film then it will in theory protect the environment from the prion exposure. However, all the films have the tendency to break at some point that makes their protection questionable. The idea of covering the grid with Epon may work because Epon is quite resistant material. However in this case the resolution can be decreased because of the thickness of epoxy film cover. The best solution to see negatively stained prion protein on the grid would be dedicated microscope. It is not cheap solution but it is safe. If you will embedd your protein in epon and then make sections then probably this protein would be inactive, but I do not have any references that prove it. You may want to contact Dr. Holger Wille from S.Prusiner lab (UCSF) who has vast experience with prion proteins in EM.
Sincerely,
-- Aleksandr Mironov Experimental Officer G450A, Stopford Building EM Unit, Faculty of Life Sciences University of Manchester Oxford Road Manchester M13 9PT UK
I've brought an old high vacuum coater back to life, and it is working well for carbon coating, but I am missing the parts to do metal coatings and set up an aperture cleaning boat.
The unit is a Jeol JEE 4B, manufactured around 1972. What I am looking for is not all that different from ring stand parts, and I wonder whether anyone out there has modified ring stand parts to use with a high vacuum coater. It seems that all I would need is a couple of insulators and a tap and die set to attach a piece of wire or moly boat and get going.
Otherwise, is there a third party company that makes parts that would fit on one of these?
Thanks for your help.
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 16 10:29:30 2005
We'd like to scratch-build an apparatus that allows us to perform voltage contrast testing in one of our SEMs. We currently have a Hitachi S-4500, a JEOL 6301, and a JEOL 6340. Does anyone on here have experience using or building such a system? If you do, could you provide a description of the components and provide hints and tips on assembling one?
Regards, Tom
Thomas Barbieri Arizona Product Development and Analysis Laboratory Freescale Semiconductor Tempe, AZ 480-413-4007
thomas.barbieri-at-freescale.com
The information contained in this email and any attachment is considered: [X] General Business Information [ ] Freescale Internal Use Only [ ] Freescale Confidential Proprietary
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 16 10:32:53 2005
Having started my professional career managing a EM lab working on CJD I have actual experience working with this fun stuff. We found that PF & Glut perfusions did not inactivate the mutant protein. Once the tissue was post-fixed in OsO4 we could not serial pass the disease again. Once in plastic it's for all intents and purposes inert. Waste materials were autoclaved for a longer length of time than normal and discarded with "normal" medical waste. In the early 80's when I was working with CJD it was a BSL2 for the most part, now all procedures should be done under BSL3. The Neuropathologist used to work with it under BSL1, removing the CJD infected brains from the cadavers. They would always be violating containment protocols in some was shape or form. They are still alive, except one (CVA). The point I'm trying to make is don't over react, take all of the prudent precautions of BSL3 you will be well protected.
Al Coritz Electron Microscopy Sciences www.emsdiasum.com
----- Original Message ----- } From: "paul r hazelton" {paul_hazelton-at-umanitoba.ca} To: "Ralph Common" {Ralph.Common-at-hc.msu.edu} Cc: {Microscopy-at-microscopy.com} Sent: Thursday, June 16, 2005 10:21 AM
Karl,
Thank you for the conversation this afternoon.
Items for the JEE-4B may be available from our Parts Department (Nancy Green). Instruction manuals (Ion Gauge) may be a bit more difficult however we may be able to provide copies.
The main number to JEOL USA Inc in Peabody Massachusetts is 978-535-5900, please ask for Nancy Green.
I will let you know when we have fixed the schedule for UC on the JEM-1230 demonstration.
Nice to have spoken with you.
Regards,
Bill Powell JEOL USA Inc Phone: 248-366-8351 Ion Beam Cross Section Polisher: http://www.jeol.com/spe/speprods/cross_section.html
-----Original Message----- } From: Karl Hagglund [mailto:hagglundk1-at-nku.edu] Sent: Thursday, June 16, 2005 10:36 AM To: microscopy-at-microscopy.com
I've brought an old high vacuum coater back to life, and it is working well for carbon coating, but I am missing the parts to do metal coatings and set up an aperture cleaning boat.
The unit is a Jeol JEE 4B, manufactured around 1972. What I am looking for is not all that different from ring stand parts, and I wonder whether anyone out there has modified ring stand parts to use with a high vacuum coater. It seems that all I would need is a couple of insulators and a tap and die set to attach a piece of wire or moly boat and get going.
Otherwise, is there a third party company that makes parts that would fit on one of these?
Thanks for your help.
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 16 12:28:25 2005
There are more then a few variables to consider when you decide to utilize voltage contrast. For example, in addition to the tooling and fixtures and SEM, the device type and fabrication process play a huge role in how successful you can be at capturing the correct electron signals. Please contact me off line and I'll share whatever experience and knowledge I have.
Regards, Evelyn
At 08:28 AM 6/16/2005, Barbieri Thomas-r53545 wrote:
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Evelyn York
Analytical Facility Scripps Institution of Oceanography University of California, San Diego 9500 Gilman Drive M/S 208 La Jolla, CA 92093-0208
(858) 534-2438
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 16 21:03:37 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (emlab-at-vet.ksu.edu) from http://www.microscopy.org/MicroscopyListserver/MLFormMail.html on Thursday, June 16, 2005 at 08:51:27 ---------------------------------------------------------------------------
Email: emlab-at-vet.ksu.edu Name: Lloyd Willard, EM Lab
A colleague has some thin metal films (5-50nm) on Si wafers and we need to know the relative film thicknesses. We don't need to know the absolute thickness. Is there any way to use the EDXS signal to get relative thickness? I.e. use the suppression of the Si signal? Or will the results be too unreliable? We have no other way here to do the job.
Any suggestions (including don't bother trying it!!!) will be welcome.
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 17 07:55:19 2005
This exercise should be rather straightforward. I've done it a number of times. If the film is thin enough, you'll pick up the Si under the metal film. The thinner the film, the stronger the Si peak. This will vary with accelerating voltage. Find a voltage that picks up both the metal film and Si with reasonably large peaks. To compare the two samples you'll need to collect a spectrum from each sample using the same accelerating voltage constant. Then compare the two spectra looking at the ratio of metal/Si peak heights within each spectrum.
To take it a step futher, you could get absolute values if you had control samples with known metal thickness for comparison.
Stu Smalinskas, P.E. SKF North American Technical Center Plymouth, MI (734) 414-6862 stu.smalinskas-at-skf.com
--- Colin.Veitch-at-csiro.au wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi, } } A colleague has some thin metal films (5-50nm) on Si } wafers and we need } to know the relative film thicknesses. We don't } need to know the } absolute thickness. Is there any way to use the } EDXS signal to get } relative thickness? I.e. use the suppression of the } Si signal? Or will } the results be too unreliable? We have no other way } here to do the job. } } Any suggestions (including don't bother trying } it!!!) will be welcome. } } Cheers. } } Colin Veitch } } Electron Microscopist } } CSIRO Textile and Fibre Technology } } PO Box 21, BELMONT, Vic. 3216. Australia. } } E-mail: colin.veitch-at-csiro.au } } Web: http://www.tft.csiro.au } } Tel: +61 (0) 3 5246 4000 } Mob: 0438 538 475 } Fax: +61 (0) 3 5246 4811 } } } } The information contained in this e-mail message may } be privileged or } confidential information. If you are not an intended } recipient, you may } not copy, distribute or take any action in reliance } on it. If you have } received this message in error, please telephone } CSIRO Textile and Fibre } Technology on +61 3 5246 4000. } } } }
__________________________________ Yahoo! Mail Stay connected, organized, and protected. Take the tour: http://tour.mail.yahoo.com/mailtour.html
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 17 08:08:03 2005
There are plenty of ways to do this. Immediately coming up in my mind is the 4-point resistivity measurement with a routine device in any chip manufacturing shop. x-section SEM should you have a hi-res SEM and be able to afford for the destruction of the sample or x-section TEM is the other
**************************************** Chaoying Ni, PhD The W.M. Keck Electron Microscope Facility Facility location: 022 Spencer Laboratory Mailing address: 201 duPont Hall Dept of Matls Sci & Eng University of Delaware Newark, DE 19716 (302) 831-2318(L); -4545(Fax) http://eml.masc.udel.edu *****************************************
On Fri, 17 Jun 2005 Colin.Veitch-at-csiro.au wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi, } } A colleague has some thin metal films (5-50nm) on Si wafers and we need } to know the relative film thicknesses. We don't need to know the } absolute thickness. Is there any way to use the EDXS signal to get } relative thickness? I.e. use the suppression of the Si signal? Or will } the results be too unreliable? We have no other way here to do the job. } } Any suggestions (including don't bother trying it!!!) will be welcome. } } Cheers. } } Colin Veitch } } Electron Microscopist } } CSIRO Textile and Fibre Technology } } PO Box 21, BELMONT, Vic. 3216. Australia. } } E-mail: colin.veitch-at-csiro.au } } Web: http://www.tft.csiro.au } } Tel: +61 (0) 3 5246 4000 } Mob: 0438 538 475 } Fax: +61 (0) 3 5246 4811 } } } } The information contained in this e-mail message may be privileged or } confidential information. If you are not an intended recipient, you may } not copy, distribute or take any action in reliance on it. If you have } received this message in error, please telephone CSIRO Textile and Fibre } Technology on +61 3 5246 4000. } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 17 10:19:21 2005
It's not too late to enter the International Metallographic Contest and Exhibit co-sponsored since 1972 by the International Metallographic Society and ASM International. The contest will be held in conjunction with the 38th annual technical meeting of the IMS and the M&M 2005 meeting this August in Honolulu. Twelve categories of competition. Best in show receives $3000 and the prestigious Jaquet-Lucas Award. Cash awards for first, second, and third place winners in eleven of the categories. Entries are prominently displayed during the M&M 2005 meeting and again in the fall during the ASM Annual Event. Deadline for entries is July 18. For additional information including rules, tips for creating a winning entry, judging guidelines, and examples of winning entries contact me or visit http://www.internationalmetallographicsociety.org/contest.html.
Jeff Stewart International Metallographic Contest Chair Metallographic Laboratory Manager Stern-Leach Co. 49 Pearl Street Attleboro, MA 02703 USA Phone: 508-222-7400 extension 1329 FAX: 508-699-4030
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 17 13:00:34 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (diller-at-stefan-diller.com) from http://microscopy.com/MLFormMail.html on Thursday, June 16, 2005 at 08:35:16 ---------------------------------------------------------------------------
Email: diller-at-stefan-diller.com Name: Stefan Diller
Question: Dear All, does anybody know of an image database in the net, preferably freely accessible, which deals with herbs and especially with the microstructure of herbs using SEM and TEM?
For years I have used a free shareware program called GMRfilm to measure submicrometer film thicknesses on substrates. It is an old DOS based program (I think there might be newer programs out now) and can be found on the MSA website through the reference and educational link or at http://www.amc.anl.gov. From the parent directory, go to 02-MMSLib/ XEDS/ GMRFILM/.
I routinely use it to calibrate coating thicknesses for our sputter coater and vacuum evaporator. As Stu mentioned, you need to choose the appropriate keV to excite both the x-ray signal from the film and the substrate. Then under identical conditions measure collect spectra from pure standards. Extract the net peak intensities and calculate the K-ratios. Once you have the K-ratios and other operating and specimen parameters are entered into GMRfilm to calculate the film thickness.
If you want, feel free to contact me off line for more information.
Lou Ross
} } Hi, } } A colleague has some thin metal films (5-50nm) on Si wafers and we need } to know the relative film thicknesses. We don't need to know the } absolute thickness. Is there any way to use the EDXS signal to get } relative thickness? I.e. use the suppression of the Si signal? Or will } the results be too unreliable? We have no other way here to do the job. } } Any suggestions (including don't bother trying it!!!) will be welcome. } } Cheers. } } Colin Veitch } } Electron Microscopist } } CSIRO Textile and Fibre Technology } } PO Box 21, BELMONT, Vic. 3216. Australia. } } E-mail: colin.veitch-at-csiro.au } } Web: http://www.tft.csiro.au } } Tel: +61 (0) 3 5246 4000 } Mob: 0438 538 475 } Fax: +61 (0) 3 5246 4811 } } } } The information contained in this e-mail message may be privileged or } confidential information. If you are not an intended recipient, you may } not copy, distribute or take any action in reliance on it. If you have } received this message in error, please telephone CSIRO Textile and Fibre } Technology on +61 3 5246 4000.
-- Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211-5120 (573) 882-4777, fax 884=2227 email: rosslm-at-missouri.edu http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 17 15:33:55 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sschmech-at-u.washington.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, June 17, 2005 at 15:22:09 ---------------------------------------------------------------------------
Email: sschmech-at-u.washington.edu Name: Steve Schmechel
Question: In a study of reovirus effects on infected cells in culture, we are finding strange cytoplasmic changes. Cells infected with some strains (and not others) have strange cytoplasmic structures. These are non-membrane bound clear spaces that appear as wide splits in the cytoplasm. They are large, approximatly the length of the nucleus with a width approximately one fifth of length. They "smile" at the middle and are curved. Although none are directly next to the nucleus, they take a curved form that generally follows a similar curve as the nuclear membrane. In most cells there are many of these. The result is that the cells have a zebra stripe appearance due to these multiple splits in the cytoplasm.
Has anyone ever seen something like this. I'm happy to email you a sample if you like.
Without an image to study, my first guess is that the 'inclusions' are cholesterol clefts. Frank Ramig, at Baylor College of Medicine, Division of Biochemical Virology has many, many years under his belt in working with reoviruses. He might be a good person to ask about these things.
Best of luck,
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
} } } by way of MicroscopyListserver {sschmech-at-u.washington.edu} 6/17/2005 4:33 PM } } }
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sschmech-at-u.washington.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, June 17, 2005 at 15:22:09 ---------------------------------------------------------------------------
Email: sschmech-at-u.washington.edu Name: Steve Schmechel
Question: In a study of reovirus effects on infected cells in culture, we are finding strange cytoplasmic changes. Cells infected with some strains (and not others) have strange cytoplasmic structures. These are non-membrane bound clear spaces that appear as wide splits in the cytoplasm. They are large, approximatly the length of the nucleus with a width approximately one fifth of length. They "smile" at the middle and are curved. Although none are directly next to the nucleus, they take a curved form that generally follows a similar curve as the nuclear membrane. In most cells there are many of these. The result is that the cells have a zebra stripe appearance due to these multiple splits in the cytoplasm.
Has anyone ever seen something like this. I'm happy to email you a sample if you like.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Gervais.Sawyer-at-bcuc.ac.uk) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, June 20, 2005 at 04:56:39 ---------------------------------------------------------------------------
Question: I am trying to SEM image an ash structure after burning pieces of wood. These are extremely beautiful and could reveal some microstructure. However, the ash is very delicate and disintegrates when I go to vacuum. The same thing happens when I try to sputter coat. Any ideas please?
I have been given the unenviable task of having to decide between a digital camera system and a digital imaging plate system (the Ditabis system) to enable our older TEMs to become fully digital imaging systems. We have Jeol 1200EX and 100CX instruments and a Phillips CM10. Has anyone had experience of the Ditabis system? In the UK there is only one currently in use, and I have contact with that person. Our samples are biological samples, some routine clinical specimens, but also research samples and cell culture samples. We currently use Kodak em film 4489 and require the replacement system to have comparable resolution. Any comments would be gratefully received.
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 20 11:40:01 2005
} We currently use Kodak em film 4489 and require the replacement } system to have comparable resolution. } Any comments would be gratefully received.
Dear Kerrie, The grain size on 4489 is extremely small (~1 um), so the resolution is much better than can be achieved with any image plate system. Image plates have many advantages over film, such as linearity and ease of digitization, so they are better than film for quantitation, but for enlargement by many times or other processes that require high resolution, film is still superior. I do not know any specifics about the Ditabis system. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 20 11:58:31 2005
Kerrie; There is an operational difficulty with imaging plate systems in that magnification and serial number are not recorded on the plate. In fact, nothing is recorded on the plate except the electron image. Think about getting one plate out of order, and how that might foul up your operations.
John Mardinly Intel
This opinion is that of the author and does not represent the opinion of Intel corporation.
-----Original Message----- } From: Kerrie Venner [mailto:k.venner-at-ion.ucl.ac.uk] Sent: Monday, June 20, 2005 6:52 AM To: Microscopy-at-microscopy.com
I have been given the unenviable task of having to decide between a digital camera system and a digital imaging plate system (the Ditabis system) to enable our older TEMs to become fully digital imaging systems. We have Jeol 1200EX and 100CX instruments and a Phillips CM10. Has anyone had experience of the Ditabis system? In the UK there is only one currently in use, and I have contact with that person. Our samples are biological samples, some routine clinical specimens, but also research samples and cell culture samples. We currently use Kodak em film 4489 and require the replacement system to have comparable resolution. Any comments would be gratefully received.
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 20 13:05:13 2005
Can anyone tell me if JB-4 embedded sample can be sectioned on an ultramicrotome (Leica Ultracut S), and what the appropriate thickness should it be cut at? Thank you in advance.
HONg Emory
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 20 13:24:48 2005
Hong, I think JB-4 is a species of methacrylate resin so sure you can cut it on an Ultra. I'd guess anywhere from 2 um semis right down to 60nm ultras (though it may be difficult to get all the way that thin) depending on what you want to do.
Happy slicing, Tobias
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I have cut 0.5 to 2 um thick JB-4 sections using a dry glass knife without a boat on many occasions. As the section starts to come off on the knife face, help it along with one of the tines of a fine forceps so that the section doesn't crumble up. Gently lift the cut section and drop in one motion on a drop of water. be sure to let go of the section before it touches the water or it will shrivel up against the forceps. Heat the section down on a hot plate as one would do for an Epon section. It works but is tedious. Furthermore, I would say the frustration is unnecessary since BMMA (butyl-methyl methacrylate/methyl methacrylate mixtures) are less expensive (generic ingredients), easier to cut (cuts on water filled boats), and can be deplasticized using acetone. We love BMMA and haven't used JB-4 ever since the esteemed Dr. Tobias Baskin told us about it. I don't have the reference to his paper but he monitors this listserver and perhaps will post it.
At 01:04 PM 06/20/05, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Please draw the following position description to the attention of any potentially interested people. We can be flexible for the right person, within usual constraints. We have just won a new WA State Government Centre of Excellence grant that will provide an additional position into the nanoSIMS laboratory later in the year or early next year.
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A CAMECA nanoSIMS 50 high-resolution ion microprobe was installed in the Centre for Microscopy and Microanalysis (CMM) at The University of Western Australia, as part of the NANO-MNRF, in June, 2003. The NANO-MNRF (www.nano.org.au) is a Major National Research Facility that links advanced nano-scale characterisation equipment at the Universities of Sydney, New South Wales, Queensland, Western Australia (UWA) and Melbourne. The range of CMM microscopy instrumentation and expertise is extensive (see http://cmm.uwa.edu.au) such that the centre and MNRF represent a premium environment for research services, research training and research programs. UWA is also a major partner in local research consortia in isotope science with a range of stable and radiogenic isotope facilities, including two SHRIMP-II.
We are seeking a highly motivated, self-guided scientist who has the ability to work with other staff within the Centre and its users. The prime responsibility will be to manage a CAMECA nanoSIMS 50 ion microprobe as a world-class National Facility. The position will have the core role in a strong team led locally by Associate Professor Brendan Griffin and nationally by Professor Simon Ringer (NANO MNRF Executive Director). This position is advertised as a Level 6 or 7 depending on qualifications and experience. The minimum qualification is a relevant degree or equivalent experience.
