Dear All This is all fun. Lost a bit of skin myself. Latex gloves does help a bit. Home made clamping devices I pressume will help, but I prefer to have a "hands on" onto the sample. For large amounts of samples automation is a option.
} Everett Ramer wrote: } } } } We have a manual metalographic grinding/polishing wheel. The sample } } } being prepared is held in the hand as it is pressed against the rotating } wheel. } } } Our safety people have asked us to provide finger protection for this } device. } } } Does anyone have any solution/suggestion? } } } Everett Ramer } } Everett, } Mary Mager's response, while funny, is actually quite accurate. There are } not too } many ways to protect fingers while properly holding a small (1-1/4", } typically) } sample for grinding/polishing. } } I preface my next remarks by pointing out that I work for a manufacturer of } } metallographic equipment and consumables, and therefore have a financial } interest in solving your problem: } } We at BUEHLER, do offer a simple grinding fixture which might help. This } fixture } is a squat, stainless steel, hollow cylinder with a carbide ring around } it's base. } The sample is clamped within another hollow cylinder seated within the } first. } The two cylinders are threaded, so that the inner can be raised or lowered } with respect to the carbide 'stop' of the outer. Engraved markings allow } material removal in increments as fine as 20microns. While this is not } actually } finger protection, per se, it will allow you to grasp something larger so } that your } fingers are not in such close proximity to the grinding wheel. We also } offer } a motor system which allows the fixture to rotate, in place, on the wheel} Mr. S H Coetzee Electron Microscope Unit Private bag X3 Wits Johannesburg 2050 Tell: +27 11 716 2419 Fax : +27 11 339 3407 E-mail stephan-at-gecko.biol.wits.ac.za
I am curious if anyone is using an automatic section stainer for TEM. If so, what brand are you using. We are using an LKB section stainer (15 years old now). I just wonder if there is anything else in the market. Thanks,
Cora Bucana ******************************************************* Corazon D. Bucana, Ph.D. Department of Cancer Biology U.T. M.D. Anderson Cancer Center 1515 Holcombe Blvd. Box 173 Houston, Texas 77030 Phone: (713) 792-8106 Email:bucana-at-audumla.mdacc.tmc.edu FAX: (713) 792-8747
May 20-22 and May 24-26, 1999 North Carolina State University Raleigh, North Carolina, USA
and
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This highly regarded hands-on course taught by expert faculty has been presented annually for more than 15 years. It deals with all phases of quantitative and computer-assisted imaging from acquisition and processing through measurement and stereological interpretation. Attendees receive The Image Processing Handbook plus a CD-ROM containing images, algorithms (Photoshop-compatible for Mac and Windows) and an extensive on-line tutorial and course notes on stereology and statistical analysis. The course is appropriate for scientists, technicians and administrators using or intending to use these techniques. Attendees typically come from materials science, geology, biological and medical sciences, pharmaceuticals, food science, industrial quality control, remote sensing, and other disciplines. You are encouraged to bring your own images for the hands-on lab sessions.
For detailed information and registration contact Cindy Allen, Dept. of Continuing and Professional Education, N. C. State University, Raleigh, NC 27695-7401, 919-515-8171, fax 919-515-7614, email: Cindy_Allen-at-NCSU.edu
Information is available on-line at the following sites:
Seadoohog-at-aol.com-at-sparc5.microscopy.com wrote: } } My next question concerns Energy Dispersive X-ray Spectroscopy (EDS). The } baseline on the EDS spectra is influenced by inelasic scattering of the } incident electron beam by the atomic nuclei of the sample, which results in a } peaked background. This is called the bremsstrahlung effect. What causes } this??? } } Thanks!
Dear Seadoohog, Brehmsstrahlung, or "braking radiation" is caused by the ac- celeration of the electron by a large mass (nucleus). Both Maxwell's equations and quantum mechanics predict that accelerated charges will give off electromagnetic radiation. The large mass is necessary so that conservation of both energy and momentum can be satisfied. Yours, Bill Tivol
There are some second hand equipment outlets here in the US who occasionally get bits and pieces for Zetopans. One is John Oren, in VT.
Re: illuminators - OptiQuip has suggested some interesting alternatives for an upgrade path. We will be working on this in Apr/May. Anyone interested is welcome to email privately for further info.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 01:41 PM 2/28/99 +0100, Yvan Lindekens wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In the process of trying to produce some Ag coated samples, I instead obtained some silver beads, which would be dandy if that's what I was after ;^ { ...
The specifics were: Denton DV502A evaporator, 4cm of 0.2mm dia Ag wire wrapped around ~1mm dia section of a standard carbon rod, 5x10^-7 Torr. Melting of the wire occured abruptly at well under 20A.
Can anyone suggest conditions or parameters to that will yield evaporation rather than melting???
Later I expect to coat with Al, which like Ag melts and boils at much lower temperatures than Pd and Pt (which are no problem with the above...) so if anyone can provide similar information regarding Al, that would also be helpful.
(This is probably pushing my luck, but if anyone knows any rule of thumb for how thick the films of the above are as a function of conditions & time, that would be super to hear....)
Thanks.
_______________________________________________________ Get your free, private email at http://mail.excite.com/
O.k., I've lost my cheat sheet which had the sizes of Tobbaco Mosaic Virus components, and I can't locate the information presently. Can anyone help me out here? I know the particles are 18.0 nm in diameter "300 nm" in length (o.k. some of them break). What is the diameter of the central core?
What is the spacing between the sprialing sub units?
Thanks!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
Silver has a melting point of 961C. Its vapor pressure is 847C = 1e-8 torr 958C = 1e-6 torr 1105C = 1e-4 torr
An equilibrium vapor pressure of 1e-6 will give ~1 monolayer per second deposition rate (using a kinetic theory of gas model with a sticking coefiicient of 1). Since this will undoubtedly not be an equilibrium situation, you can this value to be an upper limit. So, at the melting point of silver, you will get much less than 0.2nm/sec deposition rate near the sample. Farther away, it will drop as 1/r^2.
Aluminum too, will melt long before you get much evaporation 660C = melting point 677C = 1e-8 torr 821C = 1e-6 torr 1010C = 1e-4 torr
Also, thermal evaporation of Ag, Au, Al will tend to produce metal islands on the sample which can interfere with high mag imaging. Sputtering often will give a smaller grain size.
Cheers, Henk
At 08:40 AM 3/1/99 -0800, you wrote: } {snip} } } Can anyone suggest conditions or parameters to that will yield evaporation } rather than melting??? } } Later I expect to coat with Al, which like Ag melts and boils at much lower } temperatures than Pd and Pt (which are no problem with the above...) so if } anyone can provide similar information regarding Al, that would also be } helpful. } } (This is probably pushing my luck, but if anyone knows any rule of thumb for } how thick the films of the above are as a function of conditions & time, } that would be super to hear....) } } Thanks.
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu An optimist believes that we live in the best of all possible worlds. A pessimist fears that this is true.
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There was a request recently for information on a new EM techniques book. = The book is scheduled to be published in mid March so there is no = information yet other than the chapter titles. I post them here.
Electron microscopy methods and protocols / edited by M.A. Nasser Hajibagheri. (Methods in molecular biology ; v. 117) ISBN 0-89603-640-5 Expected publication date is mid March.
Contents: 1 General Preparation of Material and Staining of Sections ................= ... 1 Heather A. Davies 2 Negative Staining of Thinly Spread Biological Particulates ..............= . 13 J. Robin Harris 3 Preparation of Thin-Film Frozen-Hydrated/Vitrified Biological Specimens for Cryoelectron Microscopy .....................................= .. 31 J. Robin Harris and Marc Adrian 4 The Production of Cryosections Through Fixed and Cryoprotected Biological Material and Their Use in Immunocytochemistry .......... 49 Paul Webster 5 High-Pressure Freezing for Preservation of High Resolution Fine Structure and Antigenicity for Immunolabelling ............................= . 77 Kent McDonald 6 The Application of LR Gold Resin for Immunogold Labeling .............. = 99 J. R. Thorpe 7 Low-Temperature Embedding in Acrylic Resins .............................= . 111 Pierre Gounon 8 Quantitative Aspects of Immunogold Labeling in Embedded and Nonembedded Sections...................................................= ..... 125 Catherine Rabouille 9 Microwave Processing Techniques for Electron Microscopy: A Four-Hour Protocol ......................................................= ............. 145 Rick T. Giberson and Richard S. Demaree, Jr. 10 Electron Microscopic Enzyme Cytochemistry...............................= .... 159 Nobukazu Araki and Tanenori Hatae 11 In Situ Molecular Hybridization Techniques for Ultrathin Sections ....................................................= ................ 167 Jean-Guy Fournier and Fran=E7oise Escaig-Haye 12 Preparation of the Fission Yeast Schizosaccharomyces pombe for Ultrastructural and Immunocytochemical Study ..................... 183 M. A. Nasser Hajibagheri, Kenneth Sawin, Steve Gschmeissner, Ken Blight, and Carol Upton 13 Preparation of Double/Single-Stranded DNA and RNA Molecules for Electron Microscopy ...................................................= ............ 209 M. A. Nasser Hajibagheri 14 Applications of Electron Microscopy for Studying Protein-DNA Complexes..................................................................= .................. 229 Maria Schnos and Ross B. Inman 15 X-Ray Microanalysis Techniques .........................................= ............. 245 A. John Morgan, Carole Winters, and Stephen St=FCrzenbaum
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/apw.htm
by nss4.cc.Lehigh.EDU (8.9.1a/8.9.1) with ESMTP id OAA158376; Mon, 1 Mar 1999 14:17:15 -0500 Message-ID: {36DAF5C8.6259645-at-lehigh.edu}
Please note the following job announcement:
ELECTRON MICROSCOPE TECHNICIAN
Lehigh University seeks an Electron Microscope Technician to perform technical duties in support of the Electron Microscopy Laboratory of the Materials Science and Engineering Department. Technician will instruct students in the operation of microscopes and other equipment; maintain and repair instruments; oversee upkeep of the lab; support research professors and students; analyze samples; give tours and demonstrations; supervise students; maintain a safe environment; and other assigned duties. Bachelors degree in physical science and/or 4+ years related work experience required. Candidates should be familiar with electron microscopes, mechanical and electronic equipment, vacuum systems, PC and/or MAC and EDS/WDS systems. Good communication and interpersonal skills are essential.
Lehigh University offers excellent benefits and vacation package. Interested candidates should forward resume to: Deanne Hoenscheid, Materials Science and Engineering, Lehigh University, 5 E. Packer Ave., Bethlehem, PA 18015. EOE. M/F/D/V.
On Mon, 1 Mar 1999 edelmare-at-casmail.muohio.edu-at-sparc5.microscopy.com wrote:
} Date: Mon, 1 Mar 1999 11:42:21 -0500 } From: edelmare-at-casmail.muohio.edu-at-sparc5.microscopy.com } To: microscopy-at-sparc5.microscopy.com } Subject: Size of TMV? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } O.k., I've lost my cheat sheet which had the sizes of Tobbaco Mosaic Virus components, } and I can't locate the information presently. Can anyone help me out here? I know the } particles are 18.0 nm in diameter "300 nm" in length (o.k. some of them break). What } is the diameter of the central core? } } What is the spacing between the sprialing sub units? } } Thanks! } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." }
TMV:
Pitch = 2.3 nm
Don't know about inner diameter of core; try: http://www.ncbi.nlm.nik.gov/ICTVdb/welcome.htm
Let me know the diameter if you find it. S
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I've used W baskets to produce thin films of Au,Cr,Al and Cu in similar evaporators. I would think it would work with Ag too. The liquid metal seems to "hang" in the basket by what, I would guess, are surface tension effects. Any of the major EM houses should have these baskets.
I've also attempted to calculate, a priori, what the film thickness should be and never been too thrilled at the agreement with the result. A quartz crystal thickness monitor can be calibrated for most metals but if you don't have one, you might try using the change in resistance across a glass coverslip (Steere approach) or cross sectioning a series of thin films generated under known evaporation conditions and looking at them in a TEM.
cheers, John John Heckman Michigan State University
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
radsci-at-excite.com writes ... } } } In the process of trying to produce some Ag coated samples, I instead } obtained some silver beads, which would be dandy if that's } what I was after } ... } The specifics were: Denton DV502A evaporator, 4cm of 0.2mm } dia Ag wire wrapped around section of a standard carbon rod, } ...
I might suggest, rather than carbon rod, you wrap the Ag wire around tungsten wire which will be wetted. I imagine the Ag on carbon was like water on a duck's back.
BTW, why did a simple reply to this post want to send a message to {"radsci-at-excite.com"-at-sparc5.microscopy.com} ???
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Everett, When I have to do a quick grind on something small that I don't want to put into a mount, I use the "rubber fingers" that secretaries wear. I have found these to be much better than finger cots, which don't last one spin of the wheel. I don't know if this solution will satisfy the "safety" people but I still have finger prints. Dorrance
} ---------- } From: EVERETT RAMER } Sent: Thursday, February 1999 8:23 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Finger Protection during Sample Prep } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have a manual metalographic grinding/polishing wheel. The sample } being prepared is held in the hand as it is pressed against the rotating } wheel. } Our safety people have asked us to provide finger protection for this } device. } Does anyone have any solution/suggestion? } Everett Ramer } }
We have been asked this question by a customer: =========================================== We are looking for a holder for the Wehnelt cap for filament adjustment. AMRAY had one when we went up there but they do not sell one nor could they tell us where to buy one. It looks like a custom job. Do you know what we are talking about? =========================================== I am sure that someone has this hidden away somewhere in their catalog but I have not been able to find it. Any help would be appreciated.
Chuck
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Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
I run a very old probe in a small Geology department. I want to buy a JEOL 733 or later (eg 840A). I would be very pleased to hear from anyone who has one currently for sale or who is planning to sell one this year.
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Dear all We do evaporation of various metals on a old Edwards evaporator. First the Tungstan (W) wire gets heated up under vacuum to clean it from all dirt. Then the requiered metal wire gets wraped around a V bent in the W wire. when heated up the wire will melt and form a melted globule. this will evaporate only after futher heating. }
} } The specifics were: Denton DV502A evaporator, 4cm of 0.2mm dia Ag wire } wrapped around ~1mm dia section of a standard carbon rod, 5x10^-7 Torr. } Melting of the wire occured abruptly at well under 20A. } } Can anyone suggest conditions or parameters to that will yield evaporation } rather than melting??? }
} } (This is probably pushing my luck, but if anyone knows any rule of thumb for } how thick the films of the above are as a function of conditions & time, } that would be super to hear....) } No it does exist.
p = density of the of the coating material in g. ( cm^-3) M = weight of coating material a source (g) R = source samle seperation, (cm) dx = film thickness.
(4 pie R^2) dx p = M
But 1) efficiency factor of evaporative method method is rarely better than 75%. thuss: (4 pie R^2) dx p = M.3/4 this will only work if the sample is directly under the evaporated source
If it is at a angle theta,
3/4.M = (4 pie R^2 dx p)/ sin theta Where theta is the angle between the gounr and the tip of the V.
since Em people do like a really thin coating (nm) for M in g/cm^3 R in cm and dx in nm
M = 4/3. ( 4pie R^2 dx p)/sin theta . 10^-7
For coating material in wire form for a diameter d cm and l lenth (cm0
M = (pie d^2 lp)/4
length of wire required for a coatint thickness of dx cm 1 = 64/3 .[( R^2 dx)/d^2 sin theta)]
hope this does help
Mr. S H Coetzee Electron Microscope Unit Private bag X3 Wits Johannesburg 2050 Tell: +27 11 716 2419 Fax : +27 11 339 3407 E-mail stephan-at-gecko.biol.wits.ac.za
Good day Listmembers A while ago there was a discussion on this list on the preparation of insect eggs for ultra-thin sectioning. Without re-opening the discussion, can somebody that collected all the replies and other experts in this regard please supply me with a general protocol if there is one?
TIA Alan N Hall Laboratory for Microscopy and Micro-Analysis NWII Building University of Pretoria Pretoria 0002 Republic of South Africa Tel: +27-12-420 3896(Office) +27-12-420 2075(Laboratory) Fax: +27-12-362 5150
Dear all, In the evaporation of metal from tungsten wire it is important not to raise the temperature of the wire too fast. If you do this, the metal will melt and drop off in a blob. The thing to do is to raise the temperature until the metal just starts to melt. Then wait until the wire is wetted - you'll see the liquid creeping up the sides of the V. The bulk of the metal will remain as a blob at the apex of the V but will not drop off. Then you can turn the heat up more until the evaporation takes place.
Eric
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
At 03:54 PM 3/1/99, shAf wrote: } } BTW, why did a simple reply to this post want to send a } message to {"radsci-at-excite.com"-at-sparc5.microscopy.com} ??? } I am not sure about the why, but I see that too for various posts. I think it has something to do with the formating of the address, perhaps the use of the quotes.
You know, it could almost be beneficial. You can reply to the sender or the whole list by removing the unwanted part. However, about half the time, I forget to do the editing and my posts get kecked back for me to fix them.
Open Position: Microscopy Characterization Scientist
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Provide polymer morphology, particle and aesthetics characterization support at GE Plastics in Mt. Vernon, IN. In particular, TEM, SEM, optical microscopy, AFM, particle sizing, surface and other more specialized techniques are used. Leadership in effective problem solving approaches is a must.
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Interested candidates should send their resume to:
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The C.M.I./A.A.B. Descriptions of Plant Viruses No. 151 (Oct. 1975) on tobacco mosaic virus (type strain) gives the following information:
..cylindrical canal of radius c. 2 nm.....
As Sarah Miller states, a pitch of c. 2.3 nm is given.
More infromatiom about particle structure is given: If anyone would like a complete copy of this bulletin, I can mail you a copy. Dont' know if the fax machine will accept this document the way it is folded.
Maureen Petersen
************************************************************************ Maureen Petersen Department of Plant Pathology 1453 Fifield Hall University of Florida
My grandson in the states recently and proudly wrote to his Grand-Dad that he NOW has a microscope and could I send him some samples to look at. He told me that .....
"the microscope is EDU-SCIENCE and the magnifications are 100x 300x 600x."
Since my grandson is 10 years old and egar, I don't want to ask him questions like - Are the objectives aprochromats?. Is anyone on the list familiar with this microscope? I specifically want to know if it's optics are good, and if they are color corrected.
Once I know if he has a decent microscope or a toy that makes things biger and the user blind or crazy, I will better know how to encourage him, and what type of samples and reading matter to send.
Thanks for you help, from a land far away.
Shalom from Jerusalem, Azriel
+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+ Azriel Gorski, Head Fibers and Polymers Laboratory Division of Identification and Forensic Science Israel National Police Jerusalem
azrielg-at-cc.huji.ac.il ICQ User ID No. - 1750739
CHOICE - The enchanted blade, with an edge that shapes lifetimes. Richard Bach RUNNING FROM SAFETY
A friend is someone who knows the song in your heart, and can sing it back to you when you have forgotten the words. +-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+
Hi! I have a reference to two stains/methods for nerves: Koelle's acetyl cholinesterase technique and Winklemann's Silver Impregnation method. I have not been able to find either one. I found a "general" reference to acetyl cholinesterase from the University of Nottingham web page in which they recommend using 'DPK' mounting resin. I have never heard of it--can anyone help me out? Thanks!
Peggy Sherwood Wellman Labs of Photomedicine (224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-3192 (fax)
} -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } We have been asked this question by a customer: } =========================================== } We are looking for a holder for the Wehnelt cap for filament adjustment. } AMRAY had one when we went up there but they do not sell one nor could they } tell us where to buy one. It looks like a custom job. Do you know what we } are talking about? } =========================================== } I am sure that someone has this hidden away somewhere in their catalog but I } have not been able to find it. Any help would be appreciated. } } Chuck
Chuck:
I believe these Wehnelt cap holders are available from Energy Beam Sciences.
Ken Bart
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323 USA kbart-at-hamilton.edu (315) 859-4715
} O.k., I've lost my cheat sheet which had the sizes of Tobbaco Mosaic Virus components, } and I can't locate the information presently. Can anyone help me out here? I know the } particles are 18.0 nm in diameter "300 nm" in length (o.k. some of them break). What } is the diameter of the central core?
The diameter of the central core is 40 A, it is packed with the nucleic acid (RNA). The protein subunits are arranged in a helix containing 16 1/3 subunits per turn ( or 49 subunits per three turns). Each subunit consists of 158 amino acids in a constant sequence.
} What is the spacing between the sprialing sub units? } It is the nucleic acid.
Best regards,
Ming
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * #1074B Dentistry Pharmacy Building * * University Of Alberta. * * Edmonton, Alberta, Canada T6G 2N8 * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Charles writes ... } } We have been asked this question by a customer: } =========================================== } We are looking for a holder for the Wehnelt cap for filament } adjustment. } AMRAY had one when we went up there but they do not sell one } nor could they tell us where to buy one. ...
Just to clarify ... this sounds like a "jig" strictly for holding the wehnelt on the workbench(??) From personal experience I've never seen one of these as a necessary component for the adjustment ... or if it were, it came with the instrument, or was available from the manufacturer. Usually, its primary use is as a heat sink so that the wehnelt assy can be removed while it is hot and to allow it to cool down in a minimum amount of time. Since wehnelts vary in design and geometry I can't imagine this being available thru any other source other than the manufacturer ... which leaves you with what would seem to be a very simple job for a machine shop. (... why do I think this isn't much help? ...)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
} From: Mriglermas-at-aol.com Return-path: {Mriglermas-at-aol.com} To: Woody.N.White-at-mcdermott.com Cc: Mriglermas-at-aol.com
I wish to purchase a picoammeter for specimen current measurement (x-ray analysis work) and am requesting information on models, prices and vendors. Please contact me directly. Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Sonneborn, T.M. (1975), "The Paramecium aurelia complex of fourteen sibling species", Transactions of the American Microscopical Society 94: 155-178.
This is to support a high school student doing a very interesting project on individual variation in protozoa. The local library ordered a copy but estimates 3 weeks delivery.
Can someone out there help me to get it quickly? I will be happy to reimburse reasonable expenses.
Posted by our evaporater, dating from who-knows-when, but certainly long ago, is a sheet of recommended procedures for evaporation different metals. For silver it recommends using a molybdenum or tantalum wire as the filament. It says to wrap a fine wire of silver around the Ta or Mo filament, and then simply evaporate. I recently did a number of successful silver evaporations by fashioning (by hand) a Mo spiral basket and putting a small bead of Ag in it. (I had some Mo wire and Ag beads). I then evaporated the Ag just as I would have evaporated Au from a W basket. It worked fine, but, as always with evaporating from a basket in this way, the thickness uniformity over distances of 2-3 cm. was not very good.
My favorite form of fingertip protection has always (20+ years) been long fingernails. Not talons by any means, but longer than fingertip length. You can feel when a nail is touching without actually losing any skin. Granted, you sometimes generate a strange-looking manicure but an emery board or some 600 grit paper will soon put that to rights. One thing to watch out for: don't let the water to your wheel get so cold as to cause numbness in your fingers. If the water's too cold, you won't know you've ground off your fingerprints until you notice blood on the wheel. And it will really smart when the feeling comes back!
Stephan Coetzee wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear All } This is all fun. Lost a bit of skin myself. Latex gloves does help a } bit. Home made clamping devices I pressume will help, but I prefer } to have a "hands on" onto the sample. For large amounts of samples } automation is a option. } } } } Everett Ramer wrote: } } } } } } We have a manual metalographic grinding/polishing wheel. The sample } } } } being prepared is held in the hand as it is pressed against the rotating } } wheel. } } } } Our safety people have asked us to provide finger protection for this } } device. } } } } Does anyone have any solution/suggestion? } } } } Everett Ramer } } } } Everett, } } Mary Mager's response, while funny, is actually quite accurate. There are } } not too } } many ways to protect fingers while properly holding a small (1-1/4", } } typically) } } sample for grinding/polishing. } } } } I preface my next remarks by pointing out that I work for a manufacturer of } } } } metallographic equipment and consumables, and therefore have a financial } } interest in solving your problem: } } } } We at BUEHLER, do offer a simple grinding fixture which might help. This } } fixture } } is a squat, stainless steel, hollow cylinder with a carbide ring around } } it's base. } } The sample is clamped within another hollow cylinder seated within the } } first. } } The two cylinders are threaded, so that the inner can be raised or lowered } } with respect to the carbide 'stop' of the outer. Engraved markings allow } } material removal in increments as fine as 20microns. While this is not } } actually } } finger protection, per se, it will allow you to grasp something larger so } } that your } } fingers are not in such close proximity to the grinding wheel. We also } } offer } } a motor system which allows the fixture to rotate, in place, on the wheel} } Mr. S H Coetzee } Electron Microscope Unit } Private bag X3 } Wits } Johannesburg } 2050 } Tell: +27 11 716 2419 } Fax : +27 11 339 3407 } E-mail stephan-at-gecko.biol.wits.ac.za
Help- We are having a problem with getting nuclear autofluorescence of tissue culture cells fixed (Paraformaldehyde) and prepped for immunofluorescence. If you fix and mount the cells in glycerol-phenylene diamine with no antibody treatment, they usually look blank immediately, then over time they develop nuclear fluorescence. The nuclear stain is primarily in the fluorescein window and somewhat variable but seems to come up over time. Is it possible that this is coming from a breakdown product of the phenylene diamine we are using as an antifade? An apparently unrelated problem is nuclear background with secondary antibodies alone. If we treat with secondary and look immediately (before the other problem seems to arise), we also get nuclear staining. This seems to vary with the batch of secondary antibody so appears to be non-specific binding of the secondary. It is not blocked by serum etc. Any ideas appreciated. THanks- Dave
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax)
I am trying to dissolve Brefeldin A in Dulbecco's MEM for use in a cell = culture experiment for TEM study. However the drug does not seem to be = readily soluble in the MEM.=20 Can someone give me some help?
Dr. A.P. Alves de Matos Curry Cabral Hospital TEM Unit Portugal
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} I am trying to dissolve Brefeldin A = in=20 Dulbecco's MEM for use in a cell culture experiment for TEM study. = However the=20 drug does not seem to be readily soluble in the MEM. {BR} Can someone = give me=20 some help? {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Dr. A.P. Alves de Matos {BR} Curry = Cabral=20 Hospital {BR} TEM Unit {BR} Portugal {/FONT} {/DIV} {/BODY} {/HTML}
I have been watching what has been going on in relation to your sputter coating problems. I did not look too hard at the early postings so what = I have set out below may be covering old ground, but I hope it may help? M= ay I say that there have been many who have tried so hard to help you, my comments below in no way are critical of their stirling efforts.
Sputter coaters need a RP vacuum of just to the left of dead centre on yo= ur coater's vacuum gauge. It does not matter what the gauge has as its calibration; this setting should work for gold, gold-palladium or platinu= m.
There are three styles of sputter coater low voltage {600volts, medium voltage 1,000 to 1,500volts and a high voltage coater at 1,000 to 3,000volts.
The low voltage coaters do not have a voltage control, simply a depositio= n control which you should set at it's mid point. Set the voltage controls=
for the other two coaters in their mid range too.
As a test specimen use a 2" square piece of white paper weighted down wit= h a stub. Set the target to specimen distance at 5cms. Set the time at tw= o minutes (much too long for normal coating but fine for this test).
Pump the system down YOU DO NOT NEED ARGON! We often run with air, the difference is that argon gas gives a constant coating rate but air, wet o= r dry, gives different coatings. If you are using argon, flush to full lef= t scale once or twice to clean out the system during the pump down.
When you have a vacuum greater than 60% of the scale bleed the gas into t= he system until the pressure holds on the position mentioned above; just to the left of dead centre. When the vacuum is at the correct level, switch=
on the plasma and increase the voltage to the correct level. You should have a plasma and by adjustment of the voltage, deposition, or gas, adjus= t the current to ~20mA. Let the system run for the two minutes.
When the system completes its cycle open it up and take a look at the paper. Using a gold target - very light blue, its working but not very efficiently, pale blue, better but still not good enough, darker blue, still not good enough, blue-grey much more like it, grey to gold tint, it= s working fine.
If you do not have any of these colours the target material may be your problem. Is it one of the metals mentioned below? Even if the target is=
gold on brass and all the gold has gone you will still sputter brass! =
Chromium, aluminium and carbon will not sputter with the very simple systems used for conventional SEM. Much higher specification systems, i= n relation to current and vacuum, are required.
Well I do hope that helps? Please remember that sputter coaters are not 100% efficient when it comes to coating non-conducting specimens. Even a=
good coater will have trouble with rough and porous specimens. You shoul= d never expect to run above 15kV if you have coated this type of specimen.
Well that's it I do hope we can sort out your problem?
Regards
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
If anyone is disposing of this Amray scope, I would appreciate knowing about it so that I could purchase some spare parts and assemblies from it before it is dumped. I am particularly interested in the main 30KV power supply and the CRT power supply, photomult assembly, and any of the circuit boards. (Specific model is 1600T without the digital controls and pushbuttons. All components and assemblies are of interest.)
I am also looking for a Robinson detector for this scope. Pls let me know if you have one and want to dispose of it.
I also have an EDAX 9100 with detector and LSI-11 console that I do not need. suggestions on its disposition are welcome.
Slightly off subject, but I am sure that someone has encountered this obstacle. We have ASCII data files on 8" floppies from various sort of data acquisision hardware/software and are seeking out a way to transfer the data to a useable media 3.5" would suffice, a 8" drive on a system w/internet access would be great, etc. etc.
Does anyone know of someone I could obtain assistance from?
__ _-==-=_,-. /--`' \_-at---at-.-- { Tim (TJ) LaFave Jr. `--'\ \ {___/. Department of Physics \ \\ " / University of North Carolina, Charlotte } =\\_/` { Charlotte, NC 28223 ____ /= | \_|/ _' `\ _/=== \___/ (704)547-3244 `___/ //\./=/~\====\ (704)509-6622 [Hm] \ // / | ===: http://www.iit.edu/~lafatim | ._/_,__|_ ==: __ \/ \\ \\`--| / \\ ---------- +*+ ---------- | _ \\: /==:-\ `.__' `-____/ |--|==: Such that the future be theirs \ \ ===\ :==:`-' to shape and direct. _} \ ===\ /==/ -----------------------------------------------------------------
Hi there there seems to be 2 versions of this book, one was pub in 1988 and one in 1995, what is the diff in them besides the ISBN number?
I have the older one I just got it today and it is a lot of fun was wondering what the newer one might offer..... The one I have has all black and white pictures in it. thanks Ed Sharpe
We are interested in quantifying the amount of sulphur in wool via EDXS analysis in the TEM.
Wool has approximately 5% sulphur within the fibre which varies for different components of the fibre. We would like to be able to mount the sample and standard in the same block and microtome them together. It is not necessary for the material to be fibrous but that would help!
To date we have tried a couple of methods without much success.
In the first instance we attempted to mix sulphur compounds in with the resin to obtain a homogenous material but found that no matter what we did there was always significant variability in the amount of sulphur throughout the resin.
The next attempt used a commercially available fibre which had a known amount of sulphur in it (0.25%) which we embedded in the resin and microtomed. This "sort of" worked OK but the level of sulphur was not quite high enough for our purpose and the uncertainties in our measurements were too high.
So, if anyone has any ideas or a source of materials we could use, I (and the rest of the team) would be most grateful.
Thanks very much in anticipation.
Colin Veitch
Instrumentation Scientist Fibre Structure & Function Group CSIRO Wool Technology PO Box 21 BELMONT Vic 3216 Australia
Tel: +61 (0)3 5246 4000 Fax: +61 (0)3 5246 4811
The information contained in this email 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 Wool Technology on +61 3 5246 4000.
If anyone is disposing of this Amray scope, I would appreciate knowing about it so that I could purchase some spare parts and assemblies from it before it is dumped. I am particularly interested in the main 30KV power supply and the CRT power supply, photomult assembly, and any of the circuit boards. (Specific model is 1600T without the digital controls and pushbuttons. All components and assemblies are of interest.)
I am also looking for a Robinson detector for this scope. Pls let me know if you have one and want to dispose of it.
I also have an EDAX 9100 with detector and LSI-11 console that I do not need. suggestions on its disposition are welcome.
I need information about SOPHIA process for aluminium castings ?
Best regards
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager of Structural and Mechanical Research Laboratory str. Zakopianska 73 Call (*48 12) 2618356 (after 8th march) 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
Hi Lesley S. Bechtold, =20 I have read with interest your contribution from 1998-11-27 where you=20 described your treatment of mouse sperm prior to SEM. I want to=20 prepare thrombocytes in a similar way and wonder whether you always=20 apply 1% solutions of poly-L-lysine to render coverslips adhesive. As=20 I understand 0.01% solutions are suitable (and much less expensive...)= =20 for other applications. I deal with moderate amounts of "precious"=20 thrombocytes from knockout mice so I can not play around with=20 conditions. You certainly have extensive experience with this.=20 Any comments would be highly appreciated! =20 All the best, Matthias
I think DPK is a misprint for DPX which is a mounting medium consisting of polystyrene and plasticizers dissolved in xylene and should be widely available from most microscopy suppliers
Dan Hill Biochemistry Department Cambridge University Cambridge UK
On Tue, 2 Mar 1999, Peggy Sherwood wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi! } I have a reference to two stains/methods for nerves: Koelle's acetyl } cholinesterase technique and } Winklemann's Silver Impregnation method. I have not been able to find } either one. I found a "general" reference to acetyl cholinesterase from } the University of Nottingham web page in which they } recommend using 'DPK' mounting resin. I have never heard of it--can } anyone help me out? Thanks! } } Peggy Sherwood } Wellman Labs of Photomedicine (224) } Massachusetts General Hospital } 50 Blossom Street } Boston, MA 02114 } 617-724-4839 (voice mail) } 617-726-3192 (fax) } } } }
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There are 3-4 sulfur standards for the carbon black industry, ASTM D-1619. The concentrations I know about are 0.82%, 1.54%, and 1.93%. These are carbon black powders so combining them in block with your fibers would be a real challenge. I do know a source for these standards if you want to try them.
Chuck Butterick Engineered Carbons, Inc. ______________________________ Reply Separator _________________________________
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Hi All,
We are interested in quantifying the amount of sulphur in wool via EDXS analysis in the TEM.
Wool has approximately 5% sulphur within the fibre which varies for different components of the fibre. We would like to be able to mount the sample and standard in the same block and microtome them together. It is not necessary for the material to be fibrous but that would help!
To date we have tried a couple of methods without much success.
In the first instance we attempted to mix sulphur compounds in with the resin to obtain a homogenous material but found that no matter what we did there was always significant variability in the amount of sulphur throughout the resin.
The next attempt used a commercially available fibre which had a known amount of sulphur in it (0.25%) which we embedded in the resin and microtomed. This "sort of" worked OK but the level of sulphur was not quite high enough for our purpose and the uncertainties in our measurements were too high.
So, if anyone has any ideas or a source of materials we could use, I (and the rest of the team) would be most grateful.
Thanks very much in anticipation.
Colin Veitch
Instrumentation Scientist Fibre Structure & Function Group CSIRO Wool Technology PO Box 21 BELMONT Vic 3216 Australia
Tel: +61 (0)3 5246 4000 Fax: +61 (0)3 5246 4811
The information contained in this email 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 Wool Technology on +61 3 5246 4000.
by smtp.uky.edu (8.8.8/8.8.8) with ESMTP id JAA09170 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 Mar 1999 09:50:05 -0500 (EST) Received: from nc.gws.uky.edu ([128.163.193.169]) by pop.uky.edu (8.8.8/8.8.8) with SMTP id JAA13068 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 3 Mar 1999 09:50:05 -0500 (EST) Message-Id: {2.2.32.19990303145459.006bcec0-at-pop.uky.edu} X-Sender: jmcclin-at-pop.uky.edu X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Woody,
We have a JEOL 35CF for sale that was made in '82. Excellent condition and has backskatter detector. Dual supply Haskris water chiller also available that cooled a TEM as well as this SEM.
We have a stock of spare parts, vacuum tubes, manuals, tools, etc., for an old RCA EMU-3G microscope. Since the scope itself is long gone, these items are destined for that mysterious place where old scope parts go to die. UNLESS someone wants them.
Preserve history! Keep your EMU going! Read the manuals! Now's your chance. All yours for the mere cost of shipping.
Randy Tindall Electron Microscope Core College of Veterinary Medicine University of Missouri - Columbia Phone: 573-882-8304
I made a makeshift but useable holder for filament adjustment for a Amray 1830. I took one of the cardboard boxes that hold the coaters for Polaroid film (about 9"X2"X3/4") and glued two metal washers(1" outer diameter and ~7/16 inner diameter) on top of one another and to the middle of the box. The height of the washers was about 1/8". I then positioned the posts of the filament in the center of the washers to mark the spots. Then I used a needle to poke holes in the cardboard. The Wehnhelt can then be upright and it is easier to center the filament.
Mike Baxter Lehman College mykkb-at-juno.com
___________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com/getjuno.html or call Juno at (800) 654-JUNO [654-5866]
If this is what I think it is it should be available from E Fjeld C., Inc. 978-667-1416. CAA-FB Filament Alignment Base is item discription in my 1985 catalog.
I am posting this on behalf of nearby hospital colleagues:
1. JEOL 1200EX TEM for sale, serviced twice yearly since installation in 1985. In excellent condition. Very light use of up to 65 cases per year.
2. Reichert (now Leica) Ultracut S ultramicrotome, new in 1993, with table, drawers plus LKB Knifemaker 7800. Hardly used and in storage over the last three years.
For information, telephone Mike Sale on England (0)1872 255006. Cash offers to Keith Atkinson on England (0)1872 255006. An e-mail address: tracey.leigh-at-rcht.swest.nhs.uk
Please do not "Reply to sender" - although I would pass replies on to the correct contact!
Keith Ryan Marine Biological Association of the UK Plymouth, UK
I have been asked to research the latest methods for colloidal gold labelling in biological tissues. The researchers would first like to do some diaminobenzidine peroxidase staining on kidney tubules embedded in paraffin to see if the cell membrane antigen and distribution can be detected at the LM-peroxidase level.
Then the project would progress to preparation, sectioning, and labelling of kidney tissue at the TEM level. I have been asked to provide a beginning point of up-to-date protocols that are presently being done.
I am currently scanning Medline and Ovid search engines. Any suggestions would be greatly appreciated.
Sincerely,
Ginger Baker EM Lab Manager OSU-College of Osteopathic Medicine 918-561-8232 lizard-at-osu-com.okstate.edu
I just wanted to say a wholehearted thank you to all of you on this listserver. With your suggestions I have managed to inject an antibody which was directly conjugated to ultrasmall gold particles (thanks to Dr. Jan Leunissen and Electron Microscopy Sciences), perfuse the mouse, and embed the tissues with the specific orientation that I wanted in airtight flat embedding molds to polymerize under UV (thanks to so many people for different aspects of the embedding that I don't have space to name them all), then cut, silver enhance and examine by TEM!
The antibody in question had failed to label anything when the tissue was fixed with ANY fixative but these results were unquestionable and the morphology great!!
My most sincere thanks to you all - I'm off to try new things as our little research lab is closing down. Thanks and Good-bye!
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-med.mcgill.ca
hi, Brefeldin A does not dissolve in media , you need first to dissolve it in either Methanol or DMSO. We dissolve the Brefeldin A in Methanol at a conc of 5mgs/ml and use this as our stock to then be diluted into our buffer or media.
I am currently looking to upgrade my EDS system. I work in a semiconductor manufacturing facility(CMOS) and I currently have a Cambridge S200 SEM with Kevex Delta II.
I have an interest in Oxford, EDAX and a company called EVEX (which I am not familiar with)? Does anyone have experience with these companies - Good or Bad?
I am looking for a light element detector with possibly a WDS for Boron quantification.
Thanks in advance.
Jim Arnold Microelectronics and Technology Center AlliedSignal Electronics and Avionics Systems 9140 Old Annapolis Road Columbia, MD 21045
I have a requirement to process liquid and agar-bound pathogenic bacteria for SEM & TEM. I've been collecting them in cacodylate-buffered 3% glutaraldehyde where the final concentration has been 1.5% for the liquid suspended bacteria. These have been kept in the refrigerator. Does anyone have a protocol(s) that would lead me to SEM and TEM specimens from this point? Have I made a mistake already? My instruments are a Philips EM-400 and JEOL 6300V.
We have an immediate opening for a field service technician in the Los Angeles area. Responsibilities include the repair and maintenance of Scanning Electron Microscopes of a number of manufacturers including JEOL, Hitachi, Leo.
Candidate should have a minimum of three years electronic or instrumentation experience. Please e-mail or fax me at:
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Hello,
I have been asked to research the latest methods for colloidal gold labelling in biological tissues. The researchers would first like to do some diaminobenzidine peroxidase staining on kidney tubules embedded in paraffin to see if the cell membrane antigen and distribution can be detected at the LM-peroxidase level.
Then the project would progress to preparation, sectioning, and labelling of kidney tissue at the TEM level. I have been asked to provide a beginning point of up-to-date protocols that are presently being done.
I am currently scanning Medline and Ovid search engines. Any suggestions would be greatly appreciated.
Sincerely,
Ginger Baker EM Lab Manager OSU-College of Osteopathic Medicine 918-561-8232 lizard-at-osu-com.okstate.edu
We bought some of those chain meshed type gloves that protect fingers pretty well when hand grinding. Our metallographers say you can easily hold the samples and mounts. They look and feel rather clumsy to me tho. But if they work....alrighty then.
There are two special issues of journals on immunogold labeling which appeared recently. Cell Vision, Vol. 4, No. 6 (1997), and Microscopy Research and Technique, Vol. 42, No. 1 (1998). If you have trouble finding a hard copy of that issue of Cell Vision (I think it was from before it was indexed by Medline), you can download all the articles from the journal's web site - if you go to
this is the table of contents for this issue, and it links to the full text articles in pdf format.
Disclaimer: we supply immunogold reagents, and both these special issues contain several articles which feature results obtained with our products (Nanogold).
A good recent reference on post-embedding immunogold labeling is "Colloidal Gold Post-Embedding Immunocytochemistry" (Progress in Histochemistry and Cytochemistry, Vol 29, No 4) by Moise Bendayan (Gustav Fischer, 1995).
Hope this helps,
Rick Powell Nanoprobes, Incorporated.
} } Hello, } } I have been asked to research the latest methods for colloidal gold } labelling in biological tissues. The researchers would first like to do } some diaminobenzidine peroxidase staining on kidney tubules embedded in } paraffin to see if the cell membrane antigen and distribution can be } detected at the LM-peroxidase level. } } Then the project would progress to preparation, sectioning, and labelling } of kidney tissue at the TEM level. I have been asked to provide a } beginning point of up-to-date protocols that are presently being done. } } I am currently scanning Medline and Ovid search engines. Any suggestions } would be greatly appreciated. } } Sincerely, } } Ginger Baker } EM Lab Manager } OSU-College of Osteopathic Medicine } 918-561-8232 } lizard-at-osu-com.okstate.edu
****************************************************************** * NANOPROBES, Incorporated | US Toll-free: (877) 808-2101 * * 25 East Loop Road, Suite 113 | Tel: (516) 444-8815 * * Stony Brook, NY 11790-3350, | Fax: (516) 444-8816 * * USA | rpowell-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
Hello Jim., Since you mentioned the word evex, I suspect you missed the past discussions on this list server regarding them. It centered around their registering URLs with the trade names of all competitors so that any web search by manufacture's name produced the evex web page. Reports were that they were willing to sell the URLs to the principles for 10s of K$, perhaps more. Seems they (someone) did try a defense with an unsigned response. Personally wouldn't have confidence in any claim they made. You can probably find these discussion in the list server archives, ~a year ago.
Bruce Brinson Rice U.
Disclaimer: I use EDS & WDS on occasion. I have absolutely no financial interest & know no one with financial interest in the sales of equipment in this industry & no qualms about bringing this issue back to light.
Arnold, Jim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am currently looking to upgrade my EDS system. I work in a semiconductor } manufacturing facility(CMOS) and I currently have a Cambridge S200 SEM with } Kevex Delta II. } } I have an interest in Oxford, EDAX and a company called EVEX (which I am not } familiar with)? Does anyone have experience with these companies - Good or } Bad? } } I am looking for a light element detector with possibly a WDS for Boron } quantification. } } Thanks in advance. } } Jim Arnold } Microelectronics and Technology Center } AlliedSignal Electronics and Avionics Systems } 9140 Old Annapolis Road } Columbia, MD 21045 } } email: Jim.arnold-at-alliedsignal.com } voice: (410) 964-4118 } fax: (410) 964-5046
I have to prepare pure Al foils for TEM investigation. What I need is (as usual :) a large thin area. What are the best electrolytes for the thinning of Al? Is there anything else to consider (e.g. temperature, ...)? As you see, I have not a lot of experience with this technique. Could anybody recommend a book concerning electrolytical thinning?
TIA,
Petra -------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu Visit our WWW site! http://www.crpcu.lu/~wahlbrin
Don't forget our annual meeting in Gainesville Florida April 8-10
Dealines are fast approaching
For details see the latest issue of the Beam or our web site at :
http://www.biotech.ufl.edu/cbr/sems/sems.html
or cal or fax me below.
Greg Erdos Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
Hi, All We are looking for a Wenhelt Block Filament for filament ( SEM Philips 505 or 501), If you got one we're interested in it ( Free, or for sale , reasonable price)) , please contact us.
Thanks
***************************************************************** Mohamed Belhaj UFR SCIENCES Laboratoir d'Analyse des Solides Surfaces et Interfaces DTI/LASSI EP CNRS 120 BP 1039 Reims 51687 Cedex 2 Tel : 03 26 05 33 27 ou 03 26 05 33 14 Fax : 03 26 05 33 12 ******************************************************************
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Hi,
I have a usual question, which was posted before and, I believe, will be posted again and again. It is about printers for SEM images. So:
- What is the best printer for monochrome SEM/STEM images? - What is the best inexpensive printer for the same images? - What is the best printer for EDS output (color maps, maps+images)?
Thank you,
J.Meen
____________________________________________________________________ Get free e-mail and a permanent address at http://www.netaddress.com/?N=3D= 1
======================================================== FOCUS ON MICROSCOPY 1999
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
We wish to remind you about the upcoming deadline for abstract submission and modification.
The abstract submission and modification deadline is:
!!! Monday, 8 March 1999 --- 15.00 hours Heidelberg Time !!!
The "abstract submission part" of the database will definitely be closed at 15.00 hours, so that it is not possible to submit further abstracts or make any further modifications to the abstracts after this date.
The "registration only part" of the database will remain open until 9 April 1999!!!
Our website now also includes informations about some highlights of the commercial exhibition. This is an extract:
Leica Microsystems GmbH: Two-Photon Confocal System Leica TCS MP L.O.T. Oriel GmbH & Co KG: Nanopositioning Unit and Fiber Laser Molecular Probes: New fluorescent reagents for Cell Biology and Imaging highly - Fluorescent Alexa dyes Olympus Optical Co. GmbH: Confocal Laser-Scanning-Microscope with two-photon excitation Omicron Vakuumphysik GmbH: Scanning Near Field Optical Microscope (SNOM) Visitron Systems GmbH: 2D/3D Fluorescence Imaging System based on Cooled Digital Camera System Wallac Distribution GmbH: Confocal Microscope
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meeting point for developers and users working in these rapidly evolving fields and play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic developments in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
I have never done any material sciences work - I've only ever done biological specimens for EM. Our engineering department has asked me to look at some ball bearings that are failing, as a favour. I know I don't need to fix or dehydrate but do I simply clean them, mount them (using double-sided tape?) and coat them as usual? Or is coating unnecessary? What whould I clean these with? I'm assuming there is grease somewhere that is not good for my vacuum!
Any help would be appreciated!! Thanks in advance.....
Lesley Bechtold
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
Ginger R Baker {lizard-at-osu-com.okstate.edu} CC: Bruce A Benjamin {benjamb-at-osu-com.okstate.edu} , Connie A Hebert {hconnie-at-osu-com.okstate.edu} , William D Meek {meekwd-at-osu-com.okstate.edu} X-Mailer: QuickMail Pro 1.5.3 (Mac) X-Priority: 3 MIME-Version: 1.0 Reply-To: Paul Webster {pwebster-at-mailhouse.hei.org} Content-Type: multipart/alternative; boundary="====50515355485450534849===1"
Reply to: RE: LM and TEM Need help locating the latest = Colloidal Gold T Dear Ginger,
The methods you are looking for can be slit into two sections; (i) = specimen preparation and (ii) colloidal gold labeling. For both, you have = many choices that will be determined by your lab equipment, lab skills = and budget. A good book to have next to you is "Fine structure = Immunocytochemistry" by G. Griffiths (published 1993 by Springer Verlag) = because it will give you advice on all your choices. First, specimen preparation: Your aim is to prepare samples that have = good morphology but also have good accessibility to antigen by the = antibodies and colloidal gold. The best way to achieve this is to obtain = sections. To section, you can either embed in resin (Lowicryl, LR White/= Gold, Unicryl, Monostep), or use cryosections. If you decide to use resin,= will you embed using the PLT method or by freeze substitution? Whatever = your choice, if you are to test the system first by light microscopy then = I would highly recommend using the same system for LM and EM. Do not try = to compare paraffin-embedded, HRP-DAB labeled material with aldehyde-fixed,= sectioned material. =
The gold labeling is also filled with many choices. It is clear that = sequential labeling (ie adding the primary antibody followed by a gold-= labeled secondary) is the best choice. However, what you choose as the = gold-labeled secondary may affect your labeleing pattern. If a = quantitative result is your aim (do you want a signal that reflects the = amount of antigen present?) then you must try not to use signal = amplification. For this either protein A-gold or immunoglobulin-gold as a = single secondary step is best. If you want to get a "strong" signal and = are not too worried about quantitative result you may want to try silver = enhancement of the gold marker.
If I were doing this study, I would fix the kidney, by perfusion, in 4% = formaldehyde (buffered in phosphate buffer). I would then cryoprotect = with sucrose, freeze in liquid nitrogen and embed part of it in Lowicryl = resin through freeze substitution. Other pieces of the cryoprotected, = frozen tissue, I would then section in a cryoultramicrotome. Both the = Lowicryl and cryo sections can be mounted onto glass substrate and labeled = with antibody and fluorescent secondary antibody. You can also label the = sections with the gold conjugated secondary you choose to use for EM and = visualize this for LM by silver enhancement. The advantage of using the = same reagents and tissues for LM is that you can easily test your system = before moving onto EM preparation. Once you have the conditions worked = out, the same blocks you used for LM can be sectioned again for EM (just = mount the sections on grids) and labeled with the same (or similar) = protocols you used for LM.
The protocols you need are all covered in the Griffiths book. Some of the = newer advances have been covered in the a book that had the contents = posted recently.
Contact me if you want more details, resources or references.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm --====50515355485450534849===1 Content-Type: text/html; charset="US-Ascii" Content-Transfer-Encoding: quoted-printable
{HTML} {HEAD} {/HEAD} {BODY} {PRE WIDTH=3D"132"} Reply to: RE: LM and TEM Need help locating the latest = Colloidal Gold T
{/PRE} {FONT FACE=3D"Geneva" SIZE=3D3 = COLOR=3D"#000000"} Dear Ginger, {BR} {BR} The methods you = are looking for can be slit into two sections; = (i) specimen preparation and (ii) colloidal = gold labeling. For both, you have many = choices that will be determined by your = lab equipment, lab skills and budget. A = good book to have next to you is "Fine = structure Immunocytochemistry" by G. = Griffiths (published 1993 by Springer Verlag) = because it will give you advice on all your = choices. {BR} First, specimen preparation: = Your aim is to prepare samples that have = good morphology but also have good accessibility = to antigen by the antibodies and colloidal = gold. The best way to achieve this is to = obtain sections. To section, you can either = embed in resin (Lowicryl, LR White/Gold, = Unicryl, Monostep), or use cryosections. = If you decide to use resin, will you embed = using the PLT method or by freeze substitution? = Whatever your choice, if you are to test = the system first by light microscopy then = I would highly recommend using the same = system for LM and EM. Do not try to compare = paraffin-embedded, HRP-DAB labeled material = with aldehyde-fixed, sectioned material. = {BR} {BR} The gold labeling is also filled = with many choices. It is clear that sequential = labeling (ie adding the primary antibody = followed by a gold-labeled secondary) is = the best choice. However, what you choose = as the gold-labeled secondary may affect = your labeleing pattern. If a quantitative = result is your aim (do you want a signal = that reflects the amount of antigen present?) = then you must try not to use signal amplification. = For this either protein A-gold or immunoglobulin-gold = as a single secondary step is best. If = you want to get a "strong" signal = and are not too worried about quantitative = result you may want to try silver enhancement = of the gold marker. {BR} {BR} If I were doing = this study, I would fix the kidney, by perfusion, = in 4% formaldehyde (buffered in phosphate = buffer). I would then cryoprotect with = sucrose, freeze in liquid nitrogen and embed = part of it in Lowicryl resin through freeze = substitution. Other pieces of the cryoprotected, = frozen tissue, I would then section in a = cryoultramicrotome. Both the Lowicryl and = cryo sections can be mounted onto glass = substrate and labeled with antibody and fluorescent = secondary antibody. You can also label = the sections with the gold conjugated secondary = you choose to use for EM and visualize this = for LM by silver enhancement. The advantage = of using the same reagents and tissues for = LM is that you can easily test your system = before moving onto EM preparation. Once = you have the conditions worked out, the = same blocks you used for LM can be sectioned = again for EM (just mount the sections on = grids) and labeled with the same (or similar) = protocols you used for LM. {BR} {BR} The protocols = you need are all covered in the Griffiths = book. Some of the newer advances have been = covered in the a book that had the contents = posted recently. {BR} {BR} Contact me if you = want more details, resources or references. {/FONT} {FONT = FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#000000"} {BR} {BR} Paul Webster, Ph.D {BR} House = Ear Institute {BR} 2100 West Third Street {BR} Los = Angeles, CA 90057 {BR} phone:213 273 8026 {BR} fax: = 213 413 6739 {BR} e-mail: pwebster-at-hei.org {BR} http://www.hei.org/htm/aemi.htm {/FONT} {/BODY} {/HTML} --====50515355485450534849===1--
Has anyone had any experience with these knives and would you like to comment on their performance relative to "standard" diamond knives?
