I'm trying to remember ... Freon 113: Peldri, yes? Or is this one that stays fluid? (And how did you get it, since Freons are mostly illegal anymore...) .
First, you might try vacuum, 1 atmosphere or so, maybe not that much, gently applied, and released.
For Peldri, I put specimens in small vials -- 4 mL Wheatons are my favorite -- and completely filled the vial, then capped, leaving no air. If there were enough alcohol:Freon intermediates, and enough changes in pure Freon, the specimens should be in pure Freon now.
Mind, I mostly did crustaceans and fish bits, not insects with their waxy epicuticles, but then some or all of that wax is lost in the alcohols anyway.
But I wouldn't use methanol or acetone. Ethanol works better for insects. Acetone is OK, but I've found ethanol better, and methanol is too extractive.
You don't have a CPD for this? Also, some insect parts can be dried from ethanol or even water. Mandibles of many species for instance, or the elytra of most beetles. Not whole insects, though.
I never got my specimens to sink in Freon 13, Back When, but they usually did sink in Peldri.
Phil
} When using F-113, I preceed it with either or both } methanol and acetone. The curious problem is that } after either a lengthy immersion in acetone or methanol, } the specimen (insect) floats when put in F-113. OK. } So it is lighter than F-113. What is a good method } of encompassing the specimen with F-113 to further } dehydrate it prior to vacuum desiccation? } } Any ideas?
}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC Dept. of Animal Health and Biomedical Sciences University of Wisconsin 1656 Linden Drive Madison, WI 53706-1581 Voice: (608) 263-4162 peoshel-at-facstaff.wisc.edu fax: (608) 262-7420 (dept. fax)
Last fall we actually fabricated one of those holders from some copper water pipe, and purchased the wavy washers from McMasters. I had seen the one Tina mentions in use and thought "heck, we could make one and save $100." Well we did just that. For less than $20, if you have a metal cutting saw, some solder and access to purchasing wavy washers, you can fabricate your own. One grad student has used it for the spring semester to dehydrate, and CPD, poly-l-lysine coated 12mm coverslips classifying the morphology and size of bacteria in the hind gut of Tipula abdomalis. I just reviewed his final work and the images looked great. He had no problems with debris or samples washing off. The advantage of making it yourself is that you can customize the size for the application for very little money. You would only be limited by the availibity of sizes of wavy washers.
-Geoff
} } I am searching some device for holding (polycarbonate)filters during } } dehydration and CPD. I am trying to view mycoplasms with SEM. Has anyone } } experience with this ? } } } We have a device from Tousimis that is designed to hold round glass } coverslips, and I have used it to hold polycarbonate filters. It is a } milled-out cylinder open along one side. Wavy washers are used to } separate the stacked coverslips. There are two sizes, 13 mm and 22 mm, } part numbers 8766 and 8767. You can contact Tousimis at 1-800-638-9558, } fax 1-301-881-5374, http://www.tousimis.com } } This device works for me! My only affiliation with Tousimis is as a } satisfied customer. } } Aloha, } Tina }
-- Geoff Williams
Electron Microscope Facility Supervisor Biology Department Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
Dear Sir, I have been working for Istanbul University, Faculty of Fisheries, as a research Assistant on aquaculture. I am a doctorate student at the present. My studies are on Aquarium Fish ( ornamental ). The topic of my doctoral thesis is " A Study on the Artificial Propagation of Discus Fish ( Symphysodon spp. ) and Effective Factors on the Propagation ". Besides, I research too my thesis for discus 's embriological-larval developing and sperm developing with scanning elecron microscope ( SEM ). With regard to my doctoral thesis: Could you send me Scanning Electron Microscope SEM )' s use-book and example's processing ? I wish success your studying. Best regards,
Esra SAVAS I.U. Su Urunleri Fak., Ordu Cad., No: 200 34470 Vezneciler / Istanbul TURKEY
MAY 11TH SYMPOSIUM AT U.C. Davis TOPIC: Electron Backscatter Patterns (EBSP)