Application Details: Interested applicants must obtain the application package and address the prerequisites and selection criteria. These essential details can be accessed from the vacancy page on http://jobs.uwa.edu.au/ or the 24 hour "hotline" on 6488 3733. To discuss or clarify any aspects of the position please contact the director of the CMM, Associate Professor Brendan Griffin on (08) 6488 2770 or email bjg-at-cmm.uwa.edu.au.
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Brendan J. Griffin Director & Assoc. Professor in Electron Microscopy Centre for Microscopy and Microanalysis (M010) Director Western Australian Centre for Microscopy Associate Director NANO-MNRF President Australian Microbeam Analysis Society The University of Western Australia First floor, Physics Building, 35 Stirling Highway CRAWLEY, WA, AUSTRALIA 6009 ph 61-8-6488-2739 fax 6488-1087 mobile 0409-104-096 bjg-at-cmm.uwa.edu.au http://cmm.uwa.edu.au/
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From MicroscopyL-request-at-ns.microscopy.com Mon Jun 20 13:57:44 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (diller-at-stefan-diller.com) from http://microscopy.com/MLFormMail.html on Monday, June 20, 2005 at 09:55:20 ---------------------------------------------------------------------------
Email: diller-at-stefan-diller.com Name: Stefan Diller
Title-Subject: [Microscopy] [Filtered] Old KEVEX detector on KEVEX DELTA 8000
Question: Dear All, maybe there is someone out there who can help me connect an old KEVEX EDS (seems to be Model 3801M?) detector with a Preamplifier Modell 2002 on my "new" KEVEX Delta 8000 unit.
I especially need the layouts of 1- the PREAMP1 connector on the rear of the KEVEX 8000 concerning voltage output and preamp output coming from the detector (see image at: http://www.stefan-diller.com/rem/Kevex1.jpg) 2- the POWER connector on the EDS detector and also the voltages needed for the preamp Modell 2002 in the detector (see image at: http://www.stefan-diller.com/rem/Kevex2.jpg and http://www.stefan-diller.com/rem/Kevex3.jpg ) 3- or the simple answer: Is it possible to use this detector on the KEVEX 8000?
Sort of a way out idea but what about vapor impregnation by Cyanoacrylates then sputter coat? Like Forensic scientists use to get fingerprints of unusual places.
Cheers!
Al Coritz Electron Microscopy Sciences www.emsdiasum.com
----- Original Message ----- } From: "by way of Ask-A-Microscopist" {Gervais.Sawyer-at-bcuc.ac.uk} To: {microscopy-at-microscopy.com} Sent: Monday, June 20, 2005 9:16 AM
Group, As Tom mentioned, mixtures of butyl and methyl and methacrylate produce easy to section blocks that can serve a variety of purposes. These mixtures predate epoxies for TEM use and they are unstable in TEM beams. But the other side of that coin is most of the resin can be extracted after sectioning with a 10 minute incubation in aceteone. This provides excellent access for an antibody probe to most of the section volume. So for light level immuno work, BMM is great stuff. My innovation was to dissolve some DTT in the resin mix which keeps those pesky free radicals from oxidizing the daylights out of the sample.
My original paper on this is: Baskin TI, Busby CH, Fowke LC. Sammut M, Gubler F (1992) Improvements in immunostaining samples embedded in methacrylate: Localization of microtubules and other antigens throughout developing organs in plants of diverse taxa. Planta 187: 405 - 413.
And a few later findings in the methods section of this paper:
Baskin TI, Wilson JE (1997) Inhibitors of protein kinases and phosphatases alter root morphology and disorganize cortical microtubules. Plant Physiology 113: 493 - 502.
If anyone wants to try this, contact me off line and I can provide a few extra resources for you.
Happy slicing, Tobias
} } I have cut 0.5 to 2 um thick JB-4 sections using a dry glass knife } without a boat on many occasions. As the section starts to come off } on the knife face, help it along with one of the tines of a fine } forceps so that the section doesn't crumble up. Gently lift the cut } section and drop in one motion on a drop of water. be sure to let go } of the section before it touches the water or it will shrivel up } against the forceps. Heat the section down on a hot plate as one } would do for an Epon section. It works but is tedious. Furthermore, } I would say the frustration is unnecessary since BMMA (butyl-methyl } methacrylate/methyl methacrylate mixtures) are less expensive } (generic ingredients), easier to cut (cuts on water filled boats), } and can be deplasticized using acetone. We love BMMA and haven't } used JB-4 ever since the esteemed Dr. Tobias Baskin told us about } it. I don't have the reference to his paper but he monitors this } listserver and perhaps will post it. } } } } At 01:04 PM 06/20/05, you wrote: } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stacey.Andringa-at-uc.edu) from http://www.microscopy.com/MLFormMail.html on Monday, June 20, 2005 at 14:35:52 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (artcollc-at-emirates.net.ae) from on Monday, June 20, 2005 at 09:36:41 ---------------------------------------------------------------------------
Email: artcollc-at-emirates.net.ae Name: k. j. dhar
Organization: artco llc.
Education: Graduate College
Location: Dubai, United Arab Emirates.
Question: We are a marketing company, distributing (since 1989)all types of Material Testing equipment.
Currently, one of our customers (who is engaged in Oil production, refining, distribution) has set up a Petroleum Institute. They have a requirement for Oversized Petrographic Thin Section Coverslips, in two sizes 42mmx25mm and 74x49mm with 0.16mm thickness.
We seek your assistance/suggestions on the likely source from whom this can be ordered.
Thanking you in anticipation and with best regards,
Some years ago, Buehler Instruments (Waukegan,IL), made a special device to mount delicate samples like this. It consisted of a specially constructed vacuum jar in which was mounted a small ladle that could pour epoxy into mounting cups. The specimen was put in the cups, the jar closed, the epoxy poured, then a vacuum drawn. The result: the epoxy penetrated the delicate structure and permitted microtoming.
I actually saw the ash on a cigar mounted this way, so think that it should be a good solution for your problem. You can probably access Buehler on the web. If not, contact me off line and I will see if I can find their contact info.
Good hunting!
Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Call us today to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts. Call us at (972)954-8011.
At 08:16 AM 6/20/2005, Gervais.Sawyer-at-bcuc.ac.uk wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Jun 21 02:45:26 2005
Hi, it depends on what microscopy you are actually doing, replies already in are for TEM. I am using a modified version of JB-4 (much less catalyst and 'B') for IHC at the light microscope level and the resin does not need to be removed. I cut 2micron sections on a dry knife (often the same one I trimmed on as the resin is so soft) but I can go lower. No I can't do TEM off the same block.. Refs: Casey et al. (1988) Am J Pathol;131:183-189 Britten et al. (1993) Biotech Histochem;68:271-280
Gill Brown Histopathology Group Asthma and Allergy Disease Biology GlaxoSmithKline Medicines Research Centre, UK
"Hong Yi" {hyi-at-emory.edu} 20-Jun-2005 19:04
To " {microscopy-at-microscopy.com} {microscopy-at-microscopy.com} " cc
Subject JB-4
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Can anyone tell me if JB-4 embedded sample can be sectioned on an ultramicrotome (Leica Ultracut S), and what the appropriate thickness should it be cut at? Thank you in advance.
HONg Emory
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 21 02:50:24 2005
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From MicroscopyL-request-at-ns.microscopy.com Tue Jun 21 05:11:26 2005
We have a Philips SEM 515, about 17 years old, available. The instrument itself is in very good working order, apart from a small electrical problem with the pumping system which we did not bother to have fixed since we moved to a newer instrument.
Anyone interested?
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 21 12:22:27 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gardnere-at-wpunj.edu) from http://www.microscopy.com/MLFormMail.html on Tuesday, June 21, 2005 at 13:50:12 ---------------------------------------------------------------------------
Question: I am the chair of a Biology department at a state university in NJ. I am planning on purchasing a confocal microscope which would be used for both teaching and research. Do you have any ideas as to which brands would be easiest to use and maintain? There will be no technician dedicated to this microscope. Thanks.
Dear Listers, We are trying to find a workable solution for a problem that I believe others may have run up against regarding digital image acquisition. We have a Tecnai T12 microscope with a high resolution bottom mount digital camera (Gatan 4K x 4K). The problem is due to the slow frame rate and small field of view of the camera makes it difficult to focus and capture low magnification survey shots. We have considered adding a side-mount CCD camera to complement the main camera but would like to hear from other users to see what our options are. Does anyone have experience with direct video rate cameras (side mounted) for the purpose of locating and focusing areas of interest?
Another option would be to add a multiple image stitching software to the current camera and merge several higher mag images. My concern here is image/ specimen drift over the time these multiple acquisitions are made. I work primarily with slot grids and support film which tends to exacerbate such problems.
Thanks in advance, Jay
Jay Campbell Research Specialist University of Wisconsin Laboratory of Molecular Biology R.M. Bock Labs 1525 Linden Drive Madison, WI 53706 jmcampbe-at-wisc.edu 608 263 8481
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 21 18:43:21 2005
I'd say there should be a technician or your going to have some trouble with getting it to work. Maybe appoint someone to take care of it and train people. Go with zeiss, they have nice confocal systems that are really easy to use and integrate well with the software for control/imaging. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Tue, 21 Jun 2005, by way of MicroscopyListserver wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gardnere-at-wpunj.edu) from http://www.microscopy.com/MLFormMail.html on Tuesday, June 21, 2005 at 13:50:12 } --------------------------------------------------------------------------- } } Email: gardnere-at-wpunj.edu } Name: Eileen Gardner } } Organization: William Paterson University } } Title-Subject: [Microscopy] [Filtered] MListserver: Confocal microscope } } Question: I am the chair of a Biology department at a state university } in NJ. I am planning on purchasing a confocal microscope which would be } used for both teaching and research. Do you have any ideas as to which } brands would be easiest to use and maintain? There will be no } technician dedicated to this microscope. Thanks. } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 21 20:36:24 2005
Jay I would think, the good solution to you would be side-mounted camera with optical coupling - they normally shows quite decent refresh rate. Perhaps, it would be wise to use the same manufacturer for 100% compatibility with existing camera. As for focusing problem - it's kind of strange. Gatan's 4x4 has view area compatible to the film. As for "slowness", yes, it's slow. You need to use at least 4x (even 8x) binning in the "view" mode. I do find that focusing with wobbler is extremely effective with my camera - BioScan 600W - it's top-mounted, so comparison is not quite adequate. They have "loupe" in DM, which is also helpful. Gatan also offered auto-focus plug-in and as far as I remember it was working nicely at their demonstration with 4x4 camera. DigitalMontage is also great - they used beam shift to extend the view, so you could take pictures quite quickly, like 3x3 or even 4x4 arrays. If you'll evaporate carbon on top of your plastic film - it significantly reduces the drift. You may do it over the sections (even better). Sergey
At 04:36 PM 6/21/2005, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
} From what I have heard, the optics of the major instruments are all comparable. The choice of instrument, then, depends on ease of operation (software), reliability, and service. There are some technical details in the design of the spectral features that might bear on the sensitivity of the instrument, though.
I would recommend that you think carefully about what you want to do with this instrument. Do you want an upright or inverted microscope? You should decide which wavelengths you need for the stains you are likely to encounter, and make a similar analysis of the lenses that you will need (long or short working distances,magnifications, etc).. You should also decide if you need spectral capabilities on both the excitation and emission detectors.
A confocal microscope is a significant instrument, with a complexity not that different from the old electron microscopes. You can't just walk in and use it without training. As Gordon suggested, you shoud make sure that there is someone with responsibility for the instrument. This person should be familiar enough to give instruction and help to individual users.
The software is critical. It isn't that the different software sets will give you different capabilities, but the ease of different operations may vary. As a result, you will find that you may have a different personal reaction to the interface. You, or the person who will be in charge of the instrument, should sit down with each of the models that you are considering, and work with the instrument for an extended period --just looking at the instruments at a meeting, for instance, will not work. You should also see whether the output files can be read by 3d party software, such as Metamorph, or ImageJ, or even Photoshop so that you don't tie up a microscope by doing image processing with it.
In order to make a decision about the brand, it seems to me that, given your support situation, you should investigate the service that is available, and its cost. Service contracts can be quite expensive, and I have heard anecdotes about the difficulty of service with most of the manufacturers (although I have also heard some praises for the same service organizations).
For more details on this, scan the archives of the confocal listserv: CONFOCAL-at-LISTSERV.BUFFALO.EDU
Hope these musings are helpful. I can give you specific experiences offlist.
Joel
Date sent: Tue, 21 Jun 2005 17:49:03 -0500 To: microscopy-at-microscopy.com } From: gardnere-at-wpunj.edu (by way of MicroscopyListserver)
On our JEM-2010 we use a LaB6 emitter with a 90 degree cone and 15 micron tip.
We are considering using a smaller cone angle and/or smaller tip radius.
Does anyone have any comments concerning whether the supposed improvement in brightness and coherence is worth the shortened lifetime?
_________________________________
Joseph Kulik Research Associate Materials Research Institute The Pennsylvania State University 194 MRI Bldg University Park, PA 16802
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (xiaohutang-at-gmail.com) from http://www.msa.microscopy.org/MicroscopyListserver/MLFormMail.html on Wednesday, June 22, 2005 at 02:27:04 ---------------------------------------------------------------------------
Email: xiaohutang-at-gmail.com Name: Xiaohu Tang
Organization: TU Delft, the Netherlands
Title-Subject: [Microscopy] [Filtered] 3D image reconstruction software for stereo SEM
Question: Hello everyone,
I am looking for a 3D image reconstruction software for stereo SEM to measure the sample volume. It looks like the MeX from Alicona is the only choice so far but the price is too expensive. Could anybody give me an idea for a cheaper software or a better method to measure the volume by SEM? Thank you.
Xiaohu Tang Microlab CiTG of Delft Technology University The Netherlands x.tang-at-citg.tudelft.nl
Just a quick calendar item and announcement regarding SCANNING 2006. The 17th annual meeting on the scanning microscopies will take place Tuesday, April 25 through Thursday, April 27 in Washington, D.C. The Welcome Reception will take place on Tuesday, April 25 at the National Press Club.
SPECIAL EVENT: On Wednesday, April 26 there will be a program day at NIST. Transportation will be provided. This is a joint program day included in your SCANNING registration fee.
Hi Xiaohu Have you looked at ImageJ with the needed plugins?
ImageJ's main page is http://rsb.info.nih.gov/ij/
3D Toolkit Plug ins http://ij-plugins.sourceforge.net/ij-3D-toolkit.html
3D Toolkit is a set of plug ins for 3D and 2D operations on images in Image/J. The first group of plug ins reads and writes images in VTKand MetaImage format. Metaimage Reader MetaImage Writer VTK Reader VTK Writer The second group of plugins are early prototypes of plugins for: Morphological dilation (max) Morphological erosion (min) Connected threshold region growing Auto volume clipping
This may be the same set of plug ins http://ij-plugins.sourceforge.net/3d-toolkit/
A Google search for: imagej 3d volume brings over 600 hits and ImageJ and most plugins are free.
ImageJ is Java port with a great deal of enhancement of Sicon Image that USA National Institute of Health had done and put in the public domain. Primarily as health sciences imaging tool but with the open source pugin, scrip an macro language many disciplines are using it for many things. It is platform independent running on any machine ruining Java, Unix, Solaris, Windows, Mac, Mac OS-10, Linux and some more. For a P code machine it amazingly fast. I would like to see it run on computer that ran Java in native mode.
It is bit harder to use that Paint shop but new functionality is available almost on a daily basis and the support from the folks that write the plug ins is on the whole much much better than any commercial product that doesn't cost a fortune for maintenance to provide that kind of service. In many ways ImageJ support is better than the best commercial service because you work with the person that wrote the code. It is not uncommon for someone on the plugin mailing list to help you modify a plugin for you needs. There are no commercial vendors that can turn out code this way.
I have worked for software companies as a programmer and letting the customer work with the programmer is just not done. It uses too much of the programmers time and the customer finds out more than management wants them to know about the product. Because the programmers don't pretend that the patched up out of date software that most products are made out of are anything else but what it is. Most of the time management doesn't have the money to write new software from scratch or look at Cisco when they tried to write a new operating system for their routers and got in disagreement with the programmers and the programmers quit and went into business as Juniper who is now Ciscoes' biggest competitor in their high end most profitable business.
Source code is available for most plugins and ImageJ will compile them giving you a complete tool for learning to write plugins if you want to. Most of use will be content to play the sorcerer's apprentice and charge a few lines to make a plugin that is almost what we do just what we want and not become full blown programmers. But that is very valuable option when you need something slightly different than the person that developed the plug in needed. A Google search for 'imagej plugin' brings up almost 10,000 hits I am sure many of those are duplicates but 3,000 to 5,000 plugins is not an unreasonable estimate.
Best Regards Gordon Gordon Couger
I collect links on information related to light microscopes. www.couger.com/microscope/links/gclinks.html Please forward anything you think might be useful to others. Microscope Documentation is at www.science-info.org
by way of MicroscopyListserver wrote: } } Email: xiaohutang-at-gmail.com } Name: Xiaohu Tang } } Organization: TU Delft, the Netherlands } } Title-Subject: [Microscopy] [Filtered] 3D image reconstruction software for stereo SEM } } Question: Hello everyone, } } I am looking for a 3D image reconstruction software for stereo SEM to measure the sample volume. It looks like the MeX from Alicona is the only choice so far but the price is too expensive. Could anybody give me an idea for a cheaper software or a better method to measure the volume by SEM? Thank you. } } Xiaohu Tang } Microlab } CiTG of Delft Technology University } The Netherlands } x.tang-at-citg.tudelft.nl }
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 22 11:50:45 2005
Gordon, thank you for the valuable information. I checked ImageJ and could not find the proper plugin for my problem: I want to use the SEM stereo image pairs by tilting the sample and find out the volume data from the 3D reconstruction. Based on my understanding (I am not so familar with this area), most of the ImageJ plugins about 3D volume depend on a series of image slices but not like my case. All the suggestions and information will be highly appreciated.