Tom
Thomas Moninger moninger-at-emiris.iaf.uiowa.edu University of Iowa Central Microscopy Research Facility http://www.uiowa.edu/~cemrf Views expressed are mine.
On Wed, 3 Mar 1999 14:32:22 -0700, jwright-at-dugway-emh3.army.mil wrote... } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
For the TEM samples, fixation in small 1.5 mL eppendorf microfuge tubes works well since you can pellet the samples between solution changes if necessary. From the point you are at right now, you should rinse in your cacodylate-buffer (I used 0.1M cacodylate with 7.5% sucrose), then move on to post-fixation with 1% OsO4 in 0.1M cacodylate (no sucrose) until pellets turn dark brown or black (usually 30-min at room temp). If the pellets are really big, separate them before osmium treatment to make sure the entire pellet is fixed. Rinse in cacodylate buffer (no sucrose), dehydrate in an acetone or ethanol series. I use ethanol because I like to embed in LR White. After 2 changes in 100%, separate the SEM run (for critical point drying and coating) from the TEM samples- move on to resin infiltration right in the eppendorf tubes. I usually do 3:1 (solvent:resin) on a rotator for 1 hr., 1:1, 1:3, 100% times two and cure overnight. Good luck.
As promised, here is a summary of the replies I received to my enquiry about who uses the MSA (formerly EMSA) spectra file format.
Of eight replies, two didn't care for the format or never used it. "My personal preference is not to use MSA format for these purposes. It's way overcomplicated".
The rest of the replies were either reluctant or enthusiastic users who use the format to transfer spectra from one EDX analyser to another, or to the DTSA program, or into a desktop computer. Three would be just as happy to have any method to convert the spectra into Excel format, but three were quite enthusiastic about the ability to exchange spectra between different labs, different EDX programs on commercial systems, even different analysers from the same company of differnet versions.
Thanks to all who replied. I can send the text of all replies to anyone who wishes to see them.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
The best quality will probably always be from a dye sublimation printer. There are Kodak and Codonics machines at the high end for full page output at a cost of about $10K. Kodak also makes a small format (4x5) printer that does 3-color for about $600 and under $1/print. I don't know that they yet offer a black ribbon for true monochrome prints.
For general purpose work, most any of the inkjets will do decent work for both color and monochrome. You may just want to try some at your local computer stores to see what resolution you can get and how fast. We use an Epson 600 ( {$200) for pretty good work at maybe 2 minutes per full page print.
1200 dpi laser printers do a pretty good job for monochrome. We have a Lexmark Optra R. I have seen similar good results from an HP LaserJet 5. It takes a while to transmit the data to the printer, but then they whip out prints fast.
In all cases, the paper can do a lot to improve appearance. You will want some good stuff on had for the best prints.
None of what I have said is peculiar to EM work. You may want to enlist a few others in your area to get a good printer. Happy hunting.
Warren S.
At 09:13 AM 3/4/99 -0600, you wrote: } Hi, } } I have a usual question, which was posted before and, I believe, will be } posted again and again. It is about printers for SEM images. So: } } - What is the best printer for monochrome SEM/STEM images? } - What is the best inexpensive printer for the same images? } - What is the best printer for EDS output (color maps, maps+images)? } } Thank you, } } J.Meen
I am more of a TEM eprson, but I have a suggestion from watching my colleagues. Take an aluminium stub, which would normally go into an SEM, and using a 5 mm drill bit just drill a small , 1-2 mm deep, hole in top so that the ball bearing will sit in it. Before you mount the bearing, wash the bearing in organic solvents first, say two acetone washes and then two ethanol washes, and ultrasonically clean the bearing in an ultrasound bath with both solvents.
Dry it off by putting it into an oven at say 150 C for ten minutes and then mount it onto the stub using silver-dag mounting paint (used by almost everyone here) to fix the bearing onto the stub. If you have access to vacuum coating system with a Radio Frequency inductive plasma ring attachment or anything that can produce an Argon plasma, then put the stub in for ten minutes so that there are no organic residues left. Once it is finished you can put it straight into the SEM knowing that there should be a clean surface to look at. The silver dag paint should earth the bearing to the stub.
I hope this helps.
Jon
-- ***************************************** Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
Yes, that's right - if you are somewhere between Baltimore, Maryland and Sarasota, Florida (with a detour to visit my son in Chapel Hill, NC) I can deliver this lovely item to you in early April. I still don't want to crate it for shipment, but I am driving down to visit my mother and can drop it off on the way if it is not too far off my path. The item in question is an LKB-Huxley Ultra Microtome (model 9801-1) in nice clean condition with the knife holder and a set of chucks and collets. I have about $500 tied up in it which I would like to recover - if you can use it, get in touch and perhaps we can work something out. It just does not fit the work I am doing now, and I need both the $$ and space.
Dear Petra, We have had good success with Al alloys with the Struers Tenupol Twin-jet polisher, using their A-7 electrolyte at about -5 degrees C and 12V. The A-7 is: 100 ml. perchloric acid 700 ml. methanol 200 ml. gycerine The usual precautions with perchloric acid solutions apply. Another I have had recommended to me is: 100 ml. perchloric acid 900 ml. ethanol The solution must be kept at less than 10 degrees C. while polishing, and a voltage range of 7.5V to 20 V, depending on the alloy. I have no experience with pure Al, only commercial Al alloys. A book I recommend for these things is: "Metallography Principles and Practices" by Vander Voort, but I don't know if it is still in print.
You wrote: } } Hello Everyone, } } I have to prepare pure Al foils for TEM investigation. What I need is (as } usual :) a large thin area. What are the best electrolytes for the thinning } of Al? Is there anything else to consider (e.g. temperature, ...)? } As you see, I have not a lot of experience with this technique. Could } anybody recommend a book concerning electrolytical thinning? } } TIA, } } Petra } -------------------------------------------------------------- } Dr. Petra Wahlbring Best of luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
We've been very pleased with the Epson Color Stylus 740 (approx. $280) and Epson Stylus Photo 700 (approx. $250) using Epson's Photo Quality Glossy Film. Image and text quality is very good to excellent.
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center
Research Scientist, Chemistry Williams College
On Thu, 4 Mar 1999, Warren E Straszheim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } The best quality will probably always be from a dye sublimation printer. } There are Kodak and Codonics machines at the high end for full page output } at a cost of about $10K. Kodak also makes a small format (4x5) printer that } does 3-color for about $600 and under $1/print. I don't know that they yet } offer a black ribbon for true monochrome prints. } } For general purpose work, most any of the inkjets will do decent work for } both color and monochrome. You may just want to try some at your local } computer stores to see what resolution you can get and how fast. We use an } Epson 600 ( {$200) for pretty good work at maybe 2 minutes per full page print. } } 1200 dpi laser printers do a pretty good job for monochrome. We have a } Lexmark Optra R. I have seen similar good results from an HP LaserJet 5. It } takes a while to transmit the data to the printer, but then they whip out } prints fast. } } In all cases, the paper can do a lot to improve appearance. You will want } some good stuff on had for the best prints. } } None of what I have said is peculiar to EM work. You may want to enlist a } few others in your area to get a good printer. Happy hunting. } } Warren S. } } At 09:13 AM 3/4/99 -0600, you wrote: } } Hi, } } } } I have a usual question, which was posted before and, I believe, will be } } posted again and again. It is about printers for SEM images. So: } } } } - What is the best printer for monochrome SEM/STEM images? } } - What is the best inexpensive printer for the same images? } } - What is the best printer for EDS output (color maps, maps+images)? } } } } Thank you, } } } } J.Meen } } }
Depending on the suspected failure mechanism, just give them a good soaking in acetone or some kind of degreasing detergent (local auto store) followed by a good rinse and dry. That should remove any grease. I would however talk to the Eng. folks to make sure that your cleaning would not disturb the failure mecahnism or worse.... enhance it. Yes, I would give them a light coat of Au or AuPd. Mounting on any kind of tape (carbon or otherwise) might present some drifting problems in the SEM.......perhaps Ag or C paint would do the trick....that's our solution around here anyway. Hope this helps.
John Staman LSI Logic, Process Analysis Lab, Colorado Springs, CO. 719-573-3282
} -----Original Message----- } From: Lesley S. Bechtold [SMTP:lsb-at-aretha.jax.org] } Sent: Thursday, March 04, 1999 8:53 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Biologist needs help from Material Scientists!! } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } I have never done any material sciences work - I've only ever done } biological specimens for EM. Our engineering department has asked me to } look at some ball bearings that are failing, as a favour. I know I don't } need to fix or dehydrate but do I simply clean them, mount them (using } double-sided tape?) and coat them as usual? Or is coating unnecessary? } What whould I clean these with? I'm assuming there is grease somewhere } that is not good for my vacuum! } } Any help would be appreciated!! Thanks in advance..... } } Lesley Bechtold } } } Lesley S. Bechtold } Supervisor, Biological Imaging } The Jackson Laboratory } 600 Main St. } Bar Harbor, ME 04609 } 207-288-6191 }
Assuming that you want to look at the bearings in an SEM and that the balls are made of steel, the solution is very simple:
Clean the balls to get rid of any grease. A few years back we used Trichlorethylene to degrease materials, but that is environmentally unsafe. There are other possibilities for safer degreasing.
Then you could just use a simple specimen holder and glue the balls to the holder with silver paint or carbon paint, mainly to fix them in place. I am not sure what you mean by double-sided sticky tape, but I would not use that. Who knows what the tape might do when the chamber is pumped down. Since they are steel, you don't have to worry about conductivity, just put them in the chamber, pump down and enjoy.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
} ---------- } From: Lesley S. Bechtold[SMTP:lsb-at-aretha.jax.org] } Sent: Thursday, March 04, 1999 8:53 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Biologist needs help from Material Scientists!! } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Hi, } } I have never done any material sciences work - I've only ever } done } biological specimens for EM. Our engineering department has asked me } to } look at some ball bearings that are failing, as a favour. I know I } don't } need to fix or dehydrate but do I simply clean them, mount them (using } double-sided tape?) and coat them as usual? Or is coating } unnecessary? } What whould I clean these with? I'm assuming there is grease } somewhere } that is not good for my vacuum! } } Any help would be appreciated!! Thanks in advance..... } } Lesley Bechtold } } } Lesley S. Bechtold } Supervisor, Biological Imaging } The Jackson Laboratory } 600 Main St. } Bar Harbor, ME 04609 } 207-288-6191 } }
I need a razor blade holder to use in an old A/O CryoCut vintage 1978. It's a weird one, the blade is mounted to the far right of the holder to accommodate the knife mounting mechanism of the cryostat.
I could also use a razor blade holder for use in an old A/O rotary microtome. This is pretty standard, with the blade in the center of the holder.
I think they are available new, but are pretty expensive given the scope of my project. Anything old but serviceable would work for me. If you have something, maybe we can workout a deal.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I have a few technical papers that deal with electropolishing aluminum using our Model 550 Single Vertical Jet Electropolisher which may be of interest:
1)" Characterization of the Microstructure, Fracture, Morphology and Toughness in Particulate and Short Fibre Reinforced Aluminum Matrix Composites", M.R. Krishnadev et al
2) "Preparation of Iron and Aluminum Samples for Ion Implantation and TEM=
Examination" J.M. McDonald
and the "Bible" of Jet Polishing:
3) "Polishing Methods for Metallic and Ceramic Transmission Electron Microscopy Specimens", B.J. Kestel
Kestel report is 60+ pages and give preparation guidelines for dozens of materials. He has recipes for pure aluminum as well as several aluminum alloys. I would recommend this report for anyone doing electropolishing.=
I have copies of all 3 of these reports as well as a bibliography of over=
200 technical reports mostly dealing with specimen preparation. I would = be pleased to mail you a copy of any or all of these if you think they will = be of use. Please let me know.
DISCLAIMER: South Bay Technology does manufacture the Model 550 Single Vertical Jet ELectropolisher and has a vested interest in promoting its use.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy. Message text written by Petra Wahlbring } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Hello Everyone,
I have to prepare pure Al foils for TEM investigation. What I need is (as=
usual :) a large thin area. What are the best electrolytes for the thinni= ng of Al? Is there anything else to consider (e.g. temperature, ...)? As you see, I have not a lot of experience with this technique. Could anybody recommend a book concerning electrolytical thinning?
J. Meen asks ... } } } ... } } - What is the best printer for monochrome SEM/STEM images?
A dye-sublimation printer with its optional gray scale media.
} - What is the best inexpensive printer for the same images?
A good 600dpi laser printer configured for random dithering, although the next printer would work well too. I would opt for $$ being spent up front on the printer ... it will still cost pennies per page.
} - What is the best printer for EDS output (color maps, maps+images)?
A photographic quality ink jet would work well here ... inexpensive and could as well serve your second purpose, 'cept for speed.
If you're concerned with archiving, fading can be a problem with color ink jets ... moreso than with dye-subs ... so the dye-sub can serve dual purposes, gray scale and color. But, it is a hassle to constantly be changing its purposes from monochrome to color. The dye-sub's color media can be used to print gray scale, but it almost always has a tint because CMY is used, therefore I would not classify a color mode dye-sub as the best printer for your first need.
my $0.02 and cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Lesley writes ... } } } ... Our engineering department has asked me to } look at some ball bearings that are failing, as a favour. I } know I don't } need to fix or dehydrate but do I simply clean them, mount them (using } double-sided tape?) and coat them as usual? Or is coating } unnecessary? } What whould I clean these with? I'm assuming there is grease } somewhere } that is not good for my vacuum! } } ...
I would clean them first with acetone and then with clean ETOH and then dry to remove the small amount of absorbed water. They shouldn't need coating, but I'd mount them with conductive double-stick tape. Should be as easy as that :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I am using a Drukker Diamond Knive at 3 years. It is a excellent knive. I did perfect sections without compression. I cut plant tissues and pollen grains with a hard cell wall.
Rinaldo Pires dos Santos Dept of Botany - Plant Anatomy Laboratory - UFRGS Porto Alegre - RS - Brazil
A student of ours is going to the Washington, D.C. area this summer. I've already told her to stay away from the White House. What she really wants to know is: are any EM/Imaging facilities in the area? She would like to use one to help her with her work while she's there. So drop me a line and I'll pass the info on to her.
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Warren E Straszheim says: } The best quality will probably always be from a dye sublimation printer. } There are Kodak and Codonics machines at the high end for full page output } at a cost of about $10K. Kodak also makes a small format (4x5) printer that } does 3-color for about $600 and under $1/print. I don't know that they yet } offer a black ribbon for true monochrome prints.
1. Several other manufacturers (JVC, Sony, Panasonic) offer dye sublimation printers, marketed for catching/printing video frames. [I'm planning to buy one of those, probably Sony.]
2. I doubt that a term "ribbon" can be applied to such a printer...
} For general purpose work, most any of the inkjets will do decent work for } both color and monochrome. You may just want to try some at your local } computer stores to see what resolution you can get and how fast. We use an } Epson 600 ( {$200) for pretty good work at maybe 2 minutes per full page } print.
Interesting. For monochrome of course inkjets hardly cut it. But for color - it suddenly becomes cost-effective and attractive.
} 1200 dpi laser printers do a pretty good job for monochrome. We have a } Lexmark Optra R. I have seen similar good results from an HP LaserJet 5. It } takes a while to transmit the data to the printer, but then they whip out } prints fast.
1. I have yet to see a laser printer (within $2K price range) that comes close to Optra in reproducing photographs. LaserJet-5 is good, but not that good.
2. I wonder what kind of printer connection you use, and how large your files are. My printers are on Ethernet, and 16MB files "fly in" in less than half-a-minute.
} In all cases, the paper can do a lot to improve appearance. You will want } some good stuff on head for the best prints.
(:-) -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
Additional to the useful replies relating to the preparation of ball bearings, I would like to add a little regarding the microscopy: Small, clean and undamaged ball bearings are a good instructive aid to explain SEM functioning. This may be of some use to Jonathan when viewing those bearings and to anybody trying to explain some SEM effects. 1 At high kV (say above 20) you may have trouble seeing anything, because fine surface structure and dirt will be invisible. 2 At low kV (say 10 and below) such surface structures will be visible. 3 At low powers, regardless of kV, the ball bearing will appear like a disk, but the outer part of the disk is brighter. This nicely shows that in secondary mode, brightness almost entirely is increased with the angle of incidence. Being a sphere, tilt has no effect on the brightness distribution over that image. 4 A BS detector mounted at an angle (whereas the Robinson and some others are vertical) will make the distinction that the specimen is not a disk but a sphere, because the BS electrons directed away from the detector leave a shadow. 5 A similar effect is produced when the bias current of the secondary detector is turned off and the condenser current is turned down. This floods the secondary detector's scintillator with backscattered electrons and produces a BS image quite suitable for low powers.
Nice teaching exercise, but its useful to know about these effects when actually looking at those bearings. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Friday, March 05, 1999 6:11 AM, Jonathan Barnard [SMTP:J.Barnard-at-bristol.ac.uk] wrote: } } I am more of a TEM eprson, but I have a suggestion from } watching my } colleagues. } Take an aluminium stub, which would normally go into an } SEM, and using a } 5 mm drill bit just drill a small , 1-2 mm deep, hole in } top so that the } ball bearing will sit in it. Before you mount the bearing, } wash the } bearing in organic solvents first, say two acetone washes } and then two } ethanol washes, and ultrasonically clean the bearing in an } ultrasound } bath with both solvents. } } Dry it off by putting it into an oven at say 150 C for ten } minutes and } then mount it onto the stub using silver-dag mounting } paint (used by } almost everyone here) to fix the bearing onto the stub. If } you have } access to vacuum coating system with a Radio Frequency } inductive plasma } ring attachment or anything that can produce an Argon } plasma, then put } the stub in for ten minutes so that there are no organic } residues left. } Once it is finished you can put it straight into the SEM } knowing that } there should be a clean surface to look at. The silver dag } paint should } earth the bearing to the stub. } } I hope this helps. } } Jon } } -- } ***************************************** } Jonathan Barnard } } Microstructural Physics, } H.H.Wills Physics Laboratory, } University of Bristol, } Tyndall Avenue, } Bristol BS8 1TL. } } 0117 928 9000 ext 8750 } } ***************************************** } }
The string concerning preparation of bacteria touched on delayed second fixation. This is worth a separate discussion on delayed second (usually Os) fixation: Sabatini, first to publish GA as a fixative, also published that Os fixation could be delayed by several months. That seems true for some tissues, which show no ill-effects when compared with the usual, immediate double fixation. However, we found in the lab that other tissues are sensitive to that delay. I used to run a couple of busy service labs and cannot remember specifically which tissues and what structures were affected. It would be interesting to know when delayed double fixation is acceptable and others may have experience to share. I believe that specimen preparation for SEM is never affected by delayed second fixation. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Thursday, March 04, 1999 11:14 PM, Tina Schwach [SMTP:tschwach-at-tc.umn.edu] wrote: } } On Wed, 3 Mar 1999 14:32:22 -0700, } jwright-at-dugway-emh3.army.mil wrote... } } I have a requirement to process liquid and agar-bound } } pathogenic } } bacteria for SEM & TEM. I've been collecting them in } } cacodylate-buffered 3% glutaraldehyde where the final } } concentration has } } been 1.5% for the liquid suspended bacteria. These have } } been kept in } } the refrigerator. Does anyone have a protocol(s) that } } would lead me to } } SEM and TEM specimens from this point? Have I made a } } mistake already? My } } instruments are a Philips EM-400 and JEOL 6300V. } } } } John Wright } } Microbiologist } } } } West Desert Test Center } } Dugway, UT } } } } } } John, } I have stored samples in primary fixative (glut-para- } ruthenium red in } cacodylate) for several weeks, even months and they appear } to be fine. } For SEM, you may want to place (dry) your samples on some } kind of surface, } ie stainless steel chips, so you'll be able to view them. } You'll have to } do this at the end anyway. You can even place them on } nucleopore filter } membranes. Depending on what you want to see, the agar } strands can get in } the way. } } For the TEM samples, fixation in small 1.5 mL eppendorf } microfuge tubes } works well since you can pellet the samples between } solution changes if } necessary. From the point you are at right now, you } should rinse in your } cacodylate-buffer (I used 0.1M cacodylate with 7.5% } sucrose), then move on } to post-fixation with 1% OsO4 in 0.1M cacodylate (no } sucrose) until pellets } turn dark brown or black (usually 30-min at room temp). } If the pellets are } really big, separate them before osmium treatment to make } sure the entire } pellet is fixed. Rinse in cacodylate buffer (no sucrose), } dehydrate in an } acetone or ethanol series. I use ethanol because I like } to embed in LR } White. After 2 changes in 100%, separate the SEM run (for } critical point } drying and coating) from the TEM samples- move on to resin } infiltration } right in the eppendorf tubes. I usually do 3:1 } (solvent:resin) on a } rotator for 1 hr., 1:1, 1:3, 100% times two and cure } overnight. } Good luck. } } } } }
The peaks have suddenly started to move around a bit, although the resolution stays good. The problem comes and goes, in its good times the standard deviation of the position of the Co Ka peak is about 0.4 eV (10 determinations), but sometimes it's about 10 or even 20 eV. My thinking is that if it were the detector, the resolution would be degrading, but it's not, so maybe the culprit is the (analog) pulse-processor. Anyone got any thoughts on how to pin it down as being either a the detector b the subsequent signal-processing stuff, eg pulse-proc? Could I successfully test the pulse-proc with a ramp from a standard signal generator, or would that signal, being relatively clean compared with that from a detector, not really check it out rigorously enough?
cheers (well, I try)
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
======================================================== FOCUS ON MICROSCOPY 1999
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
We wish to remind you about the upcoming deadline for abstract submission and modification.
The abstract submission and modification deadline is:
!!! Monday, 8 March 1999 --- 15.00 hours Heidelberg Time !!!
The "abstract submission part" of the database will definitely be closed at 15.00 hours, so that it is not possible to submit further abstracts or make any further modifications to the abstracts after this date.
The "registration only part" of the database will remain open until 9 April 1999!!!
Our website now also includes informations about some highlights of the commercial exhibition. This is an extract:
Leica Microsystems GmbH: Two-Photon Confocal System Leica TCS MP L.O.T. Oriel GmbH & Co KG: Nanopositioning Unit and Fiber Laser Molecular Probes: New fluorescent reagents for Cell Biology and Imaging highly - Fluorescent Alexa dyes Olympus Optical Co. GmbH: Confocal Laser-Scanning-Microscope with two-photon excitation Omicron Vakuumphysik GmbH: Scanning Near Field Optical Microscope (SNOM) Visitron Systems GmbH: 2D/3D Fluorescence Imaging System based on Cooled Digital Camera System Wallac Distribution GmbH: Confocal Microscope
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meeting point for developers and users working in these rapidly evolving fields and play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic developments in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
Others have offered their suggestions but may I add a friendly warning, o= r offer a bit of fun?
Ball bearings are a great eye opener for the novice SEM operator. Fix a small ball bearing to a stub with a NON conducting adhesive. Run the microscope at 20 to 25kV for a few minutes and the ball will charge like=
mad. let it charge { one of the very few reasons for using such high kV := -) }. NOW drop to 2kV and wonder of wonders you will see the inside of the=
specimen chamber. Move the ball around and change the magnification and you will be able to view the chamber in detail. Take a look at the aperture of the final lens, the EDX detector face, or the backscattered detector surface. Great fun if you know what is happening, totally confusing if you did not do it deliberately.
This "problem" may occur in other circumstances where, after using high k= V and charging a specimen, moving to a low kV the charge does not go away b= ut simply acts as a reflector of the beam. Double sided tape which is non conducting is the biggest culprit. It should never be used in SEM where = we now have available carbon double sided media which is great for a quick f= ix with lighter weight specimens.
Have fun?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
As an assistance to people who have day to day problems with their equipment the listserver provides a very valuable service and I am often pointing people in this direction to solve their problems.
My area of knowledge is in the operation and maintenance of scanning and transmission electron microscopes so I take an interest in and discussion= s in this area.
Of late I have noticed a tendency for replies to become very complicated,=
so much so that the person asking the question would in my mind only beco= me bemused by the reply. Should we not build the knowledge base step by ste= p rather than leap in with solutions far distant from the actual question/problem?
As a made-up example -
Q - What do I do if I do not have a good quality image on the screen in m= y TEM?
A - Change the phosphor.
Have you seen answers like this?
I like many of you reading this I would not like to inhibit anyone sendin= g in a reply to a question, but should they not place their reply on a leve= l?
Example
A - Assuming that you have prepared the specimen correctly, have the correct alignment, kV and apertures, have the condenser lenses set correctly and are in a well darkened room and have become dark adapted - = if you notice your screen is not as bright as a colleagues TEM perhaps you should consider changing the screen as they fade due to beam damage and contamination?
I look forward to being put in my place! =
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The string concerning preparation of bacteria touched on delayed second fixation. This is worth a separate discussion on delayed second (usually Os) fixation: Sabatini, first to publish GA as a fixative, also published that Os fixation could be delayed by several months. That seems true for some tissues, which show no ill-effects when compared with the usual, immediate double fixation. However, we found in the lab that other tissues are sensitive to that delay. I used to run a couple of busy service labs and cannot remember specifically which tissues and what structures were affected. It would be interesting to know when delayed double fixation is acceptable and others may have experience to share. I believe that specimen preparation for SEM is never affected by delayed second fixation. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Thursday, March 04, 1999 11:14 PM, Tina Schwach [SMTP:tschwach-at-tc.umn.edu] wrote: } } On Wed, 3 Mar 1999 14:32:22 -0700, } jwright-at-dugway-emh3.army.mil wrote... } } I have a requirement to process liquid and agar-bound } } pathogenic } } bacteria for SEM & TEM. I've been collecting them in } } cacodylate-buffered 3% glutaraldehyde where the final } } concentration has } } been 1.5% for the liquid suspended bacteria. These have } } been kept in } } the refrigerator. Does anyone have a protocol(s) that } } would lead me to } } SEM and TEM specimens from this point? Have I made a } } mistake already? My } } instruments are a Philips EM-400 and JEOL 6300V. } } } } John Wright } } Microbiologist } } } } West Desert Test Center } } Dugway, UT } } } } } } John, } I have stored samples in primary fixative (glut-para- } ruthenium red in } cacodylate) for several weeks, even months and they appear } to be fine. } For SEM, you may want to place (dry) your samples on some } kind of surface, } ie stainless steel chips, so you'll be able to view them. } You'll have to } do this at the end anyway. You can even place them on } nucleopore filter } membranes. Depending on what you want to see, the agar } strands can get in } the way. } } For the TEM samples, fixation in small 1.5 mL eppendorf } microfuge tubes } works well since you can pellet the samples between } solution changes if } necessary. From the point you are at right now, you } should rinse in your } cacodylate-buffer (I used 0.1M cacodylate with 7.5% } sucrose), then move on } to post-fixation with 1% OsO4 in 0.1M cacodylate (no } sucrose) until pellets } turn dark brown or black (usually 30-min at room temp). } If the pellets are } really big, separate them before osmium treatment to make } sure the entire } pellet is fixed. Rinse in cacodylate buffer (no sucrose), } dehydrate in an } acetone or ethanol series. I use ethanol because I like } to embed in LR } White. After 2 changes in 100%, separate the SEM run (for } critical point } drying and coating) from the TEM samples- move on to resin } infiltration } right in the eppendorf tubes. I usually do 3:1 } (solvent:resin) on a } rotator for 1 hr., 1:1, 1:3, 100% times two and cure } overnight. } Good luck. } } } } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Additional to the useful replies relating to the preparation of ball bearings, I would like to add a little regarding the microscopy: Small, clean and undamaged ball bearings are a good instructive aid to explain SEM functioning. This may be of some use to Jonathan when viewing those bearings and to anybody trying to explain some SEM effects. 1 At high kV (say above 20) you may have trouble seeing anything, because fine surface structure and dirt will be invisible. 2 At low kV (say 10 and below) such surface structures will be visible. 3 At low powers, regardless of kV, the ball bearing will appear like a disk, but the outer part of the disk is brighter. This nicely shows that in secondary mode, brightness almost entirely is increased with the angle of incidence. Being a sphere, tilt has no effect on the brightness distribution over that image. 4 A BS detector mounted at an angle (whereas the Robinson and some others are vertical) will make the distinction that the specimen is not a disk but a sphere, because the BS electrons directed away from the detector leave a shadow. 5 A similar effect is produced when the bias current of the secondary detector is turned off and the condenser current is turned down. This floods the secondary detector's scintillator with backscattered electrons and produces a BS image quite suitable for low powers.
Nice teaching exercise, but its useful to know about these effects when actually looking at those bearings. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Friday, March 05, 1999 6:11 AM, Jonathan Barnard [SMTP:J.Barnard-at-bristol.ac.uk] wrote: } } I am more of a TEM eprson, but I have a suggestion from } watching my } colleagues. } Take an aluminium stub, which would normally go into an } SEM, and using a } 5 mm drill bit just drill a small , 1-2 mm deep, hole in } top so that the } ball bearing will sit in it. Before you mount the bearing, } wash the } bearing in organic solvents first, say two acetone washes } and then two } ethanol washes, and ultrasonically clean the bearing in an } ultrasound } bath with both solvents. } } Dry it off by putting it into an oven at say 150 C for ten } minutes and } then mount it onto the stub using silver-dag mounting } paint (used by } almost everyone here) to fix the bearing onto the stub. If } you have } access to vacuum coating system with a Radio Frequency } inductive plasma } ring attachment or anything that can produce an Argon } plasma, then put } the stub in for ten minutes so that there are no organic } residues left. } Once it is finished you can put it straight into the SEM } knowing that } there should be a clean surface to look at. The silver dag } paint should } earth the bearing to the stub. } } I hope this helps. } } Jon } } -- } ***************************************** } Jonathan Barnard } } Microstructural Physics, } H.H.Wills Physics Laboratory, } University of Bristol, } Tyndall Avenue, } Bristol BS8 1TL. } } 0117 928 9000 ext 8750 } } ***************************************** } }
We lost our Ar/Kr laser very early in its life. (it was still under warranty) A bit of advice was given to us from Zeiss regarding our Ar/Kr laser. That is to run it at 50% power rather than full blast all the time. It seems to be much better now.
} Just to give an opposite example about the laser, } } I'm running an LSM 410 equipped with an Omnichrome Ar/Kr 488/568/647 laser } since 5 years (1750 hours) and i had no problem yet ! Maybe it's just luck } though... } } } At 05:01 PM 25/2/99 -0800, you wrote: } } As for my own experience, I found the 568nm Kr laser sadly unreliable, we } had to } change it twice in the past year. } } Mr. S H Coetzee Electron Microscope Unit Private bag X3 Wits Johannesburg 2050 Tell: +27 11 716 2419 Fax : +27 11 339 3407 E-mail stephan-at-gecko.biol.wits.ac.za
A number of suggestions have been made about the preparation and observation of failed ballbearings. These will certainly give good images and may well be enough to solve the problem, but if the fault lies in the alloy rather than in the physical state of the balls, it will not show up. You might have to consider embedding the bearing in resin and polishing a flat on it. A thin coat of carbon is needed unless you use a conductive resin. Then the BSE image (and EDX if possible) will reveal the internal structure of the alloy.
Eric
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
} The peaks have suddenly started to move around a bit, although the } resolution stays good. } The problem comes and goes, in its good times the standard deviation } of the position of the Co Ka peak is about 0.4 eV (10 } determinations), but sometimes it's about 10 or even 20 eV. } My thinking is that if it were the detector, the resolution would be } degrading, but it's not, so maybe the culprit is the } (analog) pulse-processor. Anyone got any thoughts on how to pin it } down as being either } a the detector } b the subsequent signal-processing stuff, eg pulse-proc? } Could I successfully test the pulse-proc with a ramp from a standard } signal generator, or would that signal, being relatively clean } compared with that from a detector, not really check it out } rigorously enough?
One thing to note is if your movement is due to gain or offset drifts. If offset drift then both a low peak and high peak will move the same amount. If gain the low will move a little, high will move much more.
Sometimes the gain and offset pots on the pulse processor get "dirty". Run them up and down a few times or clean with electronic cleaner spray.
Disconnect and reconnect every connector. The contacts can get "dirty" over time. The physical active of disconnecting/reconnecting will clean the contacts.
You can test your MCA by using a sliding pulser. We use a Berkley Nucleonics Corp (BNC) GL-3 pulse generator to test every MCA that we ship. It's the only way to really know how well the MCA is working. It's about $5k, not cheap. You really need a very good pulser to test linearity. 10 to 20eV is a really small voltage difference.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
Is it OK to (thin) section the Falcon cell culture inserts in cross section, and would anyone who's done this please contact me directly? I have a question or two about the best way to embed to maximize cell numbers. (I think this has been discussed before.) Thank you. Grace
In the spirit recently suggested by Steve Chapman, try disturbing the cables and processor box. If this affects the peak positions or resolution, there's a good chance you have a loose or dirty connector. Try reseating the PC boards in the processor box.
Larry Thomas Washington State University
---------- From: Ritchie Sims Sent: Friday, March 5, 1999 1:28 PM To: microscopy-at-Sparc5.Microscopy.Com Subject: EDS trouble-shooting
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Dear Experts
The peaks have suddenly started to move around a bit, although the resolution stays good. The problem comes and goes, in its good times the standard deviation of the position of the Co Ka peak is about 0.4 eV (10 determinations), but sometimes it's about 10 or even 20 eV. My thinking is that if it were the detector, the resolution would be degrading, but it's not, so maybe the culprit is the (analog) pulse-processor. Anyone got any thoughts on how to pin it down as being either a the detector b the subsequent signal-processing stuff, eg pulse-proc? Could I successfully test the pulse-proc with a ramp from a standard signal generator, or would that signal, being relatively clean compared with that from a detector, not really check it out rigorously enough?
cheers (well, I try)
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
American Soc List {microscopy-at-Sparc5.Microscopy.Com} X-Mailer: QuickMail Pro 1.5.3 (Mac) X-Priority: 3 MIME-Version: 1.0 Reply-To: Paul Webster {pwebster-at-mailhouse.hei.org} Content-Type: multipart/alternative; boundary="====56495250495150514856===1"
Reply to: RE: Complications With regard to this comment By Steve Chapman, I agree that the discussion = aspect of the listserver is slowly being erroded and would sometimes like = to see the more detailed, carfully considered answers being posted again. = I use the server as a source of assistance but also find that by reading = discussions on subjects I know little about, I can actually learn = something. This sort of passive learning is useful in a busy lab environme= nt.
I am not sure we can hold the solution providers totally responsible for = this errosion either. It is sometimes difficult to know exactly the = problem, the skill of the poster or the environmentally limiting factors = from such brief postings as in the example given by Steve ("What do I do = if I do not have a good quality image on the screen in my TEM?"). Why is = it that some questions are posted in almost annotated form from posters = who do not even provide us with an identification? =
The tendency for discussions to be carried out "off-line" in more detail = is not very beneficial either. It is true that some of the really = detailed discussions, and perhaps some of the comments about particular = products, are best covered in a more private setting, but it would be = great to see edited summaries and final results.
I know writing is tough and it takes some time to write but by putting = more effort into this skill we could improve our list. I am not getting = at non-English writers here but at anyone who writes messages which are so = brief that relevant information is omitted. The more we practice our = writing, the better we get (hopefully). =
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm http://www.hei.org/htm/apw.htm
Steve Chapman wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
{HTML} {HEAD} {/HEAD} {BODY} {PRE WIDTH=3D"132"} Reply to: RE: Complications
{/PRE} {FONT FACE=3D"Geneva" SIZE=3D3 = COLOR=3D"#000000"} With regard to this comment By Steve = Chapman, I agree that the discussion aspect = of the listserver is slowly being erroded = and would sometimes like to see the more = detailed, carfully considered answers being = posted again. I use the server as a source = of assistance but also find that by reading = discussions on subjects I know little about, = I can actually learn something. This sort = of passive learning is useful in a busy = lab environment. {BR} {BR} I am not sure we can = hold the solution providers totally responsible = for this errosion either. It is sometimes = difficult to know exactly the problem, the = skill of the poster or the environmentally = limiting factors from such brief postings = as in the example given by Steve ("What = do I do if I do not have a good quality = image on the screen in my TEM?"). Why = is it that some questions are posted in = almost annotated form from posters who do = not even provide us with an identification? = {BR} {BR} The tendency for discussions to be = carried out "off-line" in more = detail is not very beneficial either. It = is true that some of the really detailed = discussions, and perhaps some of the comments = about particular products, are best covered = in a more private setting, but it would = be great to see edited summaries and final = results. {BR} {BR} I know writing is tough and = it takes some time to write but by putting = more effort into this skill we could improve = our list. I am not getting at non-English = writers here but at anyone who writes messages = which are so brief that relevant information = is omitted. The more we practice our writing, = the better we get (hopefully). {BR} {BR} Regards, {BR} {/FONT} {FONT = FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#000000"} {BR} {/FONT} {FONT FACE=3D"Geneva" SIZE=3D3 COLOR=3D"#000000"} Paul = Webster, Ph.D {BR} House Ear Institute {BR} 2100 = West Third Street {BR} Los Angeles, CA 90057 {BR} phone:213 = 273 8026 {BR} fax: 213 413 6739 {BR} e-mail: pwebster-at-hei.org {BR} http://www.hei.org/htm/aemi.htm {BR} http://www.hei.org/htm/apw.htm {/FONT} {FONT = FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#000000"} {BR} {BR} {BR} {/FONT} {FONT FACE=3D"Geneva" = SIZE=3D3 COLOR=3D"#000000"} Steve Chapman wrote: {/FONT} {FONT FACE=3D"Geneva"= = SIZE=3D1 COLOR=3D"#000000"} {BR} >-----------------------------------------------------------------------= - {BR} >The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America {BR}
id xma019702; Fri, 5 Mar 99 13:37:30 -0500 Received: by eastman.com id AA1958416 (5.67b/IDA-1.5 for microscopy-at-Sparc5.Microscopy.Com); Fri, 5 Mar 1999 13:37:57 -0500 Received: from ntmcon02.emn.com by eastman.com with SMTP id AA2032906 (5.67b/SMI-4.1 for {microscopy-at-Sparc5.Microscopy.Com} ); Fri, 5 Mar 1999 13:37:57 -0500 Received: by ntmcon02.emn.com with Internet Mail Service (5.5.2232.9) id {FZ4RM9PM} ; Fri, 5 Mar 1999 13:38:06 -0500 Message-Id: {E788D998AAC8D1119EC10000F8CD1E8A5B42BA-at-ntmail23.emn.com}
Regarding moving peaks in EDS, we once experienced a problem caused by the video monitor on the EDS unit being too close to the wires coming from the detector. Our problem became quite obvious when the resolution began to degrade. Moving the wires fixed the problem. Perhaps your problem is being caused by electromagnetic fields.
Dennis B. Barr (dennbarr-at-eastman.com) Physical Chemistry Research Laboratory Physical & Analytical Chemistry Research Division Eastman Chemical Company Kingsport, TN 37662-5150
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} The peaks have suddenly started to move around a bit, although the } resolution stays good. } The problem comes and goes, in its good times the standard deviation } of the position of the Co Ka peak is about 0.4 eV (10 } determinations), but sometimes it's about 10 or even 20 eV. } My thinking is that if it were the detector, the resolution would be } degrading, but it's not, so maybe the culprit is the } (analog) pulse-processor. Anyone got any thoughts on how to pin it } down as being either } a the detector } b the subsequent signal-processing stuff, eg pulse-proc? } Could I successfully test the pulse-proc with a ramp from a standard } signal generator, or would that signal, being relatively clean } compared with that from a detector, not really check it out } rigorously enough?
One thing to note is if your movement is due to gain or offset drifts. If offset drift then both a low peak and high peak will move the same amount. If gain the low will move a little, high will move much more.
Sometimes the gain and offset pots on the pulse processor get "dirty". Run them up and down a few times or clean with electronic cleaner spray.
Disconnect and reconnect every connector. The contacts can get "dirty" over time. The physical active of disconnecting/reconnecting will clean the contacts.
You can test your MCA by using a sliding pulser. We use a Berkley Nucleonics Corp (BNC) GL-3 pulse generator to test every MCA that we ship. It's the only way to really know how well the MCA is working. It's about $5k, not cheap. You really need a very good pulser to test linearity. 10 to 20eV is a really small voltage difference.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
I have a couple of questions w/r to EELS output in other formats. I am acquiring EELS data on a GIF at another institution, but would like to take the files back to my lab. I saved the files as ASCII X,Y, tagged ASCII, and EMSA/MAS format. I can plot the X,Y data with no problem. However, in the tagged ASCII and the EMSA/MAS format, the data are written in 5 wide rows that wrap the data. I typically use Excel to plot stuff like this, but I don't know how to unwrap the data. What I am doing is going into a Wordprocessor and getting rid of the hard returns and commas and then resaving as a text file. I can automate it a little with macros, but it still takes time.
1. Are there ways to unwrap the data directly into Excel?
2. Is there a public domain EELS program that will plot the data and perhaps do some work with the EMSA/MAS format that will run on a PC or a Mac? (I would prefer PC because our Macs are old and there is not much chance of getting new ones here.)
Thanks in advance.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Lesley S. Bechtold wrote: ============================================ I have never done any material sciences work - I've only ever done biological specimens for EM. Our engineering department has asked me to look at some ball bearings that are failing, as a favour. I know I don't need to fix or dehydrate but do I simply clean them, mount them (using double-sided tape?) and coat them as usual? Or is coating unnecessary? What whould I clean these with? I'm assuming there is grease somewhere that is not good for my vacuum! ================================================== It is interesting how so many different persons, skilled in the art, would approach the same kind of problem so many different ways. Of course, not much information was given either so we could all be those proverbial blind men feeling a different part of the elephant.
We ourselves approach bearing failures somewhat differently. For one thing, at the onset, it is rarely clear whether it is a straight SEM job. If corrosion is involved, for example, the ability to analyze corrosion product is lost if vigorous washing/cleaning procedures are used.
No mention was made of the examination of the raceway, but that is also something of importance in any kind of a failure analysis of this type.
Our approach is to liquid wash in cold solvent, such as acetone and/or heptane, perhaps even xylene, but you don't want a solvent that is "too good ." You want to leave some organic layer on the metal surfaces. We then remove the last vestiges of the lubricant system with exposure to an oxygen plasma such as in our Plasma Prep™ II plasma etcher. This is a dry process approach, corrosion product is left in situ in place, right where it is, and with EDS, sometimes complimented with Auger, one can learn information about the failure mechanism that would not be learned by straight topographical examination.
For the mounting of the bearing "balls", depending on size, we would always use one of the conductive double sided adhesive products, either carbon sheets or if larger, then Tempfix™.
We ourselves believe one should use caution before washing away the information that could be contained in corrosion product.
Disclaimer: SPI Supplies manufactures the Plasma Prep II plasma etcher and supplies the carbon based adhesives. Our Structure Probe services laboratory performs failure analysis as a service on these kinds of samples.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Ritchie Sims wrote: } Dear Experts } } The peaks have suddenly started to move around a bit, although the } resolution stays good. } The problem comes and goes, in its good times the standard deviation } of the position of the Co Ka peak is about 0.4 eV (10 } determinations), but sometimes it's about 10 or even 20 eV. } My thinking is that if it were the detector, the resolution would be } degrading, but it's not, so maybe the culprit is the } (analog) pulse-processor. Anyone got any thoughts on how to pin it } down as being either } a the detector } b the subsequent signal-processing stuff, eg pulse-proc? } Could I successfully test the pulse-proc with a ramp from a standard } signal generator, or would that signal, being relatively clean } compared with that from a detector, not really check it out } rigorously enough? } } cheers (well, I try) } } Ritchie } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } }
Dear Ritchie! In similar cases the first I do is the following: I take off the signal cable from preamplifier and processor and measure its resistance, which should be near zero, and then check stability of this resistance during curving this cable. Two times in my practice I have met the case when after several working years the copper core of the cable was corroded as result of chemical interaction with material of cable insulation, and the resistance between many wires of core became rather big and unstable. After change to cable with Teflon insulation the peaks became very (!) stable. Regards.
I have run across the same problems with EMSA format and with other files that store multiple points of data per row.
I have developed macros that insert the necessary number of blank lines (5 for EMSA) following a given row, I select the data in the row, Copy-Special using the transpose function into the blank rows, delete the original row, and move down to the next row, and repeat. It isn't pretty, but it gets the job done.
If I was doing it regularly, I think I would write a program (standalone or Word Basic) that would incorporate the smarts enough to do it all automatically given the file name. As of yet, I have not been so driven. But maybe someone else has already done one.
Good luck.
At 03:54 PM 3/5/99 -0500, you wrote: } I have a couple of questions w/r to EELS output in other formats. I am } acquiring EELS data on a GIF at another institution, but would like to take } the files back to my lab. I saved the files as ASCII X,Y, tagged ASCII, and } EMSA/MAS format. I can plot the X,Y data with no problem. However, in the } tagged ASCII and the EMSA/MAS format, the data are written in 5 wide rows } that wrap the data. I typically use Excel to plot stuff like this, but I } don't know how to unwrap the data. What I am doing is going into a } Wordprocessor and getting rid of the hard returns and commas and then } resaving as a text file. I can automate it a little with macros, but it } still takes time. } } 1. Are there ways to unwrap the data directly into Excel? } } 2. Is there a public domain EELS program that will plot the data and } perhaps do some work with the EMSA/MAS format that will run on a PC or a } Mac? (I would prefer PC because our Macs are old and there is not much } chance of getting new ones here.) } } Thanks in advance. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc.
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
On Fri, 5 Mar 1999, Walck. Scott D. wrote: | |2. Is there a public domain EELS program that will plot the data and |perhaps do some work with the EMSA/MAS format that will run on a PC or a |Mac? (I would prefer PC because our Macs are old and there is not much |chance of getting new ones here.) |
I've been using XlispStat {http://www.xlispstat.org/} for both plotting and analysis ( Mostly EDS but some EELS ).
It's a free, cross-platform (Mac,Windows,Unix) version of Lisp enhanced with vector and matrix arithmetic, math and statistical function, linear and non-linear regression, smoothing, FFTs, and simple object oriented graphics.
It's extensible with function written either in Lisp or C: I've written functions to read & write EMSA/MAS format (roughly -- the specs are somewhat ambiguous), read spectra from a 4pi SpectraEngine on a Mac, convolution and digital filtering and other utilities. ( Haven't quite gotten it all wrapped up into a complete EDS/EELS package yet. )
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| University of Virginia Health Sciences Center |--- ---| P.O. Box 10011 Charlottesville, VA 22906-0011 |---
Caldera Open Linux: "Powerful and easy to use!" -- Microsoft(*) (*) {http://www.pathfinder.com/fortune/1999/03/01/mic.html}
Dear Lim (S'pore), There are many infrared camera in the market. You might need to specify = what is the usage are for. By the way, how is Singapore nowadays? Still M= oney no enough?
Thank you for the replies so far. I should have included the info that it's a gain problem ie the zero peak stays in the right place. I guess it boils down to:
given that the resolution stays good, does anyone think that it could be a detector problem, or does everyone think that it's the pulse-processor?
rtch
} From: Self {GLGNOV2/RSIMS} } To: microscopy-at-sparc5.microscopy.com } Subject: EDS trouble-shooting } Date: Fri, 5 Mar 1999 21:28:27 GMT+1200
} Dear Experts } } The peaks have suddenly started to move around a bit, although the } resolution stays good. } The problem comes and goes, in its good times the standard deviation } of the position of the Co Ka peak is about 0.4 eV (10 } determinations), but sometimes it's about 10 or even 20 eV. } My thinking is that if it were the detector, the resolution would be } degrading, but it's not, so maybe the culprit is the } (analog) pulse-processor. Anyone got any thoughts on how to pin it } down as being either } a the detector } b the subsequent signal-processing stuff, eg pulse-proc? } Could I successfully test the pulse-proc with a ramp from a standard } signal generator, or would that signal, being relatively clean } compared with that from a detector, not really check it out } rigorously enough? } } cheers (well, I try) } } Ritchie }
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
As one of the committee members of the MSA/MAS Format I'm perplexed about your comment on the 5 wide rows. That sounds very odd indeed and I recall nothing in the format that calls out that type of coding. Perhaps there is an error somewhere or a misinterpretation of the spectral file format. .
Send me a "private" copy of the 3 files at my ANL address (Zaluzec-at-aaem.amc.anl.gov) and I'll have a look at them when I get back to the US.
Nestor
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I think that the main problem here is that we have a defined M&M=20 =46ormat (MSA/MAS in old parlance, let's face it we are a Microscopy=20 and Microanalysis community and the sooner we realize it the=20 better!), but the XEDS and EELS manufacturers don't fully support it.=20 They either write it but don't read it or read it but don't write it.=20 I think the format should be refined so that the manufacturers will=20 use it and let them have tags in the format similar to the TIFF.
Just my two cents worth (probably start a war, but just my opinion)
Jfm.
John Mansfield PhD CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 936-3352 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
Ritchie Sims wrote: } Dear Experts } } The peaks have suddenly started to move around a bit, although the } resolution stays good. } The problem comes and goes, in its good times the standard deviation } of the position of the Co Ka peak is about 0.4 eV (10 } determinations), but sometimes it's about 10 or even 20 eV. } My thinking is that if it were the detector, the resolution would be } degrading, but it's not, so maybe the culprit is the } (analog) pulse-processor. Anyone got any thoughts on how to pin it } down as being either } a the detector } b the subsequent signal-processing stuff, eg pulse-proc? } Could I successfully test the pulse-proc with a ramp from a standard } signal generator, or would that signal, being relatively clean } compared with that from a detector, not really check it out } rigorously enough? } } cheers (well, I try) } } Ritchie } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } }
Dear Ritchie! In similar cases the first I do is the following: I take off the signal cable from preamplifier and processor and measure its resistance, which should be near zero, and then check stability of this resistance during curving this cable. Two times in my practice I have met the case when after several working years the copper core of the cable was corroded as result of chemical interaction with material of cable insulation, and the resistance between many wires of core became rather big and unstable. After change to cable with Teflon insulation the peaks became very (!) stable. Regards.
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Hello
Ritchie Sims wrote: } Dear Experts } } The peaks have suddenly started to move around a bit, although the } resolution stays good. } The problem comes and goes, in its good times the standard deviation } of the position of the Co Ka peak is about 0.4 eV (10 } determinations), but sometimes it's about 10 or even 20 eV. } My thinking is that if it were the detector, the resolution would be } degrading, but it's not, so maybe the culprit is the } (analog) pulse-processor. Anyone got any thoughts on how to pin it } down as being either } a the detector } b the subsequent signal-processing stuff, eg pulse-proc? } Could I successfully test the pulse-proc with a ramp from a standard } signal generator, or would that signal, being relatively clean } compared with that from a detector, not really check it out } rigorously enough? } } cheers (well, I try) } } Ritchie } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } }
Dear Ritchie! In similar cases the first I do is the following: I take off the signal cable from preamplifier and processor and measure its resistance, which should be near zero, and then check stability of this resistance during curving this cable. Two times in my practice I have met the case when after several working years the copper core of the cable was corroded as result of chemical interaction with material of cable insulation, and the resistance between many wires of core became rather big and unstable. After change to cable with Teflon insulation the peaks became very (!) stable. Regards.
Arnold, Jim wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am currently looking to upgrade my EDS system. I work in a semiconductor } manufacturing facility(CMOS) and I currently have a Cambridge S200 SEM with } Kevex Delta II. } } I have an interest in Oxford, EDAX and a company called EVEX (which I am not } familiar with)? Does anyone have experience with these companies - Good or } Bad? } } I am looking for a light element detector with possibly a WDS for Boron } quantification. } } Thanks in advance. } } Jim Arnold } Microelectronics and Technology Center } AlliedSignal Electronics and Avionics Systems } 9140 Old Annapolis Road } Columbia, MD 21045 } } email: Jim.arnold-at-alliedsignal.com } voice: (410) 964-4118 } fax: (410) 964-5046
Jim, One of my customers sent their Kevex detector to Evex for repair. They didn't complete the repair, they bent the dewar, and they never returned. A very bad bet to do business with them.
Ken Converse owner Quality Images third party SEM service Delta, PA
Lesley S. Bechtold wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } } I have never done any material sciences work - I've only ever done } biological specimens for EM. Our engineering department has asked me to } look at some ball bearings that are failing, as a favour. I know I don't } need to fix or dehydrate but do I simply clean them, mount them (using } double-sided tape?) and coat them as usual? Or is coating unnecessary? } What whould I clean these with? I'm assuming there is grease somewhere } that is not good for my vacuum! } } Any help would be appreciated!! Thanks in advance..... } } Lesley Bechtold } } Lesley S. Bechtold } Supervisor, Biological Imaging } The Jackson Laboratory } 600 Main St. } Bar Harbor, ME 04609 } 207-288-6191
Lesley, All of the suggestions have been pretty much on the mark. Just two things 1) Get a degausser so you can demagnetize the sample, because if you don't, your resolution will have you swearing at your microscope for its poor performance, 2) a ball bearing that has a good surface may require up to 20kx magnification to see any surface detail. Some dirt on the surface can make finding that surface much easier. Materials failure analysis is a lot of fun!
Ken Converse owner Quality Images third party SEM service Delta, PA
Dear listserved all, I wish to know the examination dates for the next cycle of MSA certification in Electron Microscopy. I know that this question should have been sent to the business office at MSA, but I hope to have a quicky answer from any of the list members. Cheers and have a good day. Mohammed Yousuf A.Rawoof.
} } 1. Are there ways to unwrap EMSA/MAS data directly into Excel? }
Scott, it's a fairly straightforward process to unwrap the data in MSExcel. I recorded a macro to do it a while ago - although we were on Macs then, not PCs, I think I kept it somewhere... If you e-mail a file to me I could write it again, if you like. It is fairly easy to do yourself - I think it's just a matter of cutting and pasting columns. If you can work out a routine to do it once, you can 'record' what you're doing as a macro. All you have to do for subsequent files is to run the macro again.