Xiaohu Tang Microlab CiTG of Delft Technology University The Netherlands x.tang-at-citg.tudelft.nl
On 6/22/05, Gordon Couger {gcc-at-couger.com} wrote: } Hi Xiaohu } Have you looked at ImageJ with the needed plugins? } } ImageJ's main page is http://rsb.info.nih.gov/ij/ } } 3D Toolkit Plug ins } http://ij-plugins.sourceforge.net/ij-3D-toolkit.html } } 3D Toolkit is a set of plug ins for 3D and 2D operations on } images in Image/J. The first group of plug ins reads and writes } images in VTKand MetaImage format. } Metaimage Reader } MetaImage Writer } VTK Reader } VTK Writer } The second group of plugins are early prototypes of plugins for: } Morphological dilation (max) } Morphological erosion (min) } Connected threshold region growing } Auto volume clipping } } This may be the same set of plug ins } http://ij-plugins.sourceforge.net/3d-toolkit/ } } A Google search for: } imagej 3d volume } brings over 600 hits and ImageJ and most plugins are free. } } ImageJ is Java port with a great deal of enhancement of Sicon } Image that USA National Institute of Health had done and put in } the public domain. Primarily as health sciences imaging tool but } with the open source pugin, scrip an macro language many } disciplines are using it for many things. It is platform } independent running on any machine ruining Java, Unix, Solaris, } Windows, Mac, Mac OS-10, Linux and some more. For a P code } machine it amazingly fast. I would like to see it run on } computer that ran Java in native mode. } } It is bit harder to use that Paint shop but new functionality is } available almost on a daily basis and the support from the folks } that write the plug ins is on the whole much much better than } any commercial product that doesn't cost a fortune for } maintenance to provide that kind of service. In many ways ImageJ } support is better than the best commercial service because you } work with the person that wrote the code. It is not uncommon for } someone on the plugin mailing list to help you modify a plugin } for you needs. There are no commercial vendors that can turn out } code this way. } } I have worked for software companies as a programmer and letting } the customer work with the programmer is just not done. It uses } too much of the programmers time and the customer finds out more } than management wants them to know about the product. Because } the programmers don't pretend that the patched up out of date } software that most products are made out of are anything else } but what it is. Most of the time management doesn't have the } money to write new software from scratch or look at Cisco when } they tried to write a new operating system for their routers and } got in disagreement with the programmers and the programmers } quit and went into business as Juniper who is now Ciscoes' } biggest competitor in their high end most profitable business. } } Source code is available for most plugins and ImageJ will } compile them giving you a complete tool for learning to write } plugins if you want to. Most of use will be content to play the } sorcerer's apprentice and charge a few lines to make a plugin } that is almost what we do just what we want and not become full } blown programmers. But that is very valuable option when you } need something slightly different than the person that developed } the plug in needed. A Google search for 'imagej plugin' brings } up almost 10,000 hits I am sure many of those are duplicates but } 3,000 to 5,000 plugins is not an unreasonable estimate. } } Best Regards } Gordon } Gordon Couger } } I collect links on information related to light microscopes. } www.couger.com/microscope/links/gclinks.html } Please forward anything you think might be useful to others. } Microscope Documentation is at www.science-info.org } } by way of MicroscopyListserver wrote: } } } } Email: xiaohutang-at-gmail.com } } Name: Xiaohu Tang } } } } Organization: TU Delft, the Netherlands } } } } Title-Subject: [Microscopy] [Filtered] 3D image } reconstruction software for stereo SEM } } } } Question: Hello everyone, } } } } I am looking for a 3D image reconstruction software for } stereo SEM to measure the sample volume. It looks like the MeX } from Alicona is the only choice so far but the price is too } expensive. Could anybody give me an idea for a cheaper software } or a better method to measure the volume by SEM? Thank you. } } } } Xiaohu Tang } } Microlab } } CiTG of Delft Technology University } } The Netherlands } } x.tang-at-citg.tudelft.nl } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 22 12:56:11 2005
Joe; I have used 60 degree cone emitters with great success on a 2000FX (don't recall the tip radius) and 60 degree cones with 5 micron radius on a Topcon 002B. They add a nice bit of crispness to the high resolution image, although they are no substitute for field emission. As for brightness; no problem. As for lifetime, they don't last quite as long at a 90 degree, so if you are budget constrained that might be an issue, but for us it was well worth the minor extra operational cost. Six months was typical, for a TEM used a lot every day. Usually the wehnelt needed cleaning once or twice during the life of the tip so maintenance effort was about the same as a 90 degree. Unfortunately, Denka, which was our preferred source for that tip on the Topcon, has stopped manufacturing that particular source.
John Mardinly
This opinion is the opinion of the auther and does not represent the opinion of Intel Corporation.
-----Original Message----- } From: Joe Kulik [mailto:juk12-at-psu.edu] Sent: Tuesday, June 21, 2005 7:10 PM To: Microscopy-at-microscopy.com
On our JEM-2010 we use a LaB6 emitter with a 90 degree cone and 15 micron tip.
We are considering using a smaller cone angle and/or smaller tip radius.
Does anyone have any comments concerning whether the supposed improvement in brightness and coherence is worth the shortened lifetime?
_________________________________
Joseph Kulik Research Associate Materials Research Institute The Pennsylvania State University 194 MRI Bldg University Park, PA 16802
I do not know just how much you need to lower the magnification but the following may just help?
Raise the eucentric stage to its highest position (moving into focus when the objective lens is weakened) which will reduce the magnification factor of the objective lens. Possibly lowering the magnification by 10% or more?
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7802 966067 Web www.emcourses.comood
----- Original Message ----- } From: "Jay Campbell" {microtomy-at-gmail.com} To: {Microscopy-at-microscopy.com} Sent: Wednesday, June 22, 2005 12:36 AM
Before you choose an instrument I think you need to reconsider your "no dedicated technician" approach. A complex piece of multi-user equipment with no one person to supervise or police its use is a recipe for diaster. It won't be much good for teaching or research if it is not working properly. While you may have a service contract, service engineers are usually spread to thinly to drop by every week for minor adjustments that no one understands or has time to learn.
Geoff
-- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
by way of MicroscopyListserver wrote:
} Email: gardnere-at-wpunj.edu } Name: Eileen Gardner } } Organization: William Paterson University } } Title-Subject: [Microscopy] [Filtered] MListserver: Confocal microscope } } Question: I am the chair of a Biology department at a state university in NJ. I am planning on purchasing a confocal microscope which would be used for both teaching and research. Do you have any ideas as to which brands would be easiest to use and maintain? There will be no technician dedicated to this microscope. Thanks. } } --------------------------------------------------------------------------- } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 22 14:24:36 2005
Kerrie Digital plate story is discussed nearly every year at ListServer. I don't want to repeat everything what was discussed (perhaps you could check archives). From my prospective, digital plates has only one positive side - the linearity is superior. It may be useful if you do a lot of diffraction work. For biological samples if you satisfied with film's linearity, then why you would use image (digital) plates? From practical point of view, image plates is quite inconvenient: you need to use nearly identical procedure of loading, unloading, pumping etc as for the film and you are limited in the number of images you could produce (same as film - 50). You have to load plates manually in the scanner. Most procedures should be performed in the dark I believe. With digital camera you have less resolution but you could generate practically unlimited amount of images (I normally produce about 100 images in 40 min) and vacuum in the microscope is great (because you don't have to vent the camera). Images available immediately (you don't need to wait for scanning - few min per plate I believe). So, I don't think, image plates much suited for biological applications keeping in mind the price etc. A few words about resolution. Somebody on ListServer calculate that single EM film contains about 7 Gbyte of information - it's enormous amount of information, which we never use 100%. We normally cropped our images and print them small (look on the Science magazine pages!), so we dramatically reduce the resolution on the final "product". Then, such "final product" becomes comparable with what we have with digital cameras. Keep in mind that even super-duper printers produce gray tones with resolution about 150 dpi (may be 300 at the best). Personally, I was surprised to see that 3x3" images from my BioScan 1Mpix camera printed by good inkjet printer with "gray inks" practically undistinguishable for the real photo-print. The disadvantage of such low-res images - you could not crop or enlarge them, so you just took many pictures at different magnification. For big field you could combine a few pictures quite easily. I hope it'll help. Have a great day, Sergey
P.S. A new argument against image plates just cames to me: laser. They used, I guess, similar laser as for phosphorimager. That laser has limited life - something like 3 years and should be replaced then. Laser itself is about $20K, so you need to keep expensive service contract or be ready to invest in a new laser 2 or 3 year later... I think, CCD chip and phosphor screen on digital cameras deteriorated as well- mine is still OK after 3 years of extensive use.
At 06:52 AM 6/20/2005, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
} Gordon, thank you for the valuable information. I checked ImageJ and } could not find the proper plugin for my problem: I want to use the SEM } stereo image pairs by tilting the sample and find out the volume data } from the 3D reconstruction. Based on my understanding (I am not so } familar with this area), most of the ImageJ plugins about 3D volume } depend on a series of image slices but not like my case. All the } suggestions and information will be highly appreciated. } Dear Xiaohu. If Sterecon is still out there, it may suit your needs. It was used in Albany NY some years ago, and some of the people there may know whether it is still available. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 22 16:24:07 2005
I have to agree with Geoff and others on this list. I have been in this business for over 25 years and I have never seen a situation where a piece of equipment with the complexity of a confocal or an SEM or a TEM or FIB or STM or you name it can be maintained at factory spec without a dedicated support person to manage the device and train users. Over and over again at this campus I have seen PIs win grants for equipment without first having thought through the idea of dedicated support. They will assume an overloaded tech or lab maanger can do it in their "spare time". That won't work and eventually the machine will not be capable of quality work or reliability. The equipement wll be used less and less and within a short time will be salvaged out. Then, in a year or three, another PI will need that same piece of equipment and we will start the process again.
Rick Harris Unniversity of California Davis, CA
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 22 16:53:03 2005
Stereo techniques usually give you a surface reconstruction, not a volume reconstruction. Whether you can get meaningful volumetric data depends on what you are looking at. For example: consider a spherical object lying on a surface. Stereo images are usually taken at a few degrees tilt apart, so the two images essentially see the same half of the sphere. You can measure half of the sphere's volume, but since you cannot see the "backside", you have to make assumptions. For a sphere, it is fairly easy, but if your object is irregular, you may not be able to measure its volume without making assumptions what the "hidden" part looks like.
I am sure you have considered this. I would also point you to our website, which has information about the stereo module in our software: http://www.soft-imaging.com/rd/english/405.htm
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: by way of MicroscopyListserver [mailto:xiaohutang-at-gmail.com] Sent: Wednesday, June 22, 2005 07:05 To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (xiaohutang-at-gmail.com) from http://www.msa.microscopy.org/MicroscopyListserver/MLFormMail.html on Wednesday, June 22, 2005 at 02:27:04 ---------------------------------------------------------------------------
Email: xiaohutang-at-gmail.com Name: Xiaohu Tang
Organization: TU Delft, the Netherlands
Title-Subject: [Microscopy] [Filtered] 3D image reconstruction software for stereo SEM
Question: Hello everyone,
I am looking for a 3D image reconstruction software for stereo SEM to measure the sample volume. It looks like the MeX from Alicona is the only choice so far but the price is too expensive. Could anybody give me an idea for a cheaper software or a better method to measure the volume by SEM? Thank you.
Xiaohu Tang Microlab CiTG of Delft Technology University The Netherlands x.tang-at-citg.tudelft.nl
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-----Original Message----- } From: "Rick A. Harris" {raharris-at-ucdavis.edu}
The perspective from the Field Service Engineer:
If I had to drop by every customer every week or so for minor "stuff", I would be: 1) out of business (broke), 2) out of my mind 3) out of customers because I can't visit all of them that often and the ones who had REAL problems expect (rightly so) to be taken care of quickly and thoroughly.
PLEASE, PLEASE, PLEASE do not expect your field service engineer to substitute for a good technician. It is unfair to ALL involved.
If you want your service organization to provide full-time service, expect to pay 10 or 15 times what you're currently paying for your service contract.
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Wednesday, June 22, 2005 2:54 PM To: by way of MicroscopyListserver Cc: microscopy-at-microscopy.com
Before you choose an instrument I think you need to reconsider your "no dedicated technician" approach. A complex piece of multi-user equipment with no one person to supervise or police its use is a recipe for diaster. It won't be much good for teaching or research if it is not working properly. While you may have a service contract, service engineers are usually spread to thinly to drop by every week for minor adjustments that no one understands or has time to learn.
Geoff
-- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
by way of MicroscopyListserver wrote:
} Email: gardnere-at-wpunj.edu } Name: Eileen Gardner } } Organization: William Paterson University } } Title-Subject: [Microscopy] [Filtered] MListserver: Confocal microscope } } Question: I am the chair of a Biology department at a state university in NJ. I am planning on purchasing a confocal microscope which would be used for both teaching and research. Do you have any ideas as to which brands would be easiest to use and maintain? There will be no technician dedicated to this microscope. Thanks. } } --------------------------------------------------------------------------- } }
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 22 21:27:40 2005
We have been having oil leak problem with our JEM-2010F desiccator. We have had 3 desiccators (EMDSC-U10A) and had leaking every 5-6 months. We were told that the parts replacement wouldn't help so much, because this model had design problem with about 40% failure possibility. Now, we are looking for new reliable type of desiccator. Does anyone have any suggestion? Thanks in advance!
With best regards, ******************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 (Office) 919-843-0974 (EM Lab) Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 22 22:12:14 2005
All, I am currently trying to make TEM samples out of very porous sintered capacitor materials (Nb and NbO) in which I need to collect stoichiometric data via EELS analysis. The problem in making these samples is that they do not polish or ion mill well due to their porous morphology. I had tried to infiltrate with M-bond, which helped considerably, but even after plasma treating the samples before analysis, a large C-K edge remained throughout the sample (which unfortunately sits right in between the elements of interest). I was wondering if anyone had any suggestions of a filler that could be used to infiltrate a porous body and then could be easily removed post ion thinning. Any help would be greatly appreciated.
Regards Matt Olszta Ph.D. Materials Research Institute Penn State University
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 01:02:01 2005
We wish to negatively stain some vaccine formulations. Does anyone know of a negative stain that you can use at acid pH that is not a uranyl salt? Does anyone know of a negative stain that you can use at basic pH that is not sodium tetraborate? We have used sodium tetraborate but it tends to "bubble" under the beam!
Ross Hamilton, Senior Scientist, E.M. Unit, Bioanalytical Sciences, CSL Limited, 45 Poplar Road, Parkville,3052 Victoria,Australia. +61 3 9389 1307
**************************************************************************************************************** This email and any attached files are intended solely for the named addressee, are confidential and may contain legally privileged information. The copying or distribution of them or any information they contain, by anyone other than the addressee, is prohibited. If you have received this email in error, please let us know by telephone or return the email to the sender and destroy all copies. Thank you.
CSL Limited A.C.N. 051 588 348 45 Poplar Road Parkville Victoria 3052 Australia Phone: +61 3 9389 1911 Fax: +61 3 9389 1434 ***************************************************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 07:35:37 2005
Sounds sort of like Pixel envy doesn't it? - since the new imaging plate systems can make images with up to 110Mega pixels - you'd need to shoot more than 100 pictures at 1Meg each to get equivalent images - and by the way the systems use diode lasers that have very long life and they don't cost anywhere near $20K
Imaging plates are not necessarily the right solution for every situation but to categorically state that they are useless for biological use is simply wrong. Like with any system there are strengths and weaknesses but if nothing else the full field of view pixels/$ is unbeatable.
Bill Miller
At 03:24 PM 6/22/2005, Sergey Ryazantsev wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 08:35:45 2005
Can someone tell me the nominal consitutents of EPON used for embedding TEM biological samples. Specifically what are it's major elemental components. (C, H, O, N, ......???)
It is not obvious to me which of the many EPONS listed in a Google search is the one typically used by the life science community in preparing TEM sections.
Thanks in advance,
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 08:35:51 2005
check out www.nanoprobes.com they sell tungstate and vanadate based negative stains. i have never used them so can't comment.
At 12:03 AM 06/23/05, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dwaugh-at-kent.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, June 23, 2005 at 08:34:28 ---------------------------------------------------------------------------
Question: I'm looking to make contact microradiographs of crab claw thin sections (~40 microns thick). We have an x-ray machine, the problem is the film. Doing a google search I have found two Kodak films that are used, but can't figure out if they are still in production. Kodak SO-343, seems to be popular, I can't tell if it is an xray or holographic film, but either way there is no mention of it on the Kodak website or from any supplier, same thing for Kodak "spectroscopic film". A film "Fuji B&W POS/71337" that I found in one paper, does not seem to exist. I need something that is very high resolution, and preferable can be developed with standard B&W chemistry. Any ideas on what the current films in use are? Would a conventional film like Kodak T-max work (we have that film and chemical already lying around in the now mostly mothballed darkroom)? Thanks, David
At Energy Beam Sciences we can rebuild any tungsten filament, including Siemens. We also provide new mounted filaments for most Microscopes. You can contact us on (800) 992-9037 or by email at ebs-at-ebsciences.com.
Michael R. Nesta Managing Director Energy Beam Sciences, Inc. Tel: 860 653-0411 Fax: 860 653-0422 MNesta-at-ebsciences.com www.ebsciences.com “ADDING BRILLIANCE TO YOUR VISION”
by way of MicroscopyListserver wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 09:08:42 2005
I am posed to finally make the leap to a digital camera (a Gatan Ultrascan 1000) after years of SO-163. One potential problem that I face, however, is digital recording of SAD diffraction patterns. My top-entry Hitachi H-9000UHR HRTEM did not come with a beam stop (that rod or pointer thing that you use to block the main beam while recording a diffraction pattern) and it is therefore my concern that I might not be able to record diffraction patterns without damaging the digital camera. Hitachi does not seam to have a beam stop thing for my vintage (1991) H-9000, so I am behind the eight ball in this regard. I hear from Gatan that I should try to avoid using film once the Ultrascan is installed, so that that flaked-off film or whatever does not fall onto the digital camera and mess it up, so using film to record diffraction patterns may not be an option either. Any suggestions?
Thanks,
Mark -- Dr. Mark E. Twigg Code 6812 Naval Research Laboratory 4555 Overlook Ave., S.W. Washington DC 20375
tel: (202) 404-8543 fax: (202) 404-7194
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 09:18:16 2005
Epon 812 was a Shell patented formulation and has now been replaced by numerous "identical chemical equivalents" according to EM suppliers but rarely are the formulations given. It is a
glycerol polyglycidyl ether (i think it is a mixture of different glycidyl ethers).
DDSA (Dodecenyl Succinic Anhydride) (about 2:1:1 for epon/nma/ddsa but this varies in different labs) plus either DMP-30 Tri-(dimethylaminomethyl) phenol
C15H29N8OH or BDMA as catalysts
Tom
At 08:35 AM 06/23/05, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
The discontinued Denka source you are referring to was the M-7, which featured a hard mounted tip. All of the tip configurations that were available on the M-7 are still available on the M-3, wire mounted tip, including the 60 degree cone angle/5 micron radius configuration you referred to. The reason Denka discontinued the M-7 was that they discovered through internal testing that the M-3 provides stability equal to or greater than the M-7. Denka feels that this discovery rendered the higher cost for the M-7 an unnecessary expense.
Disclaimer - Energy beam Sciences, Inc. is the US distributor for Denka LaB6 and Thermal Field Emission Cathodes.
Michael R. Nesta Managing Director Energy Beam Sciences, Inc. Tel: 860 653-0411 Fax: 860 653-0422 MNesta-at-ebsciences.com www.ebsciences.com “ADDING BRILLIANCE TO YOUR VISION”
Mardinly, John wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 10:00:02 2005
I routinely use our US-1000 for single crystal diffraction patterns with no beam stop. This is because the central beam is actually fairly weak in this situation, as so much intensity is getting diffracted. If this is the large majority of your diffraction work you will be fine.
On the other hand, when collecting polycrystalline ring patterns or patterns from amorphous samples, you will have a problem. The rings are typically weak and the central spot is strong. To get decent signal to noise in the rings you need to up the exposure time or dose. This is where I use the beam stop so if you do a lot of this you should consider making one. Without the stop you'll pretty heavily saturate the CCD, producing a large streak across the central spot.
Talk to Gatan about conditions which would damage the phosphor on the CCD, but I think they are rather extreme - typically these are pretty robust for normal TEM and diffraction conditions.
Regards, Wharton
************************************************************* Wharton Sinkler, PhD. UOP LLC 50 E. Algonquin Rd. Des Plaines IL 60017-5017 mailto: wharton.sinkler-at-uop.com tel 847-391-3878 fax 847-391-3719
-----Original Message----- } From: twigg-at-estd.nrl.navy.mil [mailto:twigg-at-estd.nrl.navy.mil] Sent: Thursday, June 23, 2005 9:09 AM To: Microscopy-at-microscopy.com
I am posed to finally make the leap to a digital camera (a Gatan Ultrascan 1000) after years of SO-163. One potential problem that I face, however, is digital recording of SAD diffraction patterns. My top-entry Hitachi H-9000UHR HRTEM did not come with a beam stop (that rod or pointer thing that you use to block the main beam while recording a diffraction pattern) and it is therefore my concern that I might not be able to record diffraction patterns without damaging the digital camera. Hitachi does not seam to have a beam stop thing for my vintage (1991) H-9000, so I am behind the eight ball in this regard. I hear from Gatan that I should try to avoid using film once the Ultrascan is installed, so that that flaked-off film or whatever does not fall onto the digital camera and mess it up, so using film to record diffraction patterns may not be an option either. Any suggestions?
Thanks,
Mark -- Dr. Mark E. Twigg Code 6812 Naval Research Laboratory 4555 Overlook Ave., S.W. Washington DC 20375
tel: (202) 404-8543 fax: (202) 404-7194
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 10:18:22 2005
I think the idea of the list server is to share experience one has made in any topic with the microscopy community. But this assumes that one has made this experience personally. Probably everybody believes and guesses a lot about instruments, companies and persons but shall such guesswork be shared on such a public experience based forum like this?
Please let me point out what up-to-date laser technology means: Sergey is right, former laser had a limited life time. Such gas lasers being produced some decades ago had this limitations whereas modern semiconductor lasers as used in the DITABIS MICRON are very stable and have a life time comparable to CCD sensors. I am disappointed to see it obviously has become more and more common to read "personal opinions" of some forum members not being based on gained knowledge.
The resolution discussion: why does anybody want to have high resolution systems when downsizing the resolution and more important loosing the information which should be the aim of any scientifical work. I know that our scientific customers do need the information and don't make high resolution images just for their personal satisfaction.
Another point mentioned by John Mardinly was that the imaging plate store only electrons. In principle he is right except that you can detect nearly every high energy radiation with imaging plates. With former imaging plate systems an identification of the imaging plate - and thus the exposure - was impossible. The Ditabis imaging plate system is able to optically scan the surface (e.g. labeled with unique numbers.....) in parallel to normal image scans. That label information is stamped into the final image in one and the same run. A software package even allows to merge TEM settings information into the header of each respective image.
Best regards, Andreas Elkeries
Dipl.-Ing. Customer Support / R&D
------------------------------------- DITABIS AG Digital Biomedical Imaging Systems AG Stuttgarter Str. 13 D-75179 Pforzheim, Germany
} Sergey Ryazantsev wrote: } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Kerrie } Digital plate story is discussed nearly every year at ListServer. I } don't want to repeat everything what was discussed (perhaps you could } check archives). From my prospective, digital plates has only one } positive side - the linearity is superior. It may be useful if you do } a lot of diffraction work. For biological samples if you satisfied } with film's linearity, then why you would use image (digital) plates? } From practical point of view, image plates is quite inconvenient: you } need to use nearly identical procedure of loading, unloading, pumping } etc as for the film and you are limited in the number of images you } could produce (same as film - 50). You have to load plates manually in } the scanner. Most procedures should be performed in the dark I } believe. With digital camera you have less resolution but you could } generate practically unlimited amount of images (I normally produce } about 100 images in 40 min) and vacuum in the microscope is great } (because you don't have to vent the camera). Images available } immediately (you don't need to wait for scanning - few min per plate I } believe). So, I don't think, image plates much suited for biological } applications keeping in mind the price etc. A few words about } resolution. Somebody on ListServer calculate that single EM film } contains about 7 Gbyte of information - it's enormous amount of } information, which we never use 100%. We normally cropped our images } and print them small (look on the Science magazine pages!), so we } dramatically reduce the resolution on the final "product". Then, such } "final product" becomes comparable with what we have with digital } cameras. Keep in mind that even super-duper printers produce gray } tones with resolution about 150 dpi (may be 300 at the best). } Personally, I was surprised to see that 3x3" images from my BioScan } 1Mpix camera printed by good inkjet printer with "gray inks" } practically undistinguishable for the real photo-print. The } disadvantage of such low-res images - you could not crop or enlarge } them, so you just took many pictures at different magnification. For } big field you could combine a few pictures quite easily. I hope it'll } help. Have a great day, Sergey } } P.S. A new argument against image plates just cames to me: laser. } They used, I guess, similar laser as for phosphorimager. That laser } has limited life - something like 3 years and should be replaced } then. Laser itself is about $20K, so you need to keep expensive } service contract or be ready to invest in a new laser 2 or 3 year } later... I think, CCD chip and phosphor screen on digital cameras } deteriorated as well- mine is still OK after 3 years of extensive use. } } At 06:52 AM 6/20/2005, you wrote: } } } } ------------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } } } I have been given the unenviable task of having to decide between a } } digital } } camera system and a digital imaging plate system (the Ditabis system) to } } enable our older TEMs to become fully digital imaging systems. We } } have Jeol } } 1200EX and 100CX instruments and a Phillips CM10. Has anyone had } } experience } } of the Ditabis system? In the UK there is only one currently in use, } } and I } } have contact with that person. Our samples are biological samples, some } } routine clinical specimens, but also research samples and cell culture } } samples. We currently use Kodak em film 4489 and require the } } replacement } } system to have comparable resolution. } } Any comments would be gratefully received. } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } 10833 Le Conte Ave, Room 33-080 } Los Angeles, CA 90095 } } Phone: (310) 825-1144 (office) } (310) 206-1029 (Lab) } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 11:11:34 2005
Michael; Thank you for this good news. When I attempted to order an M7 a few months ago from another vendor of Denka emitters, that information was not available.
John Mardinly
-----Original Message----- } From: Mike Nesta [mailto:mnesta-at-ebsciences.com] Sent: Thursday, June 23, 2005 7:20 AM To: Mardinly, John Cc: Joe Kulik; Microscopy-at-microscopy.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 11:18:08 2005
We're planning to study bacteria suspension (Staphylococcus aureus) and bone marrow with TEM. Does anybody help us for bacteria and bone marrow (about 1 ml) processing protocol for TEM?
Thanks...
Dr. Necat Yilmaz
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 11:34:31 2005
Andreas Elkeries; As Nestor has pointed out, it is inappropriate to make disparaging remarks about individuals in the MSA Listserver, especially as to whether their comments come from personal experience or hallucination. It is especially surprising to see such obstreperousness coming from a member of any 'Customer Support' staff. In my particular case, I conducted an evaluation of Ditabis imaging plates through an authorized US dealer of your product, as Mr. Miller can attest to. The full results of that evaluation cannot be shared on the Listserver, but I can say that no microscope data that would normally be printed on film was present on the images I received, and in discussions with Mr. Miller, there was never communicated any expectation that the microscope data could ever be imprinted on imaging plates. If there is a new development that would allow that, I think we would all be interested in hearing more details about it.
John Mardinly
-----Original Message----- } From: Technik [mailto:technik-at-ditabis.de] Sent: Thursday, June 23, 2005 8:19 AM To: Microscopy-at-microscopy.com; sryazant-at-ucla.edu
Dear Community ,
I think the idea of the list server is to share experience one has made in any topic with the microscopy community. But this assumes that one has made this experience personally. Probably everybody believes and guesses a lot about instruments, companies and persons but shall such guesswork be shared on such a public experience based forum like this?
Please let me point out what up-to-date laser technology means: Sergey is right, former laser had a limited life time. Such gas lasers being produced some decades ago had this limitations whereas modern semiconductor lasers as used in the DITABIS MICRON are very stable and have a life time comparable to CCD sensors. I am disappointed to see it obviously has become more and more common to
read "personal opinions" of some forum members not being based on gained knowledge.
The resolution discussion: why does anybody want to have high resolution
systems when downsizing the resolution and more important loosing the information which should be the aim of any scientifical work. I know that our scientific customers do need the information and don't make high resolution images just for their personal satisfaction.
Another point mentioned by John Mardinly was that the imaging plate store only electrons. In principle he is right except that you can detect nearly every high energy radiation with imaging plates. With former imaging plate systems an identification of the imaging plate
- and thus the exposure - was impossible. The Ditabis imaging plate system is able to optically scan the surface (e.g. labeled with unique numbers.....) in parallel to normal image scans. That label information is stamped into the final image in one and the same run. A software package even allows to merge TEM settings information into the header of each respective image.
Best regards, Andreas Elkeries
Dipl.-Ing. Customer Support / R&D
------------------------------------- DITABIS AG Digital Biomedical Imaging Systems AG Stuttgarter Str. 13 D-75179 Pforzheim, Germany
} Sergey Ryazantsev wrote: } } ------------------------------------------------------------------------ ------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } } Kerrie } Digital plate story is discussed nearly every year at ListServer. I } don't want to repeat everything what was discussed (perhaps you could } check archives). From my prospective, digital plates has only one } positive side - the linearity is superior. It may be useful if you do } a lot of diffraction work. For biological samples if you satisfied } with film's linearity, then why you would use image (digital) plates? } From practical point of view, image plates is quite inconvenient: you } need to use nearly identical procedure of loading, unloading, pumping } etc as for the film and you are limited in the number of images you } could produce (same as film - 50). You have to load plates manually in
} the scanner. Most procedures should be performed in the dark I } believe. With digital camera you have less resolution but you could } generate practically unlimited amount of images (I normally produce } about 100 images in 40 min) and vacuum in the microscope is great } (because you don't have to vent the camera). Images available } immediately (you don't need to wait for scanning - few min per plate I
} believe). So, I don't think, image plates much suited for biological } applications keeping in mind the price etc. A few words about } resolution. Somebody on ListServer calculate that single EM film } contains about 7 Gbyte of information - it's enormous amount of } information, which we never use 100%. We normally cropped our images } and print them small (look on the Science magazine pages!), so we } dramatically reduce the resolution on the final "product". Then, such
} "final product" becomes comparable with what we have with digital } cameras. Keep in mind that even super-duper printers produce gray } tones with resolution about 150 dpi (may be 300 at the best). } Personally, I was surprised to see that 3x3" images from my BioScan } 1Mpix camera printed by good inkjet printer with "gray inks" } practically undistinguishable for the real photo-print. The } disadvantage of such low-res images - you could not crop or enlarge } them, so you just took many pictures at different magnification. For } big field you could combine a few pictures quite easily. I hope it'll
} help. Have a great day, Sergey } } P.S. A new argument against image plates just cames to me: laser. } They used, I guess, similar laser as for phosphorimager. That laser } has limited life - something like 3 years and should be replaced } then. Laser itself is about $20K, so you need to keep expensive } service contract or be ready to invest in a new laser 2 or 3 year } later... I think, CCD chip and phosphor screen on digital cameras } deteriorated as well- mine is still OK after 3 years of extensive use. } } At 06:52 AM 6/20/2005, you wrote: } } } } ------------------------------------------------------------------------ ------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------ ------- } } } } } } I have been given the unenviable task of having to decide between a } } digital } } camera system and a digital imaging plate system (the Ditabis system) to } } enable our older TEMs to become fully digital imaging systems. We } } have Jeol } } 1200EX and 100CX instruments and a Phillips CM10. Has anyone had } } experience } } of the Ditabis system? In the UK there is only one currently in use,
} } and I } } have contact with that person. Our samples are biological samples, some } } routine clinical specimens, but also research samples and cell culture } } samples. We currently use Kodak em film 4489 and require the } } replacement } } system to have comparable resolution. } } Any comments would be gratefully received. } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } 10833 Le Conte Ave, Room 33-080 } Los Angeles, CA 90095 } } Phone: (310) 825-1144 (office) } (310) 206-1029 (Lab) } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 12:25:31 2005
} } From: gilles_grondin-at-uqtr.ca } } ************************************************************* } Cutting sections from methacrylate-embedded blocks (non-plastination) } } Hi, } Anyone want to cut some plastic sections for us? } } We have a number of tissue blocks embedded for light microscopy in hpma, and } we would like them sectioned at 1.5 - 2 microns, and mounted on slides. We } aren't currently set up for cutting plastic sections, and if there is anyone } with some spare microtome capacity who would like to earn some pocket money } here's your chance. I would like an estimate of the per-slide cost of cutting } sections and mounting them on a glass slide. Just guessing, initially we } would } like about 100 sections from about 20 different blocks. } } As an aside, anyone tried to embed blocks with P40 and then cut them for } light } microscopy? } } Sincerely, } Dave Griffiths } } David Griffiths } Section of Anatomy and Pathology } Norwegian School of Veterinary Science } P. O. Box 8146 Dep } N-0033 Oslo } Norway } Tel: 22964542/Fax: 22964764 } E-mail: david.griffiths-at-veths.no } **************************************************** } } ------------------------------------------------- } Courriel expédié via https://courriel.uqtr.ca
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 18:56:17 2005
As rightly pointed out by John Mardinly, a drawback to the imaging plates has always been the lack of foolproof data and image correlation. There is now a solution that for some TEMs will automatically read the microscope parameters and link it with bar coded plates which are read during the scan process.
A complete description of the tracking system is available though ElectroImage.
Best Regards - Bill Miller
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 23 20:18:28 2005
Bill This is very common mistake thinking that more pixels gives you 'more' resolution. If your sample has, let say 10 nm details and nothing smaller, I would expect that quite decent 'resolution' on this particular sample would be if you utilize something like 3 pixels per 10 nm =} 3.3 nm/pixel. You simply don't need more pixels if there is nothing smaller on your sample. Adding more pixels you will basically add more noise to the image and that's it. Ultrathin sections of biological material, which I called 'biological samples' have quite poor resolution because of fixation, dehydratation plastic embedding and so on. Therefore, you will just spoil those 110M of pixels... It's like some people expected 3A resolution (resolution of the scope itself) on 80 nm thick poorly infiltrated chattered plastic sections of biological material, no way: 100A may be...
Again, even on my 3.3GHz Pentium 5 with 2 Gb 3400 RAM computer, I do see deterioration in performance if I am working on couple 50Mb pictures, so you really need super computer to handle your 110 Mb pictures. And I am not talking about printing those images: on my networked color Xerox Phaser 7700GX with a lot of memory (maximum permitted) it takes about 20-30 sec to sent 10-20 Mb image (and more than minute to print at 1200 dpi). Your files will simply kill all our UCLA network! I think, you guys have better network, probably dedicated to handle such giant files.... So, I am just reasoning, why from my prospective, there is not much use of image plates in biological applications. I am glad that Ditabis is using solid state lasers - it definitely will add some juice to the system. By the way, how long it takes to scan one plate at maximum resolution?
I do belive that image plate technology has a great potential in material science applications when huge dynamic range is critical. Have a great day, Sergey
At 05:34 AM 6/23/2005, you wrote: } Sounds sort of like Pixel envy doesn't it? - since the new imaging plate } systems can make images with up to 110Mega pixels - you'd need to shoot } more than 100 pictures at 1Meg each to get equivalent images - and by the } way the systems use diode lasers that have very long life and they don't } cost anywhere near $20K } } } Imaging plates are not necessarily the right solution for every situation } but to categorically state that they are useless for biological use is } simply wrong. Like with any system there are strengths and weaknesses but } if nothing else the full field of view pixels/$ is unbeatable. } } } Bill Miller } } At 03:24 PM 6/22/2005, Sergey Ryazantsev wrote: } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
Dear All, .. I might add for those interested in getting data from a Philips EM 400, 410, 420 and 430 that there is a interface card you can install without much knowledge. It will get you High tension value Magnification / diffraction camera length (dependent on chosen camera: TV, sheet film, 224x36mm port) Filmcode Filmnumber Exposure time on filmilm Please see http://www.stefan-diller.com/rem_tem3_en.htm.
For Zeiss EM 10 A, B, C there is the same possibility but without transmission of filmnumbers (but I might develop such an interface if there is the need). Values are only High tension Magnification 0.4x Magnifying switch Please see http://www.stefan-diller.com/rem_tem4_en.htm Installation should be by service technician.
Values are send via RS 232 interface to the computer. There should be no problem to incorporate them in any imaging software.
Best regards, Stefan Diller
----- Original Message ----- } From: "Bill Miller" {microbill-at-mohawk.net} To: {Microscopy-at-microscopy.com} Sent: Friday, June 24, 2005 1:55 AM
Hello Mark,
Diffraction beam stop is a standard option of SIA TEM CCD cameras. It is available separately (without a camera) as well.
Vitaly Feingold SIA 2773 Heath Lane Duluth, GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com
----- Original Message ----- } From: {twigg-at-estd.nrl.navy.mil} To: {Microscopy-at-microscopy.com} Sent: Thursday, June 23, 2005 10:09 AM
Sergey - I would normally ignore this kind of thing but you have missed the point entirely and I hope this helps to clarify it. The number of pixels does not necessarily define the resolution of an image - in this case it defines the field of view. The pixels can be as small as 7.5um in the image plane but what counts is that the image can be as large as ~11,000 x 10,000 pixels. The high pixel count allowing you to take MANY fewer pictures to get the same information; so one picture contains the information (pixels) of 100 1mega pixel images (if both sensors have 7.5um pixel in the image plane) - I've responded to your points individually below..
Bill
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Bill This is very common mistake thinking that more pixels gives you 'more' resolution.
} No one ever said that more pixels gave more resolution - resolution is dependent upon the pixel size in the image plane as well as point spread and MTF of the system - more pixels give a larger field of view not more resolution.
If your sample has, let say 10 nm details and nothing smaller, I would expect that quite decent 'resolution' on this particular sample would be if you utilize something like 3 pixels per 10 nm =} 3.3 nm/pixel. You simply don't need more pixels if there is nothing smaller on your sample.
} Resolution is one thing but being able to actually visualize a structure requires significant oversampling - how confident would you be of a structure represented by 3 pixels - would you ever enlarge the image enough to even really see 3 pixels? The Nyquist sampling limit is fine for mathematically determining the resolution limit of an image but over sampling by a factor of 5 or 10 will give you enough pixels to actually see the structure. But resolution is not the relavent point to all of this , it is field of view.
Adding more pixels you will basically add more noise to the image and that's it.
} More pixels = more noise – an interesting, but an obviously flawed concept or there would be NO market for high end digital cameras of any sort. It might be true if the pixels were smaller rather than spread over a larger area. In this case the idea of more pixels is to expand the field of view not to increase the resolution. The pixel size determines the resolution not the pixel number - you are confusing the two.
Ultrathin sections of biological material, which I called 'biological samples' have quite poor resolution because of fixation, dehydratation plastic embedding and so on. Therefore, you will just spoil those 110M of pixels... It's like some people expected 3A resolution (resolution of the scope itself) on 80 nm thick poorly infiltrated chattered plastic sections of biological material, no way: 100A may be...
} So with more pixels you can get the 100A resolution in an image at lower magnification and a concomitantly larger field of view - again fewer pictures to shoot -- seems like simple enough concept
Again, even on my 3.3GHz Pentium 5 with 2 Gb 3400 RAM computer, I do see deterioration in performance if I am working on couple 50Mb pictures, so you really need super computer to handle your 110 Mb pictures. So is it easier and faster to process over 100 images or a single image that does not have to be stitches to get all of the information? And I am not talking about printing those images: on my networked color Xerox Phaser 7700GX with a lot of memory (maximum permitted) it takes about 20-30 sec to sent 10-20 Mb image (and more than minute to print at 1200 dpi).
} Are the people who are scanning their negatives doing so at 300dpi for fear thet the file would otherwise be too big? Absolutely not. People pay a great deal of money for scanners with 7um pixels so that they get as much information as possible form their TEM negatives. Big files are a fact of life in science - how big is a confocal image made of 100’s of serial images?
simply kill all our UCLA network!
} So don't use it, but that doesn't mean that someone else might not "feel" like you do.
I think, you guys have better network, probably dedicated to handle such giant files.... So, I am just reasoning, why from my prospective, there is not much use of image plates in biological applications.