} 2. Is there a public domain EELS program that will plot the data and } perhaps do some work with the EMSA/MAS format that will run on a PC or a } Mac? (I would prefer PC because our Macs are old and there is not much } chance of getting new ones here.) } } Thanks in advance. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } PO Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } } } } }
I am alarmed by your statement. It is Evex Analytical's policy to "always" perform an on-site installation of "any" new detector install or detector repair.
Please be more specific on your customer's name, location, detector serial number, date of service. .
Are you absolutely sure the customer you mentioned sent a "Kevex" detector to "Evex Analytical"? Your prompt reply is appreciated.
Thank you Evex Analytical Peter Tarquinio
-----Original Message----- } From: Kenneth Converse {qualityimages-at-netrax.net} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} ; Arnold, Jim {Jim.Arnold-at-alliedsignal.com}
I am working with self-assembled microstructures that are composed of a sterol and a phosphatidylcholine. I need to be able to attach these structures to glass or plastic walls of a chamber they grow in. What would you recommend? I have tried various commercially coated glass slides, super glue, jewel glue, leather glue, and almost any other glue I could think of. The problem is that my structures grow in an aqueous solution, and I was not able to find an adhesive which would not only glue the structures (super glue did the job actually), but also not dissolve in water.
I would greatly appreciate your suggestions and comments.
Thank you in advance -- Yevgeniya Zastavker.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Yevgeniya V. Zastavker Massachusetts Institute of Technology, Biophysics 77 Massachusetts Ave, Room 13-2038 Cambridge, MA 02139 (617) 253-4826
Sorry to bombard you with questions. This could be the wrong list, but I thought to try anyway. I am looking for crystallographic data of various sterols, and in particular (MAJORLY) I need information on the crystal angles of various sterols. Does anybody know of a good source for this information? I would greatly appreciate your advice.
Thank you very much in advance -- Yevgeniya Zastavker
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Yevgeniya V. Zastavker Massachusetts Institute of Technology, Biophysics 77 Massachusetts Ave, Room 13-2038 Cambridge, MA 02139 (617) 253-4826
The Electron Microscopy Laboratory at New Mexico State University has an open position for an Electron Microscopy Specialist. The laboratory provides transmission and scanning electron microscopy and some light microscopy services for the university research community and a few external organizations, in biological, physical and materials sciences fields. Qualifications: bachelor's degree minimum, master's degree desirable, with at least four years of electron microscopy experience. The preferred candidate will have experience with energy-dispersive x-ray analysis. Experience with digital image capture and analysis, fluorescence microscopy, immunocytochemistry (including immunogold labeling), and laser scanning confocal microscopy is desirable. The candidate must be competent with sample preparation techniques, including vacuum evaporation, sputter coating, critical point drying, support film production, low temperature embedding, and photographic film processing and printing. The successful candidate must be able to work well with researchers, staff, and students, and be able to train graduate and undergraduate students for independent work with relevant techniques and equipment. Duties: operations and routine maintenance of transmission and scanning electron microscopes and associated equipment; fixation, embedding, semi-thin and ultrathin sectioning, staining, and coating of samples; supervision of facility users; record keeping, including billings, budgets, maintenance of instrument and research logs, and researching and designing specimen preparation protocols as required. Salary: DOQ Website: www.nmsu.edu/~personal/postings/professional/ Screening of applicants will begin May 1, 1999 and continue until a candidate is chosen. Applications should include a resume, letter of application and three letters of recommendation.
Apply to: Dr. Reed Dasenbrock Associate Dean/Director Arts and Sciences Research Center New Mexico State University MSC RC, Box 30001 Las Cruces, NM 88003-8001 rdasenbr-at-nmsu.edu
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We have an ETEC SEM available if anyone wants it. (Stockton, CA) It is = in working condition. We bought it new in 1973 and it has been under = contract since that time until Feb. 1, 1999. We also have extra ETEC = parts (power supplies, modules, etc.) We must take it out by March 24, 1999. If anyone wants it, please let me = know or we start chopping it on that day. Thanks Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
id {1DCJVQ86} ; Mon, 8 Mar 1999 12:10:10 -0800 Message-ID: {3D55EE50922DD21192CC00A0C9A9D8270120F8-at-cas.csudh.edu}
Dear subscribers,
I want to purchase a microtome that sections both paraffin and plastic embedded tissue. I have not purchased a rotary microtome before and I would like your suggestions as to what companies handle rotary microtomes at the present time. I have an ultramicrotome and I know that both types of media can be cut on it but I need a second microtome and have about $12,000 to spend.
Laura J. Robles
Laura J. Robles, PhD. Associate Dean, Student Academic Advancement Professor of Biology MBRS Program Director College of Arts and Sciences California State University, Dominguez Hills 1000 East Victoria Street, Carson CA 90747 310 243-3389, FAX 310 516-4268 lrobles-at-cas.csudh.edu
I think an inexpensive inkjet is the way to go. However, you must try to set the DPI of the printer to match the resolution setting of your digital images. If you capture a digital image at 1024 X800 for instance, you have about 820K of information. Now lets say you plan to print a 4X5 image similar to a Polaroid, then your printer should be no less than 300dpi {4"x5" x (300)2=1.8Mpixel} to accommodate the amount of pixel information from the capturing rate of the image. Hence, a 2048X1600 resolution setting captures 3.2 Mpixels of info, so a 400DPI setting should be used. The idea being to match the capturing info with the amount of pixels the printer can resolve to minimize interpolation...be it upwards or downwards. Not sure what the human eye can resolve tho.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have an ETEC SEM available if anyone wants it. (Stockton, CA) It is = in working condition. We bought it new in 1973 and it has been under = contract since that time until Feb. 1, 1999. We also have extra ETEC parts (power supplies, modules, etc.) We must take it out by March 23, 1999. If anyone wants it, please let me = know or we start chopping it on that day. Thanks Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
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Dear List, Can anyone explain what does "Reflective" means in Laser Terminology .
I read through a article in a Magazine regarding Laser Marker and in the = article the mentioned that Wood and Paper is 100% reflective . I am lost = and confuse
M.E.Lim Sr Regional Support Engineer QES(Asia Pacific) Sdn Bhd Tel : 603-7241188 ext 214 Fax : 603-7244488 Emails : melim-at-qes.po.my
Can anyone give me tips how to prepare micro-injected cells for TEM. Which cell culture support is best for detaching the cells? Is there any compound which we could micro-inject as a marker?
Try gap-filling super glue (crazy glue), available at any hobby store or it should be at a hardware store. No particular brand, they all work--it's the gap-filling that seems to be important. I used this to glue gelatin specimen blocks to the stage of a Vibratome which was then flooded with phosphate buffer. It holds under water if you let it set. This doesn't take long.
The problem may be the gap-filling property--it may cover your microstructures.
Phil
} I am working with self-assembled microstructures that are composed of a } sterol and a phosphatidylcholine. I need to be able to attach these } structures to glass or plastic walls of a chamber they grow in. What } would you recommend? I have tried various commercially coated glass } slides, super glue, jewel glue, leather glue, and almost any other glue I } could think of. The problem is that my structures grow in an aqueous } solution, and I was not able to find an adhesive which would not only glue } the structures (super glue did the job actually), but also not dissolve in } water. } } I would greatly appreciate your suggestions and comments. } } Thank you in advance -- Yevgeniya Zastavker. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Yevgeniya V. Zastavker } Massachusetts Institute of Technology, Biophysics } 77 Massachusetts Ave, Room 13-2038 } Cambridge, MA 02139 } (617) 253-4826
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 Voice: (608) 833-2885 Fax: (608) 836-1969 (please make sure my name is on any fax) oshel-at-terracom.net
May I offer a word of caution about the approach below?
While you're correct in saying you need to optimize printing conditions for your image and printer, there will probably be a little more to it than setting up comparable number of pixels. Your image is probably either a 256-level gray scale or a 16 million color (256 levels of red, green and blue). An inkjet can't print that kind of color depth in each pixel. The concepts of "halftoning" or "dithering" need to be considered. As I'm far from the expert, and we don't need a complete textbook on the listserver anyway, so I'll refer you to chapter 2 of "The Image Processing Handbook" by John Russ. As we would expect from Dr. Russ, the material is excellent.
That does bring up a question for me, though. The new HP inkjets have a "PhotoREt" technology which, I believe is supposed to be able to vary the size of the dots it produces, therefore producing better photo-printing results. Has anyone determined whether this is true, or just hype?
Jim Passmore Sr. Analytical Chemist Cryovac Division Sealed Air Corporation
---------- } From: Harry.Ekstrom } To: Microscopy } Subject: FW: Printers for SEM Images } Date: Monday, March 08, 1999 5:11PM } ------------- } } I think an inexpensive inkjet is the way to go. However, you must try to } set the DPI of the printer to match the resolution setting of your digital } images. If you capture a digital image at 1024 X800 for instance, you have } about 820K of information. Now lets say you plan to print a 4X5 image } similar to a Polaroid, then your printer should be no less than 300dpi } {4"x5" x (300)2=1.8Mpixel} to accommodate the amount of pixel information } from the capturing rate of the image. Hence, a 2048X1600 resolution setting } captures 3.2 Mpixels of info, so a 400DPI setting should be used. The idea } being to match the capturing info with the amount of pixels the printer can } resolve to minimize interpolation...be it upwards or downwards. Not sure } what the human eye can resolve tho. } } Good Luck, } Harry Ekstrom } }
Does anyone have or know of a good reference on EM lab safety? I thought there was a book called "Safety in the EM Laboratory"...but I haven't been able to find it. Probably imagined it.
FEI Company Celebrates 50 Years Philips Electron Microscopy with an Anniversary Image Contest (Calling all microscopists!)
Fifty years ago, we delivered the first Philips electron microscope. Since then, our TEMs and SEMs are used for all kinds of applications.
To celebrate the occasion, we're inviting all Philips electron microscope users all over the world to join our special Anniversary Image Contest.
To enter, simply submit prints of one or two of your best images made with a Philips electron microscope, showing the original data bar. Prints only, please! Closing date: 31 July 1999
WIN 1,000 EURO*!
Our jury panel of experts will select the best ten images, five in Life Science and five in Material Science. All ten winners will each be awarded a prize of 1000 Euro*. The winners will be announced in August 1999 on the FEI website at www.feic.com.
The following details must accompany each entry: * Name and contact address of owner * Category: Life Science or Material Science * Description of subject * Type of instrument used * Electron optic magnification * Magnification of print
Please submit your entry to: FEI Company 50 Years Philips EM Celebration P.O. Box 218 5600 MD EINDHOVEN The Netherlands
Digital images cannot be accepted for practical reasons. All submissions must be free of any legal obligations. All entries remain property of their original owners, but contestants consent to the use of their entries for promotional purposes by FEI Company without further compensation. Prizes are not transferable. Taxes are the sole responsibility of the winners. Contest rules are available on the company's website (www.feic.com) or can be requested by fax to +31 40 276 6587. FEI Company will not enter into any other correspondence regarding this contest.
*Actual prize will be the equivalent value in the winner's local currency
Yev, You might try Cell-Tak from Collaborative Biomedical Products, Two Oak Park, Bedford, Mass. 617-275-0004. This is the isolated mussel adhesion protein (map) the marine mussels and barnacles use to attach themselves to rocks and boats etc. It is commonly used by people perform atomic force microscopy to attach their molecules to a surface.
Images & Info at http://www.molbio.princeton.edu/confocal {http://www.molbio.princeton.edu/confocal}
Joseph Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University jgoodhouse-at-molbio.princeton.edu {mailto:Jgoodhouse-at-molbio.princeton.edu}
609-258-5432
-----Original Message----- From: Yevgeniya Zastavker [mailto:zhenya-at-critical.mit.edu] Sent: Monday, March 08, 1999 10:43 AM To: microscopy-at-Sparc5.Microscopy.Com Cc: Yevgeniya Zastavker Subject: adhesive for lipids
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I am working with self-assembled microstructures that are composed of a sterol and a phosphatidylcholine. I need to be able to attach these structures to glass or plastic walls of a chamber they grow in. What would you recommend? I have tried various commercially coated glass slides, super glue, jewel glue, leather glue, and almost any other glue I could think of. The problem is that my structures grow in an aqueous solution, and I was not able to find an adhesive which would not only glue the structures (super glue did the job actually), but also not dissolve in water.
I would greatly appreciate your suggestions and comments.
Thank you in advance -- Yevgeniya Zastavker.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Yevgeniya V. Zastavker Massachusetts Institute of Technology, Biophysics 77 Massachusetts Ave, Room 13-2038 Cambridge, MA 02139 (617) 253-4826
I think an inexpensive inkjet is the way to go. However, you must try to set the DPI of the printer to match the resolution setting of your digital images. If you capture a digital image at 1024 X800 for instance, you have about 820K of information. Now lets say you plan to print a 4X5 image similar to a Polaroid, then your printer should be no less than 300dpi {4"x5" x (300)2=1.8Mpixel} to accommodate the amount of pixel information from the capturing rate of the image. Hence, a 2048X1600 resolution setting captures 3.2 Mpixels of info, so a 400DPI setting should be used. The idea being to match the capturing info with the amount of pixels the printer can resolve to minimize interpolation...be it upwards or downwards. Not sure what the human eye can resolve tho.
I just scanned through postings about processing tissue cultures and coverslips for TEM, but I didn't see anything on processing cells grown on 8 well Permanox slides. I will be embedding in PolyBed 812, with a transition through propylene oxide. Unfortunately, the wells don't survive the p.o. step. I would appreciate hearing about any experiences with Permanox slides. Thank you very much.
Sandy Perkins
Laboratory for Neurotoxicity Studies Virginia-Maryland Regional College of Veterinary Medicine Virginia Tech
I agree that you must match the print resolution to the image resolution, but take some exception with your math.
At the crux of the issue is how many printer pixels are required to represent a single image pixel with a satisfactory level of gray or color scale resolution. While a dye sub printer conceptually requires only a single pixel to render the whole range of colors or gray scales, an inkjet printer may require multiple pixels based on the technology used. If an ink jet can only be tunred on and off (like a laser printer), then dithering will be required over a number of pixels to give the appearance of shades of color. More pixels will be needed for smoother or finer gradations. Now I think I heard that new inkjet printers can control the amount of ink at each pixel so that they approach the dye-subs in using only one printer pixel per one image pixel. However, I think better results would be had by allowing something like an 6x6 printer pixel pattern for each image pixel.
Using those assumptions, a 1024x800 image would require 6144x4800 pixels in a 5x4 inch space which requires 1200 dpi printer resolution. Doubling the image resolution to 2048 requires doubling the printer resolution to 2400 dpi. However, the limited resolution (both spatial and color) of the human eye may permit decent images with much less printer resolution, but there would be some loss of image detail.
At 03:11 PM 3/8/99 -0700, Ekstrom, Harry wrote: } } I think an inexpensive inkjet is the way to go. However, you must try to } set the DPI of the printer to match the resolution setting of your digital } images. If you capture a digital image at 1024 X800 for instance, you have } about 820K of information. Now lets say you plan to print a 4X5 image } similar to a Polaroid, then your printer should be no less than 300dpi } {4"x5" x (300)2=1.8Mpixel} to accommodate the amount of pixel information } from the capturing rate of the image. Hence, a 2048X1600 resolution setting } captures 3.2 Mpixels of info, so a 400DPI setting should be used. The idea } being to match the capturing info with the amount of pixels the printer can } resolve to minimize interpolation...be it upwards or downwards. Not sure } what the human eye can resolve tho. } } Good Luck, } Harry Ekstrom
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011-3232
I have an address for Electron Microscopy Safety Handbook. 2nd Edition. 1994. $15.00 Barber, V.C. and J.A. Mascorro (Eds)
San Francisco Press, Box 428600 San Francisco, CA 94142-6800
Hopefully it is still in print, Sally -- Sally Burns Center for Electron Optics B5 Center for Integrated Plant Systems E. Lansing, MI 48824 (517) 355-5004 burnssal-at-pilot.msu.edu
We are electropolishing a two-phase alloy containing a Laves phase and a bcc solid solution. The bcc phase is mainly Vanadium and the Laves phase is mainly HfV2. We are experiencing preferential polishing of the bcc phase leading to marginal TEM specimen quality.
Reply to: RE: Electropolishing of Hf alloy Try adding about 5% acetic acid, polish at -40 C, 100 volts.
Alternate polish: (worked on V-20Ti) = 5.3 g lithium chloride Temp=3D -50 C 11.1 g magnesium perchlorate voltage=3D 190-200 500 ml methanol current=3D 40-50 mA 100 ml butyl cellosolve
Above done with a South Bay 550-B single jet polisher in 1985. A notation mentions even grain boundaries. =
Bernie Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439 E-mail {kestel-at-anl.gov} 100 ml butyl cellosolve David E. Luzzi wrote: } We are electropolishing a two-phase alloy containing a Laves phase and a = bcc } solid solution. The bcc phase is mainly Vanadium and the Laves phase is } mainly HfV2. We are experiencing preferential polishing of the bcc phase } leading to marginal TEM specimen quality. } } Our solution is H2SO4 / Methanol / HF / butyl cellusolve } } Does anyone have any suggestions? Thanks in advance. } } David E. Luzzi } Department of Materials Science } University of Pennsylvania } 3231 Walnut Street } Philadelphia, PA 19104-6272 } } 215-898-8366 } 215-573-2128 - fax } luzzi-at-lrsm.upenn.edu {mailto:luzzi-at-lrsm.upenn.edu} } } } } RFC822 header } ----------------------------------- } } Received: from dns2.anl.gov (dns2.anl.gov [146.139.254.3]) by = } horus.et.anl.gov (8.6.11/8.6.11) with ESMTP id NAA23584 for {kestel-at-horus.= et.anl.gov} ; Tue, 9 = } Mar 1999 13:42:50 -0600 } Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com = } [206.69.208.10]) by dns2.anl.gov (8.9.1a/8.6.11) with SMTP id NAA21456; = Tue, 9 Mar 1999 = } 13:42:50 -0600 (CST) } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.= 11) = } id NAA12717 for dist-Microscopy; Tue, 9 Mar 1999 13:07:10 -0600 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id NAA12703 for = } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 9 Mar 1999 13:06:39 -= 0600 } Received: from sol1.lrsm.upenn.edu (SOL1.LRSM.UPENN.EDU [130.91.56.35]) = by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id NAA12692 for = } {Microscopy-at-sparc5.microscopy.com} ; Tue, 9 Mar 1999 13:06:24 -0600 } Received: from Luzzi.sol1.lrsm.upenn.edu (LRSM221PC.LRSM.UPENN.EDU = } [130.91.56.249]) } by sol1.lrsm.upenn.edu (8.8.5/8.8.4) with SMTP } id OAA08509 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 9 Mar 1999 = } 14:23:08 -0500 (EST) } From: "David E. Luzzi" {luzzi-at-sol1.lrsm.upenn.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Subject: Electropolictropoliing of Hf alloy } Date: Tue, 9 Mar 1999 14:22:45 -0500 } Message-ID: {001001be6a62$36779b20$f9385b82-at-Luzzi.sol1.lrsm.upenn.edu} } MIME-Version: 1.0 } boundary=3D"----=3D_NextPart_000_0011_01BE6A38.4DAABAE0" } X-Priority: 3 (Normal) } X-MSMail-Priority: Normal } X-Mailer: Microsoft Outlook 8.5, Build 4.71.2232.26 } X-MimeOLE: Produced By Microsoft MimeOLE V4.72.2106.4 } Importance: Normal } X-MS-TNEF-Correlator: 00000000CB8AF9838A20D111B1C200C04FDF38E4A42E5600 } Errors-to: Microscopy-request-at-sparc5.microscopy.com } Content-Type: multipart/mixed; } boundary=3D"----=3D_NextPart_000_0011_01BE6A38.4DAABAE0" } Content-Length: 3968 } Status: = } --====48555149575755524855===1 Content-Type: text/html; charset="US-Ascii" Content-Transfer-Encoding: quoted-printable
{HTML} {HEAD} {/HEAD} {BODY} {PRE = WIDTH=3D"132"} Reply to: RE: Electropolishing of Hf alloy
{/PRE} {FONT FACE=3D"Geneva" SIZE=3D3 COLOR=3D"#000000"} Try = adding about 5% acetic acid, polish at -40 = C, 100 volts. {BR} {BR} Alternate polish: = (worked on V-20Ti) {BR} {BR} 5.3 = g lithium chloride = Temp=3D -50 C {BR} 11.1 g magnesium perchlorate = voltage=3D 190-200 {BR} 500 ml methanol = current=3D 40-50 = mA {BR} 100 ml butyl cellosolve {BR} {BR} = Above done with a South Bay 550-B single = jet polisher in 1985. {BR} A notation mentions = even grain boundaries. {BR} {BR} Bernie = Kestel {BR} Materials Science Division {BR} = Argonne National Laboratory {BR} Argonne, = Il., 60439 E-mail = <kestel-at-anl.gov> {BR} 100 ml = butyl cellosolve {BR} David E. Luzzi wrote: {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >We are electropolishing = a two-phase alloy containing a Laves phase = and a bcc {BR} >solid solution. The bcc = phase is mainly Vanadium and the Laves phase = is {BR} >mainly HfV2. We are experiencing = preferential polishing of the bcc phase {BR} >leading = to marginal TEM specimen quality. {BR} > {BR} >Our = solution is H2SO4 / Methanol / HF / butyl = cellusolve {BR} > {BR} >Does anyone have = any suggestions? Thanks in advance. {BR} > {BR} >David = E. Luzzi {BR} >Department of Materials Science {BR} >University = of Pennsylvania {BR} >3231 Walnut Street {BR} >Philadelphia, = PA 19104-6272 {BR} > {BR} >215-898-8366 {BR} >215-573-2128 = - fax {BR} > {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} luzzi-at-lrsm.= upenn.edu {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} < {/FONT} {FONT FACE=3D"= Geneva" SIZE=3D1 = COLOR=3D"#0000FF"} {U} mailto:luzzi-at-lrsm.upenn.edu {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} > {BR} > {BR} > {BR} > {BR} >RFC822 = header {BR} >----------------------------------- {BR} > {BR} > = Received: from dns2.anl.gov (dns2.anl.gov = [146.139.254.3]) by {BR} >horus.et.anl.gov = (8.6.11/8.6.11) with ESMTP id NAA23584 for = < {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} kestel-at-= horus.et.anl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} >; Tue, 9 {BR} >Mar = 1999 13:42:50 -0600 {BR} > Received: from = Sparc5.Microscopy.Com (sparc5.microscopy.com = {BR} >[206.69.208.10]) by dns2.anl.gov = (8.9.1a/8.6.11) with SMTP id NAA21456; Tue, = 9 Mar 1999 {BR} >13:42:50 -0600 (CST) {BR} > = Received: (from {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = daemon-at-localhost {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} ) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) {BR} >id NAA12717 for dist-Microscopy; = Tue, 9 Mar 1999 13:07:10 -0600 {BR} > Received: = from no_more_spam.com (Sparc5 [206.69.208.10]) = by {BR} >Sparc5.Microscopy.Com (8.6.11/8.6.11) =
Arizona Imaging and Microanalysis Society Annual Spring Meeting Thursday, March 11, 1999 University of Arizona Student Union Sr. Ballroom
There is no registration fee.
8:30 - 9:00 Registration
9:00 - 9:15 Welcome Dr. Clark Lantz, President AIMS
9:15 - 10:15 Microanalysis Society Tour Speaker - Applications of SEM/EDX to forensic cases and research related to food product and pharmaceutical tampering and counterfeiting. Dr. Frank Platek, US Food and Drug Administration
10:15 - 10:30 Break
10:30 - 10:55 Metals as documents: some uses of imaging and microanalysis in African history. Dr. David Killick, Anthropology, University of Arizona
10:55 - 11:20 Visualizing surfactant aggregation with atomic force microscopy Jon Wolgemuth, Physics, University of Arizona
11:20 - 11:45 Multi-parametric analysis of cell function in 2- and 3- dimensions by spectral imaging microscopy Dr. Ron Lynch, Physiology, University of Arizona
11:35 - 1:30 Lunch AIMS Business Meeting
1:30 - 1:55 In situ molecular imaging of stress proteins and oxidative damage Dr. Claire Payne, Microbiology & Immunology University of Arizona
1:55 - 2:20 Determining the functional signficance of cytoskeletal proteins using microinjection and transfection techniques Dr. Carol Gregorio, Cell Biology and Anatomy University of Arizona
2:20 - 2:45 Fiberoptic Confocal Microscope for In Vivo Imaging Dr. Art Gmitro, Radiology, University of Arizona
2:45 - 3:00 Break
3:00 - 5:00 Student Presentations
5:30 - 7:30 Banquet ($12 for dinner, contact Suzanne Kelly {suekelly-at-ag.arizona.edu} to reserve dinner)
Microscopy Society of America Tour Speaker - Digital manipulation of acquired images: What is possible vs what is ethical Dr. Jack Kinnamon, University of Denver
.................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
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Reply to: RE: Processing of micro-injected cells for TEM Dear Raija,
If you are to microinject cells, then the best support is one that gives = access for your needles. I am sure you would prefer to use coverslips. = The material you use will depend on your cells but glass usually works = well. Get the coverslips that have locator grids etched onto them. It is = important to know if you plan to examine the cells for morphology or if = you want to immunolabel them. I will assume you only want to look at = their morphology.
Once you have micro-injected cells and know where they are on the grid (so = don't plate then at too high a density), fix with aldehyde, post fix with = osmium tetroxide, dehydrate and infiltrate with resin (as you would any = piece of tissue for TEM). During the final stages of infiltration (= acetone or propylene oxide) it is wise to transfer the cells, still on the = glass coverslip, to a glass or metal dish. Plastic will dissolve. =
Embed the glass in resin, cells up, at the bottom of an aluminum weigh = boat. Push the glass to the bottom of the dish and pour unpolymerized = resin over. Polymerize by heat and remove from the aluminum dish. Cut = the thin layer of resin away from the back of the coverslip to expose the = glass. Now you can remove the glass by plunging into liquid nitrogen and = rapidly warming few times. It will cause the block to crack but it does = work. Alteratively you can heat the resin, glass-down, on a hot plate and = slide the glass of.. Either way the cells will remain in the resin, as = will the grid locator lines. You should then be able to locate your cells = and cut sections. This is not as easy as I make it seem but it all has = been done before.
If you don't like the idea of embedding at the bottom of a dish, it is = also possible to embed the glass coverslip, cells to the resin, over BEEM = or gelatin capsules, or over flat embedding molds. You will still have to = remove the glass. You could use plastic coverslips and section them too, = but finding your cells will not be easy.
The second part of your question - is there anything you can inject to = identify the cells by EM. Yes, colloidal gold particles can be prepared = that can be microinjected into cells. However, if the gold suspension is = too concentrated it will block the needle. If it is too dilute, it will be= more difficult to detect by EM. =
Immunocytochemistry by request.
Regards,
Paul Webster
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm http://www.hei.org/htm/apw.htm
Raija Sormunen wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
by arl-img-10.compuserve.com (8.8.6/8.8.6/2.18) id DAA22626; Wed, 10 Mar 1999 03:04:50 -0500 (EST)
Hi,
The Royal Microscopical Society (Oxford England) produced a series of safety notes for its members that included - the microscopes, embedding a= nd fixation.
I have had use of some of these recently so I do know that they are still=
available.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Dear all, thanks for the tips preparing liposomes for TEM observations. In summary, most answers dealed with negativ staining of the liposomes. We' ve tried so and got good results for an overview. I added all replies to this meeages for all who're interested in this topic. Bernward 1.Don Gantz wrotes: To all who desired more info on fixing and staining of liposomes-- I apologize for the delay in my response. We had a major snowstorm and I lost a day. The initial query by Bernward Laube was about evaluating homogeneity of liposome suspensions ( {100nm diam) and estimating particle volumes. We have been using a osmium fix/negative stain technique for a number of years to look at size distribution of VLDL, chylomicrons, and synthetic lipid emulsions. The osmium fix firms up these particles nicely so that flattening is greatly reduced. In fact, based upon metal shadowing exps. the slight enlargement of diameter due to flattening after fixation is negated by slight shrinkage of particles during processing. The result is an excellent estimate of diameter and volume.
In regard to lipid vesicles/liposomes, our experience with smaller ones of 50nm or so prepared by sonication and processed with only negative stain is that they are preserved pretty well. We would expect some flattening although we have not shadowed these. However, with larger liposomes of 100nm+ diam, we believe OsO4 will aid in preservation and stability and presumably reduce flattening.
If one requires differentiation of lamellar structure (uni vs multi) among particles within a population, negative staining is unreliable because of variability of stain penetration. For this purpose, we use vitreous ice cryomicroscopy as we are fortunate to have it available.
In general our fixation/negative staining procedure is as follows:
1) Add 1 volume of 2% OsO4 in Cacodylate buffer to 2 volumes of emulsion or liposome prep. Fix for 30 mins.+
2) Place small aliquot on freshly flow-discharged carbon formvar-coated grid for a few seconds.
3) Remove and blot off excess fluid and immediately stain with 1% NaPT.
4) After a few seconds, remove and blot excess fluid. Air dry.
Optimum concentration is trial and error. Fixed suspension can be diluted as needed. Hope this helps. 2.Sheila Garcia wrotes:I've just finished my Doctorate Thesis, and I worked preparing liposome. I used phosphotungstic acid in 2% aq solution, neutralized with KOH 1M (PTK). But, before it, I used bacitracin 0,1mg/ml aq sol., as a wetting agent ( D.W.Gregory and B.J.S. Pirie-Journal of Microscopy, v.99,pt.3, dec. 1973,.261-265). Take a coated grid (carbon, formvar), put a drop of bacitracin sol. for 2 min. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the liposome (a dilueted solution), draw off again and add a drop of PTK for 2 min. I hope it could help you. Profa. Dra. Sheila Garcia
Check out the following paper:
"Sequential treatment by phosphotungstic acid and uranyl acetate enhances the adherence of lipid membranes and membrane proteins to hydrophobic EM grids", A. N. Barnakov, Journal of Microscopy. Vol 175, Pt 2, August 1994, pp. 171-174. 3.Joe Neilly:Don,
Could you elaborate on how you would fix liposomes w/ Osmium tetroxide. We have tried the negative stain approach with some success but have to live with the obvious artifacts such as flattening. There are likely other artifacts caused by ionic or chemical changes of the stain. The chemical fixation sounds promising but I'm not sure how this could be done on a liquid sample. 4.Charles Garber:For your information, we have never been successful in quenching a sample fast enough (the larger sample used for SEM work) in order to keep the vesicles from rupturing. So we do this solely by freeze fracture TEM.
Do you know the name of Richard Banfield in the UK? He pretty much was the first person to really commercialize a cryo-SEM fracture system, when he owned the former Hexland Ltd. company and which was purchased by Oxford Instruments.
As of about three or four years ago, he confirmed to me that he too, had never been able to visualize liposomes via cryo SEM because of the quench rate and rupturing problem.
Should you figure out a way to quench and see the structures by SEM, a lot of us would like to know the secret! 5.Ming Chen:The easiest way to do is by negative staining technique. A 1-2% PTA (phosphotungstic acid) soultion is commonly used. It only take a few minutes to do and you can examine it under TEM to see the distribution of liposomes right away. 6. L.R. Melsen:We have looked at liposome using routine negative staining protocol. The vesicles will flatten upon drying, but simple math can reconstruct the volume of the sphere. 7. Charles Butterick: Try negative staining with a 2% ammonium molybdate (aq). Take a coated grid (carbon, formvar, etc.), put a drop of liposomes on top of the grid for a minute. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the ammonium molybdate. After a minute, draw off drop as before and allow the grid(s) to dry. Take it to the TEM. The above procedure is only a starting point. The concentration, stain, and times can all be varied to achieve optimum results. You might check out some EM texts on negative staining for other ideas. Good luck. 8. Maria Ericsson: When I've prepared liposome suspensions for TEM I've just done a simple negative staining and it's worked fine. I adsorb the suspension to a carbon coated copper grid for about 1 minute, blot off excess liquid with a filterpaper and stain with 1-2% aqueous Uranyl Acetate for 1 minute and blot on a filterpaper again. (If there is phosphate in your buffer, you will need to wash in a drop of water before the Uranyl Acetate staining) 9. Marc Schmutz:To observe liposomes without cryo systems is quit a difficult purpose. Pure lipid systems can not be easily seen in negative staining (try always Uranyl acetate and PTA or other stains). And they generaly undergo severe structural changes during the staining process. So to calculate the volume I will not try to do it and furthermore I will not believe in a volume calculate from negative staining images. Myself I'm observing routinely liposomes pure or with proteins or DNA associated and I always use cryo TEM. It's really a easy approach and also very rapid. (You don't need more than half a day per specimen) As you said you don't have access to such a apparatus but why you don't consider collaborating with a lab equiped in cryo ? If you need some more infos about cryo you can ask me and I will try to help you.
Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit„tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
This is a multi-part message in MIME format. --------------B73E5AA6C975623FC057F918 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
"...reflector may be either a highly polish metal surface or ...coated. The coating consists of a highly brilliant metallic deposit or a dielectric material." Principles & Practice of Laser Technology.
While both wood & paper are dielectric, their fine porosity would preclude 100% reflective. Typical light reflective number for paper would be 70%. Special paper grades can be much higher. In the case of a coherent laser beam, a porous surface or a surface comprised of fine particles would cause significant scattering.
'Laser marker' refers to a family of products that are used to put bar codes and other identifying markings on products. Since they mark by burning/evaporating away the surface, a marking laser would not work on a 100% reflective surface.
J. Roy Nelson Material Testing Laboratory jrnelson-at-nj1.aae.com
"melim-at-qes.po.my"-at-sparc5.microscopy.com wrote: } } Dear List, } Can anyone explain what does "Reflective" means in Laser Terminology . } } I read through a article in a Magazine regarding Laser Marker and in the article the mentioned that Wood and Paper is 100% reflective . I am lost and confuse } } M.E.Lim } Sr Regional Support Engineer } QES(Asia Pacific) Sdn Bhd } Tel : 603-7241188 ext 214 } Fax : 603-7244488 } Emails : melim-at-qes.po.my --------------B73E5AA6C975623FC057F918 Content-Type: text/x-vcard; charset=us-ascii; name="jrnelson.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for jrnelson Content-Disposition: attachment; filename="jrnelson.vcf"
Dear all, thanks for the tips preparing liposomes for TEM observations. In summary, most answers dealed with negativ staining of the liposomes. We' ve tried so and got good results for an overview. I added all replies to this message for all those who're interested in this topic. Bernward 1.Don Gantz wrotes: To all who desired more info on fixing and staining of liposomes-- I apologize for the delay in my response. We had a major snowstorm and I lost a day. The initial query by Bernward Laube was about evaluating homogeneity of liposome suspensions ( {100nm diam) and estimating particle volumes. We have been using a osmium fix/negative stain technique for a number of years to look at size distribution of VLDL, chylomicrons, and synthetic lipid emulsions. The osmium fix firms up these particles nicely so that flattening is greatly reduced. In fact, based upon metal shadowing exps. the slight enlargement of diameter due to flattening after fixation is negated by slight shrinkage of particles during processing. The result is an excellent estimate of diameter and volume.
In regard to lipid vesicles/liposomes, our experience with smaller ones of 50nm or so prepared by sonication and processed with only negative stain is that they are preserved pretty well. We would expect some flattening although we have not shadowed these. However, with larger liposomes of 100nm+ diam, we believe OsO4 will aid in preservation and stability and presumably reduce flattening.
If one requires differentiation of lamellar structure (uni vs multi) among particles within a population, negative staining is unreliable because of variability of stain penetration. For this purpose, we use vitreous ice cryomicroscopy as we are fortunate to have it available.
In general our fixation/negative staining procedure is as follows:
1) Add 1 volume of 2% OsO4 in Cacodylate buffer to 2 volumes of emulsion or liposome prep. Fix for 30 mins.+
2) Place small aliquot on freshly flow-discharged carbon formvar-coated grid for a few seconds.
3) Remove and blot off excess fluid and immediately stain with 1% NaPT.
4) After a few seconds, remove and blot excess fluid. Air dry.
Optimum concentration is trial and error. Fixed suspension can be diluted as needed. Hope this helps. 2.Sheila Garcia wrotes:I've just finished my Doctorate Thesis, and I worked preparing liposome. I used phosphotungstic acid in 2% aq solution, neutralized with KOH 1M (PTK). But, before it, I used bacitracin 0,1mg/ml aq sol., as a wetting agent ( D.W.Gregory and B.J.S. Pirie-Journal of Microscopy, v.99,pt.3, dec. 1973,.261-265). Take a coated grid (carbon, formvar), put a drop of bacitracin sol. for 2 min. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the liposome (a dilueted solution), draw off again and add a drop of PTK for 2 min. I hope it could help you. Profa. Dra. Sheila Garcia
Check out the following paper:
"Sequential treatment by phosphotungstic acid and uranyl acetate enhances the adherence of lipid membranes and membrane proteins to hydrophobic EM grids", A. N. Barnakov, Journal of Microscopy. Vol 175, Pt 2, August 1994, pp. 171-174. 3.Joe Neilly:Don,
Could you elaborate on how you would fix liposomes w/ Osmium tetroxide. We have tried the negative stain approach with some success but have to live with the obvious artifacts such as flattening. There are likely other artifacts caused by ionic or chemical changes of the stain. The chemical fixation sounds promising but I'm not sure how this could be done on a liquid sample. 4.Charles Garber:For your information, we have never been successful in quenching a sample fast enough (the larger sample used for SEM work) in order to keep the vesicles from rupturing. So we do this solely by freeze fracture TEM.
Do you know the name of Richard Banfield in the UK? He pretty much was the first person to really commercialize a cryo-SEM fracture system, when he owned the former Hexland Ltd. company and which was purchased by Oxford Instruments.
As of about three or four years ago, he confirmed to me that he too, had never been able to visualize liposomes via cryo SEM because of the quench rate and rupturing problem.
Should you figure out a way to quench and see the structures by SEM, a lot of us would like to know the secret! 5.Ming Chen:The easiest way to do is by negative staining technique. A 1-2% PTA (phosphotungstic acid) soultion is commonly used. It only take a few minutes to do and you can examine it under TEM to see the distribution of liposomes right away. 6. L.R. Melsen:We have looked at liposome using routine negative staining protocol. The vesicles will flatten upon drying, but simple math can reconstruct the volume of the sphere. 7. Charles Butterick: Try negative staining with a 2% ammonium molybdate (aq). Take a coated grid (carbon, formvar, etc.), put a drop of liposomes on top of the grid for a minute. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the ammonium molybdate. After a minute, draw off drop as before and allow the grid(s) to dry. Take it to the TEM. The above procedure is only a starting point. The concentration, stain, and times can all be varied to achieve optimum results. You might check out some EM texts on negative staining for other ideas. Good luck. 8. Maria Ericsson: When I've prepared liposome suspensions for TEM I've just done a simple negative staining and it's worked fine. I adsorb the suspension to a carbon coated copper grid for about 1 minute, blot off excess liquid with a filterpaper and stain with 1-2% aqueous Uranyl Acetate for 1 minute and blot on a filterpaper again. (If there is phosphate in your buffer, you will need to wash in a drop of water before the Uranyl Acetate staining) 9. Marc Schmutz:To observe liposomes without cryo systems is quit a difficult purpose. Pure lipid systems can not be easily seen in negative staining (try always Uranyl acetate and PTA or other stains). And they generaly undergo severe structural changes during the staining process. So to calculate the volume I will not try to do it and furthermore I will not believe in a volume calculate from negative staining images. Myself I'm observing routinely liposomes pure or with proteins or DNA associated and I always use cryo TEM. It's really a easy approach and also very rapid. (You don't need more than half a day per specimen) As you said you don't have access to such a apparatus but why you don't consider collaborating with a lab equiped in cryo ? If you need some more infos about cryo you can ask me and I will try to help you.
Bernward Laube University of Bielefeld =46aculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=D1tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
We use 4 and 8 well Labtek=A9 Chamber Slides (permanox slides).... substituting Ethanol for the Propylene Oxide. They are my favorite slides for processing cell cultures.
See the following publication for more information:
"Subcellular Localization of SV2 and Other Secretory Vesicle Components in PC12 Cells by an Efficient Method of Preembedding EM Immunocytochemistry for Cell Cultures", Tanner, Ploug and Tao-Cheng, Journal of Histochemistry and Cyrtochemistry, Vol. 44, No. 12, pp. 1481-1488, 1996.
If you have any questions.. feel free to contact me.
Virginia Tanner Crocker
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******************************************************************* Virginia Tanner Crocker Biologist NIH, NINDS EM Facility, Bldg 36, Room 3B24 Bethesda, MD 20892
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Thanks to all who replied. The ETEC has been spoken for. If for some = reason it does not leave as scheduled I will keep the names of those who = responded, just in case, since it MUST GO.
Thanks again, Judy Murphy
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
by mail-ewr-3.pilot.net (Pilot/8.8.8) with ESMTP id MAA19999; Wed, 10 Mar 1999 12:27:37 -0500 (EST) Received: from ridexch1.rid.com ([148.189.116.16]) by mailgw.bi-pharm.com with ESMTP id MAA24037; Wed, 10 Mar 1999 12:28:05 -0500 (EST) Received: by RIDEXCH1 with Internet Mail Service (5.5.2232.9) id {F437PDQP} ; Wed, 10 Mar 1999 12:27:36 -0500 Message-ID: {5063A0AB7328D211BCAA0008C7A4467704758B-at-RIDMSG05}
Sandy:
The p.o. step is not necessary. Dehydration with any of the 812 replacements can be done through EtOH alone. I use at least 3x 100%, then grade thru the EtOH:epoxy at 2:1, 1:1 and 1:2, then into pure resin. The only caveat is that the EtOH and resin must be very carefully mixed--both to ensure complete mixing and to avoid the creation of bubbles in the mix. I also use only freshly prepared resin once I reach the 1:1 stage, but have had no problem using older batches that were stored at -20 C and thawed just prior to use.
Roger Moretz Toxicology & Safety Assessment
} -----Original Message----- } From: Sandy Perkins [SMTP:skperkin-at-vt.edu] } Sent: Tuesday, March 09, 1999 11:01 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM-cells on Permanox slides } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi- } } I just scanned through postings about processing tissue cultures and } coverslips for TEM, but I didn't see anything on processing cells grown on } 8 well Permanox slides. I will be embedding in PolyBed 812, with a } transition through propylene oxide. Unfortunately, the wells don't } survive } the p.o. step. I would appreciate hearing about any experiences with } Permanox slides. Thank you very much. } } Sandy Perkins } } Laboratory for Neurotoxicity Studies } Virginia-Maryland Regional College } of Veterinary Medicine } Virginia Tech } }
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Dear all, thanks for the tips preparing liposomes for TEM observations. I= n summary, most answers dealed with negativ staining of the liposomes. We' ve tried so and got good results for an overview. I added all replies to this message for all those who're interested in this topic. Bernward 1.Don Gantz wrotes: To all who desired more info on fixing and staining of liposomes-= - I apologize for the delay in my response. We had a major snowstorm and I lost a day. The initial query by Bernward Laube was about evaluating homogeneity of liposome suspensions ( {100nm diam) and estimating particle volumes. We have been using a osmium fix/negative stain technique for a number of years to look at size distribution of VLDL, chylomicrons, and syntheti= c lipid emulsions. The osmium fix firms up these particles nicely so that flattening is greatly reduced. In fact, based upon metal shadowing exps. the slight enlargement of diameter due to flattening after fixation is negated by slight shrinkage of particles during processing. The result i= s an excellent estimate of diameter and volume.
In regard to lipid vesicles/liposomes, our experience with smaller ones of 50nm or so prepared by sonication and processed with only negative stain is that they are preserved pretty well. We would expect some flattening although we have not shadowed these. However, wit= h larger liposomes of 100nm+ diam, we believe OsO4 will aid in preservation and stability and presumably reduce flattening.
If one requires differentiation of lamellar structure (uni vs multi) among particles within a population, negative staining is unreliable because of variability of stain penetration. For this purpose, we use vitreous ice cryomicroscopy as we are fortunate to have it available.
In general our fixation/negative staining procedure is as follows:
1) Add 1 volume of 2% OsO4 in Cacodylate buffer to 2 volumes of emulsion or liposome prep. Fix for 30 mins.+
2) Place small aliquot on freshly flow-discharged carbon formvar-coated grid for a few seconds.
3) Remove and blot off excess fluid and immediately stain with 1% NaPT.
4) After a few seconds, remove and blot excess fluid. Air dry.
Optimum concentration is trial and error. Fixed suspension can be diluted as needed. Hope this helps. 2.Sheila Garcia wrotes:I've just finished my Doctorate Thesis, and I worked preparing liposome. I used phosphotungstic acid in 2% aq solution, neutralized with KOH 1M (PTK). But, before it, I used bacitracin 0,1mg/ml aq sol., as a wetting agent ( D.W.Gregory and B.J.S. Pirie-Journal of Microscopy, v.99,pt.3, dec. 1973,.261-265). Take a coated grid (carbon, formvar), put a drop of bacitracin sol. for 2 min. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the liposome (a dilueted solution), draw off again and add a drop of PTK for = 2 min. I hope it could help you. Profa. Dra. Sheila Garcia
Check out the following paper:
"Sequential treatment by phosphotungstic acid and uranyl acetate enhances the adherence of lipid membranes and membrane proteins to hydrophobic EM grids", A. N. Barnakov, Journal of Microscopy. Vol 175, Pt 2, August 1994, pp. 171-174. 3.Joe Neilly:Don,
Could you elaborate on how you would fix liposomes w/ Osmium tetroxide. W= e have tried the negative stain approach with some success but have to live with the obvious artifacts such as flattening. There are likely other artifacts caused by ionic or chemical changes of the stain. The chemical fixation sounds promising but I'm not sure how this could be done on a liquid sample. 4.Charles Garber:For your information, we have never been successful in quenching a sample fast enough (the larger sample used for SEM work) in order to keep the vesicles from rupturing. So we do this solely by freeze fracture TEM.
Do you know the name of Richard Banfield in the UK? He pretty much was the first person to really commercialize a cryo-SEM fracture system, when he owned the former Hexland Ltd. company and which was purchased by Oxfor= d Instruments.
As of about three or four years ago, he confirmed to me that he too, had never been able to visualize liposomes via cryo SEM because of the quench rate and rupturing problem.
Should you figure out a way to quench and see the structures by SEM, a lo= t of us would like to know the secret! 5.Ming Chen:The easiest way to do is by negative staining technique. A 1-2% PTA (phosphotungstic acid) soultio= n is commonly used. It only take a few minutes to do and you can examine it under TEM to see the distribution of liposomes right away. 6. L.R. Melsen:We have looked at liposome using routine negative staining protocol. The vesicles will flatten upon drying, but simple math can reconstruct the volume of the sphere. 7. Charles Butterick: Try negative staining with a 2% ammonium molybdate (aq). Take a coated grid (carbon, formvar, etc.), put a drop of liposomes on top of the grid for a minute. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the ammonium molybdate. After a minute, draw off drop as before and allow the grid(s) to dry. Take it to the TEM. The above procedure is only a starting point. The concentration, stain, and times can all be varied to achieve optimum results. You might check out some EM text= s on negative staining for other ideas. Good luck. 8. Maria Ericsson: When I've prepared liposome suspensions for TEM I've just done a simple negative staining and it's worked fine. I adsorb the suspension to a carbon coated copper grid for about 1 minute, blot off excess liquid with a filterpaper and stain with 1-2% aqueous Uranyl Acetate for 1 minute and blot on a filterpaper again. (If there is phosphate in your buffer, you will need to wash in a drop of water before the Uranyl Acetate staining) 9. Marc Schmutz:To observe liposomes without cryo systems is quit a difficult purpose. Pure lipid systems can not be easily seen in negative staining (try always Uranyl acetate and PTA or other stains). And they generaly undergo severe structural changes during the staining process. So to calculate the volume I will not try to do it and furthermore I will not believe in a volume calculate from negative staining images. Myself I'm observing routinely liposomes pure or with proteins or DNA associated and I always use cryo TEM. It's really a easy approach and also very rapid. (You don't need more than half a day per specimen) As you said you don't have access to such a apparatus but why you don't consider collaborating with a lab equiped in cryo ? If you need some more infos about cryo you can ask me and I will try to help you.
Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=D1tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
Hello, Does anyone have a suggesstion on how to fix/reset the advance mechanism to a reichert e ultramicrotome. The mechanism on the left side (0.5 um to 2 um advance) does not work, I have to get close with the coarse knife advance then use the electronic advance which makes alignment tedious. Thanks
We are thinking of purchasing a negative scanner for use with TEM negatives from our Jeol 2000-FX, and also for SEM negatives. A scanner has been recommended to us: the Agfa Duoscan T2500, which has a resolution (hardware) of 1250 x 2500 dpi and a density of 3.4D. (The scans would be output to a Kodak DS 8650 PS printer)
We need the quality of the scans to match the quality of the standard darkroom enlarger if possible, as we would like to 'go digital' at least for routine work. Does anyone have experience of routine negative scanning for TEM prints, with this or other scanners, and if so, is it realistic to expect such high quality?
Also, what additional image processing software would people recommend we got to go along with this?
Any advice would be appreciated.
Caspar
Caspar McConville, Ph.D. Technical Specialist New York State College of Ceramics Alfred University
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
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Dear all, thanks for the tips preparing liposomes for TEM observations. I= n summary, most answers dealed with negativ staining of the liposomes. We' ve tried so and got good results for an overview. I added all replies to this message for all those who're interested in this topic. Bernward 1.Don Gantz wrotes: To all who desired more info on fixing and staining of liposomes-= - I apologize for the delay in my response. We had a major snowstorm and I lost a day. The initial query by Bernward Laube was about evaluating homogeneity of liposome suspensions ( {100nm diam) and estimating particle volumes. We have been using a osmium fix/negative stain technique for a number of years to look at size distribution of VLDL, chylomicrons, and syntheti= c lipid emulsions. The osmium fix firms up these particles nicely so that flattening is greatly reduced. In fact, based upon metal shadowing exps. the slight enlargement of diameter due to flattening after fixation is negated by slight shrinkage of particles during processing. The result i= s an excellent estimate of diameter and volume.
In regard to lipid vesicles/liposomes, our experience with smaller ones of 50nm or so prepared by sonication and processed with only negative stain is that they are preserved pretty well. We would expect some flattening although we have not shadowed these. However, wit= h larger liposomes of 100nm+ diam, we believe OsO4 will aid in preservation and stability and presumably reduce flattening.
If one requires differentiation of lamellar structure (uni vs multi) among particles within a population, negative staining is unreliable because of variability of stain penetration. For this purpose, we use vitreous ice cryomicroscopy as we are fortunate to have it available.
In general our fixation/negative staining procedure is as follows:
1) Add 1 volume of 2% OsO4 in Cacodylate buffer to 2 volumes of emulsion or liposome prep. Fix for 30 mins.+
2) Place small aliquot on freshly flow-discharged carbon formvar-coated grid for a few seconds.
3) Remove and blot off excess fluid and immediately stain with 1% NaPT.
4) After a few seconds, remove and blot excess fluid. Air dry.
Optimum concentration is trial and error. Fixed suspension can be diluted as needed. Hope this helps. 2.Sheila Garcia wrotes:I've just finished my Doctorate Thesis, and I worked preparing liposome. I used phosphotungstic acid in 2% aq solution, neutralized with KOH 1M (PTK). But, before it, I used bacitracin 0,1mg/ml aq sol., as a wetting agent ( D.W.Gregory and B.J.S. Pirie-Journal of Microscopy, v.99,pt.3, dec. 1973,.261-265). Take a coated grid (carbon, formvar), put a drop of bacitracin sol. for 2 min. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the liposome (a dilueted solution), draw off again and add a drop of PTK for = 2 min. I hope it could help you. Profa. Dra. Sheila Garcia
Check out the following paper:
"Sequential treatment by phosphotungstic acid and uranyl acetate enhances the adherence of lipid membranes and membrane proteins to hydrophobic EM grids", A. N. Barnakov, Journal of Microscopy. Vol 175, Pt 2, August 1994, pp. 171-174. 3.Joe Neilly:Don,
Could you elaborate on how you would fix liposomes w/ Osmium tetroxide. W= e have tried the negative stain approach with some success but have to live with the obvious artifacts such as flattening. There are likely other artifacts caused by ionic or chemical changes of the stain. The chemical fixation sounds promising but I'm not sure how this could be done on a liquid sample. 4.Charles Garber:For your information, we have never been successful in quenching a sample fast enough (the larger sample used for SEM work) in order to keep the vesicles from rupturing. So we do this solely by freeze fracture TEM.
Do you know the name of Richard Banfield in the UK? He pretty much was the first person to really commercialize a cryo-SEM fracture system, when he owned the former Hexland Ltd. company and which was purchased by Oxfor= d Instruments.
As of about three or four years ago, he confirmed to me that he too, had never been able to visualize liposomes via cryo SEM because of the quench rate and rupturing problem.