} That’s like asking why everybody doesn't drive a Ferrari!!! And just what direct knowledge of the use of plates in biology do you have? Just like with megapixel CCD cameras, price, not performance is the bigger barrier. How many people do you think are stuck with a 1meg small field of view camera for biological work - forced to shoot many hundreds of images to get a field of view comparable in size and resolution to what they have been getting with film - just because they couldn't afford the 16meg CCD camera? Would you have bought the 1meg camera if you could have afforded a 4 or 16meg one? So, I wonder why there is not much use of 8k x 8k cameras in biological applications? Increased noise? Nope, it's the Money!
I am glad that Ditabis is using solid state lasers - it definitely will add some juice to the system. By the way, how long it takes to scan one plate at maximum resolution?
} a few unattended minutes - far less time than it takes to shoot and stitch the comparable 100+ 1 meg images required to cover the same field of view.
I do belive that image plate technology has a great potential in material science applications when huge dynamic range is critical. Have a great day, Sergey
At 05:34 AM 6/23/2005, you wrote:
Sounds sort of like Pixel envy doesn't it? - since the new imaging plate systems can make images with up to 110Mega pixels - you'd need to shoot more than 100 pictures at 1Meg each to get equivalent images - and by the way the systems use diode lasers that have very long life and they don't cost anywhere near $20K
Imaging plates are not necessarily the right solution for every situation but to categorically state that they are useless for biological use is simply wrong. Like with any system there are strengths and weaknesses but if nothing else the full field of view pixels/$ is unbeatable.
Bill Miller
At 03:24 PM 6/22/2005, Sergey Ryazantsev wrote:
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Kerrie Digital plate story is discussed nearly every year at ListServer. I don't want to repeat everything what was discussed (perhaps you could check archives). From my prospective, digital plates has only one positive side - the linearity is superior. It may be useful if you do a lot of diffraction work. For biological samples if you satisfied with film's linearity, then why you would use image (digital) plates? From practical point of view, image plates is quite inconvenient: you need to use nearly identical procedure of loading, unloading, pumping etc as for the film and you are limited in the number of images you could produce (same as film - 50). You have to load plates manually in the scanner. Most procedures should be performed in the dark I believe. With digital camera you have less resolution but you could generate practically unlimited amount of images (I normally produce about 100 images in 40 min) and vacuum in the microscope is great (because you don't have to vent the camera). Images available immediately (you don't need to wait for scanning - few min per plate I believe). So, I don't think, image plates much suited for biological applications keeping in mind the price etc. A few words about resolution. Somebody on ListServer calculate that single EM film contains about 7 Gbyte of information - it's enormous amount of information, which we never use 100%. We normally cropped our images and print them small (look on the Science magazine pages!), so we dramatically reduce the resolution on the final "product". Then, such "final product" becomes comparable with what we have with digital cameras. Keep in mind that even super-duper printers produce gray tones with resolution about 150 dpi (may be 300 at the best). Personally, I was surprised to see that 3x3" images from my BioScan 1Mpix camera printed by good inkjet printer with "gray inks" practically undistinguishable for the real photo-print. The disadvantage of such low-res images - you could not crop or enlarge them, so you just took many pictures at different magnification. For big field you could combine a few pictures quite easily. I hope it'll help. Have a great day, Sergey
P.S. A new argument against image plates just cames to me: laser. They used, I guess, similar laser as for phosphorimager. That laser has limited life - something like 3 years and should be replaced then. Laser itself is about $20K, so you need to keep expensive service contract or be ready to invest in a new laser 2 or 3 year later... I think, CCD chip and phosphor screen on digital cameras deteriorated as well- mine is still OK after 3 years of extensive use.
At 06:52 AM 6/20/2005, you wrote:
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have been given the unenviable task of having to decide between a digital camera system and a digital imaging plate system (the Ditabis system) to enable our older TEMs to become fully digital imaging systems. We have Jeol 1200EX and 100CX instruments and a Phillips CM10. Has anyone had experience of the Ditabis system? In the UK there is only one currently in use, and I have contact with that person. Our samples are biological samples, some routine clinical specimens, but also research samples and cell culture samples. We currently use Kodak em film 4489 and require the replacement system to have comparable resolution. Any comments would be gratefully received.
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
Here is the July 2005 Microscopy Today table of contents. I will close the subscription list for this issue on Thursday July 7, 2005.
Microscopists in North America and MSA members anywhere may have free subscriptions. Anyone else may subscribe for US$35 per year (to PARTIALLY cover postage). All subscriptions at http://www.microscopy-today.com Thank you.
Ron Anderson, Editor =============================== A Closer Look at the Nuclear Pore Complex Stephen W. Carmichael, Mayo Clinic
An Innovative Method for Imaging and Chemical Analysis of Wet Samples in Scanning Electron Microscopes Irit Ruach-Nir, QuantomiX Ltd, Rehovot, Israel
An Aberration Corrected (S)TEM Microscope for Nanoresearch S. Kujawa, B. Freitag, D. Hubert, FEI Company, Eindhoven, The Netherlands
Chemical Imaging with Infrared Microscopy William J. McCarthy, Thermo Electron Corporation, Madison, WI
A Novel High-Speed Optical Scanning Microscope for Routine Clinical Applications D. Frekers1,2, I. Aksit1,2, V. Pilipenko1, K. Bünger;2 1Westfälische Wilhelms-Universität Münster, 2MedXP-GmbH, Gelsenkirchen, Germany
Easy Guide to Calibrating TEM’s and STEM’s John McCaffrey, Norrox Scientific Ltd. Ottawa, Canada
Embedding Small Specimens for TEM Examination Paul Webster, House Ear Institute
The Total Release Method for FIB In-Situ TEM Sample Preparation T.M. Moore, Omniprobe, Inc., Dallas, TX
EBSD Sample Preparation: Techniques, Tips, and Tricks Matthew M. Nowell1, Ronald A. Witt2, and Brian W. True1 1EDAX-TSL, Draper, UT, 2EBSD Analytical, Lehi, UT
Funding Opportunities for Acquiring Equipment from Federal Granting Agencies – Part III: IMR Session Chair: Debby Sherman, Purdue University
M&M 2004 Core Facility Management – Part IV: Q & A Moderator: Debbie Sherman
Industry News
NetNotes
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 24 12:46:38 2005
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Is there anyone out there in the Texas Medical Center area or near vicinity that you know who can help repair a MT5000 Sorvall ultramicrotome? It was received as a gift but is obviously not working right now. The person in charge of this thinks it might be just a belt.
Anyone you can recommend?
T.I.A.,
Claire
Claire Haueter EM Tech, Sr. Integrated Microscopy Core Baylor College of Medicine
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From MicroscopyL-request-at-ns.microscopy.com Fri Jun 24 13:15:41 2005
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7802 966067 Web www.emcourses.com
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 24 17:36:28 2005
On Jun 23, 2005, at 6:15 PM, Sergey Ryazantsev wrote:
} This is very common mistake thinking that more pixels gives you 'more' } resolution. If your sample has, let say 10 nm details and nothing } smaller, I would expect that quite decent 'resolution' on this } particular sample would be if you utilize something like 3 pixels per } 10 nm =} 3.3 nm/pixel. You simply don't need more pixels if there is } nothing smaller on your sample. Adding more pixels you will basically } add more noise to the image and that's it. Ultrathin sections of } biological material, which I called 'biological samples' have quite } poor resolution because of fixation, dehydratation plastic embedding } and so on. Therefore, you will just spoil those 110M of pixels... } It's like some people expected 3A resolution (resolution of the scope } itself) on 80 nm thick poorly infiltrated chattered plastic sections } of biological material, no way: 100A may be... } } Again, even on my 3.3GHz Pentium 5 with 2 Gb 3400 RAM computer, I do } see deterioration in performance if I am working on couple 50Mb } pictures, so you really need super computer to handle your 110 Mb } pictures. And I am not talking about printing those images: on my } networked color Xerox Phaser 7700GX with a lot of memory (maximum } permitted) it takes about 20-30 sec to sent 10-20 Mb image (and more } than minute to print at 1200 dpi). Your files will simply kill all our } UCLA network! I think, you guys have better network, probably } dedicated to handle such giant files.... So, I am just reasoning, why } from my prospective, there is not much use of image plates in } biological applications. I am glad that Ditabis is using solid state } lasers - it definitely will add some juice to the system. By the way, } how long it takes to scan one plate at maximum resolution? } } I do belive that image plate technology has a great potential in } material science applications when huge dynamic range is critical. } Have a great day, Sergey } Dear Sergey, I agree that just having more pixels does not show smaller details than are present in the specimen; however, if the mag is adjusted so that the smallest details present are, as you suggest, about 3 pixels across, then the more pixels you have, the larger the field of view--which may be important. Another way of thinking about this is that for larger pixels, as occurs with either image plates or CCDs, you need a higher mag to see the same details, so you will have a smaller field of view. For a conventional fixed and stained biological plastic section, there is no point whatever in having a magnification so large that you can see details within the clumps of stain (typically ~2 nm), and there may be no reason to have a mag high enough to distinguish individual stain clumps (so even the smallest details in such a specimen may be of no interest), but you still may want to see small gold clusters, e.g., on antibodies or other labels, over a large piece of tissue, in which case one would need high enough mag to identify the gold (or even to distinguish among several sizes of gold) and a large enough field to cover the entire region of interest in the specimen. It is in this type of study that film is still ahead of IPs and CCDs due to the higher resolution--especially with 4489. Again, when you are talking about the file size that your computer can handle (today; who knows about tomorrow), this may not be relevant to the kinds of qualitative work for which film has an advantage. If one wants to digitize the image, the advantages of film, even for the kind of qualitative work I mentioned above, become much less pronounced, since the scanner has larger pixels than the film, and it introduces noise. In order to produce a 60,000 by 90,000 pixel image from a 6.5 x 9 cm film, the pixel size of the scanner would have to be 1um, and I have never seen a 25,000 dpi scanner. In fact, it would not surprise me at all if a CCD or IP with a 15 um pixel size gave a better quality image than film scanned with a 10 um pixel size or even a 6 um (=4000 dpi, which is common) pixel size. Finally, there is at least one biological application for which the large dynamic range of IPs is useful: quantitative electron diffraction. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 24 19:32:09 2005
Dear Bill I completely agree with you: there are a number of application when large view field is important and then old friend film or image plate may be a solution. My point was that because of very low actual resolution on ultrathin sections in particular, one don't need too much pixels to show sample's details properly. For instance, my BioScan 600W camera being 1 Mpix has a view field size bigger than film - it was a whole point for purposing that particular camera. So, I actually, have bigger than on the film field size on my digital images. The price for that - low pixel conts. But - I don't need much pixels to represent trustfully what I have on my sections. It includes immuno-staining. I need to clarify: top mount cameras have lover magnification than on the film, therefore they have visible larger view area. In my particular case, my microscope do not permits to take pictures at lower than x3K magnification, but I am able to take pictures at x1.5K on digital camera, so I have more "field". Keeping in mind your arguments in favor of really large view field I still keep my dark-room running and 50 sheets of film is in the microscope. They are sitting in the scope 3rd year: nobody wants to use film since digital camera arrived. This is just practical observation: my users don't need large field and high pixels count! What they need - to generate images quickly and as many as they need, to be able to print them immediately, transfer them through existing network (not 110 Mb) - digital camera suites their need perfectly. For some rare work, I still have film available. This is my practical solution. I think, purposing digital camera was my best investment by the way.
As for scanning EM films - you absolutely right - film is superior! Personally, I prefer old friend - film. Have a great weekend, Sergey
At 03:35 PM 6/24/2005, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry and UCLA Institute of Nanotechnology 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
A postdoctoral position is available at Northwestern to work on a couple of projects, one involving low-loss energy loss spectroscopy of surface plasmons (aloof spectroscopy) in collaboration with a number of other faculty, the other a range of electron microscopy on oxide materials related to fuel cells. There will also be opportunities to interact with students/postdocs working on several other projects including oxide surfaces, atomic-scale tribology, heterogeneous catalysis and precession electron diffraction.
A strong general background in transmission electron microscopy and energy loss spectroscopy is required, and some experience in the specific research areas would be useful.
Applicants should apply by email only to L - marks -at- northwestern.edu including the names and emails of three referees and a CV not more than 3 pages in length including refereed publications and presentations. Large attachments will invoke the delete button.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2220 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 email: L - marks -at- northwestern . edu http://www.numis.northwestern.edu -----------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 26 09:24:20 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (david.griffiths-at-veths.no) from http://www.microscopy.com/MLFormMail.html on Sunday, June 26, 2005 at 03:40:39 ---------------------------------------------------------------------------
Email: david.griffiths-at-veths.no Name: David Griffiths
Organization: Norwegian School of Veterinary Science
Title-Subject: [Microscopy] [Filtered] Need LM sections from hpma blocks
Question: We have a number of tissue blocks embedded in Technovit 8100 and other hpma's, but we aren't set up for sectioning yet. Is there anyone with with spare capacity who can cut sections of these blocks for us, for payment of course? If so could you please indicate your interest and we can sort out the details. The typical block would be about 10mm x 10mm, and we would like sections at 1.5 - 2.0 microns, mounted on a glass slide. About 10 - 15 sections per block at the moment. Thanks, Dave Griffiths Section of Anatomy Norwegian School of Veterinary Science
Sergey- Actually, there are two additional cases where the 3.3 nm/pixel wouldn't help you with a problem. One is where you are trying for automated extraction of the images of 5-nm gold particles, in order to overlay them on the image. We have a procedure for this, and you do have to have 3 pixels across the 5-nm image (minimum). It is unpublished as yet (I am trying to get my former student to add it to our on-line protocols). Another case where one would need } 3 pixels across a feature is if the curvature of a contour is to be measured. Here, pixellation artifact prevents any meaningful measurements from being taken. To be specific, with a square array, one can only measure angles at 90 degrees apart (straight to right and left, up and down) and at 45 degrees apart (the corners). It is not possible to derive a meaningful measure under these circumstances, and the solution is to take more pixels and drop two between those subjected to the final measurement. We published this solution in 1980 (if I recollect correctly).
In any case, there are problems where lots of pixels in the array are essential. The problem happen to be in the arena where the image processing can be automated to effect, and that is the main advantage of taking a digital image i.m.h.o. So the deck is not stacked against the Ditabis technology -- the field just hasn't caught up with the potential applications of the digital technology. Carol P.S. No commercial interest in Ditabis or its competitors.
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-- __ Carol A. Heckman, Ph.D. Professor of Biological Sciences Director, Center for Microscopy & Microanalysis Bowling Green State University, Bowling Green, OH 43403 fax: (419) 372-2024 email: heckman-at-bgnet.bgsu.edu http://www.bgsu.edu/departments/biology/facilities/MnM ___________________________________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 27 08:13:15 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (morten.laanre-at-imbv.uio.no) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 27, 2005 at 05:20:55 ---------------------------------------------------------------------------
Question: Does anyone have a Zeiss 40x "antiflex" objective for 160mm tube length for sale.Old catalog no. 46 17 54 (or 46 17 53) ? The lens is needed for studies of cell attachment to substrate (surface).
Sincerely: C. Morten Motzfeldt Laane Institute of Molecular Bioscience (IMBV) University of Oslo, Norway P.o.Box 1041, N-0316 Blindern, Norway
Carol I absolutely agree: there are lot of cases when you need more pixels. From another hand, if you need more pixels for automatization - image plate technology is not the best: you need manually load/unload plates, load them into scanner, remove from scanner, load into cassette, load in microscope etc... and after that you'll have 50 images without proper identification (not everyone has last model microscopes). Keep in mind that scanning itself at max resolution will take a while. I was asking Bill Miller a few times how long it would take to scan a single plate. No answer. I would imagine, it's comparable with scanning time on phosphorimager- about 20 min 3x4" area. If so (correct me Bill if I wrong) - 16 hours for 50 plates... Good digital camera is much, much better for automatization. I am not even speaking about such wonderful features as "auto-tune", when microscope automatically aligned at degree human eye could not. You also could combine digital camera with energy filters etc - this is a way for automatization. I used to work on new Hitachi microscope (don't remember the model, 7200?) - they integrated 3D reconstruction from tilted series into hardware - you just press knob and microscope (with digital camera) made tilt series and reconstruct 3D image from it basically in real time - you may not do anything like that with image plates, I am sorry Bill. Have a great day. Sergey
At 01:43 PM 6/26/2005, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Jun 27 16:05:22 2005
I have some very tiny spheres (3-5 microns in diameter) that I need to obtain a thin section for TEM analysis, they are water soluable and crumble easily. What I have done already is to mount them on blank stubs with Extec and microtomy with limited results. Since they can't be floated onto water I have been catching them dry onto carbon coated copper TEM grids. They move quite a bit under the beam and the crystalline interior crumbles. Any thoughts would be appreciated.
Al Harmon MVA Scientific Consultants, Inc. "aharmon-at-mvainc.com"
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 27 16:12:37 2005
Sergey - you must have missed the scanning time e-mail, I sent it to you personally as you requested - we've had so many with the same subject it may just gotten lost in the clutter... 2.5 minutes at 15 um and ~10minutes at 7.5um or about 1 hour to load, read and unload the full load of 20 plates - less plates or lower resolution means less time, of course..
A digital camera is certainly faster but if you need the resolution, as she has indicated, they are quite costly - 4k x 4k, 14um pixel cameras (only half the pixels of the 15um plate scanner) cost upwards of $200k. I sell cameras for tomography as well and NOBODY does tomography fast enough to have it qualify as "real time" . Even the TVIPS 1k camera with } 12 frames per second takes quite some time to do a full tilt series ( usually over 100 images). That said, I'd never sell a plate system to anyone who was doing tilt series topography - it's totally the wrong tool. Any application that requires hundreds of images is best done with a CCD camera rather than plats or film.
} ------------------------------------------------------------------------------- } } Carol } I absolutely agree: there are lot of cases when you need more } pixels. From another hand, if you need more pixels for } automatization - image plate technology is not the best: you need } manually load/unload plates, load them into scanner, remove from } scanner, load into cassette, load in microscope etc... and after } that you'll have 50 images without proper identification (not } everyone has last model microscopes). Keep in mind that scanning } itself at max resolution will take a while. I was asking Bill } Miller a few times how long it would take to scan a single } plate. No answer. I would imagine, it's comparable with scanning } time on phosphorimager- about 20 min 3x4" area. If so (correct me } Bill if I wrong) - 16 hours for 50 plates... Good digital camera is } much, much better for automatization. I am not even speaking about } such wonderful features as "auto-tune", when microscope } automatically aligned at degree human eye could not. You also could } combine digital camera with energy filters etc - this is a way for } automatization. I used to work on new Hitachi microscope (don't } remember the model, 7200?) - they integrated 3D reconstruction from } tilted series into hardware - you just press knob and microscope } (with digital camera) made tilt series and reconstruct 3D image from } it basically in real time - you may not do anything like that with } image plates, I am sorry Bill. Have a great day. Sergey
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 27 16:35:31 2005
} They move quite a bit under the beam and the crystalline interior } crumbles. } Dear Al, You could try carbon coating after the particles have been picked up on the carbon-coated grids, and you could try a lower beam current. The movement under the beam indicates either charging, heating, both, or some other process--beam-induced specimen movement has not been completely characterized--and increasing electrical and thermal conductivity, or lowering the rate at which charge and/or heat accumulates on the specimen will, to some extent, ameliorate the problem. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 27 16:37:50 2005
Stating that it takes 20 minutes to scan a negative at high resolution without specifics of what you mean or without comparisons to actual modern data does not do justice to the purpose of this forum or the technological advancements that are in the scientific market today.