Should you figure out a way to quench and see the structures by SEM, a lo= t of us would like to know the secret! 5.Ming Chen:The easiest way to do is by negative staining technique. A 1-2% PTA (phosphotungstic acid) soultio= n is commonly used. It only take a few minutes to do and you can examine it under TEM to see the distribution of liposomes right away. 6. L.R. Melsen:We have looked at liposome using routine negative staining protocol. The vesicles will flatten upon drying, but simple math can reconstruct the volume of the sphere. 7. Charles Butterick: Try negative staining with a 2% ammonium molybdate (aq). Take a coated grid (carbon, formvar, etc.), put a drop of liposomes on top of the grid for a minute. Draw off drop with with a piece of torn filter paper. Before the grid dries, add a drop of the ammonium molybdate. After a minute, draw off drop as before and allow the grid(s) to dry. Take it to the TEM. The above procedure is only a starting point. The concentration, stain, and times can all be varied to achieve optimum results. You might check out some EM text= s on negative staining for other ideas. Good luck. 8. Maria Ericsson: When I've prepared liposome suspensions for TEM I've just done a simple negative staining and it's worked fine. I adsorb the suspension to a carbon coated copper grid for about 1 minute, blot off excess liquid with a filterpaper and stain with 1-2% aqueous Uranyl Acetate for 1 minute and blot on a filterpaper again. (If there is phosphate in your buffer, you will need to wash in a drop of water before the Uranyl Acetate staining) 9. Marc Schmutz:To observe liposomes without cryo systems is quit a difficult purpose. Pure lipid systems can not be easily seen in negative staining (try always Uranyl acetate and PTA or other stains). And they generaly undergo severe structural changes during the staining process. So to calculate the volume I will not try to do it and furthermore I will not believe in a volume calculate from negative staining images. Myself I'm observing routinely liposomes pure or with proteins or DNA associated and I always use cryo TEM. It's really a easy approach and also very rapid. (You don't need more than half a day per specimen) As you said you don't have access to such a apparatus but why you don't consider collaborating with a lab equiped in cryo ? If you need some more infos about cryo you can ask me and I will try to help you.
Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=D1tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
I'm trying to cobble together a new probe. So far, I have one WDS spectrometer which used to be on an 840. Can anyone tell me exactly which JEOL SEMs it will fit on to? I am pretty sure it will go onto 6300 and 6400, and maybe also the 733. So far I've had no luck getting this info from JEOL, but if someone within their organisation can help, great. Also, does anyone know whether a two-crystal spectro can be transformed into a 4-crystal type? I've been told that it can't be done in the field, but can the factory do it?
Anybody got any more of these spectros that they'd be willing to sell? And maybe also an 840 or 840A?
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I have been very happy with the results from my Polaroid Sprintscan 45 (~$8500). For a TEM neg, it will digitize at 2000dpi and it will output 12 bits. I can't remember the optical density but I think it is similar to what you are quoting here, but you can look it up on their web site. For a 35 mm slide, it will do 4000 dpi. You have to ask them for a special TEM negative carrier that fits into their 4x5 holder. I use 300 dpi as the standard for what kind of enlargement I can get because my HP 890C (which does a great job on photo deluxe paper) is 300 dpi and the two sub-dye printers that I have access to are both 300 dpi. This gives a usable enlargement factor of 2000/300 = 6.7x printing to these printers. It is also very quick. I think that Polaroid could definitely improve their user control interface a bit, but for the most part it works well. I have had some problems with the computer recognizing the scanner on two systems, (one with a Sprintscan 35 and the other with a Sprintscan 45) that have flatbed scanners attached and that are on. The solution is simply to turn the flatbed power off and reopen the program.
I think that the Duoscan is a flatbed. You have to be careful with putting the negatives on the glass because you can get Newton rings in you images. I have a Umax Powerlook II flatbed that is 600 x 1200 dpi that I sometimes use and you can sometimes see them. You need a mask to lift it off the glass. I have asked the listserver in the past if that defocuses the scanned image, but I did not get a satisfactory answer. I use the flatbed as a contact printer by scanning 6 images at once held in Neg-a-file sheets and digitizing to 150 dpi. This is the minimum value that I can still read the numbers on the JEOL negatives. This is actually better than making contacts prints in the darkroom, because I can select areas to adjust the contrast and brightness independently. This makes BF, DF, and diffraction images come out well even if they are on one sheet.
I highly recommend Adobe Photoshop (version 4 is what I have) coupled with John Russ' Image Processing Toolkit Plug-ins. I Import the images as 12 bits (16bit), adjust the levels to what I want, and then convert the images to 8 bits. You can then use all of Photoshop's features.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Caspar McConville To: Microscopy List Server -----------------------------------------------------------------------.
We are thinking of purchasing a negative scanner for use with TEM negatives from our Jeol 2000-FX, and also for SEM negatives. A scanner has been recommended to us: the Agfa Duoscan T2500, which has a resolution (hardware) of 1250 x 2500 dpi and a density of 3.4D. (The scans would be output to a Kodak DS 8650 PS printer)
We need the quality of the scans to match the quality of the standard darkroom enlarger if possible, as we would like to 'go digital' at least for routine work. Does anyone have experience of routine negative scanning for TEM prints, with this or other scanners, and if so, is it realistic to expect such high quality?
Also, what additional image processing software would people recommend we got to go along with this?
Any advice would be appreciated.
Caspar
Caspar McConville, Ph.D. Technical Specialist New York State College of Ceramics Alfred University
One approach might be to plunge freeze on filmed grids and then clamp these to a standard cryoSEM specimen support. The support could have a hole drilled through it for STEM. Because of the water film thickness, it could be sublimed by freeze drying, although any dissolved salts would be left behind.
A similar approach would be to quench the liposomes on something like aluminium foil, in a size & shape which could then be clamped under a thin ring which is drilled to screw to a standard suppport. These attachments can be done quite easily under a shallow depth of liquid nitrogen in a polystyrene container.
Hoping these ideas are useful to someone
Keith Ryan Marine Biological Association Plymouth, UK
Hello, The Newtonian Ring problem might be corrected the way it was done on anti-Newton glass slides, in days gone by. They used glass which was slightly etched on the side which went next to the film. I am not suggesting you etch the glass face of your scanner but it might be worth experimenting on a spare piece of glass. The etch is ever so slight; maybe just acid fumes would do the trick. This didn't seem to degrade projected slides. Just a suggestion. Good luck.
Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. Number 499, Post Office Box 19400 Austin, Texas 78760 Phone 512/282-5507 Fax 512/280-0702
TEM & SEM Maintenance -----Original Message----- } From: Walck. Scott D. {walck-at-ppg.com} To: Caspar McConville {mcconville-at-olsen.alfred.edu} ; Micro {microscopy-at-Sparc5.Microscopy.Com}
Tamara, The copy I have of "Safety in the EM Lab..." is from San Francisco Press, ISBN 0-911302-56-5, 1985. You should be able to get it ordered with that info. Good luck.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor 09 Mar 1999 4:30 PM CRC-Electron Microscopy Lab Ofc: 704-355-1267 Carolinas Medical Center Lab: 704-355-7220 P.O. Box 32861 Fax: 704-355-7648 Charlotte, NC 28232-2861 USA Eml: WWiggins-at-Carolinas.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
} -----Original Message----- } From: Tamara Howard [SMTP:howard-at-cshl.org] } Sent: Tuesday, March 09, 1999 9:49 AM } To: Microscopy Listserver } Subject: EM safety book? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone have or know of a good reference on EM lab safety? I thought } there was a book called "Safety in the EM Laboratory"...but I haven't been } able to find it. Probably imagined it. } } Thanks for any help anyone can give! } } Tamara Howard } CSHL } }
I think you are making an excellent choice with the new Agfa DuoScan T2500. You will appreciate the 3.4 D on this unique flatbed scanner. The glass less negative carriers fit into the lower tray, which slides into a precise position. The negatives will not contact any surface. I have just ordered also a DuoScan T2500. You will learn more about it at:
One of the most versatile imaging software is CorelDraw8 and CorelPHOTO-PAINT8. It offers the highest image format manipulation with Plug-ins, for most applications. Highly recommended.
We are looking for commercially available options for being able to transfer a metallographic type sample from a sample preparation workstation and into an SEM while under vacuum or inert atmosphere. We have a number of make and model SEMs therefore just any general information is fine on this subject.
thanks,
Mark A. Wall
Mr. Mark A. Wall Sr. Scientific Assoc. L-350 Chemistry & Materials Science Directorate Lawrence Livermore National Laboratory Livermore, CA USA 94550
I think you are making an excellent choice with the new Agfa DuoScan T2500. You will appreciate the 3.4 D on this unique flatbed scanner. The glass less
negative carriers fit into the lower tray, which slides into a precise position. The negatives will not contact any surface. I have just ordered also a DuoScan T2500. You will learn more about it at:
One of the most versatile imaging software is CorelDraw8 and CorelPHOTO-PAINT8. It offers the highest image format manipulation with Plug-ins, for most applications. Highly recommended.
Laszlo J. Veto
Caspar McConville wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are thinking of purchasing a negative scanner for use with TEM } negatives from our Jeol 2000-FX, and also for SEM negatives. A } scanner has been recommended to us: the Agfa Duoscan T2500, which } has a resolution (hardware) of 1250 x 2500 dpi and a density of 3.4D. } (The scans would be output to a Kodak DS 8650 PS printer) } } We need the quality of the scans to match the quality of the standard } darkroom enlarger if possible, as we would like to 'go digital' at } least for routine work. Does anyone have experience of routine } negative scanning for TEM prints, with this or other scanners, and if } so, is it realistic to expect such high quality? } } Also, what additional image processing software would people } recommend we got to go along with this? } } Any advice would be appreciated. } } Caspar } } Caspar McConville, Ph.D. } Technical Specialist } New York State College of Ceramics } Alfred University
I just want to say THANK YOU to all who responded to my question regarding the electrolytical thinning of Al. Now I own a collection of serveral different recipes! If anybody else is interested in it, just let me know.
Cheers,
Petra -------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu Visit our WWW site! http://www.crpcu.lu/~wahlbrin
My first posting was placed whilst away from the office, now I am back I can give you the full details of the safety data mentioned.
1. Safety in the Electron Microscope Room - S. K. Chapman, Microscopy &=
Analysis, March 89, 27-29, (only two pages of text)
2. Routine Handling of Resins, RMS E.M. Safety Committee, Single sheet handout
OR from one of the same group
3. Resins: Toxicity, Hazards and Safe Handling - B. E. Causton, Proceedings RMS, Vol 16/4, June 81, 265-269
4. Routine Handling of Fixatives, RMS E.M. Safety Committee, Single shee= t handout.
I am prepared to scan in part of 1, plus 2 and 3 and send as attachments direct to those who ask.
As you can see I was not quite correct they are not all Royal Microscopic= al Society publications. Their address is 37/38 St. Clements, Oxford OX4 1AJ= , England
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
I believe the two scanners are both excellent, although I have a preference for the Polaroid, that I am also going to buy (if can get the money).
As far as being capable of being able to use the scanner instead ot the darkroom, I strongly believe that both scanner technology, and computer technology is not yet capable of completely replacing a darkroom, unfortunately, for all its applications.
Would appreciate very much a feedback on this subject.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
May I offer a word of caution about the approach below?
While you're correct in saying you need to optimize printing conditions for your image and printer, there will probably be a little more to it than setting up comparable number of pixels. Your image is probably either a 256-level gray scale or a 16 million color (256 levels of red, green and blue). An inkjet can't print that kind of color depth in each pixel. The concepts of "halftoning" or "dithering" need to be considered. As I'm far from the expert, and we don't need a complete textbook on the listserver anyway, so I'll refer you to chapter 2 of "The Image Processing Handbook" by John Russ. As we would expect from Dr. Russ, the material is excellent.
That does bring up a question for me, though. The new HP inkjets have a "PhotoREt" technology which, I believe is supposed to be able to vary the size of the dots it produces, therefore producing better photo-printing results. Has anyone determined whether this is true, or just hype?
Jim Passmore Sr. Analytical Chemist Cryovac Division Sealed Air Corporation
---------- } From: Harry.Ekstrom } To: Microscopy } Subject: FW: Printers for SEM Images } Date: Monday, March 08, 1999 5:11PM } ------------- } } I think an inexpensive inkjet is the way to go. However, you must try to } set the DPI of the printer to match the resolution setting of your digital } images. If you capture a digital image at 1024 X800 for instance, you have } about 820K of information. Now lets say you plan to print a 4X5 image } similar to a Polaroid, then your printer should be no less than 300dpi } {4"x5" x (300)2=1.8Mpixel} to accommodate the amount of pixel information } from the capturing rate of the image. Hence, a 2048X1600 resolution setting } captures 3.2 Mpixels of info, so a 400DPI setting should be used. The idea } being to match the capturing info with the amount of pixels the printer can } resolve to minimize interpolation...be it upwards or downwards. Not sure } what the human eye can resolve tho. } } Good Luck, } Harry Ekstrom } }
The M & M '99 Golf tournament will be Sunday, Aug. 1, 1999 just outside Portland, OR. Hole sponsorships are available for $65.00 each on a first come, first serve basis. Prizes for longest drive, longest putt etc. can also be donated. In addition, Logo gifts to be distributed to all participants will also be most welcome. To reserve your holes or to supply gifts/prizes etc. please notify me as soon as possible.
Thank you,
John Arnott Chairman
-- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
We have an aging BioRad Confocal (1991!)with the associated computer and software (COMOS ver.7.0a). Has anyone replaced/upgraded their computer on this system other than just purchasing an upgrade from BioRad? What problems were encountered and what were some solutions? Thanks in advance for any info.
History of unit is below: Like an old car, different parts of the computer are beginning to need replacement, but I understand that several of the boards are specialized propietary boards (control of scan head and frame grabber) and cannot be replaced with conventional boards. We want to "upgrade" the computer, ie new mother board, more memory, etc... The existing mother board is not compatible with any device drivers other than SCSIs.
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu ********************************************
I need to know the wavelength of the main ~ 441.6nm emission line from our He-Cd laser source to at least 5 significant figures, if possible (wavelength in air atmosphere). Any references containing other related data (emission lines) would be useful too.
-- ***************************************** Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
I have recently taken over an SEM with an AN 10000 analyser and I was wondering if anyone out there kows if it is possible to get the results files out to a PC ?
Any help greatly appreciated.
******************************************* Robert McDonald EPMA & SEM Laboratories Dept Geography - Earth Sciences Division Gregory Building LilyBank Gardens University of Glasgow Glasgow G12 8QQ Scotland, UK email: robert-at-earthsci.gla.ac.uk Tel:- +44 (0)141 330 5505/5442 FAX:- +44 (0)141 330 4817 ********************************************
Reply to: RE: SEM vacuum transfer = For many years I used a simple permanent magnet to lift and transfer = samples mounted on metallographic mounts with a steel washer epoxied onto = the back surface. A hole in the circular magnet allowed a push rod to = release the mount from the magnet. Possibly a small electromagnet could = be made that would be easier to operate in your system-or even a vacuum = powered suction cup to eliminate magnetic fields. Most likely you will = need to make one. = Bernard Kestel Materials Science Division Argonne National Laboratory Argonne, Il., 60439
Phone: (630) 252-4945 E-mail {kestel-at-anl.gov} Mark Wall wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
{HTML} {HEAD} {/HEAD} {BODY} {PRE = WIDTH=3D"132"} Reply to: RE: SEM vacuum transfer =
{/PRE} {FONT FACE=3D"Geneva" SIZE=3D3 COLOR=3D"#000000"} For = many years I used a simple permanent magnet = to lift and transfer samples mounted on = metallographic mounts with a steel washer = epoxied onto the back surface. A hole in = the circular magnet allowed a push rod to = release the mount from the magnet. Possibly = a small electromagnet could be made that = would be easier to operate in your system-or = even a vacuum powered suction cup to eliminate = magnetic fields. Most likely you will need = to make one. {BR} {BR} Bernard Kestel {BR} = Materials Science Division {BR} Argonne = National Laboratory {BR} Argonne, Il., = 60439 {BR} {BR} Phone: (630) 252-4945 = E-mail <kestel-at-anl.gov> {BR} Mark = Wall wrote: {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >-----------------------------------------------------------------------= - {BR} >The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America {BR}
Peggy, Can you please contact me with your address, at your convenience? My computer crashed and I lost your e-mail, and snail mail addresses. I'll send the nerve staining info out when you reply. Best wishes Ronnie Houston Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children 2222 Welborn Street Dallas, TX 75219
I am looking for any references that can give the emission wavelength of a HeCd laser to five significant figures (in air/vacuum). The line inparticular is the 441.6 nm line, but a table of up to date measurements would be useful too.
-- ***************************************** Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
} We are thinking of purchasing a negative scanner...
Check out the Imacon FlexTight II. This is an affordable drum scanner with magnetic carriers for various size media (2x2 skides up to at least 8x10 sheeets) that make it as easy to use as a flatbed scanner. The unit has 5,760 dpi optical resolution (although this may drop to 4,800 dpi for something the size of an EM negative), 12-bit grayscale, 24, 32 and 48-bit color, and 14 bits (4.1 OD) of dynamic range, and it is fast. For image editing, Photoshop is the best. Period.
} That does bring up a question for me, though. The new HP } inkjets have a "PhotoREt" technology which, I believe is } supposed to be able to vary the size of the dots it produces, } therefore producing better photo-printing results. Has anyone } determined whether this is true, or just hype?
HP's statement, in itself, is explicid, so I doubt they could say it if it weren't true. I own one of these printers (720C) and looking closely it is hard to tell just how small the dots are, or when and where there being used. Appearance is very close to "random dithering". A couple of clues which would lead you to believe the technology is not just hype: (1) the suggested DPI for the printer is 300dpi which is beyond what would be calculated for a normal 600dpi dithering printer, and (2) dithering can not be seen at all for the primary colors reb, green & blue as created with cyan, magenta and yellow.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
A researcher would like to be able to tell the difference between apototic cells and necrotic and/or other degenerating cells in an invertebrate nervous system, using TEM. So far the only general statement I've come across is that apototic cells will undergo autophagy within their plasma membranes whereas necrotic cells tend to spill their contents and get cleaned up by other cells.
Any additional tips will be appreciated!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I've just found a neat screen capture (and more) utility called HyperSnap-DX, a free trial version can be had from www.hyperionics.com, and registration is only $25!
cheers
rtch
} Have a couple of avi movies here and we need some stills printed from them. } Anyone have any ideas/shareware/freeware? } } } Scott D. Whittaker 218 Carr Hall } EM Technician Gainesville, FL 32610 } University Of Florida ph 352-392-1184 } ICBR EM Core Lab fax 352-846-0251 } sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ } The home of " Tips & Tricks " }
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I'm not a microscopy expert, but a retired computer systems engineer.
An inkjet can produce high color if it uses CMYK inks (vs RGB). Photo-realistic inkjet printers don't dither (screen) or halftone such as with b&w printing. They attempt to produce a 1-1 relationship with the input color data. } } That does bring up a question for me, though. The new HP } inkjets have a "PhotoREt" technology which,
H-P's RET technology has been around for quite some time. I find it introduces distracting patterns into the hardcopy. H-P sat on its laurels for quite some time, and most experts say that they have fallen behind. Unless they've changed, H-Ps use RGB ink.
I find that the Epson and newer Cannon printers do a credible job of photo-color, both are CMYK.
However, you will find that the proper software will make a bigger difference in the results than the printer. If you want high color fidelity, then I recommend Adobe Photoshop, (even if you only use it for printing), calibrated for the printers ink that you use. (Epson and Cannon ink files are provided for Photoshop.) If you expect to manipulate color on the computer screen and then print the same colors on the printer, your computer has to have a "color system" such as Kodak ColorSync (adjusts for your scanner, screen and printer). Color Systems are pretty much free on the Macintosh, but you are may be out of luck if you have a Windows box (I've not found one that works very well).
Your Color System has to "know" your hardware unless you have a means of calibrating. (UMAX scanners provide a means to calibrate from a Kodak target.)
The confusion arises out of the difference between the well established printing industry and the newer technology of photorealistic inkjet printing.
In short: make sure you can calibrate your scanner for color and that it comes with a color system,
that ink files are available for your printer's ink for use by photoshop, (even if you don't use Photoshop, other programs use the PS as a standard),
I will agree wholehearted that Adobe PhotoShop is best, but I'll bet that most people never learn it properly to get their $600 worth. A very impressive PhotoShop clone is Ulead Photo Impact ( http://www.ulead.com {http://www.ulead.com} ) for ~$90.00 which does what most people use PhotoShop for (TWAIN compliant, Image resizing and level adjustments), plus Web-based image production is built in, not an add-on.) Try their 15-day fully functional demo. Enough said on image editors.
Just passing along some unbiased information.
Walt Bobrowski Subcellular Pathology Parke-Davis Research 2800 Plymouth Road Ann Arbor, MI 48105
Over the years our workforce has dwindled to the point that I find myself = working alone in the lab (metalographic sample prep/optical microscopy + = image analysis) almost all the time. The lab doesn't have windows to the = hallway, so it is difficult for people walking by to see if I am in the = lab, and if I am OK. My manager is concerned about my safety and is asking = me for suggestions. What do other labs do to make working alone safe? Everett Ramer Federal Energy Technology Center
I have replaced the original Compaq 386 with a Dell 486. I had no problems with the BioRad MRC 600 system and comos software. BioRad people have told of problems with some computers but I haven't seen any.
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
This announcement is for colleagues of Don W. Fawcett, M.D. ("A textbook = of histology", Bloom & Fawcett and "The Cell"), Professor Emeritus, = Harvard University. =
This coming Sunday (3/14/99) he will be celebrating his 82nd birthday. A = few e-mail greetings might surprize and please him. His e-mail address is = {DFawc20586-at-aol.com} . =
I am sure he would appreciate good wishes from anyone inspired by his = books and papers too.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm --====48525552525549495651===1 Content-Type: text/html; charset="US-Ascii" Content-Transfer-Encoding: quoted-printable
{HTML} {HEAD} {/HEAD} {BODY} {FONT FACE=3D"Monaco" = SIZE=3D1 COLOR=3D"#000000"} Dear Microscopists and Histologists, {BR} {BR} This = announcement is for colleagues of Don W. = Fawcett, M.D. ("A textbook of histology", = Bloom & Fawcett and "The Cell"), = Professor Emeritus, Harvard University. = {BR} {BR} This coming Sunday (3/14/99) he will = be celebrating his 82nd birthday. A few = e-mail greetings might surprize and please = him. His e-mail address is < {/FONT} {FONT FACE=3D"Monaco" = SIZE=3D1 COLOR=3D"#0000FF"} {U} DFawc20586-at-aol.com {/U} {/FONT} {FONT FACE=3D"= Monaco" = SIZE=3D1 COLOR=3D"#000000"} >. {BR} {BR} I am sure he would = appreciate good wishes from anyone inspired = by his books and papers too. {BR} {BR} Paul Webster, = Ph.D {BR} House Ear Institute {BR} 2100 West Third = Street {BR} Los Angeles, CA 90057 {BR} phone:213 = 273 8026 {BR} fax: 213 413 6739 {BR} e-mail: {/FONT} {FONT = FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#0000FF"} {U} pwebster-at-hei.org {/U} {/FONT} {= FONT = FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#000000"} {BR} {/FONT} {FONT FACE=3D"Monaco" SIZE=3D1 COLOR=3D"#0000FF"} {U} http://www.hei.= org/htm/aemi.htm {/U} {/FONT} {/BODY} {/HTML} --====48525552525549495651===1 Content-Type: application/quickmail Content-Transfer-Encoding: base64
by newton.wadsworth.org (8.8.8/8.8.8) with SMTP id QAA21360 for {microscopy-at-msa.microscopy.com} ; Thu, 11 Mar 1999 16:59:34 -0500 (EST) Sender: tivol-at-wadsworth.org Message-ID: {36E839DF.41C6-at-wadsworth.org}
Jonathan Barnard wrote: Dear Johnathan,
} I need to know the wavelength of the main ~ 441.6nm emission } line from our He-Cd laser source to at least 5 significant figures, } if possible (wavelength in air atmosphere). Any references } containing other related data (emission lines) would be useful too. } Do such variables as pressure and composition (e.g., humidity) of the air allow the wavelength to be determined to 5 sig figs? The refractive index of air is related to pressure, and I think the cor- rect expression is n-1 = kP. Since P can easily vary by ~3% at sea level, depending on k, n could vary by more than 10^-5. The humidity might be even more important (especially for any other emission lines where water has n very different from that of air). Yours, Bill Tivol
We're trying to visualize meiotic spindle microtubules using LM and TEM. = Our methods for fixing and embedding insect testes are fairly = standard, however we are using 8% tannic acid in our fixative. This = appears to be what others have used, but we were wondering if anyone has = any warnings, hints or suggestions regarding microtubule preservation = and TEM.
We are also experimenting with different methods of preparing insect = testes for immunocytochemistry and LM. We are using various antibodies = to microtubules and nucleoproteins. We would like to optimize our = methods for the preservation of these structures. Is "live" tissue best = or will fixed tissue suffice? What is the best way to get the cells = spread onto slides? We've tried thumb "squashing" and cytospinning, but = are not satisfied with the results. Any suggestions would be greatly = appreciated!
Thanks,
Laura K. Garvey Dept. of Molecular and Cell Biology University of Connecticut
} } I have recently taken over an SEM with an AN 10000 analyser and I was } wondering if anyone out there kows if it is possible to get the results } files out to a PC ? } Bob-
Several years ago I wrote a nunber of utilities that would allow you to do just what you describe. You can write the files to an AN 10000 3.5" floppy (I hope yours *is* a 3.5" system? - if not, my programs won't help), which my utility will then read on a PC, copying the files to the PC in a byte-for-byte format. Then I wrote other utilities that would convert spectra and linescan files into tab-separated text files, and would extract images from studies and convert them, or individual image files, to baseline TIFF 6.0 files. I still use these utilities on a daily basis.
For a long while these were available on an FTP server here, but after aeons of no hits, and in the general progression of computer hardware, this went by the wayside. It would be the work of a few minutes to re-establish this, if there is interest. In the meantime, Bob, you may try contacting Pat Nicholson in the Dept. of Physics and Astronomy at Glasgow. I'm not sure, but I may well have given him copies of these files. In any case, I'll post the URL when I have re-established it.
If you have Windows, you should have everything you need already.
In DOS, the PrintScrn key used to dump a copy of the screen on the dot-matrix printer. In Windows nothing apparently happens. However, if you check the clipboard, PrintScrn snaps a copy of the desktop and stores it for pasting. Try pressing PrintScrn and then try paste into Word, or elsewhere. You will get a bitmap of the screen. Perhaps you don't want the whole screen - then Alt-PrintScrn copies just the active application window to the clipboard.
Using this process you end up with a lot or a little extra window junk around the sides. Then I use MS Imager that came with Office 6.0 (and was available on the MS website) to crop the image down to what I want.
I tried this using the AVI player from MS. I stopped the video, pulled the slider to the frame I wanted, and pressed Alt-PrintScrn. I opened up MS Imager, selected the File, New, Clipboard option and up came my AVI viewer window as a bitmap. I saved copies of it before and after cropping. I will send those to you directly. They are from the PICTURE.AVI movie that came on the Windows 98 disk.
You can use your imagination to apply this technique for other things, like producing your own instructions with snapshots showing what your EDS screen looks like at various stages.
Hope this Tip and Trick helps.
Warren
At 01:03 PM 3/11/99 +0000, you wrote: } } Have a couple of avi movies here and we need some stills printed from them. } Anyone have any ideas/shareware/freeware? } } } } } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { } GO GATORS } Scott D. Whittaker 218 Carr Hall } EM Technician Gainesville, FL 32610 } University Of Florida ph 352-392-1184 } ICBR EM Core Lab fax 352-846-0251 } sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ } The home of " Tips & Tricks "
A Mac application that delivers the most commonly used tools in Photoshop (e.g. adjusting levels, assembling RGB images and montages) is Color-It!, from MicroFrontiers. It retails for about $50.
Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
On Thu, 11 Mar 1999, Bobrowski, Walter wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I will agree wholehearted that Adobe PhotoShop is best, but I'll bet that } most people never learn it properly to get their $600 worth. A very } impressive PhotoShop clone is Ulead Photo Impact ( http://www.ulead.com } {http://www.ulead.com} ) for ~$90.00 which does what most people use } PhotoShop for (TWAIN compliant, Image resizing and level adjustments), plus } Web-based image production is built in, not an add-on.) Try their 15-day } fully functional demo. Enough said on image editors. } } Just passing along some unbiased information. } } Walt Bobrowski } Subcellular Pathology } Parke-Davis Research } 2800 Plymouth Road } Ann Arbor, MI 48105 } } TEL: (734) 622-7814 } FAX: (734) 622-3478 } Mailto:Walter.Bobrowski-at-WL.COM {mailto:Walter.Bobrowski-at-WL.COM} } } } }
The nice thing about HyperSnap is that you can copy just any selected rectangular portion from your screen, and then either print it, fool around with it, or save it in an astonishing number of formats.
Ritchie
} } If you have Windows, you should have everything you need already. } } In DOS, the PrintScrn key used to dump a copy of the screen on the } dot-matrix printer. In Windows nothing apparently happens. However, if you } check the clipboard, PrintScrn snaps a copy of the desktop and stores it } for pasting. Try pressing PrintScrn and then try paste into Word, or } elsewhere. You will get a bitmap of the screen. Perhaps you don't want the } whole screen - then Alt-PrintScrn copies just the active application window } to the clipboard. } } Using this process you end up with a lot or a little extra window junk } around the sides. Then I use MS Imager that came with Office 6.0 (and was } available on the MS website) to crop the image down to what I want. } } I tried this using the AVI player from MS. I stopped the video, pulled the } slider to the frame I wanted, and pressed Alt-PrintScrn. I opened up MS } Imager, selected the File, New, Clipboard option and up came my AVI viewer } window as a bitmap. I saved copies of it before and after cropping. I will } send those to you directly. They are from the PICTURE.AVI movie that came } on the Windows 98 disk. } } You can use your imagination to apply this technique for other things, like } producing your own instructions with snapshots showing what your EDS screen } looks like at various stages.
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Everett, if the door to your lab is of a "simple" configuration, it would be easy to replace it with a "windowed" door as long as your boss is willing to accept the cost as a price towards improved safety. Obviously, not a complete answer to the problem but a first step of common sense.
Mike Bucker Feed Microscopy Consolidated Labs of Va
} } } "EVERETT RAMER" {Everett.Ramer-at-fetc.doe.gov} 03/11 4:23 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Over the years our workforce has dwindled to the point that I find myself working alone in the lab (metalographic sample prep/optical microscopy + image analysis) almost all the time. The lab doesn't have windows to the hallway, so it is difficult for people walking by to see if I am in the lab, and if I am OK. My manager is concerned about my safety and is asking me for suggestions. What do other labs do to make working alone safe? Everett Ramer Federal Energy Technology Center
Everett and all: I work at Dow Chemical and we have a fairly rigorous "lone operator" system. If we are working in an isolated area or at a time when there are few people in the building (evenings, weekends, etc.) we carry little alert transmitters which call out to plant security (as well as within the building). The building receivers are location sensitive and we have a "lone-operator" login book at the building entrance with a building map. The lone operator marks the map as s/he signs in. Between the map and the alert location, they can find us pretty fast.
This is pretty elaborate, but maybe a mini version using something like the "First Alert" products would work ("Help, I've fallen and I can't get up") - have it tied into a siren or flashing light outside your lab so that anyone along the corridor would know there was a problem. Maybe even carry a cordless phone with an autodial button to your site security - caller ID would get them to you pretty quick.
This is definitely NOT a trivial matter. It is always disconcerting to me when I am working in an area with nobody else around - I hope you get an effective solution soon.
there's lots of stuff on t.e,m, of apoptosis in vertebrate cells (it's very popular in HIV, cancer, inflammation response etc) and much of it indicates visible nuclear changes but relative stability of cytoplasm compared with necrosis. There was a review article (as a good starting point): Microscopical Study of Cell Death via Apoptosis by S. Verhaegen in MIcroscopy and Analysis, January 1998 pp5-7
but you could do a reference or citation 'trawl' on the authors: Kerr, J.F.; Wyllie, A.H. or Currie
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk ---------- } From: Tina Carvalho To: Microscopy Listserver
Hi, All-
A researcher would like to be able to tell the difference between apototic cells and necrotic and/or other degenerating cells in an invertebrate nervous system, using TEM. So far the only general statement I've come across is that apototic cells will undergo autophagy within their plasma membranes whereas necrotic cells tend to spill their contents and get cleaned up by other cells.
Any additional tips will be appreciated!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
by ahmler1.mail.eds.com (8.9.1/8.9.1) with ESMTP id JAA30032; Fri, 12 Mar 1999 09:51:43 -0500 (EST) Received: from SYS1.BSCO.COM (sys1.bsco.com [198.132.68.84]) by nnsa.eds.com (8.9.1/8.9.1) with SMTP id JAA27069; Fri, 12 Mar 1999 09:51:12 -0500 (EST) Received: from rfurdanowiczw [10.224.0.46] by SYS1.BSCO.COM (IBM VM SMTP Level 310) via TCP with SMTP ; Fri, 12 Mar 1999 09:51:21 EST Reply-To: {rwafu-at-bsco.com} {microscopy-at-Sparc5.Microscopy.Com}
Yes, on my AN10000 there is a program called MSDOSCV.SV which converts files between the Link (now Oxford) and PC DOS operating systems. It writes files to 720kB 3.5" floppies that previously have been formatted to that density on a PC.
An alternative that I sometimes resort to is to capture on a PC output meant to go to the Facit printer over the serial connection.
If you need more details, contact me directly or seek support from Oxford (they are very helpful), for example Ruth Murray ( ruth-at-oxford.usa.com ).
Valdemar Furdanowicz Research Labs Bethlehem Steel Co. valdemar-at-fast.net or rwafu-at-bsco.com
-----Original Message----- } From: Robert McDonald [mailto:R.McDonald-at-geology.gla.ac.uk] Sent: Thursday, March 11, 1999 10:08 AM To: microscopy-at-sparc5.microscopy.com
Hi All:
I have recently taken over an SEM with an AN 10000 analyser and I was wondering if anyone out there kows if it is possible to get the results files out to a PC ?
Any help greatly appreciated.
******************************************* Robert McDonald EPMA & SEM Laboratories Dept Geography - Earth Sciences Division Gregory Building LilyBank Gardens University of Glasgow Glasgow G12 8QQ Scotland, UK email: robert-at-earthsci.gla.ac.uk Tel:- +44 (0)141 330 5505/5442 FAX:- +44 (0)141 330 4817 ********************************************
Differentiating apoptosis and necrosis morphologically is based primari= ly upon nuclear changes, although there are characteristic cytoplasmic cha= nges as well. In general (note the wiggle words), necrotic cells swell and = lyse, whereas apoptotic cells shrink and fragment. Chromatin in apoptotic ce= lls forms electron-dense crescents at the nuclear envelope, then breaks up.=
Apoptotic cells fragment into "apoptotic bodies" that may contain bits = of chromatin. A good place to see characteristic ultrastructural changes = of apoptosis is in lymphoid tissues, where lymphocytes die via apoptosis a= nd then are phagocytosed by resident macrophages (the ones that are someti= mes called "tingible body macrophages" because of the staining properties o= f the apoptotic cell remnants in them.)
Differenting apoptosis from necrosis is a tricky deal. Apoptotic cells= may undergo secondary necrosis, during which they swell and lyse. So, just=
because you see necrotic cells doesn't mean that they didn't die apoptotically. Like everything else, it's complicated; there is a cont= inuum of change with apoptosis and necrosis at opposite poles and a lot of st= uff in between!
There are lots of good reviews on this topic. The Aug 28, 1998 issue o= f Science had a special section on apoptosis, and on page 1302, there is = a series of three electron micrographs of neurons undergoing apoptosis. = One of the first reviews of the subject contains the best collection of micrographs I've found - "Cell death: the significance of apoptosis" in= the International Review of Cytology 68:251-306, 1980. Another good revie= w was in Amer J of Pathol 146:3-15, 1995. The title is "Apoptosis, oncosis, = and necrosis: an overview of cell death."
Hope this helps!
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064-6202 =
In the past I have been on jobs where a great deal of the time was spent isolated. Now days I think it is a huge safety liability. One time I had appendicitis and had to drive myself to 50 miles on back roads to a clinic with my knees up on the stearing wheel. The point here is, even though you work in a laboratory complex, if something happened you probably wouldn't get help until the cleaning crew found you. Bad news! the other issue is: Life is short and work is long, and working alone sucks. It's not emotionally healthy. Make a change. Consolidate in with other workers.
Bob Derm Imaging Center Microscopist Ex-wood cutter
On Thu, 11 Mar 1999, EVERETT RAMER wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Over the years our workforce has dwindled to the point that I find myself working alone in the lab (metalographic sample prep/optical microscopy + image analysis) almost all the time. The lab doesn't have windows to the hallway, so it is difficult for people walking by to see if I am in the lab, and if I am OK. My manager is concerned about my safety and is asking me for suggestions. What do other labs do to make working alone safe? } Everett Ramer } Federal Energy Technology Center } }
The New York State Center for Advanced Thin Film Technology University at Albany - State University at New York announces the following positions:
Senior Analytical Specialist (Two Positions Available) The New York State Center for Advanced Thin Film Technology at the University at Albany - SUNY is a fast growing, high technology research and development program with a mission of supporting industry and creating new jobs. This position will serve as a primary, materials characterization, support person for the Center's scientific and technical staff in our advanced surface science facilities.
Job responsibilities include: performing day to day operation and support of the advanced surface science laboratories; assisting in data acquisition and analysis; participating in selected research projects to ensure contractual deliverables are met; providing materials characterization of microelectronic, optoelectronic and photonic samples for faculty and staff; providing instrumentation training to staff and student; and working with students on the advanced analytical tools.
The position requires: a Ph.D. in materials science or related field such as physics or chemistry and a minimum of three years experience in the characterization of microelectronic, optoelectronic or photonic materials or a bachelors degree with 10 years of relevant work experience; demonstrated ability to work in a high energy, team oriented environment; and excellent communication and analytical skills. Preference will be given to those candidates with the required work experience with Auger Electron Spectroscopy, X-Ray Photoelectron Spectroscopy, Scanning Electron Microscopy, X-Ray diffractometry, Atomic and Scanning Tunneling Microscopy, or Transmission Electron Microscopy. Salary and Benefits are highly competitive and dependent upon experience.
Please submit a resume and cover letter to:
Jacqueline DiStefano NYS Center for Advanced Thin Film Technology CESTM B110 251 Fuller Road Albany, NY 12203
The SUNY/Research Foundation is an Equal Opportunity/Affirmative Action Employer.
Well, the best method for presevering wonderfully empheral / delicate structures is cryo-preservation and freeze subsitution. I see you are a little far from us to come and try some freezing, but maybe you have some ready access to some rapid freezing.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
I'll second the opinion about getting your money's worth out of Adobe Photoshop. I've used PhotoImpact (we have v 3 here at work; I've not used v 4 which is out already) and agree it's good. My favorite, though, is Paintshop Pro (Jasc, Inc.) which I bought for home after trying a download version. I find it is a little more intuitive than PhotoImpact (at least for me!). It handles layered images like Photoshop, and runs most Photoshop plugins. I even use it with the Image Processing Toolkit (Dr. John Russ); most of the plugins run without problem, although a few tend to crash. PhotoImpact also handles layers, I believe, but with an object-oriented approach. I haven't tried the IP Toolkit plugins in PhotoImpact.
Paintshop Pro can probably be had for a little less than Photoimpact. List price is probably $90 or $100, but I've seen it on some of the on-line computer stores for much less. I think one may have even had it for something like $58 (?). Check out info & demo at http://www.jasc.com
Disclaimer: I have no ties to any of these software packages!
Jim Passmore Cryovac Division Sealed Air Corp.
---------- } From: Walter.Bobrowski } To: Microscopy } Subject: RE: Photo Editors } Date: Thursday, March 11, 1999 3:52PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think you are making an excellent choice with the new Agfa DuoScan T2500. You will appreciate the 3.4 D on this unique flatbed scanner. The glass less
negative carriers fit into the lower tray, which slides into a precise position. The negatives will not contact any surface. I have just ordered also a DuoScan T2500. You will learn more about it at:
One of the most versatile imaging software is CorelDraw8 and CorelPHOTO-PAINT8. It offers the highest image format manipulation with Plug-ins, for most applications. Highly recommended.
Laszlo J. Veto
Caspar McConville wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are thinking of purchasing a negative scanner for use with TEM } negatives from our Jeol 2000-FX, and also for SEM negatives. A } scanner has been recommended to us: the Agfa Duoscan T2500, which } has a resolution (hardware) of 1250 x 2500 dpi and a density of 3.4D. } (The scans would be output to a Kodak DS 8650 PS printer) } } We need the quality of the scans to match the quality of the standard } darkroom enlarger if possible, as we would like to 'go digital' at } least for routine work. Does anyone have experience of routine } negative scanning for TEM prints, with this or other scanners, and if } so, is it realistic to expect such high quality? } } Also, what additional image processing software would people } recommend we got to go along with this? } } Any advice would be appreciated. } } Caspar } } Caspar McConville, Ph.D. } Technical Specialist } New York State College of Ceramics } Alfred University
I am pseudocolorizing some data we have of cellular responses from striatum of rats learning a T-maze from Ann Graybiel's tetrode recording project. What I am looking for is any "standard" pseudocolor tables used by imagers to colorize 0-255 grey levels into RGB values.
I can apply a standard spectrum from 0 to 255 running from magenta (255 0 255 RGB) through blue, cyan, green, yellow to red (255 0 0) or start with blue and go to red in a linear way. What this produces is colors that are mostly in the middle. What I think imagers must be using are tables weighted toward the red and blue, so transitions show up better. I can play with my tables in Excel to boost the red and blue ends and then apply them using Paint Shop Pro - what I was wondering was if you have or know where I can find any standard tables, preferably of numerical RGB values used by the pros in the field.
Hi, A student in the lab is looking at museum samples of dinosaur bones and teeth on the SEM. He could get access to more samples if he could restore them to their original condition ( ie, remove the gold). Is there a good nondestructive way to do this?
Kim DeRuyter Histology and Electron Microscopy Labs University of Alaska Fairbanks
In a message dated 3/12/99 9:55:46 AM Hawaiian Standard Time, underwoo-at-u.washington.edu writes:
{ { Bad news! the other issue is: Life is short and work is long, and working alone sucks. It's not emotionally healthy. } }
Perhaps it is true for some that working alone is not emotionally healthy. However, if you ARE emotionally healthy, then working alone can be emotionally healthy and spiritually healthy also. It can be a time for focussed energy, contemplative thought, creative thought - all without interruption. Personally I love working alone and after the work I enjoy the company of my family and friends.
} From: Robert Underwood {underwoo-at-u.washington.edu}
} } In the past I have been on jobs where a great deal of the time was spent } isolated. Now days I think it is a huge safety liability. One time I had } appendicitis and had to drive myself to 50 miles on back roads to a clinic } with my knees up on the stearing wheel. The point here is, even though } you work in a laboratory complex, if something happened you probably } wouldn't get help until the cleaning crew found you. Bad news! the other } issue is: Life is short and work is long, and working alone sucks. It's } not emotionally healthy. Make a change. Consolidate in with other workers.
I spent most of my life farming and ranching. Both rather dangerous occupations. You spend almost all your time alone and in the busy season I might be alone 48 hours at a time so no one would even start looking for me for a couple of days.
Not many people are killed because of the isolation. Your best protection' is to think before you do something dangerous.
Gordon
Gordon Couger gcouger-at-couger.com Owner PRAG-L PRactical AGriculture List www.couger.com/prag-l Stillwater, OK 405 624-2855 GMT -6:00
Can anyone help me acquire an instruction manual for a Reichert-Jung FC4E cyroultramicrotome attachment for a Reichert-Jung Ultracut E ultramicrotome? I would be able to pay copy and mailing expenses.
Thanks
Damian Neuberger Research Scientist damian_neuberger-at-baxter.com
There are some two way pagers that have a man down feature. If the pager is in a horozonal postion for 10 seconds it beeps. If the wearer doesn't acknoledge the beep it sends an alarm to a central station. These pagers have a panic button as well and will serve as regular alpha numeric pagers that allow yes/no acknoledgement from the wearer.
Disclaimer: I own a substantial share of Datalink System that manufactures and selling these. See www.rfdata.net. Gordon
Gordon Couger gcouger-at-rfdata.net Datalink Systems www.rfdata.net Stillwater, OK 405 624-2855 GMT -6:00 =================================
Everett Ramer's question regarding working-alone has ellicited thoughtful and poignant reply. Useful reply as well, by Jerry Heeschen.
To level this solemn feeling, here is mine originally just shared with Everett.
Nathan Haese - at work, alone in a garage, on a new microscope, perhaps too alone.
Everett,
At a large chemical company that I once worked for, we had a badge with a radio alert button that we wore when we worked alone on weekends. Pressing the button would alert the gate guard to come find us in the lab=
Does any one have an instruction/operations manual for an American Optical microtome knife sharpener? Or if it's basic enough to enlighten me just put it in an email? Thanks in advance Steve D'Angelo
Let's join and pay our tribute to Drs Richard Henderson & Nigel Unwin for their Aminoff Award.. Cheers!
The Royal Swedish Academy of Sciences has decided to award the Gregori Aminoff prize in crystallography for 1999 to dr. Richard Henderson and dr. Nigel Unwin, MRC Laboratory of Molecular Biology, Cambridge, England, for their development of methods for structure determination of biological macromolecules using electron diffraction. The prize is presented at the Annual Meeting of the Academy 31. March 1999. The Aminoff symposium 29 - 30. March is organised to the honour of the prize-winners.
The symposium is supported by the Academy through its Nobel Institute for Chemistry.
Aminoff Symposium Structure Determination of Macromolecules with Electron Diffraction 29 - 30. March 1999
Monday 29. March
13.00 - 13.15 Opening of the symposium: Erling Norrby Introduction: Ivar Olovsson
General and non-biological Systems
13.15 - 14.15 Atomic Resolution Electron Microscopy in Biology Richard Henderson
14.15 - 15.00 Imaging Individual Atoms by Electron Microscopy Sven Hovmller, Stockholm
15.00 - 15.30 Coffee/Tea
Diffraction Studies of Membrane Proteins
15.30 - 16.30 Different methods for the study of membrane structures. Matti Saraste, EMBL, Heidelberg
16.30 - 17.15 Electron Crystallography of Membrane-bound Enzymes Hans Hebert, Stockholm
17.15 - 18.00 Structural Studies on the Cytochrome bc1 Complex So Iwata, Uppsala
18.00 - 18.30 General discussion
18.30 Dinner in the Club House of the Academy
Tuesday 30. March
Studies of Virus Structures
09.00 - 10.00 Combination of Different Methods in Virus Studies Michael Rossmann, Purdue
10.00- 10.45 Virus Studies, Essence of Supermolecular Symmetry Holland Cheng, Stockholm
10.45 - 11.15 Coffee/Tea
Non-crystalline Materials
11.15 - 12.00 Visualization of Single Protein Molecules by Electron tomography. Ulf Skoglund, Stockholm
12.00 - 13.00 0 - dimensional Crystallography Marin van Heel, Imperial College
13.00 - 14.30 Lunch in the Club House
Concluding talks
14.30 - 15.30 Making Light Work: The Membrane Proteins of Plant Photosynthesis Werner Khlbrandt, Frankfurt
Getting gold off museum fossil pieces is nay impossible. Al would be easier but basically its the same problem. C coating is good enough for low powers, but again, Curators do not like that dark coating, To view these "hard, dry and non-conducting" specimens uncoated, the best solution is a poor vacuum SEM; a fully fledged Environmental SEM would also do well, but its a more expensive instrument for that job. Poor vacuum SEM's (I believe at least a couple of the major manufacturers make instruments with that facility) use only mechanical pump vacuum in the specimen chamber and because of a vacuum limiting aperture retain high vacuum in the gun chamber. Secondary mode is impossible, but a Robinson detector gives excellent images for this type of work. Magnifications under these conditions are limited to about 2000x, but details in fossils do not warrant higher magnifications; its the SEM's superior depths of field that wins out over light microscopy. Kim - all you require now is one of those scopes! Years ago I modified an Etec Autoscan to function reversibly as a poor vacuum instruments. It worked well but it was a fair bit of trouble to accomplish the required modifications. Our online contain a link to an archive collated by Scott Wight. This contains listserver contributions concerned with Environmental and Poor Vacuum SEM. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Saturday, March 13, 1999 6:55 AM, Kim DeRuyter [SMTP:fnksd1-at-uaf.edu] wrote: } } Hi, } A student in the lab is looking at museum samples of } dinosaur bones and } teeth on the SEM. He could get access to more samples if } he could restore } them to their original condition ( ie, remove the gold). } Is there a good } nondestructive way to do this? } } Kim DeRuyter } Histology and Electron Microscopy Labs } University of Alaska Fairbanks }
Due to the response (and fun) of our contest at last years MSA/MAS Conference, we will repeat the contest this year in Portland. The concept of the contest is based on composite micrographs, each made up from two or more images - one of which must be microscopical in nature. Prizes of value will be awarded and one will not have to be present to win. If of interest, kindly advise by return email and I will see that you receive full contest detail. Don Grimes, Microscopy Today
Many years ago I worked in a hospital basement EM lab. One day my boss and I came out of the inner TEM room and wondered why the hallways were deserted. Then a fire marshall came by demanding to know why we hadn't vacated the building during the fire drill!
The following email notice arrived from our university safety office: ------------
Kim
I seem to remember a story - a long time ago - about about dipping the sample in liquid mercury - the gold is taken into the liquid as an amalgam and leaves the specimen clean. I have never tried it. I do not know what any Safety person would say about that these days!
Keith Ryan Marine Biological Association of the UK Citadel Hill Plymouth Devon PL1 2PB England
In a message dated 3/12/99 9:45:34 PM Mid-Atlantic Standard Time, fnksd1-at-uaf.edu writes:
{ { Hi, A student in the lab is looking at museum samples of dinosaur bones and teeth on the SEM. He could get access to more samples if he could restore them to their original condition ( ie, remove the gold). Is there a good nondestructive way to do this?
Kim DeRuyter Histology and Electron Microscopy Labs University of Alaska Fairbanks } } Good Morning! A less destructive way to analyze these samples would be to coat them with carbon instead of gold, and then ashing the carbon off with O2. Gold is tricky to remove on most materials (I work mostly with semiconductors), but would be very difficult to remove on dinosaur bones. (This is assuming your samples are small enough to fit into an available asher or RIE tool). There are several labs that could perform both the coating and ashing - let me know if you have trouble locating one close to your area. Lisa Montanaro Consultant, MME
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
im, A good way to handle specimens of this sort is to make casts of the surface and examine the casts in the SEM. This way there is no damage to the original artifact. I had an anthropology grad student do this with human teeth for his thesis research. He worked out a very inexpensive, low tech, but reliable method to do the casting. He can be reached at the following for full details of his method: Dr. Chris Schmidt Indianapolis University cschmidt-at-indy.edu
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Hi, A student in the lab is looking at museum samples of dinosaur bones and teeth on the SEM. He could get access to more samples if he could restore them to their original condition ( ie, remove the gold). Is there a good nondestructive way to do this?
Kim DeRuyter Histology and Electron Microscopy Labs University of Alaska Fairbanks
RFC822 header -----------------------------------
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by dj.stud.ntnu.no (8.9.1/8.9.3) with ESMTP id QAA03224; Mon, 15 Mar 1999 16:01:30 +0100 (MET)
I have the same problem with the Microm-Heidelberg, Heavy duty Microtome HM 350. We would also be able to pay copy and mailing expenses.
Gary. NTNU Norway. } } } Can anyone help me acquire an instruction manual for a Reichert-Jung FC4E } cyroultramicrotome attachment for a Reichert-Jung Ultracut E } ultramicrotome? I would be able to pay copy and mailing expenses. } } Thanks } } Damian Neuberger } Research Scientist } damian_neuberger-at-baxter.com } } }
Two postdoctoral fellowships are available immediately in the Supramolecular Structure and Function group in the Bioengineering & Physical Science Program at the NIH. Our laboratory is looking for a physical scientist (e.g., physics or materials) and biological scientist (e.g., biophysics) to help develop and apply new methods based on electron microscopy and spectroscopy. There is considerable flexibility in the scope of research which includes the following:
(i) development of EELS spectrum-imaging in the STEM to map phosphorus, calcium and other elements in macromolecular assemblies and cells.
(ii) development of energy-filtered TEM to establish elemental detection limits and to map elemental distributions.
(iii) development of x-ray microanalysis and cryo-preparation techniques for studies in cell biology.
(iv) image processing techniques to determine the structures of large macromolecular assemblies using cryo-EM and STEM.
(v) applications of any of the above methods to biomedical research in collaboration with investigators in other NIH laboratories.
Our laboratory is equipped with field-emission scanning transmission electron microscopy (STEM), electron energy loss spectroscopy (EELS), energy-filtered electron microscopy (EFTEM), x-ray microanalysis (EDXS), cryo-electron microscopy, and UNIX-based image processing.
Preference will be given to candidates with less than five years of relevant postdoctoral experience. Candidates from the United States or from overseas are welcome to apply.
For additional information see: http://www.nih.gov/od/ors/beps/ssfr/
Please send curriculum vitae and bibliography to:
Dr. Richard Leapman
Biomedical Engineering & Physical Sciences Program
- Eye on Imaging - MSC/SMC Conference May 26-28, 1999
Sponsored by the Microscopical Society of Canada
We are pleased to annouce the 26th annual meeting of the Microscopical Society of Canada. This spring meeting and exhibition will be taking place for three days, May 26-28, 1999, on the campus of the University of Guelph in Guelph, Ontario. Many interesting speakers have agreed to participate including Dr. John Russ (author of the Image Processing Handbook, Dr. P.C. Cheng (multi-photon microscopy and microscope construction) , Dr. Chris Yip (AFM), Dr. Nestor Zaluzec (Tele-Presence microscopy), Dr. Nick White (3-D quantitative analysis and multi-photon microscopy) and Dr. Brian Kaye (Fractal analysis) amongst others. We are also offering a variety of afternoon workshops as well as a commercial exhibition offering a full range of Microscopy and Imaging equipment and supplies. Please visit our web site for information and registration packages:
http://www.uoguelph.ca/botany/rootlab/msc99.htm
Please pass this link along to anybody that may be interested. Deadline for submission of abstracts and pre-registration is April 6, 1999. Hope you can make it,
George Harauz Microscopical Society of Canada Chairman, Local Organizing Committee University of Guelph Guelph, Ontario gharauz-at-uoguelph.ca
You might look into utilities that allow you to adjust the color LUT and data separately of each other. Then you should be able to stretch the colors to fit like you want. If that is not easily possible with your software, then you might try playing with the gamma, contrast and brightness on your image before applying the pseudo-color and you should be able to get your weighting as you like it.
If you absolutely need help, I might be able to fabricate a color table here if you can send me a typical image and your color table.
At 03:46 PM 3/12/99 -0500, you wrote: } Hi computer imagers,, } } I am pseudocolorizing some data we have of cellular responses from striatum of } rats learning a T-maze from Ann Graybiel's tetrode recording project. What I } am looking for is any "standard" pseudocolor tables used by imagers to colorize } 0-255 grey levels into RGB values. } } I can apply a standard spectrum from 0 to 255 } running from magenta (255 0 255 RGB) through blue, cyan, green, yellow to red } (255 0 0) or start with blue and go to red in a linear way. What this produces } is colors that are mostly in the middle. What I think imagers must be using } are tables weighted toward the red and blue, so transitions show up better. I } can play with my tables in Excel to boost the red and blue ends and then apply } them using Paint Shop Pro - what I was wondering was if you have or know where } I can find any standard tables, preferably of numerical RGB values used by the } pros in the field. } } } ------------------------------------------------------------------ } |Glenn Holm {mailto:karuzis-at-wccf.mit.edu} | } |Graybiel Lab (617)253-5780;fax (617)253-1599 | } |M.I.T Dept. of Brain + Cog. Sci. Cambridge, MA 02139 | } ------------------------------------------------------------------
Kim DeRuyter wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } A student in the lab is looking at museum samples of dinosaur bones and } teeth on the SEM. He could get access to more samples if he could restore } them to their original condition ( ie, remove the gold). Is there a good } nondestructive way to do this? } } Kim DeRuyter } Histology and Electron Microscopy Labs } University of Alaska Fairbanks Kim, One thing that no one has mentioned, yet, is low kV operation. If your instrument will operate at 5kV or lower (perferably around 1kV) and you limit your beam current, you should be able to view uncoated specimens at at least a couple of kX. Some intruments will go much higher at that voltage range. Grains of quartz may still present a problem because SiO2 is such a good insulator, but many other minerals will work fine under those conditions.
Ken Converse owner Quality Images third party SEM service Delta, PA
I have set up an FTP site at prism.mit.edu, port 2101, with the files I mentioned that will convert LINK/Oxford AN10/eX/L files. Apologies if the documentation is sparse!
To access a non-standard port in FTP you will need to know how your FTP program works. In Netscape, use the following URL:
ftp://prism.mit.edu:2101
In WS_FTP you have to change the port number in the Advanced tab. From a command line ftp program (e.g. Unix, or DOS from Win 95/98/NT), first invoke the program without a server name, i.e. just enter
ftp
The program responds with the ftp prompt ftp} . Then you enter:
open prism.mit.edu 2101
The user is anonymous, and the password is unimportant. I don't know how you would do it with other FTP programs. Sorry about the difficulty, but I am already using the standard FTP port for another, non-public, purpose!
I think you are making an excellent choice with the new Agfa DuoScan T2500. You will appreciate the 3.4 D on this unique flatbed scanner. The glass less
negative carriers fit into the lower tray, which slides into a precise position. The negatives will not contact any surface. I have just ordered also a DuoScan T2500. You will learn more about it at:
One of the most versatile imaging software is CorelDraw8 and CorelPHOTO-PAINT8. It offers the highest image format manipulation with Plug-ins, for most applications. Highly recommended.
Laszlo J. Veto
Caspar McConville wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are thinking of purchasing a negative scanner for use with TEM } negatives from our Jeol 2000-FX, and also for SEM negatives. A } scanner has been recommended to us: the Agfa Duoscan T2500, which } has a resolution (hardware) of 1250 x 2500 dpi and a density of 3.4D. } (The scans would be output to a Kodak DS 8650 PS printer) } } We need the quality of the scans to match the quality of the standard } darkroom enlarger if possible, as we would like to 'go digital' at } least for routine work. Does anyone have experience of routine } negative scanning for TEM prints, with this or other scanners, and if } so, is it realistic to expect such high quality? } } Also, what additional image processing software would people } recommend we got to go along with this? } } Any advice would be appreciated. } } Caspar } } Caspar McConville, Ph.D. } Technical Specialist } New York State College of Ceramics } Alfred University
} From: Self {miller.TEX.TAFA} To: Microscopy-at-MSA.Microscopy.Com
Howdy all, There is a graduate student here who is trying to look at a possible stem cell line under TEM. The problem is that they are quite small, don't seem to pellet very well, and he has only been able to get me a few thousand cells at a time. We have tried to embed the cells in 2% Agar after fixation but this hasn't worked. Is it possible to filter the media and cells, and then process the filter with the cells stuck onto it? Maybe use a cytospin to spin the cells into a filter? Is there anyone with experience with this? Any suggestions or ideas would be greatly appreciated.
Thanks in advance Frank Herbert Technician Integrated Microscopy Core Department of Cell Biology Baylor College of Medicine
Somewhere in the deep receses of my mind I recall a cytochemical test for arsenic. I think it was for EM but I am not sure. Does anyone out there know it??
Or was it all just a bad dream?
Greg Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
We are trying to equip our lab with a reflected light BF/DF microscope (+ trinocular head, B&W video system, 4X5" Polaroid system), for two basic needs: 1. viewing and photographing our fine metal, ceramic, and carbide powders for QA purposes (e.g. visual standards), and 2. viewing and photographing mounted/polished thermal spray powder and coating samples prepared at our parent company's facility.
Our budget allows for a used (reconditioned or demo) top-name microscope (e.g. Nikon, Olympus, Leitz, Zeiss) or a new lesser- name scope (e.g. Meiji...) with the same basic features. Not being professional microscopists ourselves, we ask you to comment on aspects of the above tradeoff, based on your experience. Our main concern is flatness of field and sharpness of image at magnifications up to 500X. Thanks for your help!
Sincerely,
Robert A. Miller TAFA Material Technologies, Inc. 1702 Mykawa Road Pearland TX 77581 USA PHONE: 281-485-7765 FAX: 281-485-0211 EMAIL: miller-at-tafa.com
by dogwood.botany.uga.edu (8.9.1/8.9.1) with SMTP id RAA17081 for {Microscopy-at-Sparc5.Microscopy.com} ; Mon, 15 Mar 1999 17:56:43 -0500 (EST) Message-Id: {199903152256.RAA17081-at-dogwood.botany.uga.edu} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Hi, I'm trying to find out if the elemental imaging system on my scope (a Zeiss 902A TEM) is working...or if it is a problem with the sample. Does anyone have a sample they could spare for elemental imaging?
In October, I had a great sample grid (30nm liver sections with no stain). We were able to get nice images of iron. The scope had its annual service in November and the elemental imaging system has not worked correctly since that time. The serviceman is suggesting that the sample is fried (no pun intended). I think the scope is whacked. So I'm trying to get another liver sample (my source for the first sample retired) or I'm willing to try something else. I need a "known" sample, cut 30 nm thick, no support film (600 mesh grids help) or staining.
Any help or suggestions would be GREATLY appreciated!
Sincerely, Beth Richardson
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm interested. Elaine
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility 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-unixg.ubc.ca
hello members. here is a probable dumb question. Maybe an impossible situation.
Suppose that I have a prepared microscope slide-- 1"x3" glass slide with specimen under cover slip. Ordinary LM analysis works fine. Is there some other analysis method besides confocal that would offer better resolution and increased depth of field? The idea is to not have to prepare TEM specimens.
Is this possible? Gary Gaugler, Ph.D. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Thanks to everyone for your fantastic responses to my request for a Reichert-Jung FC4E manual. The folks from Leica have been especially helpful and quick to respond and I'm most grateful for the help.
As Valdemar Furdanowicz wrote you, you can transfer LINK files to the PC = with their MSDOSCV program. However, you can also use the serial port of the = LINK=20 computer and it can be a little bit faster. What I don't know, however, = is whether you can connect the the two machines directly. When I worked in the electron = beam lab of Hungalu Engineering, we had a little box between the serial ports=20 of the machines, made by Oxford. You can try any terminal program on the PC if it has an option for the = well-known XMODEM protocol. Once transferred, the files have to be converted from one format to = another , because the numbers are formatted differently on the LINK and the PC. With fixed point numbers the only difference is the order of the bytes, = but with floating point numbers the problem is more serious.=20 Your opportunities for file conversion are very limited with the MSDOSCV = program.=20 Of course you have to know the structure of the files, if you want to = convert them.=20 You can find this information in the Oxford manuals.
I wrote some programs to solve the above mentioned problems: 1. File transfer: with our XMODEM program we can transfer multiple files = using wildcards in=20 the file specifications. For this purpose the PC program prepares the = necessary LINK macros, which are sent as a first step. 2. File inspection: with a little utility program one can look at the = files obtained from the LINK computer and compare their content with the documentation.=20 3. File conversion: I have worked out simple conversion programs for = image files, DIGISCAN image anal. result files and X-ray spectra.
With a private letter I can send you anything from these programs, but = there are still some minor=20 problems with that. When I worked for Hungalu, I was the only user of my = programs, and I didn't have enough time for a better user interface. They are written = for the DOS platform, and they send Hungarian messages to the screen. Now I work as a computer programmer in a very different field, and =
in our group we are using formvar-coated grids to stabilize ultrathin sections for immunogold-labelling. Sometimes the formvar is *heavily* labelled by gold particles. This is no= t due to binding of the gold grains, but depends only on the primary antibo= dy that had been used. It can happen both with Protein A-gold or goat anti rabbit-gold, but using other primary antibodies the formvar is clean. What is the reason for this binding of antibodies to formvar? Is there any blocking reagent that would provide this? Thanks in advance
Birgit
-------------------------------------------------------------------------= ----- Dr. Birgit Neubohn Institut fuer Pflanzengenetik und Kulturpflanzenforschung (IPK) (Institute of Plant Genetics and Crop Plant Research) Corrensstr. 3 D-06466 Gatersleben, Germany
there might be a number of other sources for your problem beside the microscope itself. You could have trouble with your camera and/or image processing system. Please give more details what equipment you are using to acquire an elemental map (SIT or CCD camera, what software, etc. ...) Due to the principle of the energy-filter of your microscope (Castaing-Henry electrostatic mirror type) you would probably not be able to see an image at all if it would malfunction. But to make things clear you could insert a fairly thick biological sample and check: 1. If you the a normal spectrum on the final screen if you are switching from image mode to spectrum mode and 2. if you are able to see the contrast change if you acquire filtered images before and above the carbon edge. If both works fine, you should search the problem elsewhere. Are the intensities of the filtered images before and on the iron edge high enough to allow the system to calculate a useful result? Did you change any other important parameters since your last successful try (slit width, energy losses, acquisition time, background correction, ...)? You can contact me directly if you like to discuss this a little more in detail.
Cheers,
Petra
beth-at-dogwood.botany.uga.edu wrote:
} Hi, } I'm trying to find out if the elemental imaging system on my scope (a Zeiss } 902A TEM) is working...or if it is a problem with the sample. } Does anyone have a sample they could spare for elemental imaging? } } In October, I had a great sample grid (30nm liver sections with no stain). } We were able to get nice images of iron. The scope had its annual service } in November and the elemental imaging system has not worked correctly since } that time. The serviceman is suggesting that the sample is fried (no pun } intended). I think the scope is whacked. So I'm trying to get another liver } sample (my source for the first sample retired) or I'm willing to try } something else. I need a "known" sample, cut 30 nm thick, no support film } (600 mesh grids help) or staining. } } Any help or suggestions would be GREATLY appreciated! } } Sincerely, } Beth Richardson -------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu Visit our WWW site! http://www.crpcu.lu/~wahlbrin
Perhaps a second-hand Reichert Zetopan equipped for reflected light could fit the bill.
These are available from time to time at +/- decent prices in the USA (about $ 1000 - 2000, depending on condition and accessories mounted on the stand). (Some) spare parts are available in Germany or Austria.
Reichert had objectives for incident light for Zetopan 5.5x/0.15; 11x/0.25; 32x/0.65, 45x/0.95. I don't know about higher magnifications... The objectives mentioned are still available trough at least one German dealer.
I don't know about their quality regarding flatness of field and sharpness: I use a Zetopan equipped for biological work/transmitted light. Contact me offline if you are interested (I have no financial interests in this).
Yvan Lindekens. ---------- } Van: Bob Miller {miller-at-tafa.com} } Aan: microscopy-at-sparc5.microscopy.com } Onderwerp: Instrument Quality vs. Budget } Datum: maandag 15 maart 1999 23:32 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We are trying to equip our lab with a reflected light BF/DF } microscope (+ trinocular head, B&W video system, 4X5" Polaroid } system), for two basic needs:
Dear Kim, an old method which I used a long while ago and now and then recently to remove gold from calcareous surfaces of REM-samples is to leave the sample in a solution of NaCN and NaOH and let air bubble through by means of a small glass pipette. The amount of NaCN is about 1-2% and NaOH should be 1 N or more. You may try it on a fresh part of bone perhaps it works. Dietmar Keyser PLEASE NOTE THE CHANGE IN TELEFON AND FAX Dr.Dietmar Keyser Zoologisches Institut und Museum Martin-Luther-King Platz 3 D-20146 Hamburg Germany Tel. +49 40/428 38 4232 Fax: +49 40/428 38 3937 E-mail: Keyser-at-zoologie.uni-hamburg.de
EUREM XII 12th European Congress on Electron Microscopy Brno, Czech Republic, July 9-14, 2000 http://www.eurem2000.isibrno.cz/ -------------------------------------------------------- Second Circular and Call for papers will be distributed in May 1999 to pre-registered scientists. DO NOT FORGET TO PRE-REGISTER YOURSELF AS SOON AS POSSIBLE! PRE-REGISTRATION FORM CAN BE COMPLETED DIRECTLY ON THE WEB SITE:
http://www.eurem2000.isibrno.cz/regform.html
Petr Schauer
+---------------------------------------------------------------------+ | Dr. Petr Schauer, Vicechairman of the Or- | tel.: (+420 5) 41514313 | | ganization Committee of the 12th EUROPEAN | fax : (+420 5) 41514404 | | CONGRESS ON ELECTRON MICROSCOPY | (+420 5) 41514337 | | (Brno, Czech Republic, July 9 - 14, 2000) | e-mail: petr-at-isibrno.cz | | Kralovopolska 147, CZ-612 64 Brno | eurem2000-at-isibrno.cz | | Czech Republic |www.eurem2000.isibrno.cz | +---------------------------------------------------------------------+
Greetings Friends, Does anyone have tips for critical point drying and SEM viewing of fresh fish samples including skin/scale layers? They seem to be VERY oily, and I am wondering if this will hamper the CPD process or muck up the vacuum in the SEM. Any food or fish scientists out there? Thanks, Linda lfox1-at-wpo.it.luc.edu
{bold} "Analyzing Materials Interfaces at Atomic Resolution" {/bold}
There will be a Materials Science symposium at Scanning 99 entitled "Analyzing Materials Interfaces at Atomic Resolution". Scanning 99 is being held at the Hyatt Regency O'Hare in Rosemont, IL from April 11-14 1999. The "Analyzing Materials at Atomic Resolution" symposium is scheduled for Tuesday April 13th. The details of the conference can be found at http://www.scanning.org or can be requested from Mary Sullivan (e-mail: scanning-at-fams.org, tel:201-818-1010). Registration for members of the Midwest Microscopy and Microanalysis Society is at the reduced rates of $150 (regular $ 235) for the whole conference or $50 (regular $ 95) for a single day (all attendees of the MMMS symposium held at UIC last May are members of MMMS).
{bold} Speakers {/bold}
9.00 {bold} M. Haider-CEOS GmbH, Germany {/bold}
"Towards sub-Angstrom resolution by correction of spherical aberration"
9.30 {bold} O. Krivanek-University of Washington {/bold}
"Towards sub-Angstrom electron probes by Cs-corrected STEM."
10.00 Break
10.30 {bold} E. M. James-University of Illinois at Chicago {/bold}
"Atomic resolution scanning transmission electron microscopy on the 200kV FEGTEM"
11.00 {bold} S. J. Pennycook-Oak Ridge National Lab {/bold}
"Probing the Origin of Interfacial Properties by STEM"
11.30 {bold} D. A. Muller-Lucent Technologies {/bold}
"The End of the Roadmap for Silicon Dioxide: The Electronic Structure of Hyper-Thin Gate
Oxides at the Atomic Scale"
12.00 {bold} D. B. Williams-Lehigh University {/bold}
"Atomic-Resolution X-ray Microanalysis in the AEM"
12.30 Lunch
2.00 {bold} L. D. Marks-Northwestern University {/bold}
"Picometer structure determination using Electron Diffraction"
2.30 {bold} J. M. Gibson -University of Illinois at Urbana-Champaign {/bold}
"Statistical Measurement of Electron Scattering Fluctuations in Amorphous
Materials - A new Structural Tool"
3.00 Break
3.30 {bold} M. Gajdardziska-Josifovska-University of Wisconsin at Milwaukee {/bold}
"Quantitative surface microscopy and diffraction over the length scales: Morphology and
crystallography of polar oxide surfaces. "
4.00 {bold} M. Tanaka-NRIM, Tsukuba, Japan {/bold}
"Nano-behavior of Small Metal Particles in the Electron Beam"
I would like to establish some contacts with the TEM comunity in Brazil and I thought that perhaps you could give me some information concerning TEM-oriented Materials Science groups in Brazil.
Regards,
LF Giles
***************************************** Luis Felipe Giles, PhD
MPI of Microstructure Physics Weinberg 2, D-06120, Halle (Saale) Germany
} Kim DeRuyter wrote: } ... } } Hi, } A student in the lab is looking at museum samples of dinosaur bones and } teeth on the SEM. He could get access to more samples if he could restore } them to their original condition ( ie, remove the gold). Is there a good } nondestructive way to do this? } } ...
Of all the suggestions, I believe you'll want to develop the casting procedure for your facility. I can give you the e-mail address for Dr Robert Pastor who can let you know which resin materials he used, but he used two ... one for the field while casting the pliable negative mold of ancient Pakastani teeth, and another casting resin for the durable positive mold in the lab. You'd then be able to coat with gold and archive these specimens for all types of future study ... and I was quite impressed with the detail of the micr-wear and lack of artifacts.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
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I sent the following message out yesterday with the incorrect Email. I talked to Chris who has gone from human teeth to mastodon teeth (a bit large to cast for SEM!!) and he would be happy to provide details and answer questions about his casting method. The correct Email is: cschmidt-at-uindy.edu
Sorry for the mistake. Debby -------------
Kim, A good way to handle specimens of this sort is to make casts of the surface and examine the casts in the SEM. This way there is no damage to the original artifact. I had an anthropology grad student do this with human teeth for his thesis research. He worked out a very inexpensive, low tech, but reliable method to do the casting. He can be reached at the following for full details of his method: Dr. Chris Schmidt Indianapolis University cschmidt-at-indy.edu
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
(SMTPD32-4.06) id A372AEA00BC; Tue, 16 Mar 1999 12:22:58 EST Message-Id: {3.0.3.32.19990316123207.00ea5d10-at-mail.map.com} X-Sender: mme-at-mail.map.com X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.3 (32)
Dear Linda,
The world class expert on fish scales is Dr. Gilbert Hartley in the UK. You can probably get his contact information by contacting the Royal Microscopical Society, 37/38 St. Clements Street, Oxford, England.
Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 07:44 AM 3/16/99 -0600, Linda Fox wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If you are growing stem cells in a culture well plate and they're fairly confluent you can do all the processing in the dish, including embedding. This saves alot of time as you dont have to spin between changes, etc. If the plates are Permanox, any resin will work as will PO. However if they're in something like Falcon or Corning you need to use Eponate 12 and skip the PO. I've tried lots of resins and most will partially dissolve the plastic. After polymerization you simply pop the capsules off the dish and section. This works very well if you have a fairly large population of cells. Otherwise you just need to cut several blocks. I can send you our protocol if you'd like. Good luck!
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
I believe that the 1 N NaOH is unecessarily strong to accomplish the purpose and would result in damage to some specimens. A very brief exposure to NaCN with a stronger oxidant such as ammonium persulfate or hydrogen peroxide would probably accomplish the result without prolonged exposure to the reagents which would lead to their becoming more deeply absorbed into capillaries that may exist.
John Twilley Conservation Scientist
KEYSER.DIETMAR wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Kim, } an old method which I used a long while ago and now and then recently } to remove gold from calcareous surfaces of REM-samples is to leave } the sample in a solution of NaCN and NaOH and let air bubble through } by means of a small glass pipette. The amount of NaCN is about 1-2% } and NaOH should be 1 N or more. You may try it on a fresh part of } bone perhaps it works. } Dietmar Keyser } PLEASE NOTE THE CHANGE IN TELEFON AND FAX } Dr.Dietmar Keyser } Zoologisches Institut und Museum } Martin-Luther-King Platz 3 } D-20146 Hamburg } Germany } Tel. +49 40/428 38 4232 } Fax: +49 40/428 38 3937 } E-mail: Keyser-at-zoologie.uni-hamburg.de
We are in the final phases of deciding which confocal microscope to purchase. We are down to the following two which are comparably equipped and priced. We are having difficulty resolving this decision and would like some input from those who may have some experience with these instruments, the software and support service.
Here are our two choices: ******* Nikon PCM - 2000 MultiLine 3 Confocal w/Compix Software
8 bit capture 2 monitors 3 PMT's Fiber Optic Image capture ******* Olympus Flouview 12 bit capture 1 monitor 2 PMT's Direct optical light *******
We are experienced with standard light microscopes, SEM and TEM but are novices with the subtleties of making a good choice in the purchase of a confocal microscope. Please ask me about any additional information I haven't listed that is pertinent to a clear comparison.
Any suggestions about these instruments and the issues we need to consider would be greatly appreciated.
Thanks in advance for your help, Gerald Harrison Biochemistry/Dental Univ. of Penn.
I am forwarding this message for a Ph. D student in our department.
"Does anyone have references for or a working procedure for differentiating between macrophages and neutrophils in tissue sections processed for TEM. I am currently trying to determine the lineage of inflammatory cells present in dog muscle. These cells harbor a parasite, H. americanum. Preliminary TEM data suggests that these cells are modified macrophages but I need to positively confirm the lineage. I would be grateful for any information or ideas as to how to positively determine that these cells are really macrophages or of a monocyte lineage".
Thanks.
Phoebe J. Doss Manager, Electron Microscope Lab Oklahoma State University
I've read with interest the comments regarding negative scanners for TEM because I'm currently starting to look for scanners for X-ray film. Questions:
1) The resolution info. I've gotten for x-ray film scanners is that they can scan to 50 micron resolution. How do I convert between the dpi resolution quoted on the microscopy listserver to this type of number? My back of the envelope calculations seems wacky when I convert.
2) The x-ray film is much thicker and darker than TEM film. Do the TEM scanner folk talk about what the scanners are capable of blasting through?
At 15:44 16/03/99 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
U.S.A. Contact Jim Hilton Advanced Database Systems 7931 S. Broadway #322 Littleton CO 80122 U.S.A. Tel: + 1 303 761-5635 Fax: + 1 303 761-592
} ***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
We are thinking about revising the user fees (including tech support) for our TEM, SEM and all related equipments. However, before doing so, we would very much appreciate some feedback about user fee structures in place and current rates. This is a centralized, university EM facility serving both biological and material people.
Secondly, the university acquired a new confocal microscope and it is going to be a part of the EM facility. I would also like to get some user fee structures for this instrument.
Thanks in advance,
Soumitra Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
} Date: Mon, 15 Mar 1999 15:01:29 -0600 } From: Frank herbert {fherbert-at-bcm.tmc.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM of Lymphocytes } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Howdy all, } There is a graduate student here who is trying to look at a possible stem } cell line under TEM. The problem is that they are quite small, don't seem } to pellet very well, and he has only been able to get me a few thousand } cells at a time. } We have tried to embed the cells in 2% Agar after fixation but this hasn't } worked. } Is it possible to filter the media and cells, and then process the filter } with the cells stuck onto it? Maybe use a cytospin to spin the cells into a } filter? } Is there anyone with experience with this? Any suggestions or ideas would } be greatly appreciated. } } } Thanks in advance } Frank Herbert } Technician } Integrated Microscopy Core } Department of Cell Biology } Baylor College of Medicine } I presume, from your description, the cells are non-adherent?
Pellet the cells. Remove media with pipet. Add small amount of 2-4% glut (depends on size of tube--?maybe 1/2 ml). Resuspend cells, place into very tiny tube with pointed end, and IMMEDIATELY pellet. Let sit in fix. Do not resuspend pellet. After an hour or so, remove most of the fix, and cut off the tip of the tube just below cells, then slice the tube above the pellet. You end up with cells in the center of a log of plastic. Gently dislodge cells with paper clip. Drain with wedge of filter paper, but don't allow to dry out--should be the consistency of thick oat meal. Cut into 1 mm cubed pieces if necessary. Coat each pile of cells with 1% molten, cooled agar (Do not fix the agar in glut). Wash in buffer. Fix in Os. Proceed as with tissue. This method was printed in Microscopy Today sometime back. Also try:
Miller SE. 1983. Electron microscopy of tissue culture. In Jones BR, Hayes RL (eds.) Techniques in Electron Microscopy: A Laboratory Workbook. Burgess Publishing Co., Minneapolis, MN. pp. 478-512.
OR reprinted as
Miller SE. 1985. Electron microscopy of tissue culture. In Jones BR (ed.) Electron Microscopy: 41 Exercises by 17 Scientists. Library Research Associates, Monroe, NY. pp. 293-315.
Good luck, Sm
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I got 12.7 um for 2000 dpi (1/2000 in x 25400 um/in) and half that if it is 4000 dpi. You should know that the Fuji plate system was originally developed for X-ray plates and was at 50 um. They had to get that down to at least 25 um for TEM usage. They key to blasting is optical density of the scanner (assuming that you have enough light and exposure control). It should be at least 3.4 or higher but costs as this goes up. See the book, Real World Photoshop (3,4,or 5). It has a very good section on scanners and buying considerations. You should also get something that outputs with a depth of 12 bits per channel or higher. It really helps in getting contrast and brightness correct in the final TEM image. I would avoid scanner software that automatically converts it to 8 bits -you can do better. You do need a software platform (NIH Image, Photoshop, Digital Micrograph, etc.) that will handle 16 bit images so that you can convert them to 8 bits.
-Scott ---------- } From: "rgriffin-at-eng.uab.edu"-at-Sparc5.Microscopy.Com To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
I've read with interest the comments regarding negative scanners for TEM because I'm currently starting to look for scanners for X-ray film. Questions:
1) The resolution info. I've gotten for x-ray film scanners is that they can scan to 50 micron resolution. How do I convert between the dpi resolution quoted on the microscopy listserver to this type of number? My back of the envelope calculations seems wacky when I convert.
2) The x-ray film is much thicker and darker than TEM film. Do the TEM scanner folk talk about what the scanners are capable of blasting through?
} Date: Tue, 16 Mar 1999 08:04:52 +0100 } From: Birgit Neubohn {neubohn-at-ipk-gatersleben.de} } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM: antibodies bind to formvar } =20 } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Hello, } =20 } in our group we are using formvar-coated grids to stabilize ultrathin } sections for immunogold-labelling. } Sometimes the formvar is *heavily* labelled by gold particles. This is no= t } due to binding of the gold grains, but depends only on the primary antibo= dy } that had been used. It can happen both with Protein A-gold or goat anti } rabbit-gold, but using other primary antibodies the formvar is clean. } What is the reason for this binding of antibodies to formvar? } Is there any blocking reagent that would provide this? } Thanks in advance } =20 } Birgit } =20 } =20 } -------------------------------------------------------------------------= ----- } Dr. Birgit Neubohn } Institut fuer Pflanzengenetik und Kulturpflanzenforschung (IPK) } (Institute of Plant Genetics and Crop Plant Research) } Corrensstr. 3 } D-06466 Gatersleben, Germany } =20 } Phone.: (+49) 039482 5447 } Fax: (+49) 039482 5139 } e-mail: neubohn-at-ipk-gatersleben.de } -------------------------------------------------------------------------= -----=9D } =20 We use Formvar routinely for ultrathin cryosections. There should be no=20 gold on the plastic. You should block your sections with a protein=20 before the primary, and use protein in the antibody solutions as well. =20 Bovine serum albumen (BSA) and fetal calf serum (FCA) are commonly used. = =20 We wash grids in 5% FCS in PBS 10 min, PBS with 100 mM ammonium=20 chloride--quenches aldehydes in cryo sections (you don't say what kind=20 of sections), PBS, then primary, then 6 washes in FCS/PBS, then=20 secondary in PBS with FCS, 6 more washes; wash off protein with water=20 before proceeding. Next step will depend on what kind of sections you=20 have. Results: no background.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710=20 Ph: 919 684-3452 FAX: 919 684-8735
I am looking for configuration details for a PCVisionplus framegrabber - = specifically, the board jumper settings. These are all listed in the = hardware manual (which we have mislaid). If anyone has this info, could = you please email or fax me, as below. The PCVisionplus card was very popular a few years ago, as one of the = preferred framegrabbers for a variety of image analysis packages (JAVA, = Optimas, etc).
Cheers,
David Vowles Senior Technical Officer Electron Microscopy Unit Australian National University PO Box 475 Canberra ACT 2601 Australia Tel: +61 2 6249 3543 Fax: +61 2 6279 8525 Email: vowles-at-rsbs.anu.edu.au
rgriffen asks ... } } ... } } 1) The resolution info. I've gotten for x-ray film scanners is that they } can scan to 50 micron resolution. How do I convert between the dpi } resolution quoted on the microscopy listserver to this type of number? My } back of the envelope calculations seems wacky when I convert.
My calculations at this late hour would imply 50 microns converts to 500 "per inch", but since "resolution" is the distance between two resolvable points (with something between), a 50u resolution scanner would need scan "optically" at 1000 pixels/inch (the distance between the centers of 2 black pixels with a white pixel between).
} } 2) The x-ray film is much thicker and darker than TEM film. Do the TEM } scanner folk talk about what the scanners are capable of blasting through?
This is a good question ... most transparency and film scanners don't do very well for dense areas. Presumably if the manufacturer advertises a relatively high "dynamic range" it will do better ... but do evaluate for detail and noise in those areas.
} I am wonder if Dietmar method could be used for beetles... } } Regards } } Ricardo } www.coleoptera.org
Hello Ricardo! Especially for insects have a look at:
C. Arno': Removal of gold coatings from biological SEM specimens, European Microscopy and Analysis, July 1998, p.13
In this suggestion bromine vapors are used. If you have problems to get this article, let me know your fax number. I then will send a fax copy -- Gerhard Frank UNIVERSITAET ERLANGEN-NUERNBERG Phone: +49 9131 85 28606 Institut fuer Werkstoffwissenschaften VII Fax: +49 9131 85 28602 Cauerstr. 6, D-91058 Erlangen, Germany frank-at-ww.uni.erlangen.de
Dear Dr. Neubohn, In our experience there is a huge variety in stickiness of primary rabbit antibodies. We have the impression that this originates from different degrees of hydrophobicity, the more hydrophobic the protein, the easier it sticks to a hydrophobic surface (formvar, resins and so on). If this problem shows itself it is almost always with rabbit species. Possible solutions are: 1. make the primary less hydrophobic. Some have done this by pre-incubating the primary antibody with free protein A, but this would make a detection with protein A/gold complexes impossible. If you need a reference I can look it up. An alternative is sometimes, but not always, to use Fab or Fab2 fragments of the primary, but then you would have to use a (goat)-anti-rabbit conjugate. 2. make the substratum less hydrophobic. In general parlodion films seem to be less sticky, but with formvar films possibilities are to use glow discharged films, and a carbon coating. An appropriate blocking step will deal efficiently with hydrophobic binding sites on the substratum. Albumin is suited and a relatively long blocking time helps, as more albumin molecules tend to share larger surface areas with the substratum, resulting in a more efficient blocking with time. If you work with resin sections, you might consider to use Tween-20 (0.1%, not less!) in PBS for an incubation buffer. At concentrations above the critical micel concentration the detergent will prevent hydrophobic interactions like the one you describe. This approach may not be suited for cryo sections, since they are much easier affected by detergents. Good luck, Jan =========================== Jan Leunissen AURION http://www.aurion.nl Costerweg 5 The Netherlands phone: 31-317-497676 fax: 31-317-415955 You will find more technical info on our web site
rgriffin, I've recently had a few attempts at using my Linotype-Hell Saphir Ultra scanner to digitise X-ray radiographs and topographs. I usually use the scanner for TEM negs, and it does a fair job for those as long as they're not too full of contrast (haven't gone in the darkroom since I bought it). I have found it woefully inadequate for X-ray images recorded on nuclear emulsion (grain size 5 or 10 um). The 'real' (supposedly non-interpolated) resolution of the scanner is 1200 dpi, even though it will interpolate up to 5000 dpi. (2.54 cm/1200 = 21 um pixel size). Nearly all of the image detail is lost in a scanned image, even at the maximum resolution setting. And yes, you have to have a fairly weak-looking negative to cope with the contrast produced by a gold wire in a radiograph. My current way of dealing with these images is to use a digital camera on a microscope, which is fine for detail of wire bonds, etc. However, I know I'll have to go into the darkroom today since I have some topographs which are several inches square. I'll scan the prints. In a couple of weeks I'll have a macro lens for the camera and may be able to take pictures from a light box. Unless scanners have improved massively since last year, I wouldn't recommend using one as your only way of getting an image. (I believe 2000 dpi is standard now, and maybe 15% better range in optical density.)
Tel. +44 1327 356363 Fax. +44 1327 356389 } ---------------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------. } } } I've read with interest the comments regarding negative scanners for TEM } because I'm currently starting to look for scanners for X-ray film. } Questions: } } 1) The resolution info. I've gotten for x-ray film scanners is that they } can scan to 50 micron resolution. How do I convert between the dpi } resolution quoted on the microscopy ListServer to this type of number? My } back of the envelope calculations seems wacky when I convert. } } 2) The x-ray film is much thicker and darker than TEM film. Do the TEM } scanner folk talk about what the scanners are capable of blasting through? } }
I have been using a system by Orion with my JEOL 840 SEM. I have been using the DOS version and am in the process of upgrading to the Windows NT version. You can visit their web site at: http://www.OrionMicroscopy.com/
Louie Kerr
At 3:44 PM -0600 3/16/99, "rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX) 508-292-0289 (Cell phone)
Hi Linda, Although I haven't tried this with oily fish scales, I've managed to defat raw food samples (at different stages of refining and production). In preparation for CPD, samples can be dehydrated through a gradient series of ethanol or acetone. If the sample still contains oil, you might try hexane followed by acetone before going to CPD. A common alternative to CPD is to use hexamethyldisilizane (HMDS) x3 following the final 100% ethanol wash. Your samples can then be air dried. Do some trial runs on samples on test samples. The HMDS is available from EMS or other EM supply companies. Let me know how this project works out. Rosemary
We have prepared fish parts including scales for SEM and TEM without problems. The standard preparation protocols of fixation, washes, dehydration and CPD should result in clean dry samples. One problem with scales is that they sometimes tend to curl up as they dry. Usually we just work around that problem but you can try to process them more slowly or try to hold them flat through the processing.
Louie
At 7:44 AM -0600 3/16/99, Linda Fox wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX) 508-292-0289 (Cell phone)
On the general subject of removing sputtered of evaporated metal from SEM samples. I am told that removing silver can be a relatively simple. The following paper describes a method using "Farmer's" reducer, a dilute aqueous solution of potassium ferricyanide and sodium thiosulphate.
"Silver as a removable conductive coating for scanning electron microscopy" by A.A. Mills Dept. Geology, University of Leicester. UK
Best regards Mike Wombwell Polaron range Business Manager V G Microtech The Birches Industrial Estate Imberhorne Lane East Grinstead West Sussex RH19 1UB UK Direct line: +44 (0)1342 310296 Switchboard: +44 (0)1342 327211 Fax: +44 (0)1342 315074 http://www.vgmicrotech.com/polaron-range E&OE
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At 03:44 PM 3/16/99 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Our company, E.L.I. Belgium has designed a very powerful image grabbing s= ystem for SEMs: Orion 5 for Windows.
This system connects to ANY SEM. Its resolution goes up to 8k x 8k (only depending on the SEM resolution). A lot of useful functions are easily available: zoom, distance measuremen= ts, image extraction, text inclusions,...=20 Orion is the only frame grabber with FULL electrical isolation between th= e SEM and the PC. Orion is fully programmable with macro commands. The Orion system is fully configurable: up to 64 image sizes stored in a driver file. Orion runs under Windows 3.1x, Windows 95 and Windows NT. Continuous software updates guarantee the evolution of the Orion system.=20 It is a fully passive device that tracks the SEM scanning signals - easy connection Orion is open to the outside world: all =93 popular =94 image file format= s (TIFF, BMP, TGA, JPG...) can be used.
Please visit our web site (orionmicroscopy.com) to get more info and also= to download a demo program to evaluate the different functions without any cnnection to an SEM or contact us directly to know our nearest dealer.
Best regards,
Paul Vanderlinden. Sales Manager.
********************************************************************* See our web site: http://www.orionmicroscopy.com
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{html} {div} At 03:44 PM 3/16/99 -0600, you wrote: {/div} {div} >----------------------------------------------------------------= -------- {/div} {div} >The Microscopy ListServer -- Sponsor: The Microscopy Society of America {/div}
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At 03:44 PM 3/16/99 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Our company, E.L.I. Belgium has designed a very powerful image grabbing s= ystem for SEMs: Orion 5 for Windows.
This system connects to ANY SEM. Its resolution goes up to 8k x 8k (only depending on the SEM resolution). A lot of useful functions are easily available: zoom, distance measuremen= ts, image extraction, text inclusions,...=20 Orion is the only frame grabber with FULL electrical isolation between th= e SEM and the PC. Orion is fully programmable with macro commands. The Orion system is fully configurable: up to 64 image sizes stored in a driver file. Orion runs under Windows 3.1x, Windows 95 and Windows NT. Continuous software updates guarantee the evolution of the Orion system.=20 It is a fully passive device that tracks the SEM scanning signals - easy connection Orion is open to the outside world: all =93 popular =94 image file format= s (TIFF, BMP, TGA, JPG...) can be used.
Please visit our web site (orionmicroscopy.com) to get more info and also= to download a demo program to evaluate the different functions without any cnnection to an SEM or contact us directly to know our nearest dealer.
Best regards,
Paul Vanderlinden. Sales Manager.
********************************************************************* See our web site: http://www.orionmicroscopy.com
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Several people in the recent past have asked about the discontinued Leafscan 45 film scanner. Leaf transferred the technology to Bremson, Inc. of Kansas. Their web page can be found at www.bremson.com . The page detailing the specs for the scanner can be found at www.bremson.com/products/45hsprod.htm .
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798 cook-at-aaem.amc.anl.gov
The biggest difference between these cell types is in the nucleus. Neutrophils have a multilobed nucleus, and in thin sections this will a= ppear as multiple small, unconnected profiles. The nucleus of macrophages ma= y be deeply indented, but it will usually not be seen as multiple, unconnect= ed profiles. The cytoplasm of neutrophils is generally more electron dens= e than macrophages, and there will be more granules in the neutrophil cytoplas= m. Macrophages will have a distinct Golgi region with lots of vesicles and=
secretory granules of many sizes.
The best way to learn to differentiate the cell types is to get a buffy= coat sample and fix it without disturbing the cell layers. Neutrophils and eosinophils will layer nearest to the erythrocyte layer, lymphocytes an= d macrophages will be next, and platelets will be at the top. I've got a= n easy procedure for processing buffy coats for TEM, if you'd like to have it.=
Jane A. Fagerland, Ph.D. Abbott Laboratories Abbott Park IL 60064-6202 =
We're new to making density measurements on digital images. I'm looking for basic papers/books on the topic. I already have John Russ's "The Image Processing Handbook" and "Computer Assisted Microscopy". Is there any other good stuff out there?
Dear Fellow Microscopists, What is the best way of determining Carbon in solution in Ni-base superalloys? Chemical analysis strikes to me as one of the best ways, but I don't have enough material. Thanks. Anita
} Date: Wed, 17 Mar 1999 15:01:15 -0600 } To: Histonet-at-Pathology.swmed.edu } From: Sue Danielson {sdaniels-at-post.its.mcw.edu} } Subject: Laboratory Technologist position } } The Department of Neurology at the Medical College of Wisconsin has an immediate opening in their Neuromuscular Diagnostics Laboratory for a Clinical Laboratory Technologist. Our fast-paced, growing laboratory receives muscle & nerve biopsies from hospitals & clinics throughout Wisconsin and Northern Illinois. } } Applicants should possess a Bachelors' degree in the Biological or related Sciences or an MT/HT certification. Laboratory experience in histology (cryosectioning and muscle enzyme histochemistry) and transmission electron microscopy techniques a plus. Excellent organizational skills and ability to work independently as well as with physicians and medical students is essential. } } Excellent salary and benefits package offered including health and dental insurance, 403B retirement plan and tuition reimbursement. This is a full-time, 40hr/wk salaried position (no weekend or evening hrs required) available immediately. } } Applicants may contact/submit resume to: } } Susan K. Danielson, MS } Neuromuscular Laboratory Coordinator } Froedtert Memorial Lutheran Hospital - West } Muscle/Nerve Lab, Rm 1132 } 9200 W. Wisconsin Ave. } Milwaukee, WI 53226 } } Ph: (414) 259-3836 } Fax: (414) 454-7905 } email: sdaniels-at-mcw.edu }
Gerald- I have worked with the Fluoview system from Olympus, and the BioRad MRC600, but not the Nikon system. I prefer the BioRad over Olympus. My advice is to sit down and work with the software, with each of the systems you are considering. Time at the scope is the only way to understand the quirks of each. If you don't have the time, assign someone you trust to do the comparison. Get both scopes set up as DEMOS for at least 2-4 weeks. Run them trough their functions, collect images, edit images, transport to various other SW programs, print to various printers, etc. All systems have bugs, most are in the software, file formats, the ability to transfer images from one platform to another is essential... find the bugs, decide if you can live with them.
Good luck -Mike
disclaimer- I have no financial interest in any of the above mentioned companies.
On Tue, 16 Mar 1999, Gerald Harrison wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear fellow microscopists, } } We are in the final phases of deciding which confocal microscope to } purchase. We are down to the following two which are comparably equipped } and priced. We are having difficulty resolving this decision and would like } some input from those who may have some experience with these instruments, } the software and support service. } } Here are our two choices: } ******* } Nikon PCM - 2000 MultiLine 3 Confocal w/Compix Software } } 8 bit capture } 2 monitors } 3 PMT's } Fiber Optic Image capture } ******* } Olympus Flouview } 12 bit capture } 1 monitor } 2 PMT's } Direct optical light } ******* } } We are experienced with standard light microscopes, SEM and TEM but } are novices with the subtleties of making a good choice in the purchase of a } confocal microscope. Please ask me about any additional information I } haven't listed that is pertinent to a clear comparison. } } Any suggestions about these instruments and the issues we need to } consider would be greatly appreciated. } } Thanks in advance for your help, Gerald Harrison } Biochemistry/Dental } Univ. of Penn. } } }
Thanks to the several contributors who posted very helpful information to me regarding our choice between a Nikon and Olympus confocal. We were restricted to these two choices by budgetary constraints.
I have just emerged from what is the last meeting for making this decision and we went over the many good questions and comments I've received. They were a significant aid in our being able to focus on the kind of unique considerations facing us.
We believed, from on-site demonstrations of both instruments and comments I received from the List, that image quality could be considered equivalent. Olympus was also including an additional remote workstation to compensate for the advantage of the Nikon's Compix compatibility with Windows NT. We were swayed by the apparently more powerful software for the Nikon, but because we will be a multi-user facility primarily doing routine visible fluorescence imaging, and the person who will work with users is already familiar with the Olympus package and believes it is easier for a novice to learn basic image acquirement, we will go with the Olympus.
Again, the department personnel involved in making this choice are very grateful for all the helpful input.
Gerald Harrison Biochemistry/Dental Univ. of Penn.
When I posted this letter for the first time, something went wrong, and = the last 10 percent were lost.
As Valdemar Furdanowicz wrote you, you can transfer LINK files to the PC = with their MSDOSCV program. However, you can also use the serial port of the = LINK=20 computer and it can be a little bit faster. What I don't know, however, = is whether you can connect the the two machines directly. When I worked in the electron = beam lab of Hungalu Engineering, we had a little box between the serial ports=20 of the machines, made by Oxford. You can try any terminal program on the PC if it has an option for the = well-known XMODEM protocol. Once transferred, the files have to be converted from one format to = another , because the numbers are formatted differently on the LINK and the PC. With fixed point numbers the only difference is the order of the bytes, = but with floating point numbers the problem is more serious.=20 Your opportunities for file conversion are very limited with the MSDOSCV = program.=20 Of course you have to know the structure of the files, if you want to = convert them.=20 You can find this information in the Oxford manuals.
I wrote some programs to solve the above mentioned problems: 1. File transfer: with our XMODEM program we can transfer multiple files = using wildcards in=20 the file specifications. For this purpose the PC program prepares the = necessary LINK macros, which are sent as a first step. 2. File inspection: with a little utility program one can look at the = files obtained from the LINK computer and compare their content with the documentation.=20 3. File conversion: I have worked out simple conversion programs for = image files, DIGISCAN image anal. result files and X-ray spectra.
With a private letter I can send you anything from these programs, but = there are still some minor=20 problems with that. When I worked for Hungalu, I was the only user of my = programs, and I didn't have enough time for a better user interface. They are written = for the DOS platform, and they send Hungarian messages to the screen. Now I work as a computer programmer in a very different field, and = microscopy remained only a hobby for me. Well, I would like to port my programs to Windows = for my previous colleagues,=20 but I can work on them only in my spare time. =20 (And maybe, I would like to earn a little money if they can afford to = pay.) Anyhow, you can try to convert the image files with Photoshop. They can = be read by Photoshop as raw bitmaps. Here you don't have byte order problems, because the = LINK machine usually stores=20 every pixel on one byte. Even black and white pictures use eight bits = for every single pixel.=20 (Such files can be packed with PKZIP on the PC with 50 to 100:1 ratios.) There is only one thing you have to do: remove the header from the beginning of the file. As far as I can remember, it is 256 bytes long.
Good luck!
******************************************* Mr. L=E1szl=F3 Varga M=C1V Informatika Ltd. H-1012 Budapest, Krisztina Krt. 37/a Hungary Tel: (36-1) 457-9387 email: varguc-at-freemail.c3.hu ********************************************=09 -------------------------------------------------------------------------= ----------- Robert McDonald wrote: Hi All:
I have recently taken over an SEM with an AN 10000 analyser and I was wondering if anyone out there kows if it is possible to get the results files out to a PC ?=20
I'm looking for spares for a Durst M800 Enlarger and a Fujimoto G70 Dichro = Enlarger. Some bright spark has dropped the negative carrier in both = cases and broken the glass plates. If I can get a replacement negative = carrier or just replacement glass plates that would be great. I'd also = like to get some specs on the plates, so if need be I can get replacement = plates made up. =20 If anyone has a complete unit that they want to get rid of rather then the = parts, let me know, we might be in a position to buy for a reasonable = price. =20
Thanks=20
George
If at first you do succeed, try not to look surprised!
George Theodossiou Dept Applied Physics RMIT GPO Box 2476V Melbourne 3001 Victoria Australia Ph: +61 3 9925 3394 Fax: +61 3 9925 5290 Email: george.theodossiou-at-rmit.edu.au
Dear All, We are in the process of mulling over the question of how much we should charge for various services on offer within the department to in-house users, external users and industry. The aim is to revise the charges at some practical level, neither frighteningly expensive nor laughably cheap. However, in order to come up with realistic values, I thought that some input on what the wider world charges would be of use. Of course, I could just sit down and calculate a figure for each of the things that we can do, but this is complicated as some of the variables are unknown - we don't get an electricity or water bill!! Also, there will be some discrepancies between different countries due to currency and the cost of living, but nevertheless, to know would be of undoubted help. So, please let me know how you price microprobe usage, XRF usage and sample preparation, and thin-section preparation. Yours gratefully, Malc. Dr MP Roberts Department of Geology Rhodes University Grahamstown 6140 South Africa Tel: +27 46 6038316 Fax: +27 46 6229715 ******************************* "A sleep is as good as a walk to a legless insomniac" Anon.
} ---------- } From: John Best[SMTP:jbest-at-elmdas.com] } Sent: Wednesday, March 17, 1999 1:53 PM } To: rgriffin-at-eng.uab.edu } Cc: Best, Christine (BEST); jbest-at-elmdas.com } Subject: Digitizing Philips 515 } } Dear Dr. Griffin, } } Your request for information regarding updating a Philips 515 with } digital imaging was recently forwarded to me. } } We manufacture just such a system, and have installed it on Philips 500 } series SEM's owned and operated at several Philips manufacturing or } research facilities. } } It has active control of the SEM probe, and will therfore add features } like frame averaging to your SEM. Perhaps the best approach is to check } out our website at http:\\www.elmdas.com. } } I look forward to hearing from you. } } Regards, } John Best } } } ELMDAS Co. ELectron } Microscopy } Data } Acquisition } Systems } } Mailing & Shipping: RD1 Box 62A, } Alexandria, PA 16611 } Phone: 814-669-4474 } Email: jbest-at-elmdas.com } Our Website: http://www.elmdas.com }
I'm looking for feedback from labs having experience with insurance companies that provide programs replacing traditional manufacturer's service contracts on scopes and other lab equipment. According to the companies' claims these programs provide substantial savings over regular contracts, with service continuing to be from the provider of choice. In other words, they seem to be saying that we can save huge amounts of money with no decline in service. This is a very attractive proposition for the cost-cutters, but can it be real?
Feel free to reply directly to me, if you like. (And thanks to those of you who have already spoken with me by phone!)
Thanks. Randy
Randy Tindall Electron Microscope Core College of Veterinary Medicine University of Missouri - Columbia Phone: 573-882-8304
I've received several requests for this protocol. Phil Oshel asked tha= t I write it up for Microscopy Today, so it will be available to subscriber= s there, as well.
PREPARATION OF BLOOD SAMPLES FOR ELECTRON MICROSCOPY
1. Collect blood in standard EDTA tube.
2. Spin blood sample in clinical centrifuge to obtain visible buffy co= at (about 3500 rpm for 20 minutes).
3. Carefully remove straw-colored plasma layer with pipette, without disturbing the buffy coat layer. It won't hurt to leave some plasma ab= ove the buffy coat.
4. Gently add buffered glutaraldehyde by running it dropwise down the = sides of the tube, again without disturbing the buffy coat layer. 3% glut in= 0.1M cacodylate or Sorensen's phosphate buffer, pH 7.2- 7.4 works fine.
5. After about 30 minutes, test the plug to see if it's fixed - it sho= uld feel rubbery with gentle prodding. If it is not firm, replace the fixa= tive with fresh and allow it to fix for another 15 - 30 minutes.
6. Remove the fixed buffy coat plug from the tube. A small spatula or=
sharpened wooden stick works well. This step can get a little messy, b= ut the idea is to get the plug out as a coin-shaped disk with only a small amo= unt of adherent erythrocytes on the underside. Erythrocytes can be washed off= with buffer, if needed.
7. Place the plug with the erythrocyte-side down on dental wax and cut= out a long transverse slice. Take the slice and slice it again crossways int= o tissue-sized blocks (1-2 mm thick). Each block will contain all cell t= ypes with erythrocytes on the bottom, then granulocytes (neutrophils, eosino= phils, and basophils), then macrophages (often mixed with large lymphocytes), = small lymphocytes, and finally platelets at the top.
8. Place the blocks into a processing vial, and fix for an additional = hour in glutaraldehyde.
9. Wash in buffer, 2 x 10 minutes each.
10. Post-fix in 1% osmium tetroxide in 0.1 M buffer (same type as used = in glut fixative).
11. Rinse several times in distilled water.
12. Dehydrate in ethanol: 50, 70, and 95%, 15 minutes in each.
13. Absolute ethanol: 2 x 15 minutes each.
14. Propylene oxide: 2 x 15 minutes each.
15. Propylene oxide: epoxy resin (1:1) - 1 hour.
16. 100% resin - 2 x 8 hours
17. Orient for embedding so that the largest face of the block will be sectioned (to get all the cell layers in the section).
18. Polymerize blocks as usual.
The dehydration and infiltration schedule is what I've used and know wo= rks. Other methods will most likely work (e.g., acetone dehydration, differe= nt infiltration schedules).
If there are any further details needed, please contact me.
Good luck!
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064-6202 =
Does the MSA have a web site or something that lists EM & LM job postings? Someone told me that it exists but I don't know where to look. So please drop me a line and I'll fish around in the listings.
With things going swimmingly well,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Dear Robin, I have had the Quartz PCI digital imaging system for several years and I am very happy with it. The software runs on Windows 95, NT and 98, is very robust (even my students can't crash it) and it can attach to any SEM. It really transforms older SEMs. You can contact them at: http://www.quartzimaging.com/ You wrote: } } I need vendors that can sell me the set-up for getting digital images from } our Phillips 515 SEM. Any suggestions? } } Robin Griffin } rgriffin-at-eng.uab.edu } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Regarding the question about charges in a centralized EM facility--We have the same situation but without sem. The federally mandated cost accounting standard has made things difficult but since all universities must now comply, we have no choice. Because of this, your rates should vary based on cost of equipment (and therefore depreciation), cost of personnel and the number of hours the facility is used by paying customers. Our facility charges $25.00 per hour for a Hitachi H7000 TEM and will be charging $35.00 for a new TEM with digital capabilities. Technical help is 35.00 per hour, an inverted multi-photon confocal is $25 per hour and a 3 laser upright confocal with nomarski is $15 per hour. We offer two hours of training at no charge. Use of the darkroom which covers chemicals and the service on our processor is $10 per hour. We charge cost plus shipping on any supplies or chemicals people use. After hours rates are the same. These rates vary yearly depending upon usage. Unfortunately we can't charge what it really cost to operate the facility or we'd have no business--so the university subsidizes us. Hope this helps,
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
It's me again. I've just been asked to speak to a group of high school kids about EM, and how I got to where I am today (I'm not gonna tell them about all the little people I squashed along the way). Actually what I wanted to know is if there are any folks out there doing Forensic EM. You know analysis of blood, guts, bullets, things like that. You know that if I gross the kids out they'll think that EM is coolest thing since ice cubes. So if anybody does that sort of stuff or knows of anybody I can talk to about it, let me know. Cuz y'all know there's nothing better than gettin' a great big EEEEWWWW out of the young'uns.