I have just gone to my older high resolution scanner (Leafscan 45) here at ANL and have digitized a 3 1/4 x 4 " negative at 1270 dpi ~ or a resolution of 20 microns/pixel. The image size was 5080 x 4128 pixels (21 Megapixels) and 16 bits deep. The total time from the point that I mounted the negative in the holder to when I had a digitized image on my screen was just under 3 minutes, not 20 minutes. This is a fairly old scanner and I know that newer ones are faster and have higher resolutions (abeit not always with as good a bit depth).
One certainly could have gotten a similar image with a CCD camera by taking multiple images (the number varying with the resolution of the camera) but not in the same amount of time or cost. It would require either a very large and expensive CCD camera or off-line stitching of multiple images to merge and align the individual images together to form an equivalent single image.
I use both technologies in my research, choosing the best technology for the image as appropriate, just as I would selecte the most appropriate microscope to analyze a specimen.
Let me remind everyone, that we all have the responsibility to make sure that we are reasonably factual, accurate and up-to-date when we point out the pros and cons of any technology. Using significantly out of date or inaccurate information does not help disseminate useful knowledge to the community, which is one of the purposes of this forum.
Nestor Your Friendly Neighborhood SysOp
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-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov NEESLab: http://neestpm.mcs.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
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From MicroscopyL-request-at-ns.microscopy.com Mon Jun 27 16:43:40 2005
Dear Lists, Thanks everyone for the many helpful comments and suggestions. After a series of investigations, we tried something that, in retrospect, was obvious. When a smaller objective aperture was inserted, the problem became less severe, leading to the conclusion that somehow the larger objective aperture was out of position. Some of the possibilities are that the aperture was not placed properly in the holder, that the foil came loose from the body of the aperture, or that the tip was being displaced when moved to the farthest-in position. In our case the we still do not know exactly what occurred, but the apertures were properly positioned and looked fine. This can occur in a microscope where the aperture is in the back focal plane, as well as one like the F30 where the aperture is not, so if you experience a similar problem with your aperture not being centered in Focus state when it is centered in Exposure state, see if this occurs just with one aperture or is present for all the apertures. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 27 18:39:18 2005
Dear Nestor In my message I was talking about image plates, not negatives. In my speculation about time necessary to scan image plate I based on estimation from scientific instrument, which is using very similar technology as a Ditabis plate reader: phosphorimager. It uses basically the same image plates and "imaging technology" is very similar if not identical. Latest model of our Departmental phosphorimager Typhoon 9410 needs about 40 min to scan 2x3" area with resolution of 10 um (see specification on instrument). For 25 um, which perhaps comparable with Ditabis it would be 15-20 min. You may not compare optical scanner with image plate technology - it's absolutely different instruments and principles! By the way, if you will use good specialized optical scanner for difractogtramm (which gives you optical density) - it will take much longer than 2-3 min to scan EM size film, so I am quite sure that my approximation is quite right and this is exactly why I was talking about that: many people would think similar to you that "scanning is not a big deal". To get correct readings in OU is basically a big deal even nowadays. "Household" scanners may not be used for quantitative analysis - they are not calibrated and sometime not linear. Their "color" sensor (depends from construction) may compromise the data. In my message, I clearly stated that I was trying to obtain technical data from Ditabis and they refuse to answer, so I have to speculate instead using real data from manufacturer. Basically, even if it takes according you just 3 min per plate - still, it will take 2.5 hours to scan the standard set of 50 plates. Not very impressive to me. I am sorry if I broke some rules AGAIN, I am naturally rule-breaker, I think most of the science progress based on breaking somebody's rules (your boss perhaps?). Have a good day, Sergey
At 02:37 PM 6/27/2005, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Jun 27 20:45:18 2005
Sergey - just for comparison sake lets try to compare apples to apples... figure out how long it would take to do 50 film images with darkroom work - developing, drying , printing, drying - it's been so long since I've had to actually do that I'd no longer feel confident in even venturing a guess - but not minutes for sure
and then with your 1k camera shot to cover the same area as the film at the same resolution as the plates with image tiling - that's roughly 35-40 CCD images per 6k x 5k image plate (don't forget the necessary overlap for the tiling) for a total of ~1800 CCD images -- you claimed that you took 100 images in 40 minutes so the math works out to roughly 720 minutes (12 hours) BEFORE tiling
Two and a half hours to viewable images with the plates doesn't look so bad from a time stand point when you look at the process objectively - particularly since for most of the time the scanner is doing all the work leaving you free to do something else. If you don't need or want the resolution then it's rather hard to make a rational comparison between low pixel count CCDs and imaging plates since the image plates would not be the proper imaging device for such an application anyway. No body is trying to talk anybody into using the plates for anything other than what they are good for. If I had to pull a 60 foot trailer I wouldn't pick a motorcycle to do it with just because it was faster than a truck.
That said, the plates may still not be the best answer for many application. When short turn around time is critical CCD cameras are the answer. There is an endless list of applications where one technology is clearly more suitable than another. Pick the one that work for you.
Regards - Bill
At 07:38 PM 6/27/2005, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Jun 27 23:53:42 2005
It is good to see factual information about scanning times for image plates. I also concur with your comments on using this technology for tomography.
The digitization times you presented are certainly consistent with my experiences using a high resolution high end negative scanners like the Leafscan 45, which is certainly not a run of the mill household scanner as the un-informed might presume.
As a followup, I would like to hear of comparisons using the older technology namely-negative drum scanners if anyone is still using this technology. I haven't used one of these used in years, and only vaguely remember using one over a decade ago here at ANL, it unfortunately has long since disappeared so I cannot test it out nor do I still have access to the original instrument specifications for comparison.
If anyone has additional real experience with plate or other high end negative scanners to add to this discussion it would be appropriate to post that data for the archives and to complete this discussion.
Finally, out of interest I checked on the specification of the Typhoon 9410 which was used as a basis for some scanning time estimates posted earlier in this discussion thread, which seemed unrealistic to me.
I was surprized to find out that the Typhoon 9410 is neither a negative nor image place scanner, but rather is a "high performance gel and blot imager ". Checking the technical specifications I see that while it has the same bit depth resolution as the Leafscan 45, it only has a uniformity of +/5% over the scan area, making this unsuitable for TEM work. The Typhoon manufacturer GE, in addition, makes absolutely no claims or has any application notes on its WWW site which indicate it is suitable for use with TEM negatives or image plates, nor is it apparently designed to be used for TEM application. It's scanning times are optimized for the detection of signal in gel/blot image data and not for data relevant to the discussion in this thread. In light of this fact, the performance characteristics estimated from an instrument like the Typhoon9410 and then extrapolated to TEM negative and/or image plate scanning are, in my opinion, a comparison which carries no technical merit.
Nestor Your Friendly Neighborhood SysOp
Disclaimer: I have no financial interests in any of the imaging/scanning technologies discussed herein.
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 09:31:27 2005
A couple of questions. First, I would like to receive information from vendors on their current models of elemental x-ray detectors. I would also like to hear a comparison between the detectors that use liquid nitrogen and those models that don't need liquid nitrogen cooling. Secondly, while we hope to get a new SEM, we may not be able to, in that event I would like to know if a EDX unit could still be fitted to our JEOL T220A SEM. As always any information is appreciated from everyone.
Tom Bargar Core Electron Microscopy Research Facility University of Nebraska Medical Center 986395 Nebraska Medical Center Omaha, NE 68198-6395 phone: 402-559-7347 tbargar-at-unmc.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 11:25:01 2005
Checking if anyone knows of any new florescent bead labeling techniques when marking specific sites on tissues. Specifically, something that will not dissolve in PO. A polystyrene replacement?
Thanks
~~~~~~~~~~~~~~~~~~~~~~~ Andrew F.M. Johnstone Deptartment of Biology Cooper Lab University of Kentucky email: johnstone-at-uky.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 13:25:49 2005
On Jun 27, 2005, at 9:52 PM, Nestor J. Zaluzec wrote:
} If anyone has additional real experience with plate or other high end } negative scanners to add to this discussion it would be appropriate } to post that data for the archives and to complete this discussion. } Dear Nestor, I have had experience with scanners ranging from a CCD used to image the light transmitted through a negative, through a linear diode array, and a HiScan drum scanner to a Perkin Elmer microdensitometer. For images, where generally the intensities do not vary rapidly over small distances, and where the OD is centered around 1 and ranges from a few tenths to, say, 2, any of the scanners will give a decent representation of the image, and the fidelity of quantitation depends on the number of pixels, the response time of the electronics, and the stability of the illumination (among other variables). For electron diffraction patterns, where the range of ODs is from ~0.001 to ~4, and can vary greatly over distances on the order of tens of um, accurate quantitation cannot be obtained unless the illuminated area of the negative is the same as the scanning pixel. This is due to the fact that only a small fraction of the light transmitted through adjacent areas and scattered in the detector housing will give a falsely low OD reading. For quantitation of ED patterns, only a microdensitometer, such as the Perkin Elmer, will give sufficiently accurate data, and then only at a fairly slow scan speed and small pixel size. I am not up to date on the latest improvements in scanning times, but back in the day such a scan would take hours for each negative. I have no relationship to Perkin Elmer except as a former satisfied user. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 13:52:43 2005
Dear Nestor You need to read my postings: I did not claim that Typhoon is a "scanner" for negatives or have something to do with EM. Nevertheless, it's the best on the market reader for "IMAGE PLATES". Image plate technology used in many areas when relatively high energy particles (alpha/beta particles etc) or gamma need to be detected. It's commonly used in molecular biology when people used radioactive (32P, 14C, 3H) labelling, DNA for instance. You separate your stuff on the gel and detect radioactivity using image plate. In this area image plate technology is well established and respected. Because (and I mentioned it in my original posting) image plate technology is the same for EM and for other application it's legit and even interesting to compare performance of different instruments- "image plate readers" (correct name for it). From another hand I could not understand why you are trying to compare optical scanners with image plate readers? I think it's simply not legit to make any bridge between (even non-household) optical scanner and image plate reader because of differences in principles of operation - you have to know better, you read manual to Typhoon. Leafscan 45 based on my quick research on Internet - it's very good professional scanner for general photography, it scans color and B&W slides and transparencies etc - it's not specialized scanner for EM, it's not a densitometer ether- similarly as Typhoon is not specialized reader for EM. If you think you may compare the data from non-EM general scanner and image plate readers, than, my comparison of Typhoon and Diabis is even more legit! Typhoon will scan Diabis image plates (as any other) perfectly, by the way. 5% uniformity - this is what you may expect from image plate readers due image plate properties, I guess, and this is another point to think twice before using this technology. I have about 10 years experience with image plates and readers (in molecular biology where this technology is proven to have numerous of advantages), so I am quite confident that information I provided is correct and it's not my fault if somebody do not read the original postings and misinterpret it: may be it's just because my broken English? I am sorry for that. Sergey
At 09:52 PM 6/27/2005, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 15:16:00 2005
We've been working with NT-MDT and Leica on a new AFM/Ultramicrotome hybrid that could provide an answer to your dilemma. The fully automated system will be announced officially later this year but the manual version was shown at Cell Bio last December and will be at the SPM '05 in Sapporo this month. It allows you to cut then image in one system, without having to catch sections. The results can be fed into a 3D reconstruction program (NT-MDT has tried, successfully, with Media Cybernetic's 3D Constructor) to generate 3D images and resulting image analysis. The AFM uses local differences in elasticity and hardness (among other modalities) to create contrast. If you would like to send some samples, we can forward them to NT-MDT and have them run them for you. You can contact me off-line for details.
Hope this is helpful.
Best regards, Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Call us today to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts. Call us at (972)954-8011.
At 04:09 PM 6/27/2005, Al Harmon wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 16:00:05 2005
Another issue to consider in negative scanning vs. image plates, is that only a fraction of the negatives would ever get scanned because examination of the negatives with a magnifier (human scan?) typically reveals that too many of the negatives are not perfect, due to focus, drift, or some other glitch. I can't quote a number, but for 30 years it always seemed too high, either due to my skill, or in later years to my being too picky (or both). With image plates, every exposure needs to get scanned to be observed.
John Mardinly Intel
This opinion is the opinion of the author, not of Intel corporation. -----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-aaem.amc.anl.gov] Sent: Monday, June 27, 2005 9:53 PM To: microscopy-at-microscopy.com
Bill
It is good to see factual information about scanning times for image plates. I also concur with your comments on using this technology for tomography.
The digitization times you presented are certainly consistent with my experiences using a high resolution high end negative scanners like the Leafscan 45, which is certainly not a run of the mill household scanner as the un-informed might presume.
As a followup, I would like to hear of comparisons using the older technology namely-negative drum scanners if anyone is still using this technology. I haven't used one of these used in years, and only vaguely remember using one over a decade ago here at ANL, it unfortunately has long since disappeared so I cannot test it out nor do I still have access to the original instrument specifications for comparison.
If anyone has additional real experience with plate or other high end negative scanners to add to this discussion it would be appropriate to post that data for the archives and to complete this discussion.
Finally, out of interest I checked on the specification of the Typhoon 9410 which was used as a basis for some scanning time estimates posted earlier in this discussion thread, which seemed unrealistic to me.
I was surprized to find out that the Typhoon 9410 is neither a negative nor image place scanner, but rather is a "high performance gel and blot imager ". Checking the technical specifications I see that while it has the same bit depth resolution as the Leafscan 45, it only has a uniformity of +/5% over the scan area, making this unsuitable for TEM work. The Typhoon manufacturer GE, in addition, makes absolutely no claims or has any application notes on its WWW site which indicate it is suitable for use with TEM negatives or image plates, nor is it apparently designed to be used for TEM application. It's scanning times are optimized for the detection of signal in gel/blot image data and not for data relevant to the discussion in this thread. In light of this fact, the performance characteristics estimated from an instrument like the Typhoon9410 and then extrapolated to TEM negative and/or image plate scanning are, in my opinion, a comparison whic! h carries no technical merit.
Nestor Your Friendly Neighborhood SysOp
Disclaimer: I have no financial interests in any of the imaging/scanning technologies discussed herein.
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 16:24:23 2005
Theoretical Question: if you were purchasing a TEM equipped with a high quality (2k x 2k) digital camera would you still opt for the film camera as well? Just wondering. -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 16:32:42 2005
I am told by someone who visits my lab that protein fibrils treated with congo red will show an apple green birefringence in crossed polars.
She has brought samples for us to look at, but we cannot see the green birefringence. She 'knows' there are fibrils present by another type of assay.
Has anybody heard of this technique? I think I can see the stained fibrils, but nothing shows with the crossed polars. Maybe I don't know what I should be looking for, maybe it is very subtle. Maybe it doesn't really work.
If you have heard of this technique and maybe know a trick or two, let me know so I can help this person.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 16:41:51 2005
I'm looking for a couple of EM texts, and having no luck with the on-line used book sources: M. A. Hayat, "Fixation for Electron Microscopy" Reimer and Hawkes "Scanning Electron Microscopy : Physics of Image Formation and Microanalysis" Anyone happen to have an extra copy in good or better condition that they might be willing to part with?
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 17:11:09 2005
Phil, If you want, for the price of mailing them, Hayat Principles and Techniques of Electron Microscopy Biological Applications Vols 1 and 3, 1970 and 1973 respectively. Kind of old but unless someone wants them, they go into the trash along with a bunch more. Damian
-----Original Message----- } From: Philip Oshel [mailto:peoshel-at-wisc.edu] Sent: Tuesday, June 28, 2005 4:41 PM To: Microscopy-at-microscopy.com
Microphiles,
I'm looking for a couple of EM texts, and having no luck with the on-line used book sources: M. A. Hayat, "Fixation for Electron Microscopy" Reimer and Hawkes "Scanning Electron Microscopy : Physics of Image Formation and Microanalysis" Anyone happen to have an extra copy in good or better condition that they might be willing to part with?
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 17:16:24 2005
Greetings, The trick here is that you have to think in terms of dichroism rather than birefringence. What happens with congo red (at least in theory) is that it binds to the substrate in an oriented way. So if you have a substrate that is oriented then the congo red molecules themselves become oriented. Now congo red happens to be good at absorbing linearly polarized light oriented along one axis of the molecule and bad along the perpendicular axis. So a congo red stained sample should act like a polarizing filter, itself. So to see your apple green signal, what you do is to remove the analyzer from the system, have just the polarizer (you can do it the other way if you want, it doesn't matter). Then rotate the sample through at least 90 degrees. You are using the sample to make "crossed polarizers". The apple green should show up when the sample fibers are at some defined orientation to the polarizer and the signal should disappear at +/- 90 degrees to that orientation (no getting more specific without knowing which way the fibers go and how the congo binds to them).
You get green presumably becaue the congo red absorbs red and blue better and so more green leaks through. Actually, polarizing filters themselves are often made by aligning really strong crystals that have dichroic abosorption, and they too have a green cast because it is difficult to have no wavelength dependence to the dichroism.
Keep in mind that congo red stained fibers are going to be a whole heck of a lot less good at being a polarizing filter than a real polarizing filter, so the green signal may be pretty faint. But its tell tale appearance and disappearence at +/- 90 gives you a story. You can play with using green light (or other wave bands) but I think one's eyes are most senstivite to seeing differences in color rather than intensity.
Hope this helps, Tobias } } } I am told by someone who visits my lab that protein fibrils treated with } congo red will show an apple green birefringence in crossed polars. } } She has brought samples for us to look at, but we cannot see the green } birefringence. She 'knows' there are fibrils present by another type of } assay. } } Has anybody heard of this technique? I think I can see the stained fibrils, } but nothing shows with the crossed polars. Maybe I don't know what I should } be looking for, maybe it is very subtle. Maybe it doesn't really work. } } If you have heard of this technique and maybe know a trick or two, let me } know so I can help this person. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
To my knowledge green birefringence after Congo Red treatment appears when your protein has a lot of b-sheets in its structure. Amyloids formed from different proteins (APP, prion etc.) show such phenomenon. So, perhaps, the sample you was looking at did not have amyloid conformation.
Regards, Aleksandr Mironov EM Unit University of Manchester
On 28 Jun 2005, at 22:23, Jon Krupp wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } I am told by someone who visits my lab that protein fibrils treated } with } congo red will show an apple green birefringence in crossed polars. } } She has brought samples for us to look at, but we cannot see the green } birefringence. She 'knows' there are fibrils present by another type of } assay. } } Has anybody heard of this technique? I think I can see the stained } fibrils, } but nothing shows with the crossed polars. Maybe I don't know what I } should } be looking for, maybe it is very subtle. Maybe it doesn't really work. } } If you have heard of this technique and maybe know a trick or two, let } me } know so I can help this person. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 18:50:11 2005
An evaluation of a Ditabis scanner a few years ago left the impression that it was slightly less convenient to use than film, although now that less film is being processed that may be changing. But the very great advantage in our applications was the dynamic range rather than spatial resolution. 32 bit is something else. Diffraction patterns easily showed features that would have had to be carefully sought out using film. For biological work, IP plates offer the possibility of much more frequent use of unstained material - looking at the real tissue rather than a heavy metal incrustation of osmium, lead or uranium, in embedded as well as frozen material. I would be interested to know if anyone has been doing much along these lines?
IP is complementary to CCD cameras at the moment, rather than a practical replacement. The immediacy of the CCD camera is such an advantage that its hard to remember how we worked without it. My take on the "2K x 2K CCD vs film camera" question is "2k by 2K...keep the film just in case. But once 4K by 4k or more comes in affordably..... time for some serious re-design of the TEM!
My two cents worth..