Gotta go, I hear my microtome calling me,
Paula :-)
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Dear Robin, We (Soft Imaging System SIS)have a few options that you may want to look at for acquiring digital images from your SEM. This is where you can find a listing of our products on the web: http://www.soft-imaging.de/products/p_one.htm
The first solution is our ADDA II, slow-scan interface for active or passive digital image acquisition from SEM/STEM. Technical data: Resolution: 4096 x 4096 pixel, 4096 gray values. 8 analog (ADC) inputs. 16 logical input and output channels. http://www.soft-imaging.de/products/hardware/h_add.htm
The second solution is our framegrabber (the grabBit) and our software (analySIS) for acquisition of standard video images generated on your microscope. The image quality, S/N ratio, is much worse for video images than it is for the slow-scan interface.
Our software is a state-of-the-art image acquisition, processing, analysis, and archiving package. We have application specific modules, like STEREO, for generating and viewing height mapped images from a stereo pair.
Please contact me if you have any further questions about either of these solution. I would be glad to send out brochures describing our products, or discuss your specific application. By telephone, (888) FIND SIS, or e-mail ac-at-soft-imaging.com
-Sincerely,
Andrew Cahill Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 web: www.soft-imaging.com, www.soft-imaging.de email: ac-at-soft-imaging.com
At 03:44 PM 3/16/99 -0600, you wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
We bought several large Plexiglas developing tanks from Energy Beam Sciences for developing TEM negatives. Several of them split their seams after a few months and we sent them out for repair. After a few months, they split again. Has anyone else had problems with these tanks? They are model DT-111 and are 1-1/2 gallon size (approx. 8-3/4" x 12" x 8").
Please contact me off-line.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Dear Listers, We are planning to purchase a cold stage for a Zeiss Axioplan and would welcome your experience with vendors in this field. It will primarily be used for imaging food products at temps from -20C to -120C. Thanks in advance! Rosemary
Randy- from my experience these Insurance policies are valid, however the biggest variable is timeliness of service. You will not be at the top of the priority list once you leave the manufacturer and use a middle man to schedule your service needs. -Mike
On Thu, 18 Mar 1999, Tindall, Randy D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } I'm looking for feedback from labs having experience with insurance } companies that provide programs replacing traditional manufacturer's service } contracts on scopes and other lab equipment. According to the companies' } claims these programs provide substantial savings over regular contracts, } with service continuing to be from the provider of choice. In other words, } they seem to be saying that we can save huge amounts of money with no } decline in service. This is a very attractive proposition for the } cost-cutters, but can it be real? } } Feel free to reply directly to me, if you like. (And thanks to those of you } who have already spoken with me by phone!) } } Thanks. } Randy } } } Randy Tindall } Electron Microscope Core } College of Veterinary Medicine } University of Missouri - Columbia } Phone: 573-882-8304 } } }
We encircle our tissue samples using a PAP pen to keep the solutions in place during immunocytochemical procedures. We just received a new PAP pen and we think they changed the formula!!
The PAP material lifts off the slides during incubation whereas with the old pens the material stayed put until we removed it.
Do you all have any suggestions regarding something else we could use in place of the PAP pen??
MSA's Placement Office is the location to look for this information. It is located on the MSA WWW Site. Look under General Society Information. at http://www.msa.microscopy.com
} } } Hidee-ho Boarders, } } Does the MSA have a web site or something that lists EM & LM job } postings? Someone told me that it exists but I don't know where to look. } So please drop me a line and I'll fish around in the listings. } } With things going swimmingly well, } } } Paula :-) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } phone: 510-642-2085 } fax: 510-643-6207 } http://biology.berkeley.edu/EML
Three years ago almost to the day we had a discussion on ENTOMO-L about confocal laser microscopes, which several contributors believed would eventually largely replace the use of electron microscopes in insect taxonomy and morphology. One huge advantage is that images are "captured" directly into a computer, where they can be stored and manipulated later. As Richard L. Brown wrote, with regard to insect genitalia, "The confocal can take images every two microns (or more) from the top to bottom and then reconstruct the photo-sections to provide a flat or 3-D image (with resolution comparable to SEM at lower magnifications). The stored data from all the sections can then be compiled and used to provide an image at any level of the structure or a movie or photo of the whole genitalia as it rotates, allowing one to see areas of the genitalia that are obscured by overlaying structures."
Our University has now purchased such a microscope system, and it has recently been installed. Can a confocal laser microscope be used to view entire objects, or just slide mounts?? I called the technician who will be training us in its use, and he had only heard of using it with microscope slides. I went through the email discussion we had on the list, and nowhere is this limitation stated. My desire is to use it with very small beetles (0.6 mm up to whatever the limit is, max. for my taxa about 12 mm) which cannot be dissected or slide mounted because they are type material or otherwise special specimens.
Perhaps we could hear generally, again, the potential uses for these instruments in entomological research. Surely much has happened since Mar. 1996?
Lawrence ..................................................................... Lawrence R. Kirkendall lawrence.kirkendall-at-zoo.uib.no Associate Professor FAX: +47 55 58 96 73 Univ. Bergen Dept. of Zoology TELEPHONE:+47 55 58 23 42 Allegaten 41, N-5007 BERGEN Norway time = GMT + 1 hour Home ph. (if you can't beat the time zone differences):+47 55 95 00 34
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptiste (surname) of Bejsak (Bayshark)-Collorado-Mansfeld
Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., P.O.Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
http://www.coleoptera.org
Home address: 32 Girrawheen Ave. Kiama NSW 2533 Australia
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
We are having problems with our Kevex 770 keyboard. They seem to = print out random letters and numbers when you press certain keys. Does = anyone have a spare that they are willing to part with. thanks.
______________________ Roberto Garcia Senior Analyst, Metallography North Carolina State University Analytical Instrumentation Facility Box 7531, Room 303 EGRC Raleigh, NC 27695-7531 rgarcia-at-unity.ncsu.com http://spm.aif.ncsu.edu/aif
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD} {META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {STYLE} {/STYLE}
{META content=3D'"MSHTML 5.00.0910.1309"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT size=3D3} Hi, {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D2} We are having problems with our = Kevex 770=20 keyboard. They seem to print out random letters and numbers when you = press=20 certain keys. Does anyone have a spare that they are willing to part = with.=20 thanks. {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D3} ______________________ {BR} Roberto Garcia {BR} Senior = Analyst,=20 Metallography {BR} North Carolina State University {BR} Analytical = Instrumentation=20 Facility {BR} Box 7531, Room 303 EGRC {BR} Raleigh, NC=20 27695-7531 {BR} rgarcia-at-unity.ncsu.com {BR} http://spm.aif.ncsu.edu/aif {/FONT= } {/DIV} {/BODY} {/HTML}
I've just been asked to speak to a group of high school kids about EM, and how I got to where I am today (I'm not gonna tell them about all the little people I squashed along the way). Actually what I wanted to know is if there are any folks out there doing Forensic EM. You know analysis of blood, guts, bullets, things like that. You know that if I gross the kids out they'll think that EM is coolest thing since ice cubes. So if anybody does that sort of stuff or knows of anybody I can talk to about it, let me know. Cuz y'all know there's nothing better than gettin' a great big EEEEWWWW out of the young'uns.
I would be glad to talk to you about forensic applications of scanning electron microscopy as that is my job, however......you should understand "forensic EM" goes far beyond the common opinion of " analysis of blood, guts, bullets, things like that". An electron microscopist performing airborne particle analysis for possible occupational/environmental exposure to harmful particulates or the pathologist using electron microscopy to identify a pathogen/toxin induced lesion or foreign object are also performing forensic EM analyses. Forensic means "Of or used in legal proceedings or in public debate" (The American Heritage Dictionary, 2nd Edition). The investigative accountant analyzing data in the investigation of corporate illegal dealings for prosecution and the forensic scientist in the laboratory matching bloody fibers or a bullet found at a crime scene are both performing forensic analyses.
Is the point of your giving this talk to "gross them out" and generate " a great big EEEEWWWW out of the young'uns" or are you teaching to really stimulate some young minds and show them other areas of science that use microscopy for possible careers? High school students are not "young'uns" and you may have a profound impact on a few futures. You can either encourage and stimulate OR you can deter further interest altogether. Many of us were originally guided to our profession by a good speaker or teacher.
As one who both instructs and lectures extensively on forensic applications of electron microscopy, I have found the audiences (high schools, colleges, forensic science and microscopist societies) all enjoy the case history (not gore) from an investigation. The myriad of television shows on the subject speak to the popularity of this current "hot" topic. I'm sure the international emphasis on the "Crime of the Century" has contributed greatly to the recent popularity in forensic science and scientific evidence. You propose to SHOCK your audience with raw crime scene photos when the students are not used to seeing "the real thing". This is counter productive as they will be discussing the shock image(s) the whole time you are trying to explain the science/EM in the investigation. The "gore" is primarily what they will remember and take away with them.
If you really want to TEACH these high school students, show them how to identify objects from two places (exa. - particles such as soil samples from the "crime scene" with particles from a suspect's shoes - SEM/TEM/EDX) or how to find specific particles that confirm a scenario (exa. - specific lake diatoms in "lung fluids" indicating respiration of water from a lake in a possible drowning victim). You can make up your own "scenarios" and supply the "evidence" from the crime scene. Make up a class exercise "crime" and lead them through an investigation of the evidence by EM. Make your SEM/TEM/EDX lecture slides in advance and discuss possible conflicts/artifacts, etc.. You won't need "blood, guts, bullets, things like that" to impress and instruct the students. Leave that for "Hollywood" and the criminal investigator in the proper arena (the court room or instructing other forensic scientists).
You may want to attend the SCANNING 99 meeting in Chicago (April 11-14, 1999) where an all day short course (FORENSIC SCIENCE AND SCANNING MICROSCOPY) will be offered followed by a second day symposium (SCANNING MICROSCOPY APPLICATIONS IN FORENSIC SCIENCE). For information on that International Meeting, call Mary K. Sullivan (201) 818-1010 , web site { http://www.scanning-fams.org } .
Again, I offer my assistance with some other ideas if you are interested. Feel free to contact me off line or by DIRECT E-mail.
Sincerely,
S. Frank Platek Research Biologist/Electron Microscopist Forensic Chemistry Center US Food and Drug Administration 6751 Steger Drive, Cincinnati, OH 45237-3097 (513) 679-2700 , [FAX] (513) 679-2761 E-mail: fplatek-at-ora.fda.gov
DISCLAIMER: THE OPINIONS EXPRESSED ARE SOLELY MY OWN AND DO NOT REPRESENT THOSE OF THE U.S. FOOD AND DRUG ADMINISTRATION OR ANY OTHER AGENCY OF THE U.S. FEDERAL GOVERNMENT OR THE FOUNDATION FOR ADVANCES IN MEDICINE AND SCIENCE (FAMS, INC.).
We purchased two of those tanks this past January. One of them had a leak, but it was suggested to use a silicon adhesive to repair the split (instead of returning the tank). So far the tank is fine. In retrospect, I should have stuck with the stainless steel developing tanks. } } } Bruce
Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124-4305
} ---------- } From: Walck. Scott D.[SMTP:walck-at-ppg.com] } Sent: Thursday, March 18, 1999 10:17 AM } To: Micro } Subject: Plexiglas developing tank -problems } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We bought several large Plexiglas developing tanks from Energy Beam } Sciences } for developing TEM negatives. Several of them split their seams after a } few } months and we sent them out for repair. After a few months, they split } again. Has anyone else had problems with these tanks? They are model } DT-111 and are 1-1/2 gallon size (approx. 8-3/4" x 12" x 8"). } } Please contact me off-line. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P.O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } }
We have been experimenting with telepresence for this purpose. One can get high quality zoom-pan cameras with over-the-web control for less than about $1000 or a simple static camera for about $100. Creating a web page with this view is not very difficult and allows coworkers to monitor each others safety from any place that has network access.
Eric B. Steel, Leader e-mail: eric.steel-at-nist.gov Microanalysis Research Group Office: 301-975-3902 N.I.S.T. FAX: 301-417-1321 100 Bureau Drive, Stop 8371 Gaithersburg, MD 20899-8371
} Over the years our workforce has dwindled to the point that I find myself working alone in the lab (metalographic sample prep/optical microscopy + image analysis) almost all the time. The lab doesn't have windows to the hallway, so it is difficult for people walking by to see if I am in the lab, and if I am OK. My manager is concerned about my safety and is asking me for suggestions. What do other labs do to make working alone safe? } Everett Ramer } Federal Energy Technology Center
When I worked at Sarastro a decade ago, applications specialists in our parent company in Sweden had taken pictures of aphids, orchid seeds, and various other 3D objects very successfully with the confocal. As long as the structures of interest are either autofluorescenct, reflective, or can be stained with a fluorescent tag so that they can be imaged, the confocal is a great instrument for this type of work.
The only caveats: (1) make sure to match the numerical aperture of the objective (which governs the depth of field) to the vertical step height so that you get continiguous information (no gaps in the image) (2) be aware of the size of the image file so that you will have enough room on your computer/tape to handle the file (3) check out various types of reconstruction to see which ones best reproduce the information you are seeking. I seem to remember that "look through projections" worked really well but this is an area which has grown tremendously in the recent past. See what is available on your particular system.
Best of luck, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 10:00 PM 3/19/99 +1100, ricardo wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Three years ago almost to the day we had a discussion on ENTOMO-L about } } confocal laser microscopes, which several contributors believed would } } eventually largely replace the use of electron microscopes in insect } } taxonomy and morphology. One huge advantage is that images are "captured" } } directly into a computer, where they can be stored and manipulated later.
Just an aside here, many SEMs can also capture images digitally. Also, (although it involves an additional step) photographs can be scanned at very high resolutions and bit depths, and subsequently digitally manipulated and stored.
} } Our University has now purchased such a microscope system, and it has } } recently been installed. Can a confocal laser microscope be used to view } } entire objects, or just slide mounts??
My $0.02, based on my previous experience...
Several years ago I worked with Prof. Dan Young of our entomology Dept. on doing some coleopteran cranial morphology studies. The samples in question were, in some cases, one-of-a-kind museum pieces; the only known examples of their species. For this reason dissection and platinum coating of the beetle heads was not a possiblility.
We had some good luck using autofluorescence of the exoskeleton to capture striking tilt-stereo images of the beetle heads, which could then be displayed as 3D stereo-pairs. The dense, opaque, and reflective nature of the exoskeletal material was such that the laser didn't penetrate it readily, so traditional confocal z-series and reconstruction was not possible. We arrived at a protocol that worked, however, in order to capture these pictures, most of the advantages of the confocal had to be circumvented. We used a 3.5x lens with the pinhole open as far as it would go so that we could get a large area and depth of field to capture the large (for microscopy; about 3 x 2 mm) specimens.
It is my opinion that we would have been much better off using a good quality dissecting scope with a and a color video camera to obtain our tilt-stereo images. For smaller and less opaque organelles, there is the possibility that the confocal would work quite well. But for 3D tilt-stereo capture of the larger external features, I would recommend that people try out a good quality, video-equipped dissecting scope.
-- Charles Thomas 4D Software Engineer 4D Imaging Coordinator Integrated Microscopy Resource Univ. of Wisconsin-Madison 1675 Observatory Dr. Madison, WI 53706 608-263-6288 Office 608-265-4076 Fax
cthomas-at-facstaff.wisc.edu
VISIT OUR WEB SITE: http://www.bocklabs.wisc.edu/imr/facility/4D/4d.htm
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A colleague is looking for the following: Leitz Accessory 10-419-613 4x5 cone for the Photomacroscopes (connection point is circular; needs to fit into an MPS 11 manual shutter)
If your lab has one and is interested in selling it, please contact Dennis O'Leary of MicroOptical Methods: 518-482-8200
Thanks Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
I am wondering how many microscopists are suffering from RSI (repetitve stress injuries). I have recently developed DeQuervain's tendonitis in my thumb and I am wondering if it is from moving the specimen drive control knob on the E.M. with my thumb for many years. Not to mention the knobs on the microtome. We work with alot of knobs, some easy to turn and some not Here at Stanford, 3 out of 5 electron microscopists ( 2 are no longer working here but not because of RSI), suffer from hand problems. I thought I would take a survey to find out the prevalence of this condition among microscopists.
Olympus has discontinued a dual output module for their AX and BX line of optical microscopes. This accessory allows two lamps (e.g., halogen and mercury) to be mounted to a vertical illuminator; either source can be used by flipping a mirror. I am told the decision to discontinue the accessory was due to liability concerns that could arise if an operator switched from a halogen to UV lamp source without engaging an appropriate fluorescence cube (with barrier filter).
Several microscope vendors have told me that third-party accessories are available, but can or will not provide any documentation. One company mentioned is 20-20 Technology.
I am seeking referrals to a company providing such an accessory, or a microscopist who wishes to part with one.
Thank you.
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center http://members.tripod.com/~James_Martin/dasrhome.htm
Research Scientist in Chemistry Williams College www.williams.edu http://members.tripod.com/~James_Martin
Does anyone know of an authoritative source explaining the deduction of quantitative information (such as void volume, etc.) from stereomicroscopy of membrane microstructures? Thanks, Sai
******************************************************************** Sai S. Prakash, M.S. Doctoral Student / Graduate Research and Teaching Assistant (Research Area: Cryo-SEM of Coating Microstructures) Chemical Engineering and Materials Science Department University of Minnesota 421 Washington Avenue SE, Box 137 Minneapolis, MN 55455. USA. Tel: 612 625 4809 Fax: 612 626 7246 Email: sprakash-at-cems.umn.edu ********************************************************************
Is anyone familiar with reasonably priced software programs that can manipulate 16-bit gray scale images? While Adobe Photoshop can read 16-bit images, you have to change the images to 8-bit before you can run filters etc. Gatan's Digital Micrograph can work with 16-bit images, but its price is a little steep for equipping several computers. What are the capabilities of other packages? I would like to use the software to work with diffraction patterns as well as images.
TIA, Henk
Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility (614) 292-0674 "An optimist believes that we live in the best of all possible worlds. A pessimist fears that this is true."
WOW!! Thanks for all the advice and info!! You folks are really wonderful!
I'm sorry to take so long to get back to you; I have been "down and out" with a killer cold. Then trying to get caught up on all the "urgent" jobs that always seem to appear just when all you really feel good for is a doorstop.
I'll try to answer everyone at once, to not clutter the list with a bunch of messages. Yes, I think it must be a Zetopan. That name rings a faint bell in the old belfrey. But it is an older model than the one in the image Yvan kindly sent. No, I don't have an image of mine; but I was able to dig up the original inventory entry for it. It was purchased in 1955! And retired to storage in 1981, apparently because it was impossible to get a bulb that would fit it. Yvan, that is interesting about the "Reichert adapter" for the bulb you mentioned. We have quite a few bulbs (apparently bought for it), but they are not long enough to reach the contact at the base of the socket. This was no doubt the problem way back then - our "Mr. Fix-it" (who keeps this place running) has now solved that problem by soldering a tiny washer onto the contact at the base of the socket so that when the bulb is screwed in all the way, it makes contact with the washer. And we have light! The bulbs we have are Osram BPA 554, 6V 5A. There is also a burned-out Osram 8100 with it (was in the socket) but it is even shorter than the BPAs., so didn't meet the contact either. Maybe someone threw out the adapter with the last bulb? Both types of bulbs are the same diameter so have the same tight fit into the bulb housing. There is no identification on the housing as to illuminator type; and the rheostat/transformer has been replaced with one from another make of microscope.
The bulb housing at the back looks like the one in Laslo's image, not Yvan's. There is no "Epi" attachment though; and no "flip-in" phase telescope. Between "6" and "7" in Yvan's image (objective lens turret and eyepiece attachment) my scope has two filter holder slides (empty of course; one with a hole about dime-sized, the other about nickle-sized) and the lever to send the light up the photo tube.
Many, many thanks to Barbara for taking the time to send such detailed instructions re phase set-up! Unfortunately... I can't get past the first step! My scope doesn't have a turret condenser with a brightfield option. I said "dedicated phase" and I should have said "phase only" to be clearer. Sorry. This condenser is strictly one-size-fits-all! It doesn't have an "aperture iris" either. But, it does have a knurled ring around its bottom and a measuring scale (like a ruler) stamped around its front. Turning the knurled ring screws the innards of the condenser up and down through a total of about 15 divisions (I don't know what units) on the scale. It also has the following stamped on its front above the scale: "Objektragerdicke 1,0 mm". I'm sure that means something important! So one can focus the condenser by the usual control that moves the entire gizmo in its fork mount, as well as this "internal" focus. I have, eons ago, set up phase on another scope with a turret condenser, so the steps you detailed are familiar. I don't recall ever using a condenser like this one. But many thanks for the procedure!
Wayne; great! I'd like to have a manual if you have one that might apply to this old microscope. Julian and Susan; I would like bits and pieces ( like a condenser with brightfield option! and brightfield objectives) if you wouldn't have a problem sending them to Canada. I'm appending my address below. Unfortunately we have no money (as usual) for servicing, or new parts. I've just been told to have the manual etc. sent COD, and we'll deal with it when it arrives. I guess it had better not exceed what's in petty cash that day!
Again, many thanks.
Sharon Godkin Canadian Food Inspection Agency Centre for Plant Health 8801 East Saanich Road Sidney, B.C. Canada V8L 1H3
I agree with Jim that the best way to deal with gold on museum specimens, = artwork, taxonomic type specimens, fossils etc is to leave it off in the = first place where possible. The material can sometimes be viewed in a = normal SEM by keeping kV and probe current low, or generally more = conveniently by using one of several types of variable pressure SEM. = Until recently the cheaper versions, working up to about 2 torr, were = restricted to non-SE detection modes (BSE, CL, absorbed current etc) but = several SE detector systems working in this range have become commercially = available. =20 We have just installed a new Robinson detector which allows convenient= BSE and/or SE detection to low kV, in high vacuum or variable pressure = conditions, on a small Hitachi NSEM. (1992) and find the combination mode = in particular gives very good results for topographic imaging. =09 beats explaining a caustic cyanide stew to your friendly curator, I should = think.
cheers
Sally
Dr Sally Stowe Facility Coordinator Australian National University EM Unit Research School of Biological Sciences Box 475, ACT 2601, Canberra, Australia FAX 06 (0)2 6279 8525
James , I don't understand how the light from the mercury illuminator could get to the observers eyes without a filter in place. If a filter position is blank and in place the light from the illuminator has no beam splitter or prisim to direct light to the eyepieces. There fore the light remains enclsoed within the illuminator module... Also there should be a stop on the illuminator to block any light coming through. Leica has the same option built into the stand of their DMR series microscope and does not seem to have a problem with light reaching the observer unfiltered.. This is probably another argument why the Leica is preferred over tthe competition.. Another is being the Lecia optics tend to absorb the harmful UV more so then its inferior Japanese competitors. Did you ever notice the Olympus has the filter shield in place so you don't see the uv light coming out of the objectives at the sample. Thats there for a reason....Harmful UV........ In answer to your question, I do not know of anyone who has one of these devices but would check with some of the larger Olympus dealers. Since it was not such a good idea maybe some of them still have units unsold on the shelf waiting to unload... Good Luck C YA CIP -----Original Message----- } From: James Martin {James.S.Martin-at-williams.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
That Olympus reasoning is absurd! We (Nikon), Zeiss and Leica all still manufacture this part! In any event, here are a few third-party sources where you can get such an item. Try Opti-Quip, Inc., Highland Mills, NY (914) 928-2254 or MicroTec, Inc. Milford, NJ (800) 724-5508. I have used the MicroTec one (product #MIT-001), just specify Olympus adapter fittings.
Good Luck,
Lawrence Kordon Nikon, Inc. Columbia, Maryland nikon-at-jagunet.com
Cono Passione wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } James , } I don't understand how the light from the mercury illuminator could get to } the observers eyes } without a filter in place. If a filter position is blank and in place the } light from the illuminator has } no beam splitter or prisim to direct light to the eyepieces. There fore the } light remains enclsoed } within the illuminator module... Also there should be a stop on the } illuminator to block any light } coming through. } Leica has the same option built into the stand of their DMR series } microscope and does not } seem to have a problem with light reaching the observer unfiltered.. This } is probably another } argument why the Leica is preferred over tthe competition.. Another is } being the Lecia optics } tend to absorb the harmful UV more so then its inferior Japanese } competitors. Did you ever } notice the Olympus has the filter shield in place so you don't see the uv } light coming out of the } objectives at the sample. Thats there for a reason....Harmful UV........ } In answer to your question, I do not know of anyone who has one of these } devices but would check } with some of the larger Olympus dealers. Since it was not such a good idea } maybe some of them } still have units unsold on the shelf waiting to unload... } Good Luck } C YA CIP } -----Original Message----- } } From: James Martin {James.S.Martin-at-williams.edu} } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } Date: Friday, March 19, 1999 10:40 PM } Subject: light microscopy: dual output monitor } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Olympus has discontinued a dual output module for their AX and BX line of } } optical microscopes. This accessory allows two lamps (e.g., halogen and } } mercury) to be mounted to a vertical illuminator; either source can be } } used by flipping a mirror. I am told the decision to discontinue the } } accessory was due to liability concerns that could arise if an operator } } switched from a halogen to UV lamp source without engaging an appropriate } } fluorescence cube (with barrier filter). } } } } Several microscope vendors have told me that third-party accessories are } } available, but can or will not provide any documentation. One company } } mentioned is 20-20 Technology. } } } } I am seeking referrals to a company providing such an accessory, or a } } microscopist who wishes to part with one. } } } } Thank you. } } } } James Martin } } Director of Analytical Services and Research } } Williamstown Art Conservation Center } } http://members.tripod.com/~James_Martin/dasrhome.htm } } } } } } Research Scientist in Chemistry } } Williams College } } www.williams.edu } } http://members.tripod.com/~James_Martin } } } } } }
A point that Cono seems to miss in the criticism of the Olympus scope (with the U.V. filter flange over the objective nosepiece) is that scattered U.V. from the sample is not a function of the quality of the optics or the manufacturer but rather a function of the type of excitation in use.
Some users, particularly biologists, use a mercury or xenon lamp but are not relying upon the U.V. output per se. In some filter combinations a high level of excitation in a certain short wavelength region of the visible spectrum is the most efficient for stimulating emission from a fluorochrome which emits in the longer wavelength region of the visible. Sometimes the cubes which are optimized for this effect limit the amount of short-wave U.V. which reaches the sample. Therefore, scattered light from the sample surface is lower in proportion of U.V. than the proportion emitted from the lamp.
However, people who make use of intrinsic U.V. fluorescence in unlabeled specimens, or cascade effects, often use cubes which pass a significant quantity of U.V. directly to the sample. The scatter or reflected light may be correspondingly high in it's proportion of U.V. Those in forensics and art conservation who rely on this mechanism for all or part of their work are dealing with a different situation by intention rather than accident and must take precautions regardless of the manufacturer.
John Twilley Art Conservation Scientist P.O. Box 2324 Sag Harbor, NY 11963-0113
Cono Passione wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } James , } I don't understand how the light from the mercury illuminator could get to } the observers eyes } without a filter in place. If a filter position is blank and in place the } light from the illuminator has } no beam splitter or prisim to direct light to the eyepieces. There fore the } light remains enclsoed } within the illuminator module... Also there should be a stop on the } illuminator to block any light } coming through. } Leica has the same option built into the stand of their DMR series } microscope and does not } seem to have a problem with light reaching the observer unfiltered.. This } is probably another } argument why the Leica is preferred over tthe competition.. Another is } being the Lecia optics } tend to absorb the harmful UV more so then its inferior Japanese } competitors. Did you ever } notice the Olympus has the filter shield in place so you don't see the uv } light coming out of the } objectives at the sample. Thats there for a reason....Harmful UV........ } In answer to your question, I do not know of anyone who has one of these } devices but would check } with some of the larger Olympus dealers. Since it was not such a good idea } maybe some of them } still have units unsold on the shelf waiting to unload... } Good Luck } C YA CIP } -----Original Message----- } } From: James Martin {James.S.Martin-at-williams.edu} } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } Date: Friday, March 19, 1999 10:40 PM } Subject: light microscopy: dual output monitor } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Olympus has discontinued a dual output module for their AX and BX line of } } optical microscopes. This accessory allows two lamps (e.g., halogen and } } mercury) to be mounted to a vertical illuminator; either source can be } } used by flipping a mirror. I am told the decision to discontinue the } } accessory was due to liability concerns that could arise if an operator } } switched from a halogen to UV lamp source without engaging an appropriate } } fluorescence cube (with barrier filter). } } } } Several microscope vendors have told me that third-party accessories are } } available, but can or will not provide any documentation. One company } } mentioned is 20-20 Technology. } } } } I am seeking referrals to a company providing such an accessory, or a } } microscopist who wishes to part with one. } } } } Thank you. } } } } James Martin } } Director of Analytical Services and Research } } Williamstown Art Conservation Center } } http://members.tripod.com/~James_Martin/dasrhome.htm } } } } } } Research Scientist in Chemistry } } Williams College } } www.williams.edu } } http://members.tripod.com/~James_Martin } } } } } }
Depends on what you mean by reasonably priced. I am a firm believer that Research Systems (RSI) IDL software is the most complete quantitative image analysis software available. It runs on UNIX, VMS, Windows, and Mac OS. I have used all versions except Windows, but I expect it to work like the others. It is not cheap; $1500 single user licence. Has been as low as $500 for .EDU institutions. The software comes with an enormous number of canned routines and its own procedural programming language. It will also interface with external routines and has been used for many years to run the Scanning X-ray Microscope (STXM) at the National Synchrotron Light Source, Brookhaven. It handles any data format you want 8, 16, 24 bit images, 3-D data sets, etc. You do have to spend a week or two learning the basics but I don't consider it difficult. Frequently we ended up hiring work study students to do any special programming. These were biology students not programmers. So if you want something that is complete, extensible, more color tables than you'd ever want, and handles any data format, get IDL. I know this sounds like a commercial but I have no financial interest in RSI or anyother imaging software company.
As an alternative I recommend NIH-Image for the Mac and its equivalent Scion-Image for PCs obtainable from the site shown below. Just remember to check the "calibrate box" and use the import file option. It cannot export 16-bit tiff but with calibrate turned on it will do calculations using 16 bit for projecting 3-D data, numerical calculations,etc. These programs will import image stacks only limited by the amount of available computer ram. For raw data, the import dialog under "custom" requires you to tell how many frames to import. Best of all these programs are free. However, they are not suitable for diffraction analysis. IDL is. Good luck.
http://www.scioncorp.com
} } Is anyone familiar with reasonably priced software programs that can } manipulate 16-bit gray scale images? While Adobe Photoshop can read 16-bit } images, you have to change the images to 8-bit before you can run filters } etc. Gatan's Digital Micrograph can work with 16-bit images, but its price } is a little steep for equipping several computers. What are the } capabilities of other packages? I would like to use the software to work } with diffraction patterns as well as images. } } TIA, } Henk } } Hendrik O. Colijn colijn.1-at-osu.edu } OSU Campus Electron Optics Facility (614) 292-0674 } "An optimist believes that we live in the best of all possible worlds. } A pessimist fears that this is true."
Mario M. Moronne, Ph.D. Material and Life Science Div. M/S 6-2100 University of California Berkeley Lab 1 Cyclotron Rd. Berkeley, CA 94720
ph (510) 486-4236 FAX (510) 528-8076 mariomm-at-ux5.lbl.gov or biostar-at-aldecoa.com
Am headed out of town and don't have time to track down the Reichert manuals, but the phase system you describe sounds amazingly like the "anopteral phase" which I have on my system. Will be back in about a week and will check back with you then.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 08:48 PM 3/19/99 -0500, Sharon Godkin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Check with Media Cybernetics. I am pretty sure that their Image Pro is at least 16 bit now. They are in Silver Spring MD and on the web.
Best of luck Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 08:16 PM 3/19/99 -0500, Hendrik O. Colijn wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The dual output accessory couples two lamp housings to the back end of the vertical illuminator. A mirror is used to reflect light from either lamp housing down the vertical illuminator to a rotating turret, situated over the nosepiece, that contains filter cubes. A variety of excitation/emission cubes would be used for fluorescence examination (UV or visible excitation) using a mercury or xenon source. A brightfield cube would be used for visible light examination using a halogen source. If a mercury or xenon source was used with a normal brightfield cube (not designed for use with a UV source), an observer would not be protected from UV wavelengths reflected or fluoresced from a sample. Of course, this sort of mistake, followed by examination, ought not to occur in practice. However, the possibility does exist with such multipurpose illumination systems. I hope this helps to answer your most reasonable question.
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center http://members.tripod.com/~James_Martin/dasrhome.htm
Research Scientist in Chemistry Williams College www.williams.edu http://members.tripod.com/~James_Martin
On Sat, 20 Mar 1999, John Twilley wrote:
} Cono Passione and James Martin, } } A point that Cono seems to miss in the criticism of the Olympus scope (with the } U.V. filter flange over the objective nosepiece) is that scattered U.V. from } the sample is not a function of the quality of the optics or the manufacturer } but rather a function of the type of excitation in use. } } Some users, particularly biologists, use a mercury or xenon lamp but are not } relying upon the U.V. output per se. In some filter combinations a high level } of excitation in a certain short wavelength region of the visible spectrum is } the most efficient for stimulating emission from a fluorochrome which emits in } the longer wavelength region of the visible. Sometimes the cubes which are } optimized for this effect limit the amount of short-wave U.V. which reaches the } sample. Therefore, scattered light from the sample surface is lower in } proportion of U.V. than the proportion emitted from the lamp. } } However, people who make use of intrinsic U.V. fluorescence in unlabeled } specimens, or cascade effects, often use cubes which pass a significant } quantity of U.V. directly to the sample. The scatter or reflected light may be } correspondingly high in it's proportion of U.V. Those in forensics and art } conservation who rely on this mechanism for all or part of their work are } dealing with a different situation by intention rather than accident and must } take precautions regardless of the manufacturer. } } John Twilley } Art Conservation Scientist } P.O. Box 2324 } Sag Harbor, NY 11963-0113 } } Cono Passione wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } James , } } I don't understand how the light from the mercury illuminator could get to } } the observers eyes } } without a filter in place. If a filter position is blank and in place the } } light from the illuminator has } } no beam splitter or prisim to direct light to the eyepieces. There fore the } } light remains enclsoed } } within the illuminator module... Also there should be a stop on the } } illuminator to block any light } } coming through. } } Leica has the same option built into the stand of their DMR series } } microscope and does not } } seem to have a problem with light reaching the observer unfiltered.. This } } is probably another } } argument why the Leica is preferred over tthe competition.. Another is } } being the Lecia optics } } tend to absorb the harmful UV more so then its inferior Japanese } } competitors. Did you ever } } notice the Olympus has the filter shield in place so you don't see the uv } } light coming out of the } } objectives at the sample. Thats there for a reason....Harmful UV........ } } In answer to your question, I do not know of anyone who has one of these } } devices but would check } } with some of the larger Olympus dealers. Since it was not such a good idea } } maybe some of them } } still have units unsold on the shelf waiting to unload... } } Good Luck } } C YA CIP } } -----Original Message----- } } } From: James Martin {James.S.Martin-at-williams.edu} } } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } } Date: Friday, March 19, 1999 10:40 PM } } Subject: light microscopy: dual output monitor } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Olympus has discontinued a dual output module for their AX and BX line of } } } optical microscopes. This accessory allows two lamps (e.g., halogen and } } } mercury) to be mounted to a vertical illuminator; either source can be } } } used by flipping a mirror. I am told the decision to discontinue the } } } accessory was due to liability concerns that could arise if an operator } } } switched from a halogen to UV lamp source without engaging an appropriate } } } fluorescence cube (with barrier filter). } } } } } } Several microscope vendors have told me that third-party accessories are } } } available, but can or will not provide any documentation. One company } } } mentioned is 20-20 Technology. } } } } } } I am seeking referrals to a company providing such an accessory, or a } } } microscopist who wishes to part with one. } } } } } } Thank you. } } } } } } James Martin } } } Director of Analytical Services and Research } } } Williamstown Art Conservation Center } } } http://members.tripod.com/~James_Martin/dasrhome.htm } } } } } } } } } Research Scientist in Chemistry } } } Williams College } } } www.williams.edu } } } http://members.tripod.com/~James_Martin } } } } } } } } } } } } }
I've been looking at purchasing a dry 60x objective for use on a Nikon E600 microscope (application is temporary and permanent mounts of soil invertebrates, particularly soil mites, nematodes, etc.). The particular lens I'm trying right now is the CFI PLAN APO Phase 60x Dry DM objective. According to the sales rep, a DIC slider is not made specifically for this objective, and Nikon suggests that users try different combinations and choose what they like best. We've mixed and matched what the rep had available, and the best image was with CD DIC slider for the Plan Fluor ELWD (extra long working distance) DLL 60x C Phase objective. This objective is for an inverted microscope and is meant to look through a lot more material than one coverslip. I'd like to know if anyone on the list uses this Plan Apo objective for DIC, and what slider they use. Does this seem right to have to mix and match DIC sliders and objectives? ______________________________________________________________________ Edmond R. Zaborski phone (217) 265-0330 Soil Invertebrate Ecology Center for Economic Entomology FAX (217) 333-8076 Illinois Natural History Survey zaborski-at-uiuc.edu 607 E. Peabody Dr. Champaign, Illinois 61820 U.S.A. http://www.inhs.uiuc.edu/ ______________________________________________________________________
I am a neophyte to the microscopy world and am curious about EBIC and Voltage Contrast. I am wondering if there are good resources on the general mechanics of setting up the system. I also am having trouble finding basic information/ uses for EBIC. The Goldstein et al book does not delve too much into the topic of EBIC and am not sure of any other good SEM resource books.
Sharon Godkin says: } I'll try to answer everyone at once, to not clutter the list with a bunch } of messages. Yes, I think it must be a Zetopan. That name rings a faint } bell in the old belfrey. But it is an older model than the one in the } image Yvan kindly sent.
The *shape* should be roughly the same. The *color* would be black in your case (the old model).
} No, I don't have an image of mine; but I was able to dig up the original } inventory entry for it. It was purchased in 1955!
(:-) [but doesn't the recond say what microscope it is?]
} And retired to storage in 1981, apparently because it was impossible to } get a bulb that would fit it.
Osram 8100 should fit. See below.
} Yvan, that is interesting about the "Reichert } adapter" for the bulb you mentioned. We have quite a few bulbs } (apparently bought for it), but they are not long enough to reach the } contact at the base of the socket. This was no doubt the problem way } back then - our "Mr. Fix-it" (who keeps this place running) has now } solved that problem by soldering a tiny washer onto the contact at the } base of the socket so that when the bulb is screwed in all the way, it } makes contact with the washer. And we have light! The bulbs we have } are Osram BPA 554, 6V 5A. There is also a burned-out Osram 8100 } with it (was in the socket) but it is even shorter than the BPAs., so didn't } meet the contact either. Maybe someone threw out the adapter with } the last bulb?
I think this is EXACTLY the case: somebody "discarded" the bulb adapter (probably because another idiot soldered the bulb TO the adapter instead of just screwing it in - Yvan told me about one such incident).
} Both types of bulbs are the same diameter so have the same tight fit } into the bulb housing.
Hmm... This is to the experts to comment on.
} There is no identification on the } housing as to illuminator type; and the rheostat/transformer has been } replaced with one from another make of microscope.
No worries - the one I got also came with a "foreign" transformer. Who cares (unless you are a collector :-).
} The bulb housing at the back looks like the one in Laslo's image, not } Yvan's. There is no "Epi" attachment though; and no "flip-in" phase } telescope.
No Epi - because it was configured for transmitted light. No flip-in Bertrand lens ("phase telescope") - because yours is an older model (mine is "younger" than yours but still doesn't have Bertrand lens).
} Between "6" and "7" in Yvan's image (objective lens turret } and eyepiece attachment) my scope has two filter holder slides (empty } of course; one with a hole about dime-sized, the other about } nickle-sized) and the lever to send the light up the photo tube.
One slot (the lower narrow one, going 45 degrees) is for a compensator (don't know the details). The wider one is of course for the filter slider. Your swivelling head is identical to mine.
} It also has the following stamped on its front above the scale: } "Objektragerdicke 1,0 mm". I'm sure that means something important!
I suspect it says "object-vertical-thickness 1mm". Might it apply to the specimen slide?
As for bits and pieces - hey, me too! Objectives, condensers, TL DIC, etc.! Please e-mail with whatever Zetopan parts you're willing to part, and how much you'd like for it. -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
Hello Frank and everybody, I hope this will be of some help:
Dr. Pickett-Heaps recently published (Journal of Phycology 34:1088) a solution that, I remember, just mesmerized me with its elegance. "A rapid, highly efficient method... for fixation, dehydration, and embedding of small (e.g. planktonic) cells dispersed in large volumes of culture medium... based on continuous filtration..." -- so that you never really pellet the cells, just gently concentrate them over a cellulose filter in a standard Millipore apparatus while gradually adding the fixatives-washes-dehydrant-resin from the top. Seems worth trying , doesn't it? And yet the setup is VERY simple.
Just contact me directly if you can't get the journal. And if you do try it, I would ask you to please comment about the outcome. I have not yet made the time to try that for myself and will be MOST INTERESTED to hear how it worked for, say, lymphocytes. Or maybe somebody else uses something similar routinely?
Sincerely yours, Vlad.
} } Howdy all, } There is a graduate student here who is trying to look at a possible stem } cell line under TEM. The problem is that they are quite small, don't seem } to pellet very well, and he has only been able to get me a few thousand } cells at a time. } We have tried to embed the cells in 2% Agar after fixation but this hasn't } worked. } Is it possible to filter the media and cells, and then process the filter } with the cells stuck onto it? Maybe use a cytospin to spin the cells into a } filter? } Is there anyone with experience with this? Any suggestions or ideas would } be greatly appreciated. } } } Thanks in advance } Frank Herbert } Technician } Integrated Microscopy Core } Department of Cell Biology } Baylor College of Medicine } } } Vladislav V. Speransky School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Phone: 207 581 2998 Fax: 207 581 2969 Email: vladis-at-maine.maine.edu
The description provided by Sharon and some educated guesses:
..."... The bulbs we have are Osram BPA 554, 6V 5A..."
I didn't found that one in older Osram catalogues, but these doesn't go back to the 50's. I can send some pages from Osram with descriptions of bulbs that could fit and their most important parameters (voltage and current, foot type, dimensions of lamp and filament, relative position of the filament....).
"Longer" bulbs are:
* 8001 (6V, 4.35A, E14 foot, filament (4 x 0.9 mm) at 57mm from the middle contact of the foot, overall dimensions the same as 8100)
* 8025 (6V, 5A, BA20d foot, filament (2.2 x 2 mm) at 54 mm from the "bottom" of the foot, overall dimensions about the same as 8100, length is 60mm instead of 65 in the 8100)
* ...
..."... My scope doesn't have a turret condenser with a brightfield option. I said "dedicated phase" and I should have said "phase only" to be clearer. Sorry. This condenser is strictly one-size-fits-all! It doesn't have an "aperture iris" either. But, it does have a knurled ring around its bottom and a measuring scale (like a ruler) stamped around its front. Turning the knurled ring screws the innards of the condenser up and down through a total of about 15 divisions (I don't know what units) on the scale. It also has the following stamped on its front above the scale: "Objektragerdicke 1,0 mm". ..."...
"Objektragerdicke 1,0 mm" means: thickness of slides (to be used with this condenser): 1mm
As Barbara suggested (unless I misunderstand that) the condenser could be a "MS1.40" condenser, usable with phase and/or Anoptral inserts (the "UV-inserts"), but the MS1.40 is a brightfield condenser usable too for fluorescence and it doesn't has a ".... knurled ring around its bottom and a measuring scale (like a ruler) stamped around its front. Turning the knurled ring screws the innards of the condenser up and down through a total of about 15 divisions...". Could the curled ring device be an insert in the condenser that can be removed? (The MS 1.40 uses magnetic inserts that can easily be removed, you just have to pull these out of the condenser...). If it's the same condenser as the one I have, it should read on the top lens "UV". Sure it's not an adjustable DF-condenser?
Can send you some pictures of different condensers offline if you like... Can't send pictures to the list as it's considered poor netiquette (I had some reactions regarding that after sending the Zetopan picture...).
There seems to be some misunderstanding regarding the 6V, 5A bulb for the "LUX FNI" illuminator for Zetopan:
Uri: "... } I think this is EXACTLY the case: somebody "discarded" the bulb adapter } (probably because another idiot soldered the bulb TO the adapter instead } of just screwing it in - Yvan told me about one such incident)....".
Ussualy the bulb + the adapter were discarded. reichert sold these bulbs soldered in their individual adapter, thus providing a fully precentered and preadjusted bulb for the Zetopan. desoldering the bulb and replace it for an osram 8100 in the original adapter is just a way of fixing the problem that the bulbs are rather rare now...
Sharon: I'm preparing a multipage tif-file containing information and pictures on bulbs, lamphouses, filterholders and condensers for Zetopan. Interested?
Chris, For a number of years I performed what we call "electrical failure analysis" using electron beam techniques on semiconductor devices and integrated circuits. My favorite was Electron Beam Induced Current measurements. I found no definitive text or reference for that or the other measurements, Voltage Contrast, Specimen Absorbed Current, Electron Bean Induced Voltage, Charge Contrast, although I do have a few publications which discuss them. I have included in-depth discussions of these techniques in classes, and have assembled a number of example analyses which help to explain them. Send me a personal note if you have an interest in any of this. Or respond with a particular question that might be of interest to the e-beam community on the list. Jerry
_______________________ Jerome D. Schick, Ph.D. Semiconductor Devices and Electron Microscopy 26 Kuchler Drive LaGrangeville, NY 12540 Bus (914)223-7393 FAX (914)227-2743 jdschick-at-worldnet.att.net
Chris Parks wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello, } } I am a neophyte to the microscopy world and am curious about EBIC and } Voltage Contrast. I am wondering if there are good resources on the } general mechanics of setting up the system. I also am having trouble } finding basic information/ uses for EBIC. The Goldstein et al book does } not delve too much into the topic of EBIC and am not sure of any other } good SEM resource books. } } Thanks } CP
JoAnn, About 5 years ago I developed a bad case of tendonitis which our occupational health adviser attributed to the magnification knob in our SEM. She advised on a modification which our worksops made up. This effected a rapid improvement in my condition, although I still get some discomfort if I do a lot of typing.
Eric
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
Jerome D. Schick, Ph.D. Semiconductor Devices and Electron Microscopy 26 Kuchler Drive LaGrangeville, NY 12540 Bus (914)223-7393 FAX (914)227-2743 jdschick-at-worldnet.att.net
March 22, 1999
We are currently doing a very small R&D project for an electrical contector company which involves SEM EBIC. I would love to read any articles or notes that you may have on the subject.
J. Roy Nelson, Ph.D Material Testing Laboratory Pennington, NJ 08534 (609) 730-0575 FAX 737-7119 jrnelson-at-nj1.aae.com
Hi, Several people have asked for a summary of the responses to this question, = and there were many. Thanks to all for some good suggestions. Everett Ramer Federal Energy Technology Center
The solutions suggested were: 1. call a coworker/secretary/boss/security every hour 2. a pendant call button worn around neck---some have location capabilities= to guide rescuers to wearer's location. 3. a pendant worn around neck that automatically calls for help when the = wearer is in the horizontal position. 4. a pager with built-in motion detector that beeps after several minutes = of inactivity. If wearer does not shut off the beeping, the pager = automatically calls security. 5. a zoom-pan camera with over-the-web control with a web page that allows = coworkers to monitor each other safety from any place that has network = access.
Question: I recently purchased an older B&L microscope w/ 3 objectives and a prism inclined eyepiece. I think this model of microscope is from the 50's or 60's. That is all that I know of the model as there are no other markings that I can find to this regard. I was wanting to get a 4X & 20X objective for it and was wondering if AO Spencer objectives would fit. I have an opportunity to get these over the Web, but cannot try them out prior to purchase. We are planning to use the microscope to augment our childrens' studies and perhaps allow my oldest who is interested in cattle use it to examine parasites.
Replies to my inquiry about vendors of dual illumination modules are summarized below:
Opti-Quip, Inc. Highland Mills, NY (914) 928-2254 Model 1030 Universal Mirror housing will adapt to just about any microscope. The price is $998.00. For the Olympus there are 3 models: Olympus IMT2 (Part number: 1030E), Olympus BH (Part number: 1030-F) and Olympus B MAX (Part number: 1030-G)
MicroTec, Inc. Milford, NJ (800) 724-5508 #MIT-001
Thank you for your assistance.
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center http://members.tripod.com/~James_Martin/dasrhome.htm
Research Scientist in Chemistry Williams College www.williams.edu http://members.tripod.com/~James_Martin
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Hi, I need to get information (specs and quotations) rather quickly on = vibration tables for ultramicrotomes, light microscopes, and an atomic = force microscope. If any manufacturers of such equipment read the list, would you = please contact my e mail directly with information. This is for our new building and we need to get updated info into the = state in a very short time.
Thanks in advance, Judy M.
Judy Murphy, PhD Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
with SMTP (Eudora Internet Mail Server 1.2); Sun, 21 Mar 1999 16:47:32 -0800 Message-ID: {n1290087840.86882-at-sjdccd.cc.ca.us}
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Hi, I am trying to find a good negative processor that will process the = following films: EM film (Kodak4489,SO163,etc.) Ektapan (4162) and Kodak 4127 (for SEM) i.e. 4 x 5 sheet film.
1. We want to go from dry film to dry film, thus do not want tank develop= ment. 2. We now have Mohr processors however in 3 yrs have not been able to = get rid of things like roller marks, etc. Believe me, we have tried = every solution that everyone has suggested. We are of course open to new = ones. 3. Does anyone know of other possibilities out there? possibly from our = photographic friends. 4. We need to specify something very quickly, but I haven't come up with = a solution.
Any help is appreciated. Thanks Judy M.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 FAX 209/954-5600 e-mail; jmurphy-at-sjdccd.cc.ca.us
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Thanks to everyone who took their time to respond to my inquiry about microscope user fees. It was a big help.
Best wishes,
Soumitra
Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003-8001 Tel: 505-646-1531/3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu
Dr. Nelson, I have FAXed you a short five page article I wrote describing VC, SAC, and EBIC. Remember, traditionally, EBIC refers to the measurement of currents generated in the specimen, usually a semiconductor with a PN junction at least, and the function of the electron beam is to excite hole-electron pairs in the sample. In this measurement, the primary electrons do not significantly contribute to the current measured. It sounds like your project for a connector company may not involve a semiconductor material as the sample, in which case the wiring connections of the sample, the current amplifier, the time constants, and the interpretation might be somewhat different. When I get a chance, I shall dig around for a few more references which may be of use. I hope the FAXed copy is legible. Jerry
jrnelson wrote: } } Jerome D. Schick, Ph.D. } Semiconductor Devices and Electron Microscopy } 26 Kuchler Drive } LaGrangeville, NY 12540 } Bus (914)223-7393 } FAX (914)227-2743 } jdschick-at-worldnet.att.net } } March 22, 1999 } } We are currently doing a very small R&D project for an electrical } contector company which involves SEM EBIC. I would love to read any } articles or notes that you may have on the subject. } } J. Roy Nelson, Ph.D } Material Testing Laboratory } Pennington, NJ 08534 } (609) 730-0575 } FAX 737-7119 } jrnelson-at-nj1.aae.com
I am working on a business project that requires an analysis of the microscopy market and am hoping someone can direct me to a good source of information. Defining Microscopy to include all techniques which employ a probe such as: photons (including x-rays), electrons, ions, mechanical and/or electromagnetic radiation to form a representation or characterization of the morphology, crystallography, elemental, chemical or electronic structure of any material in either physical and/or life sciences applications. The questions I am trying to answer are: How large is the microscopy community in the US and worldwide? What technology has the greatest number of users? (Optical microscopy, x-ray microscopy, scanning electron microscopy, transmission electron microscopy, atomic force microscopy, scanning tunneling microscopy, scanning ion microscopy, analytical electron microscopy, electron microprobe, x-ray energy dispersive spectroscopy, electron energy loss spectroscopy, electron diffraction, convergent beam diffraction, high resolution electron microscopy, high voltage electron microscopy, etc.) Thank you for your assistance. Dave Akerson Tektronix CPID 682-7471 800-825-6100, ext. 7471 david.l.akerson-at-tek.com {mailto:david.l.akerson-at-tek.com}
Last week, there was a posting regarding the preparation of buffy coats for the TEM. Unfortunately, I didn't save the message and I now have a student who would benefit from the protocol. If you saved the protocol (or are the author of the email) could you please forward it to me offline.
Thanks for your assistance!
Steve
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
Biology Dept., University of Saskatchewan http://www.usask.ca.biology
The individual will be responsible for the departemntal Electron Microscope facility, consisting of 2 Philips TEMs, an SEM and related vacuum and preparatory equipment. Duties will include alignment and maintenance of the instuments, assisting and training students, faculty and outside users in their use, and providing general electronics and instrumentation expertise to the Department.
Required qualifications: Grade XII, post-secondary technical qualifiactions in electronics, and five years of relevant experience, including experience in EM maintenance. A strong interest in computer technology is also desireable. Salary range $36,336 - $46,320 depending on qualifications, with good fringe benefits. Starting date May 1, 1999. Forward resume with names, addresses and phone numbers of references to:
Dr. L.C. Fowke, Head Department of Biology University of Saskatchewan Saskatoon, Sask., S7N-5E2 phone: (306) 966-4400
The posting is directed in the first instance to Canadian citizens and permanent residents. The University is committed to employment equity.
I would second Franks comments. As he points out not all "Forensic" labs do criminal case analysis for law enforcement agencies that you think of when the phrase comes up. I happen to work in one that does. To a great extent our work is not "gore" related. Though you do have to deal with it on a routine basis, the science is the interesting part not the "gore". After a few years you hardly notice it is there most of the time, the tissue on a bullet removed from a body, for example, (I guess that is what you'd classify as "gore") is just something you have to get past to get to the fibers that may tell you something when you turn them over the hair and fiber specialist.
I am a Firearm and Toolmark Examiner that also does some SEM work. As such I work mostly Homicide and other crimes aginst people type cases. Light microscopy makes up the majority of Firearms analysis (The bullets or cartridge case comparison to a given firearm) or Toolmark Analysis. Before they go to the comparison scope, stereo microscopy is the majority of the non comparative examination of bullets and cartridge cases, (though this information is still comparitive in nature). Bullets are looked at for trace evidence and GRC (General Rifling Charicteristics) on the stereo microscope then taken to the comparison scope. Most firearms comparison is carried out between 10 and 40 power on a forensic comparison microscope. The unknown bullet being mounted on one scope (stage) with a standard (known) bullet fired from the firearm in question on the other, the two fields of view being observed through the comparison bridge (though this is a highly intagrated unit in modern firearms scopes and thought of as a single scope). Stereo scopes are also used in examining clothing items for bullet holes, gunpowder, etc.. The SEM is a good tool for the examination of trace evidence, but the application of the SEM to the comparison of bullets is unnecessary and provides no additional information (the James E. Ray appeal claims not withstanding) beyond that seen on the stereo and comparison scopes if you are considering comparison to a firearm only. Only SEMs designed for direct comprison of two fields are sutable for bullet or toolmark comparison. Manipulation of captured images is dificult compared to the Comparison microscope and not considered appropriate without the real time control of the two images. As no properly trained firearms examiner would make an identification from a photograph you would also run into problems with acceptability in court if not using a comparison SEM (only a few exist that I am aware of). The SEM can be highly useful for examination of trace evidence on bullets.
LMs of various types plays a big part in trace evidence examinations, as can SEM/EDS. Blood and other physiologic fluid analysis (serology) relies on LM and other techniques but older methods are quickly being replaced by DNA analysis (this is out of may area). Guts? Our toxicologists do stomach contents for toxicological materials at times using some LM I think (also out of my area). Gun Shot Residue is one of the heavy uses of SEM in the Forensic Science Labs of police agencies (firearms examiners tend to stay clear of this work -do to our contamination from shooting and handling firearms all the time).
War Stories have their place but usually are only interesting if in answer to a question or from the speakers personal experience. Show them the interesting information they can learn with EM and maybe you'll get a different kind of "EEEEWWWW ."
I hope this fills in some of the gaps for you but don't think its really went where you wanted to go. You can probably get some "war storys" (we all have them) at Scanning '99 if your there, but the real information will be more interesting (to you and the students at the HS I suspect).
Jim Roberts Firearm and Toolmark Examiner
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Dear Paula:
I have read with some concern your request for forensic EM assistance.
I've just been asked to speak to a group of high school kids about EM, and how I got to where I am today (I'm not gonna tell them about all the little people I squashed along the way). Actually what I wanted to know is if there are any folks out there doing Forensic EM. You know analysis of blood, guts, bullets, things like that. You know that if I gross the kids out they'll think that EM is coolest thing since ice cubes. So if anybody does that sort of stuff or knows of anybody I can talk to about it, let me know. Cuz y'all know there's nothing better than gettin' a great big EEEEWWWW out of the young'uns.
I would be glad to talk to you about forensic applications of scanning electron microscopy as that is my job, however......you should understand "forensic EM" goes far beyond the common opinion of " analysis of blood, guts, bullets, things like that". An electron microscopist performing airborne particle analysis for possible occupational/environmental exposure to harmful particulates or the pathologist using electron microscopy to identify a pathogen/toxin induced lesion or foreign object are also performing forensic EM analyses. Forensic means "Of or used in legal proceedings or in public debate" (The American Heritage Dictionary, 2nd Edition). The investigative accountant analyzing data in the investigation of corporate illegal dealings for prosecution and the forensic scientist in the laboratory matching bloody fibers or a bullet found at a crime scene are both performing forensic analyses.
Is the point of your giving this talk to "gross them out" and generate " a great big EEEEWWWW out of the young'uns" or are you teaching to really stimulate some young minds and show them other areas of science that use microscopy for possible careers? High school students are not "young'uns" and you may have a profound impact on a few futures. You can either encourage and stimulate OR you can deter further interest altogether. Many of us were originally guided to our profession by a good speaker or teacher.
As one who both instructs and lectures extensively on forensic applications of electron microscopy, I have found the audiences (high schools, colleges, forensic science and microscopist societies) all enjoy the case history (not gore) from an investigation. The myriad of television shows on the subject speak to the popularity of this current "hot" topic. I'm sure the international emphasis on the "Crime of the Century" has contributed greatly to the recent popularity in forensic science and scientific evidence. You propose to SHOCK your audience with raw crime scene photos when the students are not used to seeing "the real thing". This is counter productive as they will be discussing the shock image(s) the whole time you are trying to explain the science/EM in the investigation. The "gore" is primarily what they will remember and take away with them.
If you really want to TEACH these high school students, show them how to identify objects from two places (exa. - particles such as soil samples from the "crime scene" with particles from a suspect's shoes - SEM/TEM/EDX) or how to find specific particles that confirm a scenario (exa. - specific lake diatoms in "lung fluids" indicating respiration of water from a lake in a possible drowning victim). You can make up your own "scenarios" and supply the "evidence" from the crime scene. Make up a class exercise "crime" and lead them through an investigation of the evidence by EM. Make your SEM/TEM/EDX lecture slides in advance and discuss possible conflicts/artifacts, etc.. You won't need "blood, guts, bullets, things like that" to impress and instruct the students. Leave that for "Hollywood" and the criminal investigator in the proper arena (the court room or instructing other forensic scientists).
You may want to attend the SCANNING 99 meeting in Chicago (April 11-14, 1999) where an all day short course (FORENSIC SCIENCE AND SCANNING MICROSCOPY) will be offered followed by a second day symposium (SCANNING MICROSCOPY APPLICATIONS IN FORENSIC SCIENCE). For information on that International Meeting, call Mary K. Sullivan (201) 818-1010 , web site { http://www.scanning-fams.org } .
Again, I offer my assistance with some other ideas if you are interested. Feel free to contact me off line or by DIRECT E-mail.
Sincerely,
S. Frank Platek Research Biologist/Electron Microscopist Forensic Chemistry Center US Food and Drug Administration 6751 Steger Drive, Cincinnati, OH 45237-3097 (513) 679-2700 , [FAX] (513) 679-2761 E-mail: fplatek-at-ora.fda.gov
DISCLAIMER: THE OPINIONS EXPRESSED ARE SOLELY MY OWN AND DO NOT REPRESENT THOSE OF THE U.S. FOOD AND DRUG ADMINISTRATION OR ANY OTHER AGENCY OF THE U.S. FEDERAL GOVERNMENT OR THE FOUNDATION FOR ADVANCES IN MEDICINE AND SCIENCE (FAMS, INC.).
Leica Microsystems, Diatome US, Electron Microscopy Sciences, and Bal-Tec announce another in a series of EM Specimen Preparation workshops. These seminars will focus on the following techniques:
Ultramicrotomy of Materials
Embedding of industrial samples Specimen trimming
Ultramicrotomy of hard materials Ultramicrotomy of polymers
Collection & handling of sections Staining of polymer sections
Low temperature ultramicrotomy
Ion Beam Milling of Materials
Ion Milling of plan view samples for TEM
Ion Milling of cross section samples for TEM
SEM preparation of interfaces using fast slope cutting technique
TEM / SEM preparation of temperature sensitive specimens
SEM / LM preparation of surface structures with ion milling
The format of our workshop is half day lecture and half day bench work. Samples will be supplied by the course instructors. Participants are encouraged to bring their own samples to work with as time allows.
Course Speakers & Instructors
Ultramicrotomy of Materials
Dr. Tom Malis CANMET Characterization Group Leader Materials Technology Laboratory Ottawa, Ontario
Mr. Bob Vastenhout DOW Chemical Polymer Microscopist Analytical Science Department Terneuzen, The Netherlands
Mr. Helmut Gnaegi Product Manager Microtomist Extaordinaire Diatome, Ltd., Switzerland
Ion Beam Milling
Dr. Wolfgang Gruenewald Bal-Tec Head of Applications Laboratory Liechtenstein
Mr. Arthur Buechel Bal-Tec Product Manager Liechtenstein
Hosted by: JoAn Hudson Clemson University Microscopy Facility Clemson, SC
When: Ion Milling June 7-8, 1999
Ultramicrotomy June 9-11, 1999
Tuition: Ion Milling $400.00
Ultramicrotomy $1,400.00
Ion Milling & Ultramicrotomy $1,700.00
Includes lodging at the lakefront Conference Center & Inn at Clemson University, continental breakfast and lunch daily, one group dinner, course supplies, and lab charges.
Contact: Mike Boykin, Leica Microsystems, Inc. 800-248-0665 X5092
I'm sorry to bother everyone, again, with a question about fixation of bacteria. I greatly appreciate the responses I received a few weeks ago. At one time Ryter-Kellenberger fixation was considered the standard. It produced an image with a relatively clear nucleoid containing fibrillar chromatin. Is this now considered to be an artifact of preparation?
John Wright West Desert Test Center (435) 831-3017
have you red the chapter about EBIC in the book of Ludwig Reimer?
Here is the reference:
Reimer, L: Scanning Electron Microscopy: Physics of Image Formation and Microanalysis. Springer Series in Optical Sciences Vol. 45 Chapter 7: Electron-Beam-INduced Current, Cathodoluminescence and Special Techniques p.272-312 (Springer-Verlag Berlin, Heidelberg, New York, Tokyo, 1985) ISBN 3-540-13530-8
Petra
} Hello, } } I am a neophyte to the microscopy world and am curious about EBIC and } Voltage Contrast. I am wondering if there are good resources on the } general mechanics of setting up the system. I also am having trouble } finding basic information/ uses for EBIC. The Goldstein et al book does } not delve too much into the topic of EBIC and am not sure of any other } good SEM resource books. } } Thanks } CP
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu Visit our WWW site! http://www.crpcu.lu/~wahlbrin
A phantastic opportunity to meet well-kmown scientists and an excellent chance to see and to compare the latest equipment for confocal and multiphoton fluorescence microscopy. Attend an excellent meeting and see the most recent technological developments.
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D FOCUS ON MICROSCOPY 1999
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
we would like to invite you to attend this years "Focus on Microscopy" conference.
Many of the well known scientists in this field will present their recent progress.
Here is a list of the plenary speakers: - G.J. Brakenhoff, Amsterdam - Christoph Cremer - Winfried Denk - Timothy Holmes - Anthony A. Hyman - Stefan Hell - Satoshi Kawata - Karsten K=F6nig - Andres Kriete - Ulrich Kubitscheck - Eric Manders - Colin Sheppard - Ernst H.K. Stelzer, Heidelberg - Kevin F Sullivan - Tony Wilson - Daniele Zink
The commercial exhibition offers an excellent overview. The modern instruments of all major microscope manufacturers as well as microscope accessories are on display. This is the 1999 event focussed on microscopy= in Europe.
Highlights of the commercial exhibition include:
Hamamatsu Photonics: ORCA Series - state of the art digital imaging =B7 Progressive scan interline transfer CCD with 1280x1024 pixel =B7 Lens-on-chip technology, TE cooling and optimized readout guarantee highest sensitivity =B7 Black & white cameras with 10-14 bit output =B7 Color cameras with RGB matrix filter or filterwheel =B7 Plug & play set incl. framegrabber and comfortable TWAIN driver avai= lable
L.O.T. Oriel GmbH & Co KG: Nanopositioning Unit and Fiber Laser
Molecular Probes: New fluorescent reagents for Cell Biology and Imaging highly - Fluorescent Alexa dyes - click here for more details
Nikon GmbH: Two compact confocal microscopes + fully automatic motorized research microscope
Olympus Optical Co. GmbH Confocal Laser-Scanning-Microscope with two-phot= on excitation
Omicron Vakuumphysik GmbH Scanning Near Field Optical Microscope (SNOM)
Visitron Systems GmbH: 2D/3D Fluorescence Imaging System based on Cooled Digital Camera System
Wallac Distribution GmbH: 1. EG&G Wallac LSR UltraView, confocal fluorescence microscope for "real time" images. It is the first presentation of this instrument in Germany. 2. EG&G Wallac Signifer, fluorescence microscope for recording images wi= th the time resolved fluorescence prinziple. 3. EG&G Berthold NightOWL, a universal imaging system for low level luminescence images.
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques = are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meet= ing point for developers and users working in these rapidly evolving fields a= nd play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic development= s in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
A phantastic opportunity to meet well-kmown scientists and an excellent chance to see and to compare the latest equipment for confocal and multiphoton fluorescence microscopy. Attend an excellent meeting and see the most recent technological developments.
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D FOCUS ON MICROSCOPY 1999
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
we would like to invite you to attend this years "Focus on Microscopy" conference.
Many of the well known scientists in this field will present their recent progress.
Here is a list of the plenary speakers: - G.J. Brakenhoff, Amsterdam - Christoph Cremer, Heidelberg - Winfried Denk, Murray Hill - Timothy Holmes, Watervliet - Anthony A. Hyman, Heidelberg - Stefan Hell, Goettingen - Satoshi Kawata, Osaka - Karsten K=F6nig, Jena - Andres Kriete, Giessen - Ulrich Kubitscheck, Muenster - Eric Manders, Amsterdam - Colin Sheppard, Sydney - Ernst H.K. Stelzer, Heidelberg - Kevin F Sullivan, La Jolla - Tony Wilson, Oxford - Daniele Zink, Munich
The commercial exhibition offers an excellent overview. The modern instruments of all major microscope manufacturers as well as microscope accessories are on display. This is the 1999 event focussed on microscopy= in Europe.
Highlights of the commercial exhibition include:
Hamamatsu Photonics: ORCA Series - state of the art digital imaging =B7 Progressive scan interline transfer CCD with 1280x1024 pixel =B7 Lens-on-chip technology, TE cooling and optimized readout guarantee highest sensitivity =B7 Black & white cameras with 10-14 bit output =B7 Color cameras with RGB matrix filter or filterwheel =B7 Plug & play set incl. framegrabber and comfortable TWAIN driver avai= lable
L.O.T. Oriel GmbH & Co KG: Nanopositioning Unit and Fiber Laser
Molecular Probes: New fluorescent reagents for Cell Biology and Imaging highly - Fluorescent Alexa dyes - click here for more details
Nikon GmbH: Two compact confocal microscopes + fully automatic motorized research microscope
Olympus Optical Co. GmbH Confocal Laser-Scanning-Microscope with two-phot= on excitation
Omicron Vakuumphysik GmbH Scanning Near Field Optical Microscope (SNOM)
Visitron Systems GmbH: 2D/3D Fluorescence Imaging System based on Cooled Digital Camera System
Wallac Distribution GmbH: 1. EG&G Wallac LSR UltraView, confocal fluorescence microscope for "real time" images. It is the first presentation of this instrument in Germany. 2. EG&G Wallac Signifer, fluorescence microscope for recording images wi= th the time resolved fluorescence prinziple. 3. EG&G Berthold NightOWL, a universal imaging system for low level luminescence images.
12th International Conference on 3D Image Processing in Microscopy 11th International Conference on Confocal Microscopy April 11th-15th, 1999 European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Confocal microscopy, multiphoton excitation and deconvolution techniques = are increasingly applied in the study of three-dimensional structures such as are encountered in biology, medicine and material sciences. Three-dimensional analysis and representation are crucial tasks in subsequent data assessment. These conferences offer a most efficient meet= ing point for developers and users working in these rapidly evolving fields a= nd play an important role in the dissemination of information about new developments. Special attention will be given to the dramatic development= s in live cell imaging and manipulation, such as the role of the green fluorescent protein. Further information:
Local Organizing Committee: Dr. Ernst H.K. Stelzer, EMBL, Heidelberg Prof. G.J. Brakenhoff, University of Amsterdam Dr. Andres Kriete, University of Giessen
Under the auspices of The International 3D Microscopy Society: Prof. Colin Sheppard, University of Sydney Dr. Andres Kriete, University of Giessen Prof. G.J. Brakenhoff, University of Amsterdam Prof. P-C. Cheng, SUNY at Buffalo Prof. Tony Wilson, University of Oxford Dr. Carol Cogswell, University of Sydney Dr. Vyvyan Howard, University of Liverpool Dr. Guy Cox, University of Sydney Dr. Ernst H.K. Stelzer, EMBL Prof. S. Kawata, Osaka University
David asks ... } } } I am working on a business project that requires an analysis of the } microscopy market and am hoping someone can direct me to a } good source of information. } ...
The best jump off point is the WWW-Virtual Library ...
http://www.ou.edu/research/electron/www-vl/ ...
... hope this helps :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
It is possible to use a variety of other fixatives to preserve bacteria. However, we find that when all else fails, the R-K procedure nearly always gives a good fixation on organisms that otherwise look poorly preserved. The buffer is somewhat cumbersome to prepare since the veronal (sodium barbitol) is a controlled substance.
} I'm sorry to bother everyone, again, with a question about fixation of } bacteria. I greatly appreciate the responses I received a few weeks } ago. At one time Ryter-Kellenberger fixation was considered the } standard. It produced an image with a relatively clear nucleoid } containing fibrillar chromatin. Is this now considered to be an } artifact of preparation?
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I am looking for a procedure to do TEM of low density polyethylene films. I would like to stain the film so I can then microtome at room temperature. For high density films we use chlorosulfonic acid at 60C but this dissolves the low density material. I can cryo-microtome the low density films and then stain the sections with ruthenium tetroxide but this is giving only marginal results. I would really like to be able to room temperature microtome the material. Any help greatly appreciated.
------------------------------ Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
Need a simplified recipe for viewing circular DNA ~5000KB in a gamish of linear DNA. No special marker or complementary sequence available. Thanks.
*Disclaimer: Whatever... is not Tulane opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web:[ http://www.tmc.tulane.edu/ferminlab/] or [http://www.tmc.tulane.edu/imaging/] Internet: [cfermin-at-mailhost.tcs.tulane.edu] 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
I've received the following query and thought I'd post it so the various types of microscopists out there besides me might like to help this fellow out. Please reply directly to his eddress since he's not on the listserver.
Many thanks,
Dee Breger
hello:
my name is francisco sandoval, and I live in Guatemala in central america.
I am a photographer, and I am interested in microscope photography, but I do not know where to begin, could you help me with it.
I guess I do not understand the kind of microscope you are using. I do not know anything about microscopes..... If you do not mind helping me, tell me where to begin..... Thanks
Francisco Sandoval
kikophoto-at-geocities.com jfse30-at-hotmail.com
www.geocities.com/paris/jardin/4108/
____________________________________________________________________________ Automatic note: Sometimes I don't receive incoming emails (with no notice to the sender). If I don't respond to your message, please send it again! ____________________________________________________________________________ _ Dee Breger Manager, SEM/EDX Facility Lamont-Doherty Earth Observatory Route 9W Palisades NY 10964 USA
As a company that produces image processing systems (see disclaimer below), I would like to invite you to check out our website at http://www.soft-imaging.com or http://www.soft-imaging.de. We routinely deal with 12 to 16 bit gray scale images as most digital cameras supply them.
If you need further information, don't hesitate to contact us at one of the URLs or numbers below.
Thank you.
Michael Bode
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
} ---------- } From: Hendrik O. Colijn[SMTP:colijn.1-at-osu.edu] } Sent: Friday, March 19, 1999 6:16 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Image processing software } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Is anyone familiar with reasonably priced software programs that can } manipulate 16-bit gray scale images? While Adobe Photoshop can read } 16-bit } images, you have to change the images to 8-bit before you can run } filters } etc. Gatan's Digital Micrograph can work with 16-bit images, but its } price } is a little steep for equipping several computers. What are the } capabilities of other packages? I would like to use the software to } work } with diffraction patterns as well as images. } } TIA, } Henk } } Hendrik O. Colijn colijn.1-at-osu.edu } OSU Campus Electron Optics Facility (614) 292-0674 } "An optimist believes that we live in the best of all possible worlds. } A pessimist fears that this is true." }
I provide third party maintenance for EM in the midwest, so take the following as a somewhat biased view. I'd also like to apologize in advance for my typically long and meandering response. I don't pull punches, so perhaps I should start adding a disclaimer that the following are my opinions - if that's not enough, go ahead and sue, I don't have anything anyway.
An insurance company's primary usefulness is to large organizations that can off-load the responsibility for a large amount of equipment. Their customers will contract with them for all of their equipment, they handle the paperwork and arrange service. In these cases, a good portion of the cost savings can be in the customer's reduction in paperwork - one contract as opposed to hundreds. To do the service, they hire the manufacturer (don't know how many manufacturers go along with this, not all do) or an independant service provider (I have been approached a number of times, haven't done it yet) on an hourly basis to do the work, usually negotiating reduced rates. I am sure that, here in the early stages, they are underestimating their costs and will raise their rates as they gain a greater understanding.
A number of direct problems. Mainly, they will have a hard time guaranteeing response time. For any manufacturer or independant, contract customers come first. You are also at their mercy for who comes to work on your equipment, it may even be a different organization each time. From my point of view, you run a risk of never getting someone who has some long term interest or commitment in your needs, even if they use the original equipment manufacturer. The OEM will no longer bear any responsibility other than providing service when needed (and the more often the better for their bottom line when hourly).
Service used to be a loss-leader, a way for a company to increase their sales by creating good will and a positive corporate image. Question this, just count up the number of JEOLs sold in the last fifteen years. They made a conscious effort to provide exceptional service and sold like crazy on the basis of those efforts. A couple of decades ago, most companies found that there are profits to be made in service and have run their service accordingly and made the industry an increasingly tempting market for new manufacturers, third party service providers and the new insurance companies.
If you only have a few pieces of equipment, you'd do better to find a local independant. You'll probably find that an independant can be very flexible in contract terms. Consider asking them to remove the 'insurance' aspects of a service contract by removing parts coverage. You could still get PMs and emergency labor, but the service provider would not have to be escrowing a good chunk of money just in case an expensive part goes. That, of course, would require you to do so. There is some insurance value to the contract labor rates, but generally not much. A good technician can do a lot to prevent future service needs and it is a lot less traumatic to a small operation to spend extra time fixing an instrument than it is to replace that $10,000 whiz-bang that blew.
You can also, of course, handle your service on a billable basis with manufacturer or independent. Watch out for the manufacturers, though. Some like to charge a premium (I've seen over $300/hour) in order to encourage you to go under contract. Independents may play this game too, but not to that extent (I don't know of any who do). While I am obviously biased towards the independents, I truely think that supporting them is the only way to keep any real competition in this or any other industry.
Frankly, the insurance companies are hoping to become the big kid on the block in the future so that they can create their own terms with the manufacturers. Instrumental service rates are bad enough already, but this squeeze play could eventually hurt the industry the same way managed care and HMOs have hurt the medical industry. Manufacturers have income goals that will be met, one way or another. If the insurance companies squeeze out their service profits, manufacturer's prices on remaining contracts will go up which will then force more users to go to the insurance companies. Manufacturers will also have to increase the selling price for their equipment. We are merely adding another layer of for-profit companies to take a bite out of your pocket.
There is a potentially insidious effect of organizations like this. In claiming to reduce costs for a few, they wind up increasing costs for all. Rather than introducing new competition, they manipulate a market to the point where some existing competitors leave the market. They bring no new instruments to market, work counter to free enterprise by attempting to be the arbitor of pricing without actually providing the service and, as the market becomes saturated by them, will go into bidding wars between themselves reducing services in order to lower their costs. There will be some manufacturers that won't be able to stay competitive with the external pressure and others that won't want to bend to those pressures. Look to a future of greater cutbacks and mergers as these new players are accommodated in the market.
We are also already seeing another aspect of this. In attempting to cut costs, manufacturers may cut back their own sales efforts, relying on others instead. The EM markets are getting large enough and there are enough manufacturers that we may eventually see 'distributers' added on the front end, yet another layer of for-profit companies taking a piece of the pie. It is common for a company to enter a new national market through marketing arrangements. ISI has for a long time had direct sales presence in the US. While this is a company that has had its share of problems, their new marketing arrangement is a cutback in its sales force that is in large part due to its poor service history and poor profits in sales and service. While not precipitated by the insurance companies, it still may offer a view of their future effects.
It will not benefit any manufacturer to take part in these plans in the long run. Doing so would be giving up all control over a very important sector of their public relations, cut into their profits and resolve them to having other companies dictate their policies and pricing. Look for some manufacturers to resist because doing so allows them to distinguish themselves in the market by maintaining their own high level of service responsiveness and performance. However, as this snowballs we may well reach a critical mass where all manufacturers will have to participate in order to survive.
Good luck, whatever you choose.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com
-----Original Message----- } From: Tindall, Randy D. {TindallR-at-missouri.edu} To: 'microscopy-at-sparc5.microscopy.com' {microscopy-at-sparc5.microscopy.com}
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Stephen,
The following procedure has been adapted from the literature (I don't have the references with me):
The method that I normally use to image LLDPE is to first embed a strip of the film with Epofix epoxy using a silicone mold. After curing the epoxy, face-off the end of the specimen block to expose the film (assuming you want a cross-section) and then attach the block to a piece of double-sided tape on a glass microscope slide. The next step involves staining the specimen block with the vapor of a mixture of 0.2 g of ruthenium trichloride trihydrate and 10 ml of 5.25% sodium hypochlorite (regular household bleach). This produces a fairly aggressive RuO4 vapor. The staining mixture and the slide should be kept in a glass jar with a loosely fitting lid for 2 to 3 hours -at- room temp. Occasionally, the temp. needs to be elevated for higher density polyolefins. Do all of the staining in a fume hood!!
After staining is complete, remove the slide/specimen, rinse well and allow to dry. The specimen block can be trimmed and microtomed at room temperature. I normally orient the film parallel to the edge of the diamond knife. This helps prevents delamination from the epoxy.
I hope this procedure helps. I've been using this technique for several years with great success.
Gene Young Microscopy/Microanalysis Group Dow Chemical USA Freeport, TX
------------------------------ Original message:
I am looking for a procedure to do TEM of low density polyethylene films. I would like to stain the film so I can then microtome at room temperature. For high density films we use chlorosulfonic acid at 60C but this dissolves the low density material. I can cryo-microtome the low density films and then stain the sections with ruthenium tetroxide but this is giving only marginal results. I would really like to be able to room temperature microtome the material. Any help greatly appreciated.
------------------------------ Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
} Date: Tue, 23 Mar 1999 16:56:22 -0600 } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: photographer needs microscopy info } } } Dear collegues, } } I've received the following query and thought I'd post it so the various } types of microscopists out there besides me might like to help this fello } out. Please reply directly to his eddress since he's not on the listserver } } } Many thanks, } } Dee Breger } } hello: } } my name is francisco sandoval, and I live in Guatemala in central } america. } } I am a photographer, and I am interested in microscope photography, but } I do not know where to begin, could you help me with it. } } I guess I do not understand the kind of microscope you are using. I do } not know anything about microscopes..... If you do not mind helping me, } tell me where to begin..... Thanks } } Francisco Sandoval } } kikophoto-at-geocities.com } jfse30-at-hotmail.com } } _________________________________ } Francisco, I don't know if it is still available but the booklet by Eastman Kodak, Photography through the Microscope, is quite good. It isKodak Publication P-2 CAT 152 8371. Published in 1980
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
} I am looking for a procedure to do TEM of low density polyethylene films. I } would like to stain the film so I can then microtome at room temperature. } For high density films we use chlorosulfonic acid at 60C but this dissolves } the low density material. I can cryo-microtome the low density films and } then stain the sections with ruthenium tetroxide but this is giving only } marginal results. I would really like to be able to room temperature } microtome the material. Any help greatly appreciated.
I'm an etcher, not a stainer, but we do have some experience of staining materials in bulk. Possibly a room temperature chlorosulphonation might help. I've never used RuO4, but it might also work in bulk on a thin enough film. I'll have to ask a colleague, when he's in his office.
If you're interested in larger scale variations in morphology, then permanganic etching followed by reflection optical microscopy (Nomarski) is very useful.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Before you do anything, including cleaning the ball bearings, find out their history! The goop on the surface may be part of the story.
My advice: try light microscopy first. Reflected light DIC may tell you alot about the surface. Also try interferometry (I'll be back in the office Monday; call or contact me directly for further info). I had a similar project for a client some years ago with ball bearings for jet engines. If the height of the asperities was too tall, they would break off, leaving metallic residue in the raceway which caused the lubricant to polymerize and freeze up. Needless to say, not a desirable outcome for an airplane at 30,000+ feet!
If the bumps on the ball bearings are not the problem, then you might want to try SEM + EDS to see if there is some problem in the alloy.
Best of luck and let me know how things turn out.
Barbara Foster 125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
At 10:53 AM 3/4/99 -0500, Lesley S. Bechtold wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I've attached a recent announcement for a workshop as an example of an item "for sale." I believe that items for sale, whether services, supplies, or instruments all require the same kind of response you gave to my posting for "instruments available" recently. Please see that workshops for profit are stricken from the Listserver so that you are consistent with your own policy or, change your posting policy.
I believe that any of us who are in this field should be able to post any kind of useful informative item that would be of interest to those on the list. The availability of microscopes is a useful piece of info for someone who has a legitimate need for a used scope. There are a lot of people looking for microscope users and owners as a trusted resource rather than the manufacture's sales reps. Please consider this in the future.
Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: from rly-yc02.mx.aol.com (rly-yc02.mail.aol.com [172.18.149.34]) by air-yc04.mail.aol.com (v58.13) with SMTP; Tue, 23 Mar 1999 00:26:21 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by rly-yc02.mx.aol.com (vx) with SMTP; Tue, 23 Mar 1999 00:25:48 -0500 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id WAA03774 for dist-Microscopy; Mon, 22 Mar 1999 22:52:10 -0600 Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id WAA03770 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 22 Mar 1999 22:51:39 -0600 Received: from smtp2.mindspring.com (smtp2.mindspring.com [207.69.200.32]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id WAA03763 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 22 Mar 1999 22:51:26 -0600 Received: from [209.86.60.157] (user-38lcf4t.dialup.mindspring.com [209.86.60.157]) by smtp2.mindspring.com (8.8.5/8.8.5) with ESMTP id UAA25474; Mon, 22 Mar 1999 20:54:55 -0500 (EST) Message-Id: {199903230154.UAA25474-at-smtp2.mindspring.com} X-Mailer: Microsoft Outlook Express for Macintosh - 4.01 (295)
TEM & SEM Sample Preparation of Materials
A Two Part Workshop & Seminar
Leica Microsystems, Diatome US, Electron Microscopy Sciences, and Bal-Tec announce another in a series of EM Specimen Preparation workshops. These seminars will focus on the following techniques:
Ultramicrotomy of Materials
Embedding of industrial samples Specimen trimming
Ultramicrotomy of hard materials Ultramicrotomy of polymers
Collection & handling of sections Staining of polymer sections
Low temperature ultramicrotomy
Ion Beam Milling of Materials
Ion Milling of plan view samples for TEM
Ion Milling of cross section samples for TEM
SEM preparation of interfaces using fast slope cutting technique
TEM / SEM preparation of temperature sensitive specimens
SEM / LM preparation of surface structures with ion milling
The format of our workshop is half day lecture and half day bench work. Samples will be supplied by the course instructors. Participants are encouraged to bring their own samples to work with as time allows.
Course Speakers & Instructors
Ultramicrotomy of Materials
Dr. Tom Malis CANMET Characterization Group Leader Materials Technology Laboratory Ottawa, Ontario
Mr. Bob Vastenhout DOW Chemical Polymer Microscopist Analytical Science Department Terneuzen, The Netherlands
Mr. Helmut Gnaegi Product Manager Microtomist Extaordinaire Diatome, Ltd., Switzerland
Ion Beam Milling
Dr. Wolfgang Gruenewald Bal-Tec Head of Applications Laboratory Liechtenstein
Mr. Arthur Buechel Bal-Tec Product Manager Liechtenstein
Hosted by: JoAn Hudson Clemson University Microscopy Facility Clemson, SC
When: Ion Milling June 7-8, 1999
Ultramicrotomy June 9-11, 1999
Tuition: Ion Milling $400.00
Ultramicrotomy $1,400.00
Ion Milling & Ultramicrotomy $1,700.00
Includes lodging at the lakefront Conference Center & Inn at Clemson University, continental breakfast and lunch daily, one group dinner, course supplies, and lab charges.
Contact: Mike Boykin, Leica Microsystems, Inc. 800-248-0665 X5092
Hi. I was recently asked of a way to determine C film thickness. I = remember that using a polished brass specimen and observing the color is = one way. Does anyone have a source in the literature for this method? = Thanks.
______________________ Roberto Garcia Senior Analyst, Metallography North Carolina State University Analytical Instrumentation Facility Box 7531, Room 303 EGRC Raleigh, NC 27695-7531 rgarcia-at-unity.ncsu.com http://spm.aif.ncsu.edu/aif
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{META content=3D'"MSHTML 5.00.0910.1309"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT size=3D2} Hi. I was recently asked of a way to determine C = film=20 thickness. I remember that using a polished brass specimen and observing = the=20 color is one way. Does anyone have a source in the literature for this = method?=20 Thanks. {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D3} ______________________ {BR} Roberto Garcia {BR} Senior = Analyst,=20 Metallography {BR} North Carolina State University {BR} Analytical = Instrumentation=20 Facility {BR} Box 7531, Room 303 EGRC {BR} Raleigh, NC=20 27695-7531 {BR} rgarcia-at-unity.ncsu.com {BR} http://spm.aif.ncsu.edu/aif {/FONT= } {/DIV} {/BODY} {/HTML}
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a student who has done confocal on tetrahymena and now wants to move on to TEM. She is primarily interested in the microtubular arrays in the cilia and also within the body of the organism, especially at division. However, we also would like to get the best possible preservation of the entire organism.
Since these two needs may conflict, I would appreciate hearing from anyone with experience with this or similar organisms.
Possible approaches could include: a) using tannic acid to stabilize the microtubules (percent?, in all solutions or only primary fix?, best buffer system?, etc) b) using a combined glutaradyhde-osmium fix followed by straight osmium (recommended by Hayat.
Thanks in advance for the assistance.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
id HMDAAJC2; Wed, 24 Mar 1999 12:54:34 -0600 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" X-Sender: schattenh-at-pop.email.missouri.edu (Unverified) Message-Id: {v04020600b31ee56f600f-at-[128.206.215.129]}
Ph.D. graduate student or post-doctoral student (D.V.M. or M.D.) position available for research in the area of protozoal parasitology. Research will involve electron microscopy and cellular biology studies utilizing in vitro derived Sarcocystis spp. , Neospora spp. and Toxoplasma gondii parasites. Individual will work with a multi-disciplinary team at the University of Missouri-Columbia, in the Department of Veterinary Pathobiology and the Molecular Biology Program Electron Microscopy Core facility. Successful candidate should have background in tissue/cell embedding and processing for light and electron microscopy, and demonstrated proficiency in written and spoken English. Molecular biology and cell culture skills are helpful, but not a necessary requirement. U.S. citizenship is not required. For further information contact Dr. A.E. Marsh at ph: 573-884-2673, fax: 573-884-5414, or email: marshae-at-missouri.edu {mailto:marshae-at-missouri.edu} .
Try The American Mineralogist, Volume 58, pages 920-925, 1973. The role of Carbon Film Thickness in Electron Microprobe Analysis - Kerrick DM, Eminhizer LB amd Villaume JF.
Hi. I was recently asked of a way to determine C film thickness. I = remember that using a polished brass specimen and observing the color is = one way. Does anyone have a source in the literature for this method? = Thanks.
______________________ Roberto Garcia Senior Analyst, Metallography North Carolina State University Analytical Instrumentation Facility Box 7531, Room 303 EGRC Raleigh, NC 27695-7531 rgarcia-at-unity.ncsu.com http://spm.aif.ncsu.edu/aif
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD} {META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {STYLE} {/STYLE}
{META content=3D'"MSHTML 5.00.0910.1309"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT size=3D2} Hi. I was recently asked of a way to determine C = film=20 thickness. I remember that using a polished brass specimen and observing = the=20 color is one way. Does anyone have a source in the literature for this = method?=20 Thanks. {/FONT} {/DIV} {DIV} {/DIV} {DIV} {FONT size=3D3} ______________________ {BR} Roberto Garcia {BR} Senior = Analyst,=20 Metallography {BR} North Carolina State University {BR} Analytical = Instrumentation=20 Facility {BR} Box 7531, Room 303 EGRC {BR} Raleigh, NC=20 27695-7531 {BR} rgarcia-at-unity.ncsu.com {BR} http://spm.aif.ncsu.edu/aif {/FONT= } {/DIV} {/BODY} {/HTML}
------=_NextPart_000_0017_01BE75E6.4E13C940--
Robert MacKay Department of Earth Sciences Dalhousie University Halifax, Nova Scotia, Canada B3H 3J5 Tel: 902 494-7087 Fax: 902 494-6889 e-mail rmackay-at-ac.dal.ca
} Subject: job opening } Priority: normal } X-mailer: Pegasus Mail for Windows (v2.42a) } Message-ID: {5586A5401D-at-dcsmserver.med.sc.edu}
} } Hi, } } We currently have a position open for a microscopist in the } University of South Carolina School of Medicine Instrumentation } Resource Facility. Primary responsibilities involve assisting } faculty, staff and students in the use of confocal and electron } microscopes and in digital preparation of images (primarily Adobe } Photoshop). Secondary responsibilities involve assistance a flow } cytometer and a BioRad phosphorimager. } } All current work in the facility is biological. Applicants should } have a masters degree or equivalent experience. The } position is listed as a Research Specialist II with a salary range } from $24,618-$35,081. Please reply directly to me at
} Price-at-med.sc.edu. } Robert L. Price } Director, Instrumentation } Resource Facility } USC School of Medicine } Garner's Ferry Road } Columbia, SC 29208 } Phone: 803-733-3393 } Fax:803-733-1533
I am one of the people involved in the "for sale" example that Mark feels to be equivalent to his "instruments available" posting, which, I must confess, I don't remember seeing, or Nestor's apparent striking of it from the Listserver. I have participated in 8 of these TEM specimen prep workshops for Leica and RMC over the years, plus I have taken part (with many others) in similar workshops or short courses that are university-based, with the famous Lehigh short courses being the most widely-known example. I think that there are 2 key points you are missing, Mark:
1) These workshops are not designed to make money. If they break even, the companies that organize them are ecstatic.
2) These workshops are highly educational, which is the justification used by the students attending to their superiors (and confirmed by attendee feedback). As a supervisor, I continually look for such workshops on the Listserver for professional upgrading and/or career changes for my scientists and technologists.
When I want a new (or used) research tool, I tend not to look to the Listserver, but talk to vendors or colleagues, walk the floor at the MSA commercial exhibition, etc, etc. Whether or not your posting meets Nestor's criteria is not up to me, but let's not confuse apples with oranges (or the difficulty in properly using a complex instrument with the initial purchase of the instrument).
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 613-992-8735 email: malis-at-nrcan.gc.ca
} ---------- } From: } "Mriglermas-at-aol.com"-at-Sparc5.Microscopy.Com[SMTP:"Mriglermas-at-aol.com"-at-Sparc } 5.Microscopy.Com] } Sent: March 24, 1999 10:49 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Cc: Mriglermas-at-aol.com } Subject: Fwd: TEM / SEM Sample Prep Workshop } } { {Message: TEM / SEM Sample Prep Workshop} } } Dear Friendly Sysop - Nestor: } } I've attached a recent announcement for a workshop as an example of an } item } "for sale." I believe that items for sale, whether services, supplies, or } instruments all require the same kind of response you gave to my posting } for } "instruments available" recently. Please see that workshops for profit } are } stricken from the Listserver so that you are consistent with your own } policy } or, change your posting policy. } } I believe that any of us who are in this field should be able to post any } kind } of useful informative item that would be of interest to those on the list. } The availability of microscopes is a useful piece of info for someone who } has } a legitimate need for a used scope. There are a lot of people looking for } microscope users and owners as a trusted resource rather than the } manufacture's sales reps. Please consider this in the future. } } } Thanks for your attention to this matter. } } Mark W. Rigler, Ph.D. } VP, MAS, Inc. }
} I'm sorry to bother everyone, again, with a question about fixation of } bacteria. I greatly appreciate the responses I received a few weeks } ago. At one time Ryter-Kellenberger fixation was considered the } standard. It produced an image with a relatively clear nucleoid } containing fibrillar chromatin. Is this now considered to be an } artifact of preparation? } } John Wright } West Desert Test Center } (435) 831-3017 }
Two of the more recent reviews by the same E. Kellenberger ("The bacterial nucleoid revisited" (1994) in Microbiol Rev 58:211-32; and "Functional consequences of improved structural information on bacterial nucleoids" (1991) in Res Microbiol 142:229-38) should answer your question in great detail. They also make a most enjoyable reading!
Briefly, the nucleoid, according to the results obtained with rapid freezing-freeze substitution, "is now more granular than fibrillar", thus, conceivably, reflecting the natural supercoiled state of the DNA. It has a "coralline" shape, and "the excrescencies reach far into the cytoplasm. Membrane contact is no longer excluded". Interestingly, but the liquid-crystalline form of the DNA also appears to be native in some cases...
The classic Ryter-Kellenberger method still makes a standard for a good chemical fixation. It is beneficial sometimes to add an aldehyde prefixation step or make other minor modifications, but treatment with AQUOEUS uranyl acetate before dehydration remains the most critical for good nucleoid preservation. Bacterial "chromosome" lacks the proteins that keep eukaryotic chromatin from collapse during dehydration, and only aqueous uranyl acetate, following conventional fixation, cross-links the nucleoid in that "liquid-crystalline" state.
I remember your original posting also generated discussion about delayed secondary fixation, or for how long the material can be left in glut. For fine ultrastructure work, I would not leave bacteria (as well as anything else) at any step longer than necessary. Even if the osmotic pressure is adjusted to minimize swelling/shrinking, you will most likely end up with a specific unpleasant granularity of the cytoplasm in your bacteria if you leave them in a glutaraldehyde fixative for too long. To split the protocol into two days, it is best to leave the material overnight in 70% ethanol in the refrigerator. But for just diagnostic, etc., purposes, it is, of course, O.K., and sometimes simply unavoidable, to store the samples for quite a while in a glutaraldehyde fixative in a refrigerator.
Please feel free to contact me directly if you can't find the journals or if other questions arise. Sincerely, Vlad.
Vladislav V. Speransky School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Phone: 207 581 2998 Fax: 207 581 2969 Email: vladis-at-maine.maine.edu
Theoretical stereology Mathematical morphology Advances in image=20 analysis, Modern techniques in microscopy /image acquisition=20 Quantitative fractography 3-D modelling and analysis dissemination of=20 stereology and image analysis applications=20
Conference language English, no translations anticipated
Conference program (proceedings will be distributed prior to the=20 conference)=20
No parallel sessions Invited lectures (25') Oral presentations (15')=20 Poster presentations combined with panel discussion=20
Deadlines=20
Preliminary registration, October 31, 1999=20 Second circular, December 31, 1999=20 Final papers, March 31, 2000=20 Final registration, May 31, 2000=20 Final circular with conference program, July 31, 2000=20
Correspondence:
STERMAT 2000
Institute of Materials Science Cracow University of Technology Al. Jana Paw=B3a II 37 31-864 Krak=F3w, Poland
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Roberto, We used a thin gold coating on the shiny side of household = aluminum foil. This works great for relative thicknesses. Russ
-----Original Message----- } From: Roberto Garcia [mailto:rgarcia-at-unity.ncsu.edu] Sent: Wednesday, March 24, 1999 11:06 AM To: MSA Microscopy
The accurate control of the thickness of evaporated carbon films.
Principle. When carbon is deposited onto gold, it forms interference colors that are well defined. They can be used to determine the thickness of the carbon.
The colors. If carbon is evaporated onto gold, as the thickness of the carbon increases, the color changes through the following sequence: gold, orange, red, blue, grey. The change of color from red to blue is particularly sharp and clear. The change of color from red to blue occurs when the thickness of the carbon is 24.0 nm +/- 0.5nm. This result was obtained by people at Balzers using a multibeam interference technique for calibration.
Details. 1 Take a glass slide (or any other suitable substrate) and evaporate onto it a layer of gold. The thickness is not critical as long as the gold is thick enough to give an opaque film that looks like gold. 2 Mount the slide in the same chamber with the specimen to be coated with carbon. the thickness of the carbon on the slide will be 24 nm so arrange the distance of the slide and the sample so that (by the inverse square law) the desired thickness on the sample will occur when the thickness on the slide is 24 nm. 3 Evaporate the carbon; stop the evaporation as the color changes form red to blue. If you are using a normal arc for the carbon evaporation, the light from the arc will allow you to see the colors. The bell jar will need to be reasonably clean.
Example. Suppose you need to deposit a carbon film of thickness T nm. Let d be the distance from the carbon arc to the gold slide; let D be the distance from the carbon arc to the specimen. Then [d/D]squared = T/24.
Reference: My thesis (1967).
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Reply to: RE: Thickness of carbon films. Re: Carbon Film Thickness When great accuracy of film thickness is not needed, we simply = place a small metal washer upon a glass slide near the "specimen area". after evaporation the step height between the shaded and coated areas is = measured optically on a bench interference microscope. Ours is a Zeiss = two beam unit that is good for 30 nanometer resolution, (+ or -), with no = physical contact with the film. It works for ANY reflective metallic film = and fairly well even on "dull" carbon coatings because it has three = different reference mirrors that can be quickly changed. Each mirror has a = different reflectivity that one merely tests to get reasonable = interference fringes which can also be photographed via polaroid or 35 mm. = film. Bernie Kestel Materials Science Division = Argonne National Laboratory 9700 South Cass Ave. Argonne, Il., 60439
E-mail {kestel-at-anl.gov} FAX: (630) 252-4289
Alwyn Eades wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
{HTML} {HEAD} {/HEAD} {BODY} {PRE = WIDTH=3D"132"} Reply to: RE: Thickness of carbon films.
{/PRE} {FONT FACE=3D"Geneva" SIZE=3D3 COLOR=3D"#000000"} Re: = Carbon Film Thickness {BR} When = great accuracy of film thickness is not = needed, we simply place a small metal washer = upon a glass slide near the "specimen = area". {BR} after evaporation the step = height between the shaded and coated areas = is measured optically on a bench interference = microscope. Ours is a Zeiss two beam unit = that is good for 30 nanometer resolution, = (+ or -), with no physical contact with = the film. It works for ANY reflective metallic = film and fairly well even on "dull" = carbon coatings because it has three different = reference mirrors that can be quickly changed. = Each mirror has a different reflectivity = that one merely tests to get reasonable = interference fringes which can also be photographed = via polaroid or 35 mm. film. {BR} Bernie = Kestel {BR} Materials Science Division {BR} = Argonne National Laboratory {BR} 9700 South = Cass Ave. {BR} Argonne, Il., 60439 {BR} {BR} = E-mail <kestel-at-anl.gov> FAX: = (630) 252-4289 {BR} {BR} Alwyn Eades wrote: {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} {BR} >-----------------------------------------------------------------------= - {BR} >The = Microscopy ListServer -- Sponsor: The Microscopy = Society of America {BR} They can be used to determine the thickness = of the carbon. {BR} > {BR} >The colors. {BR} >If = carbon is evaporated onto gold, as the thickness = of the carbon {BR} >increases, the color = changes through the following sequence: = gold, orange, {BR} >red, blue, grey. The = change of color from red to blue is particularly {BR} >sharp = and clear. The change of color from red = to blue occurs when the {BR} >thickness = of the carbon is 24.0 nm +/- 0.5nm. {BR} >This = result was obtained by people at Balzers = using a multibeam {BR} >interference technique = for calibration. {BR} > {BR} >Details. {BR} >1 Take = a glass slide (or any other suitable substrate) = and evaporate onto {BR} >it a layer of gold. = The thickness is not critical as long as = the gold is {BR} >thick enough to give an = opaque film that looks like gold. {BR} >2 Mount = the slide in the same chamber with the specimen = to be coated with {BR} >carbon. the thickness = of the carbon on the slide will be 24 nm = so arrange {BR} >the distance of the slide = and the sample so that (by the inverse square {BR} >law) = the desired thickness on the sample will = occur when the thickness on {BR} >the slide = is 24 nm. {BR} >3 Evaporate the carbon; = stop the evaporation as the color changes = form red {BR} >to blue. If you are using = a normal arc for the carbon evaporation, = the {BR} >light from the arc will allow = you to see the colors. The bell jar will {BR} >need = to be reasonably clean. {BR} > {BR} >Example. {BR} >Suppose = you need to deposit a carbon film of thickness = T nm. Let d be the {BR} >distance from = the carbon arc to the gold slide; let D be = the distance from {BR} >the carbon arc to = the specimen. Then [d/D]squared =3D T/24. {BR} > {BR} >Reference: = My thesis (1967). {BR} > {BR} >Alwyn Eades {BR} >Department = of Materials Science and Engineering {BR} >Lehigh = University {BR} >5 East Packer Avenue {BR} >Bethlehem {BR} >Pennsylvannia = 18015-3195 {BR} > Phone 610 758 4231 {BR} > Fax = 610 758 4244 {BR} > {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 = COLOR=3D"#0000FF"} {U} jae5-at-lehigh.edu {/U} {/FONT} {FONT FACE=3D"Geneva" = SIZE=3D1 = COLOR=3D"#000000"} {BR} > {BR} > {BR} > {BR} >RFC822 = header {BR} >----------------------------------- {BR} > {BR} > = Received: from dns2.anl.gov (dns2.anl.gov = [146.139.254.3]) by {BR} >horus.et.anl.gov = (8.6.11/8.6.11) with ESMTP id IAA26354 for = < {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} kestel-at-= horus.et.anl.gov {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} >; Thu, 25 {BR} >Mar = 1999 08:32:30 -0600 {BR} > Received: from = Sparc5.Microscopy.Com (sparc5.microscopy.com = {BR} >[206.69.208.10]) by dns2.anl.gov = (8.9.1a/8.6.11) with SMTP id IAA03123; Thu, = 25 Mar 1999 {BR} >08:32:29 -0600 (CST) {BR} > = Received: (from {/FONT} {FONT FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#0000FF"} {U} = daemon-at-localhost {/U} {/FONT} {FONT = FACE=3D"Geneva" SIZE=3D1 COLOR=3D"#000000"} ) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) {BR} >id HAA19493 for dist-Microscopy; = Thu, 25 Mar 1999 07:53:13 -0600 {BR} > Received: = from no_more_spam.com (Sparc5 [206.69.208.10]) = by {BR} >Sparc5.Microscopy.Com (8.6.11/8.6.11) =
for {Microscopy-at-Sparc5.Microscopy.Com} id QAA26644 (ESMTP). Thu, 25 Mar 1999 16:50:44 +0100 (MET) Received: by tnt1.chem.tue.nl with Internet Mail Service (5.5.2448.0) id {GXT3GKNM} ; Thu, 25 Mar 1999 16:50:44 +0100 Message-ID: {23285647B6CFD111B783002048083698174AC4-at-tnt1.chem.tue.nl} {Microscopy-at-Sparc5.Microscopy.Com}
Hallo Roberto,
In the first edition of "Electron Microprobe Analysis" of S.B.J. Reed (Cambridge University Press 1975) you can find the next table. for Carbon on polished brass
Thickness in nm Colour
15 Orange 20 Indigo red 25 Blue 30 Bluish green 35 Green blue 40 Pale green 45 Silver gold
We determined the thickness with our thin layer program and found out that the values were very good. The table is not found in newer editions of Reed's book.
Ir.Hans Heijligers Solid State and Materials Chemistry Lab. STO 2.45, Eindhoven University of Technology POBox 513 NL-5600 MB Eindhoven E-mail: H.J.M.Heijligers-at-TUE.NL Tel.: +31 (0)402473051 Fax.: +31 (0)402445619
Hi. I was recently asked of a way to determine C film thickness. I remember that using a polished brass specimen and observing the color is one way. Does anyone have a source in the literature for this method? Thanks.
______________________ Roberto Garcia Senior Analyst, Metallography North Carolina State University Analytical Instrumentation Facility Box 7531, Room 303 EGRC Raleigh, NC 27695-7531 rgarcia-at-unity.ncsu.com http://spm.aif.ncsu.edu/aif
I'm in the market for a good quality, but inexpensive, film scanner- e.g., for scanning 35mm slides into my PC. I've heard Nikon makes a good one (model LS2500), but it runs a hefty $2,500 or so (too expensive for me). Got any suggestions?
-- Steven J. Fliesler, Ph.D. Professor Dept. of Ophthalmology Saint Louis Univ. School of Med. 1755 S. Grand Blvd. St. Louis, MO 63104-1540 Phone: (314) 577-8259 Fax: (314) 771-0596 E-mail: Fliesler-at-slu.edu
I am thinking of purchasing a microwave tissue processor. I would appreciate your opinion as to the pro's and con's of this type of tissue preparation.
If you have access to an AFM, accurate measurement of C thickness takes about 4 minutes. If I'm looking at flat samples, I use a toothpick to remove a thin line o= f carbon. I then can bring the sample to the AFM and directly measure the height of the step betwwen the sample and the top of the carbon film. Alternatively, if I can't work directly on the sample I coat a plain glas= s slide (making sure it is the same distance from the arc as the sample) an= d measure the C thinckness on it. Hope this helps
Hello everyone, A friend of mine has informed me that there is a young boy in the south of England who has a terminal cancer and is close to the end of his suffering. Apparently, he wishes to enter the Guiness Book of Records for the largest collection of buisness cards. Could you all take the time to help boost this young man's collection. I was asked to tell everyone else to forward this information to as many other people as possible. His particulars are: Master Gary Richard, 30 Selby Road, Carshalton, Surrey, England, U.K.