Cheers Sally
Dr SJ Stowe Facility Coordinator ANU Electron Microscopy Unit ANU CRICOS#00120C
} -----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] } Sent: Wednesday, 29 June 2005 4:24 AM } To: microscopy-at-msa.microscopy.com } Subject: [Microscopy] Re: TEM Negative & Image Plate digitization times } } } } } --------------------------------------------------------------- } --------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver
} } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 19:05:37 2005
} } A couple of questions. First, I would like to receive information } from vendors on their current models of elemental x-ray detectors. I } would also like to hear a comparison between the detectors that use } liquid nitrogen and those models that don't need liquid nitrogen } cooling. Secondly, while we hope to get a new SEM, we may not be } able to, in that event I would like to know if a EDX unit could still } be fitted to our JEOL T220A SEM. As always any information is } appreciated from everyone. }
Whatever you do, think carefully about the detailed mechanical design of whatever detector you end up buying, and, if possible, the manufacturer's reputation for standing behind their products.
I have an approximately 5-year-old PGT Prism 2000, the sort which has an adjustable dewar angle to allow it to be used at a variety of takeoff angles.
At 2 1/2 years old, its LN2 consumption had steadily increased to the point where it had to be refilled every second day, so I returned it to PGT to have the leak fixed and to be re-evacuated, for over a thousand bucks.
When I (eventually) got it back, the LN2 consumption was OK, but over the next two years it steadily deteriorated again, quite spontaneously (no warmups or other mishaps), and, to my extreme disappointment, PGT's non-negotiable attitude is "it was OK when it left here, it must have developed another leak".
This defies common sense --- we're talking here about a detector which twice experienced a dewar vacuum deterioration, and whose leak was apparently not fixed when it was returned to the factory.
I don't think it's a leaky window, as it's been on my SEM under vacuum 24/7 all the time, so my suspicion falls on the O-ring-sealed elbow.
Unfortunately, the PGT vacuum port is non-standard and is on the top surface right underneath the dewar, and, even more ridiculously, the PGT adaptor (US$500!!!) won't fit onto it if the dewar is tilted to be vertical for a 40 degree takeoff angle, so, you guessed it, AFTER evacuating the detector, the bolts holding the elbow joint have to be slackened so that the elbow can be adjusted to the correct angle! This, of course, placed the bright shiny new vacuum at risk.
The guys in the excellent workshop to which I have access have now made me an adaptor which will fit onto the untilted detector, and when I work up the courage I will repump it myself.
Of course, everyone makes their own purchase decisions, but I will never again buy a detector which:
- has an adjustable elbow - has a non-standard vacuum port - is made by a manufacturer who won't guarantee the vacuum integrity of the dewar for five years following manufacture or factory repair (barring accidents) - is made by PGT.
In the meantime I fill the damned thing every day, including Saturdays and Sundays, with considerable resentment.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 19:06:17 2005
YES 2x2 is good, but film is still better for some things. I just wrote a proposal for a 'scope with both. David
On Jun 28, 2005, at 2:23 PM, John J. Bozzola wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Theoretical Question: if you were purchasing a TEM equipped with a } high quality (2k x 2k) digital camera would you still opt for the film } camera as well? Just wondering. } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Email: bozzola-at-siu.edu } ############################################################## }
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 19:09:57 2005
Well, you've got most of this right, but not entirely.
Your procedure is correct: use just the polarizer in fixed position and rotate the sample. (Actually, either could be fixed, so if you don't have a rotating stage, you can leave the sample in place and rotate a polarizing filter over the light port).
The sample doesn't really act as the second polarizer (analyzer). This is the classic experiment for either dichroism (materials which exhibit two different refractive indices) and pleochroism (materials which exhibit 3 different refractive indices).
First, a few definitions: Refractive index is an optical property which expresses the relative interaction between the electric field in light and the electric field in matter. The stronger the interaction, the more light passing through a material will slow down and the higher the refractive index. (Mathematically, RI = velocity of light in air or vacuum/velocity of light as it interacts with a material where V air = 300,000 km/sec).
If you dissect the word "birefringence", you can see that it refers to materials * having the property of ("gence") * two (bi) * refractive indices (ref in ). In actuality, materials can have three different RI's, all at right angles to each other, but will only exhibit 2 at a time (think of either films, fibers, or crystal faces), so we never refer to them as "tri-refringent".
Polarized light is different from ordinary light in the following way: All light has a direction of travel. You see things in the world around you because light is traveling from them to you. Light is electromagnetic radiation. As microscopists, we are most typically interested in the electrical field part (yes, I know that there have been magnetic microscopes built). The sine wave we use to describe light is actually the tracing of the tip of that electrical vector, building up then dropping off in a positive direction then building up and dropping off in negative direction. The motion of that sine wave gives light a direction of vibration. The direction of vibration is always at right angles to the direction of travel.
Ordinary light contains all directions of vibration (imagine little vectors vibrating N-S, E-W, and all angles in between; all at right angles to the direction of travel). To convert ordinary light to polarized light, you simply have to impose some sort of interaction (reflection, specific types of absorption, beam splitting) which absorbs all but one permitted direction of vibration. The result is "Plane Polarized Light", which is not necessarily always the same as linearly polarized light (tune in another time for the discussion of "States of Polarization"). However, for the sake of simplicity, plane polarized light does indeed vibrate linearly.
So what happens when you rotate any birefringent material over a polarizer? I can't draw diagrams for you here, but imagine a rectangle with one RI oriented along the short edge and the other along the long edge. As you rotate the sample, the short edge will eventually align with the permitted direction of the light coming from the polarizer. In this position, the light only "sees" one refractive index (not the most scientific explanation, but accurate). Rotating 90 degrees presents the other RI. Anywhere in between, contributions from each of the RI's will be visible. In practice, we use these unique positions to isolate each of the RI's to actually measure them.
And what's the story with things that are dichroic or pleochroic? In addition to being birefringent, they are also colored in normal illumination (ex: Congo red). To properly observe them, first set up Koehler illumination and just observe the normal color. Then, insert just a single polarizer and rotate. The colors that you see will not be the typical magentas, golds, and turquoises characteristic of polarized light interactions. Rather they will be absorption colors (browns, reds, yellows, etc.), derived from the unique property that, for these materials, not only does refractive index vary with direction, absorption does too. The result - changes in color and intensity. ... and yes, one of these directions can be very subtle.
In normal polarized light analyses, the next step would be to insert the analyzer and rotate. It is also important to note that the normal Polarization colors may be affected by the absorption colors. (For those of you who are interested, I have a "polarized light road map" which outlines all these test in flow-chart form).
Here are some interesting examples of materials which are dichroic or pleochroic: Everyone cites tourmaline which is dark brown in one orientation and pale yellow in another,. as well as biotite (a type of mica) and cordierite. Hartshorne and Stuart (Crystals and the Polarizing Microscope, Arnold, 1970) cite magnesium platino-cyanide which oscillates between bluish-red (parallel to "c" axis) and carmine red; hypersthene (ferro-magnesium silicate): brownish red to green; remind us that some biological materials also exhibit this property. Patzett (Polarized Light Microscopy: Principles, Instruments, Applications, Leitz - now out of print) reminds that the phenomenon is widespread in organic materials and is responsible for the circular dichroism we chemists routinely test for in levo and dextro rotatory compounds. And finally, I still have fond memories of doing experiments in grad school from Chamot & Mason (Handbook of Chemical Microscopy, Vol. I - now available through McCrone Associates) on a slew of materials. (For those of you who are teaching, they recommend viscose rayon dyed with congo red; crystals of o-nitrophenol, azobenzene, iodoquinine sulphate, silver chromate, copper acetate, red ammonium picrate and the magnesium platinocyanide - mentioned above).
Well, we are back at Congo red, so I'll stop here. For those of you interested in the physics of this process, I encourage you to read Hartshorne and Stuart further. For the rest of you, find some dichroic or pleochroic materials and just have fun.
After many years of teaching, I realized that all the beauty and complexity of polarized light boils down to one simple underlying concept: refractive index. And here it is again.
Hope this is helpful. Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Call us today to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts. Call us at (972)954-8011.
At 05:15 PM 6/28/2005, Tobias Baskin wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America more specific without knowing which way the fibers go and how the congo binds to them). } } You get green presumably becaue the congo red absorbs red and blue better and so more green leaks through. Actually, polarizing filters themselves are often made by aligning really strong crystals that have dichroic abosorption, and they too have a green cast because it is difficult to have no wavelength dependence to the dichroism. } } Keep in mind that congo red stained fibers are going to be a whole heck of a lot less good at being a polarizing filter than a real polarizing filter, so the green signal may be pretty faint. But its tell tale appearance and disappearence at +/- 90 gives you a story. You can play with using green light (or other wave bands) but I think one's eyes are most senstivite to seeing differences in color rather than intensity. } } Hope this helps, } Tobias } } } } } } I am told by someone who visits my lab that protein fibrils treated with } } congo red will show an apple green birefringence in crossed polars. } } } } She has brought samples for us to look at, but we cannot see the green } } birefringence. She 'knows' there are fibrils present by another type of } } assay. } } } } Has anybody heard of this technique? I think I can see the stained fibrils, } } but nothing shows with the crossed polars. Maybe I don't know what I should } } be looking for, maybe it is very subtle. Maybe it doesn't really work. } } } } If you have heard of this technique and maybe know a trick or two, let me } } know so I can help this person. } } } } Thanks } } } } Jonathan Krupp } } Microscopy & Imaging Lab } } University of California } } Santa Cruz, CA 95064 } } (831) 459-2477 } } jmkrupp-at-cats.ucsc.edu } } } -- } _ ____ __ ____ } / \ / / \ / \ \ Tobias I. Baskin } / / / / \ \ \ Biology Department } /_ / __ /__ \ \ \__ 611 N. Pleasant St. } / / / \ \ \ University of Massachusetts } / / / \ \ \ Amherst, MA, 01003 } / / ___ / \ \__/ \ ____ } http://www.bio.umass.edu/biology/baskin/ } Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243
At 05:15 PM 6/28/2005, Tobias Baskin wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America more specific without knowing which way the fibers go and how the congo binds to them). } } You get green presumably becaue the congo red absorbs red and blue better and so more green leaks through. Actually, polarizing filters themselves are often made by aligning really strong crystals that have dichroic abosorption, and they too have a green cast because it is difficult to have no wavelength dependence to the dichroism. } } Keep in mind that congo red stained fibers are going to be a whole heck of a lot less good at being a polarizing filter than a real polarizing filter, so the green signal may be pretty faint. But its tell tale appearance and disappearence at +/- 90 gives you a story. You can play with using green light (or other wave bands) but I think one's eyes are most senstivite to seeing differences in color rather than intensity. } } Hope this helps, } Tobias } } } } } } I am told by someone who visits my lab that protein fibrils treated with } } congo red will show an apple green birefringence in crossed polars. } } } } She has brought samples for us to look at, but we cannot see the green } } birefringence. She 'knows' there are fibrils present by another type of } } assay. } } } } Has anybody heard of this technique? I think I can see the stained fibrils, } } but nothing shows with the crossed polars. Maybe I don't know what I should } } be looking for, maybe it is very subtle. Maybe it doesn't really work. } } } } If you have heard of this technique and maybe know a trick or two, let me } } know so I can help this person. } } } } Thanks } } } } Jonathan Krupp } } Microscopy & Imaging Lab } } University of California } } Santa Cruz, CA 95064 } } (831) 459-2477 } } jmkrupp-at-cats.ucsc.edu } } } -- } _ ____ __ ____ } / \ / / \ / \ \ Tobias I. Baskin } / / / / \ \ \ Biology Department } /_ / __ /__ \ \ \__ 611 N. Pleasant St. } / / / \ \ \ University of Massachusetts } / / / \ \ \ Amherst, MA, 01003 } / / ___ / \ \__/ \ ____ } http://www.bio.umass.edu/biology/baskin/ } Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 28 20:40:06 2005
Ours is a happier story........... We purchased a Link detector some fifteen years. It has Be, thin window and windowless positions. I am sure it is considered obsolete now, but having the turret allows us to easily replace the thin window in the field. Also, I believe having the permeable window positions allows us "pump down in situ" as our LN consumption seems about the same as when it was new. Other than very occasionally allowing the LN to evaporate out and cleaning out any ice, the detector has never been serviced.
We did have some early problems with the AN10000 electronics. Link's response was that we should buy a whole new system. Our in-house tech found a blown fuse which was undocumented fuse and well hidden beneath a circuit board.
We have since replaced all of the electronics and software with a WinEDS system, but we will hold onto this detector as long as it keeps chugging along.
Alan Stone ASTON
At 07:05 PM 6/28/2005, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 29 08:27:08 2005
Dear Roman, The V7 hardware button you are referring to is a switch which not only closes V7/V4 but disables the valves. If you try to open the valve after pressing the button the HT unfortunately shuts down trying to protect itself since the valve has been disabled for some reason. One must press this button again to re-enable the valve before you will be able to open it. Unfortunately there is no way to know whether you are disabled or not and we have had similar problems when changing cameras and a new user accidentally presses the button which on our F20 and Polara are in close proximity. These saftey buttons are in place in case the PC dies and the valves need to be shut directly. There is a similar arrangement with the HT as far as I understand but at least that button lights up when it is enabled. Regards, Bob
At 09:46 AM 6/29/2005 +0200, Koning, R.I. \(MCB\) wrote: } Dear all, } } I am sorry to hear about your tip. Thia happening is always the biggest } fear of a FEG user (after floodings and fires). I must say that we for } safety reasons only open the FEG valve when we look at the specimen. In } any other situation the lock is closed. } } We experienced major problems and many bugs after upgrading our TECNAI FEG } 20 software to version 2.1.5. The most important being not to be able to } get the negatives exposed in low dose. It appeared that having the plate } number counter and/or exposure number in the window of our TUI created } major communication problems with the TEM server. This resulted in a } multitude of (irreproducable) bugs and mistakes. Getting rid of the plate } number counter and exposure number corrected for all of these problems. } } Not related to this, it also appeared that the hardware button to close V7 } (which I normally never use but for educational purposes did once) not } only closed V7 but also shut down the HT. Something that a FEI engeneer } experienced before. } } With kind regards, } } Roman } } R.I.Koning (Roman) Ph.D. } Leiden University Medical Center } Department of Molecular Cell Biology } Center for Electron Microscopy } Wassenaarseweg 72 2333 AL } P.O.Box 9503 2300 RA } Leiden } The Netherlands } tel. (+31) 71 527 6463 } fax. (+31) 71 527 6440 } r.i.koning-at-lumc.nl } } } -----Original Message----- } From: owner-tecnai-at-wadsworth.org [mailto:owner-tecnai-at-wadsworth.org]On } Behalf Of Angel M Paredes } Sent: Tuesday, June 28, 2005 20:40 } To: Bill Tivol } Cc: microscopy-at-msa.microscopy.com; tecnai-at-wadsworth.org; 3dem-at-ucsd.edu } Subject: [Microscopy] tecnai version 2.1.5 } } } Hi all, } } Sunday we experienced a rather bad malfunction that people should be aware } of that destroyed our FEG tip. On our Polara, the user working with the } scope decided to remove the MSC to exchange the specimen. To do this you } have to vent the airlock. He pushed the "vent airlock" button. Since he } had not closed the column valves, the software I believe was supposed to } have detected this and closed the column valves for him to protect the } FEG. The software unfortunately did not. What the user did not know was } that there was an electrical short in valve B that told the software that } valve B was closed when it was in fact opened. Instead of telling him to } close valve B as it should have done, the software detected that valve B } was already closed and told the user instead to close valve A... which he } did. The software then vented the airlock but with valve B opened instead } of closed, the column was vented and since the software had not closed the } column valves when the user pushed "vent airlock", the FEG was vented too. } FEG tip gone. } } We are currently using version 2.1.5 of the user interface. Has anyone } out there been experiencing any other bugs with 2.1.5? Thorsten had } problems with his system rebooting when a log file was full and I went } through the same thing during a bugcheck the system decided to perform. } Another bug we've seen is in low dose. We use diffraction while in } search. Every now and again the search mode goes from looking normal to } looking really strange and becoming very difficult to align. When I go } from diffraction back to regular image mode in search, I find that the } microscope has gone from a magnification of 52,000x to 52x while in } diffraction and this is the reason why the search mode goes from looking } okay to looking strange. FEI's response is that we are not supposed to be } using diffraction and experiments in the factory have shown that if one } switches in and out of diffraction too quickly, it causes the mag to } change. All I know is that version 2.1.3 did not have this problem. Are } there any other problems I should be made aware of? } } Thanks, } angel
******************************** Robert Grassucci Howard Hughes Medical Institute Wadsworth Center Empire State Plaza Albany, NY 12201-0509
Barbara and group, This is a complex area and I am far from optical crystallagrapher (!). But there are a couple of things I want to say, based on what I learned in school.
Birefringence describes a material that has two (or three) refractive indices. In contrast, dichroism describes a material that has two (or three) different absorption behaviors. (I guess we don't call them absorption indecies).
It is true that the aligned congo red molecules on the sample would probably also be birefringent, but this would presumably be so weak that unless the user has a really sensitive microscope, he or she would be unlikely to be able to detect it. In contrast, it is a lot easer to detect differences in light intensity and color, so there is a chance to pick up the dichroic absorption. And the way you do that is to illuminate the sample with linearly polarized light and rotate the sample (you are correct, you can rotate the polarizer instead) without any analyzer, and look for the changes in color as a function of the polarized light angle.
Now it is true that you can also add an analyzer and with cryastals this gives you a chance to see further and different (and beautiful) colors as the analyzer is rotated. It might increase the contrast of a weak signal so it might be worth trying for the congo stained protein fibers. What you would need to do in that case would be to uncross the polars by a certain amount, say 5 degrees, rotate the sample through at least 90, then uncross by 5 more degrees, rotate the sample, and so on.