Regards Martin Roe Macaulay Land Use Research Institute Aberdeen Scotland AB15 8QH U.K.
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Reply to: RE: Thickness of carbon films. Re: Carbon Film Thickness When great accuracy of film thickness is not needed, we simply place a small metal washer upon a glass slide near the "specimen area". After evaporation the step heigth between the shaded and coated areas is measured optically on a bench interference microscope. Ours is a Zeiss two beam unit that is good for 30 nanometer resolution, (+ or -), with no physical contact with the film. It works for ANY reflective metallic film and fairly well even on "dull" carbon coatings because it has three different reference mirrors that can be quickly changed. Each mirror has a different reflectivity that one simply tests to get reasonable interference fringes which can also be photographed via polaroid or 35 mm. film. Bernie Kestel Materials Science Division Argonne National Laboratory 9700 South Cass Ave. Argonne, Il., 60439
E-mail {kestel-at-anl.gov} FAX: (630) 252-4289
Alwyn Eades wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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We have a Cambridge S240 scanning electron microscope that is available, that needs a new lab home. The S240 was wrapped up recently when we unexpectedly received a field emission SEM. The S240 is a digital frame store SEM; has a LaB6 source; and a second backplate that adapts to a Microspec WDS spectrometer. The S240 also includes a mating Noran thin window EDS detector. The S240 was well maintained under service contract for 11 years, and still would be in use if the FESEM has not suddenly become available from another Raytheon facility.
We also can include a used Kevex 8000 EDS analyzer with the interfaces to the Noran detector. In addition, I know where that Microspec spectrometer can be obtained.
We would like to trade the Cambridge SEM/Noran detector, and if desired, the Kevex analyzer, for a new Noran Vantage upgrade to our Noran EDS analyzer. We would like to hear from anyone who is interested.
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Reply to: RE: Thickness of carbon films.
Re: Carbon Film Thickness When great accuracy of film thickness is not needed, we simply place a small metal washer upon a glass slide near the "specimen area". after evaporation the step height between the shaded and coated areas is measured optically on a bench interference microscope. Ours is a Zeiss two beam unit that is good for 30 nanometer resolution, (+ or -), with no physical contact with the film. It works for ANY reflective metallic film and fairly well even on "dull" carbon coatings because it has three different reference mirrors that can be quickly changed. Each mirror has a different reflectivity that one merely tests to get reasonable interference fringes which can also be photographed via polaroid or 35 mm. film. Bernie Kestel Materials Science Division Argonne National Laboratory 9700 South Cass Ave. Argonne, Il., 60439
E-mail {kestel-at-anl.gov} FAX: (630) 252-4289
Alwyn Eades wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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by darkwing.uoregon.edu (8.9.3/8.9.3) with SMTP id NAA13960; Thu, 25 Mar 1999 13:51:51 -0800 (PST)
Alwyn writes ...
} The accurate control of the thickness of evaporated carbon films. } } ... } } The colors: } If carbon is evaporated onto gold, as the thickness of the carbon } increases, the color changes through the following sequence: } gold, orange, red, blue, grey. The change of color from red to } blue is particularly sharp and clear. The change of color from } red to blue occurs when the thickness of the carbon is } 24.0 nm +/- 0.5nm.
We also find the red-} blue transistion the most easily recognized and the most consistent, even for a multiple-user facility ... and altho it may be considered a bit thick, the transistion is sharp enough to use for EPMA without need for coating standards and unknowns at the same time. However, for the sake of clarification ... the Mineralogy reference would imply "blue" is 24nm ... are you claiming the red-} blue transistion (i.e., "purple") is 24nm?? We have been writing our technique up as 22(+/-1)nm.
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Lawrence Kordon Nikon, Inc. Columbia, MD nikon-at-jagunet.com
"Steven J. Fliesler" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I'm in the market for a good quality, but inexpensive, film scanner- } e.g., for scanning 35mm slides into my PC. I've heard Nikon makes a } good one (model LS2500), but it runs a hefty $2,500 or so (too } expensive for me). Got any suggestions? } } -- } Steven J. Fliesler, Ph.D. } Professor } Dept. of Ophthalmology } Saint Louis Univ. School of Med. } 1755 S. Grand Blvd. } St. Louis, MO 63104-1540 } Phone: (314) 577-8259 } Fax: (314) 771-0596 } E-mail: Fliesler-at-slu.edu
Can anyone out there help us out with a current phone number for Dunaway - or with anyone else that might have replacement heaters for a diffusion pump?
Many thanks in advance
Gill Bond Department of Materials & Met. Eng. New Mexico Tech
Steve, It depends on the resolution you need. Microtek sells a scanner with a 35 mm film attachment (X6EL) for under $200. You can also get better ones for $600-1000 with a second scan bed designed for slides. If you only have a few slides, we have even taken slides apart and scanned them directly on a flat bed scanner with a transparency adapter.
Best regards,
Chris Baumann
"Steven J. Fliesler" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I'm in the market for a good quality, but inexpensive, film scanner- } e.g., for scanning 35mm slides into my PC. I've heard Nikon makes a } good one (model LS2500), but it runs a hefty $2,500 or so (too } expensive for me). Got any suggestions? } } -- } Steven J. Fliesler, Ph.D. } Professor } Dept. of Ophthalmology } Saint Louis Univ. School of Med. } 1755 S. Grand Blvd. } St. Louis, MO 63104-1540 } Phone: (314) 577-8259 } Fax: (314) 771-0596 } E-mail: Fliesler-at-slu.edu
Dear all, Does anyone have an old Hitachi H450 SEM lying around that they would like to donate or sell cheaply for parts? Eric Hines Microscopy Centre CSIRO Entomology Canberra.
Gil Bond wrote: =============================================== Can anyone out there help us out with a current phone number for Dunaway - or with anyone else that might have replacement heaters for a diffusion pump ? =============================================== The information for Duniway is the following: Duniway Stockroom Corporation 1305 Space Park Way Mountain View, California USA 94043 Toll Free: 800-446-8811 Phone: 650-969-8811 Fax: 650-965-0764 E-MAIL: info-at-duniway.com WEB: www.duniway.com
If they don't have it, one place that seems to always have the odd-ball item others don't have is the following:
TORR International, Inc. 12 Columbus Street New Windsor, NY USA 12553 Ph: 1-914-565-4027 Fax:1-914-561-7731 E-mail: torr.intl-at-juno.com WEB: www.torr.com Ask for Dr. Masud Naraghi
We have no interest in either of these firms, however we have done some amount of business as a satisfied customer with both.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Glenn Poirier and Bernard Kestel (both messages appended)=20 advocate very accurate means of determining C film=20 thickness. No doubt these means have some applications.=20 However, for most applications it is rather more convenient=20 to determine thickness at the time of coating with fairly=20 good accuracy. It is no use to an analyst to break the vacuum to determine=20 that another 3nm of carbon are required. For these reason=20 the polished brass slide method and the still more=20 convenient, auto-terminating thickness monitors are the=20 preferred means to determine coating thickness. Incidentally, for WDS/EDS I used indigo red - 20nm; its=20 enough C to prevent charging on flat specimens and absorbs=20 fewer light X-rays. Cheers Jim Darley ProSciTech Microscopy=20 PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Friday, March 26, 1999 4:28 AM, Glenn Poirier=20 [SMTP:glennp-at-eps.mcgill.ca] wrote: } =20 ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } =20 ---------------------------------------------------------- } -------------. } } } If you have access to an AFM, accurate measurement of C } thickness takes } about 4 minutes. } If I'm looking at flat samples, I use a toothpick to } remove a thin line of } carbon. I then can bring the sample to the AFM and } directly measure the } height of the step betwwen the sample and the top of the } carbon film. } Alternatively, if I can't work directly on the sample I } coat a plain glass } slide (making sure it is the same distance from the arc=20 as } the sample) and } measure the C thinckness on it. } Hope this helps } } Glenn } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} } Glenn Poirier Tel: (514) 398 6774 } MicroAnalysis Laboratrory Fax: (514) 398 4680 } Earth and Planetary Sciences email: glennp-at-eps.mcgill.ca } Rm. 238, 3450 University St.=09 } http://castaing.eps.mcgill.ca } Montr=E9al, Qc } H3A 2A7 } Millennium hand and shrimp } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}
When great accuracy of film thickness is not needed, we=20 simply place a small metal washer upon a glass slide near=20 the "specimen area". after evaporation the step height between the shaded and=20 coated areas is measured optically on a bench interference=20 microscope. Ours is a Zeiss two beam unit that is good for=20 30 nanometer resolution, (+ or -), with no physical contact=20 with the film. It works for ANY reflective metallic film=20 and fairly well even on "dull" carbon coatings because it=20 has three different reference mirrors that can be quickly=20 changed. Each mirror has a different reflectivity that one=20 merely tests to get reasonable interference fringes which=20 can also be photographed via polaroid or 35 mm. film. Bernie Kestel Materials Science Division Argonne National Laboratory
John, you are possibly right; a similar thought did occur to me before sending it to the list (what if it's bogus and is really someone trying to get marketing, company information, or even a non-cancer patient who is building up a collection for himself etc.) and I questioned my friend as to how reliable her information was. Of course she could not guarantee that this wasn't a hoak but in the absence of any evidence to the contary I was prepared to take this more or less at face value and I thought well what if it is true? It's not much of an effort to help someone out and nothing has been lost in doing so even if it turns out not to be genuine.
Regards
Martin Roe Macaulay Land Use Research Institute Aberdeen Scotland AB15 8QH
John Mansfield wrote: } } I hate to pour cold water on this, but it looks like another of } } those net myths.
Steven writes ... } } } I'm in the market for a good quality, but inexpensive, film scanner- } e.g., for scanning 35mm slides into my PC. I've heard Nikon makes a } good one (model LS2500), but it runs a hefty $2,500 or so (too } expensive for me). Got any suggestions? } } ...
I believe you mean the LS-2000 ... and it can be found for less $$. For example, http://www.pricewatch.com will show you many places for which it can be purchased for near $1600. There are lesser expensive scanners and I've seen a comparison of the Nikon with the new HP Photosmart scanner with will only leave you with why would you pay so much more for the Nikon. Still, the Nikon comes with "dust removal" software which works very well, and you also have an option for 4X and 16X multiple scans for removing LED noise in the dense areas of the slide or negative. Both the Nikon and new HP will deliver 48bit files to image editors which can handle that color depth (e.g., Photoshop).
I agree with Jim Darley when he says=20 =93However, for most applications it is rather more convenient=20 to determine thickness at the time of coating with fairly=20 good accuracy.=94 What I find hard to understand is the assumption he and other contributor= s to this thread have made that the interference color method allows you to evaporate carbon of only one thickness. The brass (or in the method I gave, gold) test sample has always the same thickness of carbon but the thickness of carbon on the sample can be what you like. The idea is that the brass/gold test piece is placed at a distance from the source which i= s different from the distance between the source and the sample to be coate= d. Then the thickness on the sample can be calculated from the inverse squa= re law. When I first used this technique, I used it to apply a coating only= 1 nm thick.
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Dear Steven, I have the top model Hewlett-Packard flat bed scanner and that comes with a 35 mm slide scanning attachment that works very well. Cost ~ $500. You wrote:
} } I'm in the market for a good quality, but inexpensive, film scanner- } e.g., for scanning 35mm slides into my PC. I've heard Nikon makes a } good one (model LS2500), but it runs a hefty $2,500 or so (too } expensive for me). Got any suggestions? } } -- } Steven J. Fliesler, Ph.D. } Professor } Dept. of Ophthalmology } Saint Louis Univ. School of Med.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Ph.D. graduate student or post-doctoral student (D.V.M. or M.D.) position available for research in the area of protozoal parasitology. Research will involve electron microscopy and cellular biology studies utilizing in vitro derived Sarcocystis spp. , Neospora spp. and Toxoplasma gondii parasites. Individual will work with a multi-disciplinary team at the University of Missouri-Columbia, in the Department of Veterinary Pathobiology and the Molecular Biology Program Electron Microscopy Core facility. Successful candidate should have background in tissue/cell embedding and processing for light and electron microscopy, and demonstrated proficiency in written and spoken English. Molecular biology and cell culture skills are helpful, but not a necessary requirement. U.S. citizenship is not required. For further information contact Dr. A.E. Marsh at ph: 573-884-2673, fax: 573-884-5414, or email: marshae-at-missouri.edu {mailto:marshae-at-missouri.edu} .
A colleage at our university needs servicing on an ISI Alpha 9 SEM. The problem is related to the manually valved vacuum system. The hand-turned knob for valving does not appear to be actually moving the vacuum cylinder (plunger) up and down. Either it has come off of the main shaft or the vacuum cylinder (plunger) is jammed.
Does anyone service these instruments commercially? Any directions available for opening the vacuum system (for example, to change the diffusion pump oil)? Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
My name is Randy Anderson and I am a senior at Northern Il University. I am currently finishing the second semester of a TEM/SEM independent study. The reason that I am writing is to find out how I can apply for an E.M. certification and to see how the best way to look for a job position in the E.M. field when I graduate in the summer of 99. Thank you for taking the time to answer my questions as the information will be a valuable assest.
I've put copies of my Reichert Zetopan manuals on http://users.skynet.be/sky95421/
Availlable manuals ? Zetopan.zip Instruction manual for the Zetopan.Large research Microscope. In Englisch= . 1 298 kb. Contains one page from an older Osram-catalogue regarding much used bulbs for older European microscopes.
? Zetopol.zip Instruction manual for the Zetopan-Pol .Large Polarization Microscope. In French. 487 kb.
? reflight.zip Instruction manual for the Universal polarization illuminator for opaque objects. In French. 179 kb.
? micflash.zip Instruction manual for the Universal microflasch equipment for Zetopan. I= n German. 523 kb.
? MS140.zip Instruction manual for the MS 1.40 multisystem condenser. In German. 319 = kb.
? Binolux.zip Instruction manual for the Binolux III twin lamp unit. In Englisch. 1 063 kb.
uri-at-watson.ibm.com has devised a procedure to treat these files when you = are using Unix (thanks Uri=85)
Nigel Browning schrieb: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } {bold} "Analyzing Materials Interfaces at Atomic Resolution" {/bold} } } } } There will be a Materials Science symposium at Scanning 99 entitled } "Analyzing Materials Interfaces at Atomic Resolution". Scanning 99 is } being held at the Hyatt Regency O'Hare in Rosemont, IL from April 11-14 } 1999. The "Analyzing Materials at Atomic Resolution" symposium is } scheduled for Tuesday April 13th. The details of the conference can be } found at http://www.scanning.org or can be requested from Mary } Sullivan (e-mail: scanning-at-fams.org, tel:201-818-1010). Registration } for members of the Midwest Microscopy and Microanalysis Society is at } the reduced rates of $150 (regular $ 235) for the whole conference or } $50 (regular $ 95) for a single day (all attendees of the MMMS } symposium held at UIC last May are members of MMMS). } } } } } {bold} Speakers {/bold} } } } } 9.00 {bold} M. Haider-CEOS GmbH, Germany {/bold} } } "Towards sub-Angstrom resolution by correction of spherical } aberration" } } } 9.30 {bold} O. Krivanek-University of Washington {/bold} } } "Towards sub-Angstrom electron probes by Cs-corrected STEM." } } } } 10.00 Break } } } } 10.30 {bold} E. M. James-University of Illinois at Chicago {/bold} } } "Atomic resolution scanning transmission electron microscopy on the } 200kV FEGTEM" } } } 11.00 {bold} S. J. Pennycook-Oak Ridge National Lab {/bold} } } "Probing the Origin of Interfacial Properties by STEM" } } } 11.30 {bold} D. A. Muller-Lucent Technologies {/bold} } } "The End of the Roadmap for Silicon Dioxide: The Electronic Structure } of Hyper-Thin Gate } } Oxides at the Atomic Scale" } } } 12.00 {bold} D. B. Williams-Lehigh University {/bold} } } "Atomic-Resolution X-ray Microanalysis in the AEM" } } } } 12.30 Lunch } } } } 2.00 {bold} L. D. Marks-Northwestern University {/bold} } } "Picometer structure determination using Electron Diffraction" } } } 2.30 {bold} J. M. Gibson -University of Illinois at } Urbana-Champaign {/bold} } } "Statistical Measurement of Electron Scattering Fluctuations in } Amorphous } } Materials - A new Structural Tool" } } } } 3.00 Break } } } } 3.30 {bold} M. Gajdardziska-Josifovska-University of Wisconsin at } Milwaukee {/bold} } } "Quantitative surface microscopy and diffraction over the length } scales: Morphology and } } crystallography of polar oxide surfaces. " } } } 4.00 {bold} M. Tanaka-NRIM, Tsukuba, Japan {/bold} } } "Nano-behavior of Small Metal Particles in the Electron Beam" } } } } } } } } } } } } ___________________________________________________________________________ } } } Nigel D. Browning, PhD } } Assistant Professor } } University of Illinois at Chicago } } Department of Physics } } 845 West Taylor Street, } } Chicago } } IL 60607-7059. USA } } } Tel: 312-413-8164 } } Fax: 312-996-9016 } } } } http://interface.phy.uic.edu } } } ___________________________________________________________________________ } } }
I work on virtually everything in EM but have not worked on that model. Is that one of ISI's integrated vacuum valves, i.e. a hand operated valve with a number of individual positions for backing, roughing, etc. (usually just marked 0, 1, 2)? Or is it an individual valve with a single function?
I suspect that it is one of their integrated models, similar to manual and electrically operated models common to many of their instruments. They do come completely apart and have a very simple mechanism. There is probably a spring or taper pin that connects the hand turned portion to the central shaft. In some models, the shaft is visible from the top. If the shaft is not turning with the knurled hand piece then that pin has sheared off. While the pin can be easily driven out and replaced, this would also be a good time to rebuild the valve as there might be a problem inside that caused the pin to shear (these pins are often used a 'fuses' to prevent excessive force being applied to a mechanism).
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com
-----Original Message----- } From: John J. Bozzola {bozzola-at-siu.edu} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
Linda asks ... } } Dear listservers: } I would like to know which brands/models of dye sub printers do people } find give the best quality images. Thanks. Linda Chicoine }
A timely question ... 6-10 months ago photographic quality and trouble-free dye-subs were $5k-$10k printers ... plus $3/page. While you can buy a dye-sub today for { $700, it is still very much a case of how well the printer's software works with your application and OS and the quality of its color profile. I believe the consensus at one time was Kodak and Fujitsu for photographic and color quality. Today Alps should be considered in any decision because it has broken the price barrier to allow it to be considered along with color ink jets. I've been keeping an eye on lists and newsgroups without a good feeling for how well Alps supports its printers (... tech support, and ICC profiles ..) ... but the demo print they sent me was definitely excellent quality. If I were you I'd indicate which computer platform you use, and with what applications ... someone may reply with specific problems associated with less expensive dye-subs.
We have a M=E4rzh=E4user MRC-3 which is used to control stage movement and focusing by a joystick. The equipment is capable of being controlled from a computer but due to the need to write Basic instructions most users are put off this option. A similar problem exists using NIH Image macros. All very sad for the stereologists who would like to be able to have random sampling, montages, etc. The main problem is we use a Power Macintosh for the image processing side of things so would like to find a stage control program compatable with a Mac. At this point things grind to a halt, has anyone any knowledge of a Mac compatable stage control program with a user friendly interface?
Thanks
Andrew McNaughton
____________________________________________________________________________= __ Andrew McNaughton South Campus Electron Microscope Unit C/-Department of Anatomy and Structural Biology School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
We have great luck with the Codonics NP1600. However, Ii have recently talked with a Tektronix rep who tells me that they are geting out of the dye sub market and focusing on color laser and solid ink technologies. According to him, with the current color laser printers on the market, you can approach dye sub quality for between 2 and 3 k but the cost per sheet is } $.30 where a Codonics print costs $1.90.
Regards,
Chris Baumann
Linda Chicoine wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear listservers: } I would like to know which brands/models of dye sub printers do people } find give the best quality images. Thanks. Linda Chicoine
You might consider the Fuji Pictrography. We have one here and it delivers very high quality prints. I believe the price is competitive with dye-subs.
Mohan Kalyanarman
Materials Characterization Catalyst Technology Group Mobil Technology Company PO Box 480 Paulsboro, NJ 08066 609-224-3989 (ph) 609-224-3608 (fax) mohan_kalyanaraman-at-email.mobil.com
My company's R&D group is interested in visualizing several of our = polymers. The M&M '99 session, "Developments in Scanned Probe = Microscopy of Polymers", will be helpful. However, we would like some = basic/general information prior to the meeting. Any suggestions of = review articles, textbooks, etc. would be greatly appreciated.
In advance, thanks!
Janet H. Woodward, Ph.D. Corporate Technical Specialist - Microscopy & Microbiology Buckman Laboratories 1256 N. McLean Street Memphis, TN 38108 (901) 272-6408 jhwoodward-at-bbuckman.com
My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly) PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration: Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr
6. Gap Juctions in Drosophila PBS = phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} DW = distilled water wash buffer = 0.5 M NaCl and 0.05% Tween-20 in PBS
Fixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C) antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. “Differential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.” Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. “Gap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.” Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. “Gap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.” Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. “Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.” Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. “The 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).” Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. “Electron Microscope Immunolocation of Gap Junctions in Drosophila.” Tissue & Cell 1989;21(6): p 835-39.
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I am thinking of purchasing a microwave tissue processor. I would appreciate your opinion as to the pro's and con's of this type of tissue preparation.
Laura Robles
Hi Laura. I have been using my Pelco microwave for over a year now and I am very happy with it. I can process tissue in 2 hours instead of 2 days. It is especially useful for dehydration and infiltration. I would highly recommend getting one. It will pay for itself in the amount of time saved very quickly. The only con is that you have to experiment to find the best parameters for your tissue since there are few published protocols. Try it!
Does anyone have tips for dimpling soft metal alloys for TEM sample prep? I'm having difficulty dimpling an aluminium composite alloy. It appears that the abrasive (2-4 um cubic BN slurry) is getting embedded into the surface of the alloy. This then attacks the dimpling wheel (phosphor bronze).
Thanks, Wharton
++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
Central Michigan University Electron Microscope Facility Supervisor Biology Department
Central Michigan University is looking for an electron microscopy technician to supervise teaching/research EM facility. Assists in teaching introductory TEM and SEM classes. Solicits externally funded contracts. Required qualifications include education equivalent to Bachelor's degree; two years qualifying work experience; working knowledge of the principles and techniques of transmission and scanning electron microscopy, ultramicrotomy, vacuum evaporation, critical point drying and biological specimen preparation; general knowledge of solid-state electronics; proficiency with TERM, SEM, EDS, digital imaging and light microscopy. Experience with routine maintenance of EM and optical microscopes desired.
Review of applications will begin on May 1, 1999, and will continue until the position is filled. Please send resume materials to Central Michigan University, Human Resources/Staff, Rowe 109, Mt. Pleasant, MI 48859. CMU provides flexible benefits, an excellent retirement program with tax deferred investment options, tuition waiver for employee and family, and competitive salaries in an environment committed to excellence and customer service.
-- Amy B. Gambrell Phone: 517.774.3339 Human Resources/Staff Fax: 517.774.3256 109 Rowe Hall E-mail: Amy.Beth.Gambrell-at-cmich.edu Central Michigan University http://www.sps.cmich.edu Mt. Pleasant, MI 48859
To find out about exciting job opportunities at CMU, click here: http://www.sps.cmich.edu/jobs.html
My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly) PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration: Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr
6. Gap Juctions in Drosophila PBS = phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} DW = distilled water wash buffer = 0.5 M NaCl and 0.05% Tween-20 in PBS
Fixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C) antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. “Differential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.” Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. “Gap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.” Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. “Gap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.” Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. “Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.” Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. “The 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).” Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. “Electron Microscope Immunolocation of Gap Junctions in Drosophila.” Tissue & Cell 1989;21(6): p 835-39.
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My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly) PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration: Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr
6. Gap Juctions in Drosophila PBS = phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} DW = distilled water wash buffer = 0.5 M NaCl and 0.05% Tween-20 in PBS
Fixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C) antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. “Differential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.” Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. “Gap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.” Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. “Gap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.” Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. “Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.” Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. “The 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).” Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. “Electron Microscope Immunolocation of Gap Junctions in Drosophila.” Tissue & Cell 1989;21(6): p 835-39.
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Hey people, Anyone have any experience with the new RMC crx cryounit that can attach to a Riechert E? It is supposedly much cheaper than Leica's but is it as good? Does anyone have any general experience with RMC. Thanks.
I am interested in finding out if there is any facility that has an ESEM Tensile Stage with Peltier capabilities for structure/function studies using ESEM.
SPM of Polymers is one of the hottest new trends in microscopy. I am in the middle of writing a review of "What's New in Microscopy at PITTCON" (May, 1999, American Lab) and this topic was well represented by new offerings from several companies. I'd suggest you visit the websites for
1. Digital Instruments - they have lots of great app notes on this topic
2. ThermoMicroscopes (previously known as Park Scientific and Topometrix) and look for information on the new Explorer PolymerSystem SPM (integrated microthermal analysis and pulsed force mode imaging)
3. JEOL - their new SPM has a full hot/cold stage which runs from 130K to 800K, for watching thermal transitions.
Other relevant technology I found at Pittcon included the new thermal stage in Philips' ESEM, the new Continuum microscope for full microscopy imaging (Including DIC) integrated with FTIR from SpectraTech, and a number of interesting systems for Raman/confocal from Renishaw and Instruments SA.
Hope this "preview" was helpful.
CAVEAT: Neither Barbara Foster nor MME has any commercial interest in any of these instruments.
Best regards,
Barbara Foster
Consortium President
Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118
MME is America's first national consortium providing
customized on-site workshops in all areas of
microscopy, sample preparation, and image analysis.
At 10:29 AM 3/29/99 -0600, Janet H. Woodward wrote:
} } } }
{excerpt} My company's R&D group is interested in visualizing several of our polymers. The M&M '99 session, "Developments in Scanned Probe Microscopy of Polymers", will be helpful. However, we would like some basic/general information prior to the meeting. Any suggestions of review articles, textbooks, etc. would be greatly appreciated.
My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Fixation: 1% paraformaldehyde-0.1M PB Sectioning: 30 mm Rinsing: PBS (24 hr) Blocking: BSA-PBS (30 min) 1st Incubation:(4 C) antibody+[BSA-PBS] {1:1000} (12 hr) Washing: (4 C) PBS (12 hr) 2nd Incubation: (4 C) Au conj. anti-mouse IgG+[BSA-PBS] {1:20} (12 hr) Post Fixation:=09 1% osmium tetroxide-0.1 M PB (1 hr) Dehydration:=09 Embedding: Epon 812 Ultra-thin Sectioning:=09 80 nm Mounting: mesh grids Total Time: 61.5 hr =09 2. Human Smooth Muscle =09 PB =3D phosphate buffer Au conj. =3D gold conjugated (post-embedding) {1993} =09 Fixation: 0.5% glutaraldehyde- 0.175 M cacodylate buffer Dehydraton:=09 Embedding: (4 C) LR White Polymerization:=09 cold+UV light (?? hr) Ultra-thin Sectioning:=09 parallell to plane of c-slip Mounting: formvar, 200 Ni mesh grids Blocking: 0.01 M glycine+1% NFDM+0.5% gelatin+0.1 M PB (?? hr) 1st Incubation: (R.T.) rabbit anti-Cx43 serum+[NFDM+gelatin+PB] {1:750} (1 hr) Washing: (entensive) PB 2nd Incubation: (R.T.) Au conj. goat anti-rabbit IgG+[NFDM+gelatin+PB] {1:10} (1hr) Washing: =09 Fixation: (brief) 2% glutaraldehyde+PB Rinsing: water Drying: air Staining: uranyl acetate and Reynold's lead citrate Total Time: ?? hr =09 3. Stratum Granulosum Cells of Mouse Follicles=09 PB =3D phosphate buffer PBS =3D phosphate buffered saline Au conj. =3D gold conjugated (pre-embedding) {1993} =09 Fixation: 1% paraformaldehyde- 0.1 M PB (2-4 hr) Sectioning: 80 mm Rinsing: PB Blocking: PBS containing 1% BSA & 0.05% sapoinin (30 min) 1st Incubation: (4 C)=09 anti-Cx43 rabbit serum+[PBS containing 1% BSA & 0.05% sapoinin] {1:200} (28-40 hr) Washing: PBS (5 hr) 2nd Incubation: 5 nm Au conj. Antibody+[PBS containing 1% BSA & 0.05% sapoinin] {1:3} (22-24 hr) Washing: PBS (4-6 hr) Fixation: sodium cacodylate buffer (pH 7.4, 0.1 M) + 2.5% glutaraldehyde (1.5 hr) Post Fixation:=09 1% osmium tetroxide (1.5 hr) Dehydration: ethanol & propylene monoxide Embedding: Epon 812 Ultra-thing Sectioning:=09 Mounting: formvar-film coated single-slot grids Staining: uranyl acetate and lead citrate solutions Total Time: 64.5 - 82.5 hr =09 4. Rat Heart Gap Junction Membranes=09 PBS =3D phosphate buffered saline (pre-embedding) {1990}
1st Incubation: (R.T.)=09 poly-L-lysine coated 96-well plates (unbouding of membranes??) (14 hr) Blocking: PBS containing 5% BSA (1 hr) 2nd Incubation: 25-to-100 fold dilution of preimmune or immune serum (1 hr) ---or--- 2nd Incubation:=09 80-115 mg ml-1 affinity purified antibodies Washing: PBS 3rd Incubation:=09 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10} (1 hr) Washing: PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration:=09 Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 15+ hr =09 5. Heart Gap Junctions=09 PBS =3D phosphate buffered saline (pre-embedding) {1989}
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: =095 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly)=09PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration:=09 Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr =09 6. Gap Juctions in Drosophila=09 PBS =3D phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} =09 DW =3D distilled water wash buffer =3D 0.5 M NaCl and 0.05% Tween-20 in PBS
Fixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C)=09 antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. =93Differential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.=94 Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. =93Gap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.=94 Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. =93Gap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.=94 Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. =93Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.=94 Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. =93The 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).=94 Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. =93Electron Microscope Immunolocation of Gap Junctions in Drosophila.=94 Tissue & Cell 1989;21(6): p 835-39.
My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Fixation: 1% paraformaldehyde-0.1M PB Sectioning: 30 mm Rinsing: PBS (24 hr) Blocking: BSA-PBS (30 min) 1st Incubation:(4 C) antibody+[BSA-PBS] {1:1000} (12 hr) Washing: (4 C) PBS (12 hr) 2nd Incubation: (4 C) Au conj. anti-mouse IgG+[BSA-PBS] {1:20} (12 hr) Post Fixation:=09 1% osmium tetroxide-0.1 M PB (1 hr) Dehydration:=09 Embedding: Epon 812 Ultra-thin Sectioning:=09 80 nm Mounting: mesh grids Total Time: 61.5 hr =09 2. Human Smooth Muscle =09 PB =3D phosphate buffer Au conj. =3D gold conjugated (post-embedding) {1993} =09 Fixation: 0.5% glutaraldehyde- 0.175 M cacodylate buffer Dehydraton:=09 Embedding: (4 C) LR White Polymerization:=09 cold+UV light (?? hr) Ultra-thin Sectioning:=09 parallell to plane of c-slip Mounting: formvar, 200 Ni mesh grids Blocking: 0.01 M glycine+1% NFDM+0.5% gelatin+0.1 M PB (?? hr) 1st Incubation: (R.T.) rabbit anti-Cx43 serum+[NFDM+gelatin+PB] {1:750} (1 hr) Washing: (entensive) PB 2nd Incubation: (R.T.) Au conj. goat anti-rabbit IgG+[NFDM+gelatin+PB] {1:10} (1hr) Washing: =09 Fixation: (brief) 2% glutaraldehyde+PB Rinsing: water Drying: air Staining: uranyl acetate and Reynold's lead citrate Total Time: ?? hr =09 3. Stratum Granulosum Cells of Mouse Follicles=09 PB =3D phosphate buffer PBS =3D phosphate buffered saline Au conj. =3D gold conjugated (pre-embedding) {1993} =09 Fixation: 1% paraformaldehyde- 0.1 M PB (2-4 hr) Sectioning: 80 mm Rinsing: PB Blocking: PBS containing 1% BSA & 0.05% sapoinin (30 min) 1st Incubation: (4 C)=09 anti-Cx43 rabbit serum+[PBS containing 1% BSA & 0.05% sapoinin] {1:200} (28-40 hr) Washing: PBS (5 hr) 2nd Incubation: 5 nm Au conj. Antibody+[PBS containing 1% BSA & 0.05% sapoinin] {1:3} (22-24 hr) Washing: PBS (4-6 hr) Fixation: sodium cacodylate buffer (pH 7.4, 0.1 M) + 2.5% glutaraldehyde (1.5 hr) Post Fixation:=09 1% osmium tetroxide (1.5 hr) Dehydration: ethanol & propylene monoxide Embedding: Epon 812 Ultra-thing Sectioning:=09 Mounting: formvar-film coated single-slot grids Staining: uranyl acetate and lead citrate solutions Total Time: 64.5 - 82.5 hr =09 4. Rat Heart Gap Junction Membranes=09 PBS =3D phosphate buffered saline (pre-embedding) {1990}
1st Incubation: (R.T.)=09 poly-L-lysine coated 96-well plates (unbouding of membranes??) (14 hr) Blocking: PBS containing 5% BSA (1 hr) 2nd Incubation: 25-to-100 fold dilution of preimmune or immune serum (1 hr) ---or--- 2nd Incubation:=09 80-115 mg ml-1 affinity purified antibodies Washing: PBS 3rd Incubation:=09 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10} (1 hr) Washing: PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration:=09 Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 15+ hr =09 5. Heart Gap Junctions=09 PBS =3D phosphate buffered saline (pre-embedding) {1989}
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: =095 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly)=09PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration:=09 Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr =09 6. Gap Juctions in Drosophila=09 PBS =3D phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} =09 DW =3D distilled water wash buffer =3D 0.5 M NaCl and 0.05% Tween-20 in PBS
Fixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C)=09 antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. =93Differential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.=94 Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. =93Gap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.=94 Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. =93Gap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.=94 Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. =93Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.=94 Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. =93The 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).=94 Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. =93Electron Microscope Immunolocation of Gap Junctions in Drosophila.=94 Tissue & Cell 1989;21(6): p 835-39.
My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly) PBS Fixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration: Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr
6. Gap Juctions in Drosophila PBS = phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} DW = distilled water wash buffer = 0.5 M NaCl and 0.05% Tween-20 in PBS
Fixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C) antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. “Differential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.” Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. “Gap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.” Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. “Gap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.” Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. “Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.” Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. “The 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).” Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. “Electron Microscope Immunolocation of Gap Junctions in Drosophila.” Tissue & Cell 1989;21(6): p 835-39.
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We are a non-profit botanical garden conducting plant anatomy research. We are looking for donations or second hand low-cost microscopy equiptment. We can pay for the shipping of the following items:
Compound research/student style microscope
We are looking for a Riechert Sliding Microtome & knives from anyone/institution wishing to dispose of one.
We are also looking for non-disposible blades for a Spencer 820 rotary microtome
& just the INSTRUCTIONS for a Sensaur microtome knife sharpener: Aloe - Cat # V60041; Model # SH 67-R; Serial # 1107; 115 Vac 50/60 C 5A
Please reply to my email: crow-at-aloha.net Thanks very much in advance
"The light which puts out our eyes is darkness to us. Only that day dawns to which we are awake. There is more day to dawn. The sun is but a morning star."
Try one, two, three or all of the following: 1. Use a stainless steel wheel instead of a bronze wheel; 2. Decrease the balance load; 3. Change slurry (size, abrasive particles etc. etc....) 4. If I were you, I would also completely skip the dimpling procedure and try a Grind Holder (such as Gatan's) or Tri-pod (such as south Bay Tech's) and put the specimen on ion-mill right after obtaining {20um thick foil.
Hope the above helps.
-cy, Rodelee-to-be
On Mon, 29 Mar 1999, Wharton Sinkler wrote:
} Does anyone have tips for dimpling soft metal alloys for TEM sample prep? } I'm having difficulty dimpling an aluminium composite alloy. It appears } that the abrasive (2-4 um cubic BN slurry) is getting embedded into the } surface of the alloy. This then attacks the dimpling wheel (phosphor } bronze). } } Thanks, } Wharton } } ++++++++++++++++++++++++++++++++++++++++++++++++++ } Wharton Sinkler } Argonne National Laboratory West } P. O. Box 2528 } Idaho Falls, ID 83403 } Tel: (208) 533-7724 } FAX: (208) 533-7863 } } mailto:wharton.sinkler-at-anlw.anl.gov } }
Politeness would suggest that you enter something a little more informative in your subject field!
To just say "I AM NOT SPAM" makes you look suspiciously like spam, but also somehow manages to suggest that you think that, compared to YOUR important message, others may be spam.
rtch
} Hello, } } My name is Aaron Rea. I am working on an undergraduate research project at } Southwest Missouri State University (Springfield, MO USA). I am starting work } with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying } to develop a specimen preparation protocol for the immunolocalization of Cx43 } within this cell line. I have outlined the prep protocols from 6 papers that } utilized immunoEM below. I am looking for any suggestions or recommendations } on which protocol might yield the greatest success. Any other thoughts or } ideas would be greatly appreciated. Thanks to all who responded back in } October when I began this project!! Please reply offlist at: } aaronrea-at-hotmail.com }
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Hello, could you please post my message below to the list with the title of the subject being, "HELP! ImmunoEM Prep for Cx43 Localization". Thank you for your time.
Sincerely, Aaron Rea 439 E. Madison #7 Springfield, MO 65806 aaronrea-at-hotmail.com ---------------- Hello,
My name is Aaron Rea. I am working on an undergraduate research project at Southwest Missouri State University (Springfield, MO USA). I am starting work with a BALB 3t3 mouse fibroblast cell line from ATCC (CCL 163). We are trying to develop a specimen preparation protocol for the immunolocalization of Cx43 within this cell line. I have outlined the prep protocols from 6 papers that utilized immunoEM below. I am looking for any suggestions or recommendations on which protocol might yield the greatest success. Any other thoughts or ideas would be greatly appreciated. Thanks to all who responded back in October when I began this project!! Please reply offlist at: aaronrea-at-hotmail.com
Blocking: 200 ml of 5% BSA in PBS (15 min) 1st Incubation: antiserum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) ---or--- 1st Incubation: preimmune serum + [5% BSA in PBS] {1:20, 1:50, or 1:100} (1.5 hr) Washing: PBS 2nd Incubation: 5 nm colloidal gold-labelled goat anti-rabbit IgG antibody+[5% BSA in PBS] {1:10, 1:20, or 1:40} (1 hr) Washing: (thoroughly) PBS =46ixation: 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) (?? hr) Post Fixation: 1% osmium tetroxide Staining: uranyl acetate Dehydration: Embedding: Epon Staining: lead citrate and uranyl acetate Total Time: 6+ hr
6. Gap Juctions in Drosophila PBS =3D phosphate buffered saline (150mM NaCl, 6mM Na2HPO4, 4mM KH2PO4) (post-embedding) {1989} DW =3D distilled water wash buffer =3D 0.5 M NaCl and 0.05% Tween-20 in PBS
=46ixation: (4 C) 2% paraformaldehyde/0.5% glutaraldehyde in PBS (3 hr) Dehydration: (4 C) 1 X 5 min in 50%, 70%, 95%, and 100% ethanol (20 min) Embedding: (-20 C) 100% ethanol + Lowicryl K4M resin {1:1} (16 hr), 2 X 3 hr in Lowicryl K4M resin in BEEM capsules (6hr) 1st Polymerization: (-20 C) UV light (16 hr) 2nd Polymerization: (R.T.) UV light (24 hr) Sectioning: 10 nm sections gathered on parlodion-coated gold grids Blocking: 3% ovalbumin in wash buffer (10 min) 1st Incubation: (4 C) antibody, preimmune serum, or non-immune IgG + 1% ovalbumin in wash buffer (16 hr) Washing: 5 X 5 min in wash buffer (25 min) Blocking: 3% ovalbumin in wash buffer (10 min) 2nd Incubation: 10 nm colloidal gold-labelled goat anti-rabbit IgG antibody + [1% ovalbumin in wash buffer] {1:25} (1 hr) Washing: 5 X 5 min in wash buffer (25 min) Washing: 1 X 5 min with DW (5 min) Staining: lead citrate and uranyl acetate Total Time: 83.5 hr ----------------------------------------------------------------------1. Muramatsu T, Hashimoto S, Shimono M. =ECDifferential Expression of Gap Junction Proteins Connexin32 and 43 in Rat Submandibular and Sublingual Glands.=EE Journal of Histochemistry and Cytochemistry 1996;44(1): p 49-56. 2. Campos de Carvalho AC, Roy C, Moreno AP, Melman A, Hertzberg EL, Christ GJ, Spray DC. =ECGap Junctions Formed of Connexin43 are Found Between Smooth Muscle Cells of Human Corpus Cavernosum.=EE Journal of Urology June 1993;149: p 1568-75. 3. Koike K, Watanabe H, Hiroi M, Tonosaki A. =ECGap Junction of Stratum Granulosum Cells of Mouse Follicles: Immunohistochemistry and Electron Microscopy.=EE Journal of Electron Microscopy 1993;42: p 94-106. 4. Laird DW, Revel JP. =ECBiochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes.=EE Journal of Cell Science 1990;97: p 109-17. 5. Yancey SB (I), John SA (II), Ratneshwar L (III), Austin BJ, Revel JP. =ECThe 43-kD Polypeptide of Heart Gap Junctions: Immunolocalization (I), Topology (II), and Functional Domains (III).=EE Journal of Cell Biology June 1989;108:p 2241-54. 6. Ryerse J. =ECElectron Microscope Immunolocation of Gap Junctions in Drosophila.=EE Tissue & Cell 1989;21(6): p 835-39.
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Barbara Foster says: } Uri, Let me see what I can dig out. I'm headed out on an extended } business trip... will be back around 3/15. Could you send me a reminder?
Barbara, since my attempts to contact you via e-mail failed so far, I decided to post the reminder here, hoping that it will reach you.
Do you (or anybody else on this list) have any Zetopan manuals, especially for Phase Contrast, Anoptral Contrast, transmitted light Interference Contrast, Fluorescence, Zetopan-POL? [Needless to say, I need those.]
Thank you. -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
} My company's R&D group is interested in visualizing several of our } polymers. The M&M '99 session, "Developments in Scanned Probe } Microscopy of Polymers", will be helpful. However, we would like some } basic/general information prior to the meeting. Any suggestions of } review articles, textbooks, etc. would be greatly appreciated.
Janet,
It strikes me that it is better not to limits oneself to SPM. This technique is powerful, but limited in its application. Etching and staining can often do better - it really depends which polymers you're lloking at. If you can get hold of "Atlas of Polymer Morphology" by Arthur E. Woodward (distributed in USA by Oxford University Press, New York, ISBN 0-19-520758-0) this would give you a lot of ideas.
If staining interests you, the following article is superb. However, the author is out of reprints, so it might be better to use some form of library loan, if you don't take the journal.
TI: Reflections on the use of microtomy for materials science specimen preparation AU: Plummer_HK NA: FORD MOTOR CO,RES LAB,MAIL DROP 3028,SRL,DEARBORN,MI,48121 JN: MICROSCOPY AND MICROANALYSIS, 1997, Vol.3, No.3, pp.239-260 IS: 1431-9276 DT: Review
If you are into polyethylene or polypropylene, and also some blends, etching might be useful. You can see a few pictures on my home page by following the link "Picture Gallery" and also on:
http://www.reading.ac.uk/~spshosir/gallery.htm
from a colleague. If you want to follow this further, please let me know.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
} Janet H. Woodward, Ph.D. Corporate Technical Specialist - Microscopy & } Microbiology Buckman Laboratories 1256 N. McLean Street Memphis, TN } 38108 (901) 272-6408 jhwoodward-at-bbuckman.com
I was having problems with my e-mail account yesterday and wasn't sure if I was getting my message through. I now have the problem figured out and this will not happen again. I am truly sorry for any inconvenience or annoyance this caused. A special thanks to those who responded with Cx43 protocol ideas!
Sincerely,
Aaron Rea Get Your Private, Free Email at http://www.hotmail.com
***************************************************** Electron Microscopy/Image Analysis Technician
An NIH-funded position is available immediately for an electron microscopist to study the molecular structure and function of muscle. Projects involve determination of the 3D molecular structure of actin and myosin filaments and the structural changes underlying contraction. Our main approaches involve electron microscopy (negative staining, cryo-EM) and 3D image analysis (helical and tomographic reconstruction) combined with biochemical, immunological and molecular biological methodology. Candidates should be motivated individuals with a good background in molecular level electron microscopy and/or image processing. Our laboratory is equipped with Philips CM120 cryo, CM10 and EM400 TEMs, Gatan cryoholder and TV system, ancillary EM equipment (freeze plungers, freeze fracture, evaporators, etc), optical diffractometer, scanning densitometer, Silicon Graphics image processing workstations, etc. Applicants should provide a CV and names and addresses of 3 references to Dr Roger Craig, Department of Cell Biology, University of Massachusetts Medical School, Worcester MA 01655. Tel: (508) 856 2474; Fax: (508) 856 6361; Email: Roger.Craig-at-ummed.edu. *****************************************************
Dear Wharton, I have successfully dimpled Al2O3-Al MMC (metal matrix composite) samples, using the VCR Dimpler. The dimpling abrasive is 3 um diamond paste diluted in water and the wheel is steel. In the notes, this problem of embedding the abrasive in a soft work piece is addressed and recommends using a softer abrasive or a diamond tool. You might contact VCR at: 650-875-1000 for advice. You wrote:
} } Does anyone have tips for dimpling soft metal alloys for TEM sample prep? } I'm having difficulty dimpling an aluminium composite alloy. It appears } that the abrasive (2-4 um cubic BN slurry) is getting embedded into the } surface of the alloy. This then attacks the dimpling wheel (phosphor } bronze). } } Thanks, } Wharton } } ++++++++++++++++++++++++++++++++++++++++++++++++++ } Wharton Sinkler } Argonne National Laboratory West
No financial interest, just a happy customer. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Barbara Foster says: } I have a general manual which outlines all the transmitted light functions. } Unfortunately, there is only a small amount on Anoptral Phase --- they } refer to a "separate manual" for that. There are also other accompanying } materials which discuss Fluorescence.
I am interested in the Fluorescence materials. The general manual - I got it, thanks to Yvan and Wayne.
} Also, Yvan Lindekens has posted his copy of the Zetopan manual at } {http://users.skynet.be/sky95421} . I am having trouble with my unzip files } so have not been able to open his manual to compare it to mine, but suggest } you see what is there.
I was probably the first who got Yvan's copy of the manual (:-). It's a pretty good one, in case anybody cares to know.
After you unzip it, you get one large TIFF file. So now you need software that (a) can deal with multi-page TIFF's, or (b) can convert TIFF to something the printers like better [such as PostScript]. See my comments on Yvan's Web site.
Yvan, please get in touch with me off-line - I need to figure out how to scan a few manuals myself, and put them on your Web site. -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
The Listserver system went bizzare today in synchronism with my attempts to check for viruses. As a result, a number of of mail messages were rejected.
I believe all is well again. Those of you that were rebuffed please accept my apology.
My name is Birna Gu=F0bj=F6rnsd=F3ttir working at Icelandic Fisheries Laboratories as a food scientist. This field is quite new to me and I am looking for information how I can prepare wood samples, both wet nad dry, for SEM. Any information would be greatly appreciated.
My name is Birna Gu=F0bj=F6rnsd=F3ttir working at Icelandic Fisheries Laboratories as a food scientist. This field is quite new to me and I am looking for information how I can prepare wood samples, both wet nad dry, for SEM. Any information would be greatly appreciated.
Birna wrote: } My name is Birna Gudbjoernsdottir working at Icelandic Fisheries } Laboratories as a food scientist. This field is quite new to me and I am } looking for information how I can prepare wood samples, both wet nad dry, } for SEM. Any information would be greatly appreciated.
Dear Birna,
If you wish to keep the wood wet, you will have to use an environmental SEM (ESEM), in which you can examine the sample with a pressure above 5 torr (the equilibrium pressure of water vapour just above 0 degree C) in the specimen chamber. If you can accept that the wood slowly dries, you may use a low-vacuum SEM (LVSEM) where you can work with a pressure of up to 1-2 torr. Both ESEM and LVSEM allows the examination of electrical insulators.
In the conventional high vacuum SEM you cannot have wet wood. Dry wood may be examined, but then the surface must be made conductive e.g. by sputtering gold onto the sample.
Best regards, Joergen.
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
I have been using Syquest 230 EZ Flier dirves on my TDL12 8000/Delta system for some time. Other than the reliability problem of the Syquest hardware and the posibility of little future support from Syquest, they work well and are rather faster than the bernoulliis. A "sort-of" special cable is needed to interconnect the drives. The interface instructions simply address the setup of DMON (nowdays that would be called a driver, I guess). The disks are formatted rt11 and instead of 230MB, hold 44Mb, but it was an overall improvement. I would guess that similar, more current external SCSI-1 drives could also be made to work.
First, thanks to everyone who responded to my question on fixation of cells grown on chambered Permanox slides. As soon as I try the suggested procedures (the cells crashed!), I will post a summary of responses to the list.
My question today has to do with the presence of "pepper" on TEM sections. We process rather large tissue blocks (4X5 mm) of nervous tissue that we have dissected from perfused animals (5% glut. in PB is routine perfusate). Post-fixation is in osmium tetroxide in PB and all buffer rinses are done in PB. We dehydrate using an ethanol series, transition through propylene oxide (although we are experimenting with ethanol with no succes so far) and infiltrate/embed in PolyBed 812. The pepper shows up most obviously on the cytoskeletal elements of nerves, mitochondria, and collagen. I have tried pretreating the sections with 0.5-1.0 N HCl or 1% EDTA (Mollenhauer, 1987)prior to staining with uranyl acetate/lead citrate.....no luck. I can visualize the pepper on unstained sections. I seem to recall that this may have something to do with phosphate buffer, but the details are hazy. I would appreciate any suggestions.
Birna Guðbjörnsdóttir wrote: =================================================== My name is Birna Guðbjörnsdóttir working at Icelandic Fisheries Laboratories as a food scientist. This field is quite new to me and I am looking for information how I can prepare wood samples, both wet nad dry, for SEM. Any information would be greatly appreciated. =================================================== Assuming you would have just a conventional SEM at your disposal, we have generally had good results in our own laboratory, when the need has arisen in the past, to examine wood, to dry it with critical point drying. A second (but we believe inferior) approach is to use the HMDS drying protocol , which tends to be favored by those without access to a CPD unit. Whatever the drying method, since the sample itself will be nonconductive, it should be made conductive via the application of a thin layer of gold, or for higher resolution work, chromium or (more recently) osmium (metal).
Information about CPD and HMDS and also metal coaters can be found on our website given below or the websites of the other major manufacturers and suppliers of sample preparation equipment and consumable supply items for SEM laboratories.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Does anyone involved in Electron Microscopy know where I can obtain = spare glass scoring wheels for an LKB 2178 Knife Maker II. Any helpful = advise would be welcome as our machine requires a replacement scorer = ASAP.
Dr Terry Robertson (PhD) Senior Research Fellow Department of Pathology University of Western Australia Nedlands 6907
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Hello,
I am interested in finding out if there is any facility that has an ESEM Tensile Stage with Peltier capabilities for structure/function studies using ESEM.
by majestic.tcs.tulane.edu (8.9.3/8.9.3) with SMTP id PAA21132; Wed, 31 Mar 1999 15:13:53 -0600 (CST) Message-Id: {199903312113.PAA21132-at-majestic.tcs.tulane.edu}
Dear Colleagues: Sometime ago I posted a request for information on microscopy fees. I received several wonderful information packages and thank those who contributed. Before submitting a proposal internally at Tulane, I would like to build a table with a comparison among several institutions. Thus, I need information on fees charged as shown in the sample below. Additional information on structure and function of morphology cores and microscopy coresis welcome as well. You do not have to be specific, and I will not disclose your name or associate it with the information outside. I will only use your name inside Tulane if you authorize it. Thus far not a single institution can recoup costs from fees. Thus, what I need to demonstrate is that such fact is not just an idea, and for that I need your input.
} Our facility charges $25.00 per hour for a Hitachi H7000 TEM and will be } charging $35.00 for a new TEM with digital capabilities. Technical help is } 35.00 per hour, an inverted multi-photon confocal is $25 per hour and a 3 } laser upright confocal with nomarski is $15 per hour. We offer two hours of } training at no charge. Use of the darkroom which covers chemicals and the } service on our processor is $10 per hour...Unfortunately we can't charge } } what it really cost to operate the facility or we'd have no business--so } } the university subsidizes us.
When all is done, I will summarize information received (leaving out names and institutions unless specifically asked to leave in the posting) and post for MSA users. Thanks in advance for your help.
*Disclaimer: Whatever... is not Tulane opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web:[ http://www.tmc.tulane.edu/ferminlab/] or [http://www.tmc.tulane.edu/imaging/] Internet: [cfermin-at-mailhost.tcs.tulane.edu] 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
We are trying to study the microstructure of cold drawn 25 =B5m gold = wires. We meet the difficulty in etching the gold wires to reveal the grain structure in SEM and optical observations. Could somebody give us some suggestions on how to etch the wires.
Thanks in advance!
Best regards,
Kun Li
Kun Li, Ph. D Institute of Materials Research and Engineering BLK S7, Level 3 Office: BLK S13, #02-13d National University of Singapore Lower Kent Ridge Road Singapore 119260