As ever, Tobias
} Hi, Tobias, } } You've got most of this right, but not entirely. } } Your procedure is right: use just the polarizer in fixed position } and rotate the sample. (Actually, either could be fixed, so if you } don't have a rotating stage, you can leave the sample in place and } rotate a polarizing filter over the light port). } } The sample doesn't really act as the second polarizer (analyzer). } This is the classic experiment for either dichroism (materials which } exhibit two different refractive indices) and pleochroism (materials } which exhibit 3 different refractive indices). } } First, a few definitions: } Refractive index is an optical property which expresses the relative } interaction between the electric field in light and the electric } field in matter. The stronger the interaction, the more light } passing through a material will slow down and the higher the } refractive index. (Mathematically, RI = velocity of light in air or } vacuum/velocity of light as it interacts with a material where V air } = 300,000 km/sec). } } If you dissect the word "birefringence", you can see that it refers } to materials } } having the property of ("gence") } two (bi) } refractive indices (ref in ). } In actuality, materials can have three different RI's, all at right } angles to each other, but will only exhibit 2 at a time (think of } either films, fibers, or crystal faces), so we never refer to them } as "tri-refringent". } } Polarized light is different from ordinary light in the following way: } All light has a direction of travel. You see things in the world } around you because light is traveling from them to you. Light is } electromagnetic radiation. As microscopists, we are most typically } interested in the electrical field part (yes, I know that there have } been magnetic microscopes built). The sine wave we use to describe } light is actually the tracing of the tip of that electrical vector, } building up then dropping off in a positive direction then building } up and dropping off in negative direction. The motion of that sine } wave gives light a direction of vibration. The direction of } vibration is always at right angles to the direction of travel. } } Ordinary light contains all directions of vibration (imagine little } vectors vibrating N-S, E-W, and all angles in between; all at right } angles to the direction of travel). To convert ordinary light to } polarized light, you simply have to impose some sort of interaction } (reflection, specific types of absorption, beam splitting) which } absorbs all but one permitted direction of vibration. The result is } "Plane Polarized Light", which is not necessarily always the same as } linearly polarized light (tune in another time for the discussion of } "States of Polarization"). However, for the sake of simplicity, } plane polarized light does indeed vibrate linearly. } } So what happens when you rotate any birefringent material over a } polarizer? I can't draw diagrams for you here, but imagine a } rectangle with one RI oriented along the short edge and the other } along the long edge. As you rotate the sample, the short edge will } eventually align with the permitted direction of the light coming } from the polarizer. In this position, the light only "sees" one } refractive index (not the most scientific explanation, but } accurate). Rotating 90 degrees presents the other RI. Anywhere in } between, contributions from each of the RI's will be visible. In } practice, we use these unique positions to isolate each of the RI's } to actually measure them. } } And what's the story with things that are dichroic or pleochroic? } In addition to being birefringent, they are also colored in normal } illumination (ex: Congo red). To properly observe them, first set } up Koehler illumination and just observe the normal color. Then, } insert just a single polarizer and rotate. The colors that you see } will not be the typical magentas, golds, and turquoises } characteristic of polarized light interactions. Rather they will be } absorption colors (browns, reds, yellows, etc.), derived from the } unique property that, for these materials, not only does refractive } index vary with direction, absorption does too. The result - } changes in color and intensity. } } The next step would be to insert the analyzer and rotate. It is } also important to note that the normal Polarization colors may be } affected by the absorption colors. (For those of you who are } interested, I have a "polarized light road map" which outlines all } these test in flow-chart form). } } Here are some interesting examples of materials which are dichroic } or pleochroic: } Everyone cites tourmaline which is dark brown in one orientation and } pale yellow in another,. as well as biotite (a type of mica) and } cordierite. } Hartshorne and Stuart (Crystals and the Polarizing Microscope, } Arnold, 1970) cite magnesium platino-cyanide which oscillates } between bluish-red (parallel to "c" axis) and carmine red; } hypersthene (ferro-magnesium silicate): brownish red to green; } remind us that some biological materials also exhibit this property. } Patzett (Polarized Light Microscopy: Principles, Instruments, } Applications, Leitz - now out of print) reminds that the phenomenon } is widespread in organic materials and is responsible for the } circular dichroism we chemists routinely test for in levo and dextro } rotatory compounds. } And finally, I still have fond memories of doing experiments in grad } school from Chamot & Mason (Handbook of Chemical Microscopy, Vol. I } - now available through McCrone Associates) on a slew of materials. } (For those of you who are teaching, they recommend viscose rayon } dyed with congo red; crystals of o-nitrophenol, azobenzene, } iodoquinine sulphate, silver chromate, copper acetate, red ammonium } picrate and the magnesium platinocyanide - mentioned above). } } Well, we are back at Congo red, so I'll stop here. For those of you } interested in the physics of this process, I encourage you to read } Hartshorne and Stuart further. For the rest of you, find some } dichroic or pleochroic materials and just have fun. } } After many years of teaching, I realized that all the beauty and } complexity of polarized light boils down to one simple underlying } concept: refractive index. And here it is again. } } Hope this is helpful. } Barbara Foster } } Microscopy/Microscopy Education } 313 S Jupiter Rd, Suite 100 } Allen, TX 75002 } P: 972-954-8011 } W: www.MicroscopyEducation.com } } P. S. } Need a good general reference or light microscopy text? Call us } today to learn more about "Optimizing LIght Microscopy". Copies } still available through MME... even for class-room lots ... and we } give quantity discounts. Call us at (972)954-8011. } } } } } } } The colors exhibited are absorption colors and differ considerably } from those seen between crossed polars (the result of the } birefringenc } } } } At 05:15 PM 6/28/2005, Tobias Baskin wrote: } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Although 90+ percent of our needs could be met by good quality digital imaging, I would still retain film capability for reasons that often come up in these discussions. For important or irreplaceable images:
1) Film is archival, if processed correctly; 2) Film is platform-independent; 3) Film doesn't care if you have a CD, DVD, Zip, 3.5" floppy, 5.25" floppy, MO drive, etc., etc., etc. (i.e., it archives itself and doesn't need to be rearchived in newer formats as they come around); 4) Film can always be scanned into whatever format you need, hardware- or software-wise; 5) Film is still the highest-quality imaging medium available, and; 6) Film is still relatively low in cost (and you don't HAVE to print it unless you want to).
On the other hand:
1) Who knows how long film will be available? (Kodak has just announced that it will stop making photo paper! Talk about the end of an era!); 2) It is a pain in the derriere to maintain film processing facilities in labs with limited space; 3) New developments in digital imaging could catch up with films resolution in the not-too-distant future (although upgrading to that would certainly be pricey); 4) You might add film capability and no one will use it after all.
In my opinion, we are still a few years away from film being completely obsolete. As Mama Tindall says, "It's better to have it and not need it, than need it and not have it."
Now the next question is: should TEM manufacturers do away with traditional viewing screens and go entirely to monitors?
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 29 09:14:48 2005
Just a quick note for those using the new EMS "Ultra Smooth Carbon Adhesive Tabs".
The surface of the tabs is much improved and very smooth and the conductivity of the tabs is also improved, there is apparently a nickel rich wire mesh support/substrate embedded in these tabs. The ends of the wires are usually only visible at the very edge of the tabs and do not appear in the area where a sample would be affixed and nickel does not show up in an EDS spectrum of the sample collection area, many of you may have noticed this.
I have encountered a problem when trying to lift particles off of surfaces using these tabs. When pressure is applied to the tab while collecting particles forces some of the wire to the surface in the sample collection area. For most applications this isn't a problem, but if you are analyzing a group of particles and are interested in metals this could pose a significant problem.
Sincerely Bryan Bandli MVA Scientific Consultants
Disclaimer: I have no interest, financial or otherwise, in the product mentioned or any competitors.
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 29 09:17:10 2005
Microscopy Facility Supervisor. Biology Department, Central Michigan University.
Established in 1892, Central Michigan University has a growing enrollment of approximately 28,000 students, including 19,800 students on the university's main campus. Recently classified by the Carnegie Foundation as a doctoral/research-intensive university, CMU is recognized for strong undergraduate education and a range of focused graduate and research programs. CMU is a student-focused university with opportunities for leaders and involvement for an energetic team.
The Microscopy technician is responsible for supervising the teaching/research microscopy facility including a transmission electron microscope, scanning electron microscope, confocal microscope, specimen preparation lab, and photographic darkroom. Responsibilities: teach and assist in teaching microscopy courses, support faculty and student research, solicit externally funded contracts, and perform routine maintenance on all microscopy equipment and departmental light microscopes.
The position requires a Bachelor's degree or equivalent; two years qualifying work experience; excellent organizational and communication skills; ability to teach principles and techniques of microscopy; working knowledge of the principles and techniques of transmission and scanning electron microscopy; and experience with routine maintenance of EM, optical microscopes.
Desired qualifications include a Master's degree, experience with confocal microscopy, EDS, digital imaging, light microscopy, ultramicrotomy, vacuum evaporation, critical point drying, and biological specimen preparation preferred. Experience with equipment associated with the microscopy facility preferred.
Applicants must apply online at www.jobs.cmich.edu
CMU, an AA/EO institution, strongly and actively strives to increase diversity and provide equal opportunity within its community. CMU does not discriminate in employment against persons based on age, color, disability, gender, familial status, height, marital status, national origin, political persuasion, race, religion, sexual orientation, veteran status, or weight (www.cmich.edu/aaeo/)
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 29 09:41:58 2005
} Now the next question is: should TEM manufacturers do away with } traditional viewing screens and go entirely to monitors?
I am afraid so. Even airplane manufactures are considering replacement of cockpit and passenger cabin windows with monitors... On the other hand, virtual controls work just fine - such as desktop and mouse. But I would keep a peep hole, just in case. Especially in the cockpit :-)
Vitaly Feingold SIA 2773 Heath Lane Duluth, GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com
----- Original Message ----- } From: "Tindall, Randy D." {TindallR-at-missouri.edu} To: {microscopy-at-microscopy.com} Sent: Wednesday, June 29, 2005 9:52 AM
Randy, You bring up an interesting point at the end of your message. I believe that some of the manufacturers are indeed thinking about if not already eliminated the projection chamber all together. I believe one of the reasons was that with all of the new things that can be added to the column i.e. in column energy filters, Cs correctors etc.. the column was getting to tall to fit into most rooms. To get back on point with the film vs digital discussion I can only relate the experience in the structural biology community where there are some labs that have switched to digital with 4K ccd cameras (expensive). For practical reasons most of us are still using the tried and true film to collect our images for high resolution ( {1.0 nm) data collection of beam sensitive frozen hydrated samples. It gives more real estate per shot at a small pixel size but the jury is still out with the all digital crowd. These are just a few thoughts from someone who has been developing film and scanning for 20 years. Regards, Bob
At 08:52 AM 6/29/2005 -0500, Tindall, Randy D. wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
******************************** Robert Grassucci Howard Hughes Medical Institute Wadsworth Center Empire State Plaza Albany, NY 12201-0509
} You bring up an interesting point at the end of your message. I } believe that some of the manufacturers are indeed thinking about if not } already eliminated the projection chamber all together. I believe one of } the reasons was that with all of the new things that can be added to the } column i.e. in column energy filters, Cs correctors etc.. the column was } getting to tall to fit into most rooms.
It's done ! Have a look at :
http://www.jeol.com/tem_/temprods/jem2200fs.html
Jacques Faerber
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 29 10:45:36 2005
Listers, I have a JEOL 2200FS. Indeed there is no viewing chamber in the conventional sense. There is a small viewing screen hidden away for alignment purposes only. Imaging is carried out solely with cameras.
Chris
Christopher J Gilpin Ph.D. Assistant Professor Director, Molecular and Cellular Imaging Facility K1.A04 UT Southwestern Medical Center at Dallas 5323 Harry Hines Boulevard Dallas, TX 75390-9039 Phone +1 214 648 2827 Fax +1 214 648 6408
-----Original Message----- } From: Faerber Jacques [mailto:Jacques.Faerber-at-ipcms.u-strasbg.fr] Sent: Wednesday, June 29, 2005 10:16 AM To: Microscopy Society of America
Bob Grassucci wrote
} You bring up an interesting point at the end of your message. } I believe that some of the manufacturers are indeed thinking about if } not already eliminated the projection chamber all together. I believe } one of the reasons was that with all of the new things that can be } added to the column i.e. in column energy filters, Cs correctors etc.. } the column was getting to tall to fit into most rooms.
It's done ! Have a look at :
http://www.jeol.com/tem_/temprods/jem2200fs.html
Jacques Faerber
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 29 11:59:23 2005
I've have similar problems with samples only slightly soluble in water. To overcome dissolution, I saturate the water with the compound. In my case, it only takes 1-2 mg of material. Seem to work great. No more Swiss cheese!
John W. Catino Analytical Investigator - Microscopy Minerals Technologies, Inc. 640 N. 13th Street Easton, PA 18042
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 29 12:13:34 2005
I have a Rontec X-Flash detector on a JEOL 5910 SEM. This is a silicon-drift detector which operates at -15C using simple thermoelectric cooling. I am very satisfied with it for our application, which is qualitative microanalysis and x-ray mapping. I can't speak for quantitative analysis, as we almost never attempt it in this system. The resolution and baseline tend to change more with countrate than with a Si(Li), which might make high-precision quantitative analysis more problematic, but I expect the electronics will improve, too, and help compensate. It does not detect the light elements (C-N-O) at higher countrates, but sees them fine at 1000cps. I would consider a SDD very seriously for a future application, but I'm not convinced yet that it would replace a Si(Li) in every case.
I have an EDAX Sapphire Si(Li) detector with a small (angular) dewar. The concept made sense, that one filled it up only when needed for an occasional analysis. The reality depends on the meaning of "occasional". The SEM it is on is very busy with imaging, and EBSD, but users use EDS once or twice a week. It is a pain to have to interrupt the other work on the SEM to chill the detector two hours before you need to use it! The detector works exactly as intended - I'm not criticizing EDAX, it was just the wrong decision for our application.
I have 3 Oxfords, of varying ages, including a windowless on the VG STEM, which was built in 1991 and has never yet had to go for repair and which still has a resolution at Mn of {135eV (using a modern pulse processor) compared with the original specification of 138eV. This detector has run with two preamplifiers, on an AN10000, an eXL Mk. 1, two Isis systems, a (short-lived) Emispec ES Vision 4, and currently an INCA. Quite a history!
Tony.
At 10:30 AM 6/28/2005, Tom W Bargar wrote:
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*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
That's a good idea but it may not deal with all the problems since the presence of moisture may allow recrystallization to take place, resulting in the substitution of one polymorph for another, even if there is no net loss of material.
John Twilley
John.Catino-at-mineralstech.com wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I've have similar problems with samples only slightly soluble in water. To } overcome dissolution, I saturate the water with the compound. In my case, } it only takes 1-2 mg of material. } Seem to work great. No more Swiss cheese! } } John W. Catino } Analytical Investigator - Microscopy } Minerals Technologies, Inc. } 640 N. 13th Street } Easton, PA 18042 } } Tel: 610.250.3363 } Fax: 610.250.3206 } john.catino-at-mineralstech.com } } ********************************************************************** } This email and any files transmitted with it are confidential and } intended solely for the use of the individual or entity to whom they } are addressed. If you have received this email in error please notify } the system manager. } } This footnote also confirms that this email message has been swept by } MIMEsweeper for the presence of computer viruses. } www.mimesweeper.com } **********************************************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 29 14:15:26 2005
I'm looking for a high frequency acoustic microscope (200+ Mhz) in pretty much any university or government lab in or near the san francisco bay area that I might be able to bargain some time on. At a minimum, this includes UC Berkeley, UC Davis, UC Santa Cruz, UC SF, Stanford, LBNL, and LLNL. Any idea where I might find one? Unfortunately, the original Stanford group that pioneered this work no longer studies or uses acoustic microscopy.
I'm trying to characterize the mechanical properties of small cells approx 10um in diameter (size, impedance, elasticity, etc).
Patrick. ---- Graduate Student UCSF/UC Berkeley Joint Graduate Group in Bioengineering 408-203-6052
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 29 20:28:04 2005
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From MicroscopyL-request-at-ns.microscopy.com Thu Jun 30 04:40:45 2005
} I have a Rontec X-Flash detector on a JEOL 5910 SEM. ...
We too have recently installed a Roentec, and we are absolutely amoazed at its speed. We measure Mn metal the other day at 150kcps (into the spectrum) at Mn Ka FWHM=165eV! However, our applications are mineral mapping. Like Dr Garratt-Reed, we would also believe with the addition of this detector on the market, potential purchasers should consider their applications relative to "quantitative analysis" versus "image or spatial analysis", and don't forget to test drive the software.
cheerios ... Michael Shaffer :o) Avalon Peninsula, Newfoundland
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 30 08:00:25 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (exploratorium-at-tiscali.it) from http://www.microscopy.org/MicroscopyListserver/MLFormMail.html on Tuesday, June 28, 2005 at 11:42:12 ---------------------------------------------------------------------------
Email: exploratorium-at-tiscali.it Name: giovanni de caro
Organization: museo laboratorio di scienze naturali per ragazzi "L. MontalbÚ" - casalciprano (Cb) - Molise - Italia
Title-Subject: [Microscopy] [Filtered] MListserver: Request for prepared microscope slides for science museum
Question: Hi all! I am the director of a small, volunteer operated and self funded science museum for youngsters based in southern Italy (see our website - translated in english : http://web.tiscali.it/exploratorium). We have here a small biology lab equipped with optical microscopes; we do organize small courses of biology for kids. If you have to donate some prepared microscope slides of: vegetal tissues, protozoa, fungi, insects or other which maybe of ineterst for us, let us know; we can pay for shipping from USA or elsewhere, if requested. We also are looking for an older, working slide projector; shipping as above.
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I would like to ask if someone of you know if uranyl acetate (generaly used in transmission electron microscopy) can be used in ESEM and if it possesses a potential radioactivity risk.
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Email: thomas.richards-at-arkemagroup.com Name: Thomas Richards
Organization: Arkema
Title-Subject: [Microscopy] [Filtered] Service for LEO 1530 FE-SEM near Philadelphia, PA.
Question: Hello,
I am looking for someone to service our LEO 1530 FE-SEM and would appreciate any help. We are located near Philadelphia, PA. Thanks.
Thomas Richards Senior Research Chemist, Systems and Materials Analysis Analytical and Systems Research Arkema Inc. 900 First Avenue King of Prussia, PA 19406 (610) 878-6309 (610) 878-6196 fax thomas.richards-at-arkemagroup.com
In our lab, we currently use a Congo red staining for amyloid fibers in spleen sections. We do use both polarizer and analyzer and have a very bright specific green birefringence, provided that the intensity of light is very high. Furthermore, we have been able to see a bright red fluorescence in these areas. For the fluorescence, though, artefacts seem to be a problem.
What protein are you trying to look at? does it display some kind of alignment as beta-sheets?
Regards,
Marie-Claude Belanger Montreal
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From MicroscopyL-request-at-ns.microscopy.com Thu Jun 30 09:11:44 2005
In our lab, we currently use a Congo red staining for amyloid fibers in spleen sections. We do use both polarizer and analyzer and have a very bright specific green birefringence, provided that the intensity of light is very high. Furthermore, we have been able to see a bright red fluorescence in these areas. For the fluorescence, though, artefacts seem to be a problem.
What protein are you trying to look at? does it display some kind of alignment as beta-sheets?
Regards,
Marie-Claude Belanger Montreal
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 30 09:35:52 2005
Hello all, I have a student wishing to do some freeze fracture of Bacteria for SEM. We are looking for a unit where we could have some samples prepared, preferably in the Great Lakes area. Any suggestions would be appreciated.
Thanks, Ron.
Ronald J. Smith Department of Biology Room 235, Biological & Geological Sciences Bldg. U.W.O., London, Ontario N6A 5B7 (519) 661-2111 ext.86486
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 30 11:38:01 2005
On Jun 30, 2005, at 6:00 AM, by way of MicroscopyListserver wrote:
} I would like to ask if someone of you know if uranyl acetate (generaly } used in transmission electron microscopy) can be used in ESEM and if } it possesses a potential radioactivity risk. } Dear Monica, I don't know what you want to examine, so I don't know what use UAc will be to you in the ESEM, but I think that you could look at a specimen to which UAc has been added without risk to the instrument. There is no particular problem related to the very small amount of radioactivity in the small quantities of UAc generally used in EM; however, you should check with your safety office about proper handling and disposal of UAc, since the laws regarding this vary from place to place. Since U is an alpha emitter, the radiation will not penetrate through the dead layer of the skin, so it is a hazard only when ingested or inhaled. Your safety office will advise you on protocols to reduce the likelihood that you will be at risk from ingestion or inhalation. In our lab, all use of UAc in confined to a region in a fume hood, and anything used with UAc that is to be disposed of is placed in a labeled container in that space and picked up by the safety office. One should always wash ones hands after working with UAc. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 30 16:14:24 2005
We are comparing dye sub printers versus the Fuji Pictrography PG4500. The price differential is a staggering $22,000 versus $6,500, respectively. It is my understanding that both produce good quality, gray-scale prints (versus dithered, inkjet prints). Why the price differential? I would welcome any comments, user experiences, etc. Does anyone know the average cost per 8.5 x 11in print?
Thank you. -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 30 18:04:09 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bberkmeyer-at-flood.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, June 30, 2005 at 11:38:55 ---------------------------------------------------------------------------
Question: What would be suggested for a microtomes on a piece of wood (pine) which has been coated with a thermoplastic polymer coating? I would probably have to freze the sample to avoid any heat generated in cutting...coating will be staned with Rhodamine 6 and viewed under UV Flour..Goal is looking at coating penetration. Also, is a microtome the correct preparation?