Featured Speakers: 1) David Dingley, TSL, "Automated Crystallography for the TEM: Recent Developments" 2) Adam Schwartz, LLNL, "Coupling Automated Electron Backscatter Diffraction with Transmission Electron and Atomic Force Microscopies". 3) Patrick Camus, Noran Instruments, "Phase Identification versus Phase Discrimination" 4) Pierre Rolland, Oxford Instruments, "Mapping Copper Interconnects for the Semiconductor Industry" 5) John Sutliff, HKL Technology, "Quantifying Polycrystalline Microstructures: The Automated-ESBP Technique and its Applications"
The symposium will be held in the AGR room in the Buehler Alumni & Visitors Center, UC Davis
Meeting starts at 3:00 No Host Social -at- 5:30 Dinner Buffet -at- 6:30 Meeting/Dinner will end by 9:00
Dinner Buffet- Rosemary & Orange Chicken Fettucine Alfredo Spring Mix Salad with Raspberry Vinaigrette Greek Salad topped with Crumbled Feta Cheese on a Bed of Greens Chef's Selection Vegetable Fresh Baked Rolls with Butter New York Cheesecake with Melba Sauce garnished with Mint Coffee, Decaf, Hot Tea & Iced Water
Please call Greg Lum at 415/338-1339 or Email: "glum-at-sfsu.edu" for dinner reservations by Monday May 8th. The symposium starts at 3:00 p.m. The dinner will start at 6:30 pm.
The Nikon D1 Digital SLR camera does not have a mirror lockup but does have an "Anti Vibration Mode" which causes the shutter opening to be delayed until mirror shock has subsided. The D1 has 2.7million pixels, ISO range 200-1600 and accepts most Nikon lenses, electronic releases,etc. It is excellent for both copystand and light microscopy applications.
George Laing National Graphic Supply 226 North Allen Street Albany, NY 12206 E-mail: scisales-at-ngscorp.com (800) 223-7130 X3109 (518) 438-8411 X3109
At 08:45 PM 4/28/00 , you wrote: } } Just returned from a course on VP SEM (which was excellent) and just now } can add to some of the digital camera discussion. } } One thing not mentioned about using a SLR digital camera like the E1, N60 } is vibration from the mirror and the shutter. If you are going to use a } camera of this type you must be able to lift the mirror independently, } like the old F1, whether it's film or CCD. I have a Kodak DCS420 that is } based on the Nikon N90s camera body and it works ok at low magnifications } but anything over about 200X just doesn't cut it. } } }
I have a Philips 501 SEM and 201 TEM that I am about to dismantle and discard. They were both operational until four months ago. If anyone needs them for spare parts please feel free to contact me.
Best Regards,
Kirk J. Czymmek, Ph.D. Director, Core Microscopy Facility Department of Biological Sciences University of Delaware Newark, DE 19716 kirk-at-udel.edu PH: (302) 831-1158
If you find them let me know I would like a copy. I will pay reasonable or slightly unreasonable charges for copying and mailing.
Gordon
G. C,. Couger 624 Cheyenne Stillwater, OK 74075-1411 405 624 2855 between 3:00 pm to 1:00 am CST 405 742 2758 cell ----- Original Message ----- } From: "John Foust" {jfoust-at-threedee.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, May 01, 2000 7:28 AM
Meeting: "PASEM 2000" - THE PRACTICAL ASPECTS SERIES OF SHORT COURSES Dates: May 22 through June 2, 2000 Topic: The Practical Aspects Series consists of intensive 3.5 to 4.5 day short courses that provide participants with a thorough coverage of the basic theory and practice of scanning electron microscopy and associated techniques. Training is achieved through the use of easy to understand lectures coordinated with supervised laboratory sessions. The laboratory class sizes are purposely kept small so that participants will have an opportunity to gain extensive hands- on experience with available SEM and EDS instrumentation. Designed for the academic, government and industrial user, the courses are beneficial to microscopists and microanalysts at all levels from novice through advanced. Course titles and dates for our Year 2000 series follow:
1. SCANNING ELECTRON MICROSCOPY - May 22 through May 26, 2000 2. ADVANCED TOPICS IN SCANNING ELECTRON MICROSCOPY - May 30 through June 2, 2000 3. X-RAY MICROANALYSIS - May 30 through June 2, 2000
Sponsor: University of Maryland Location: College Park, Maryland, USA Interests: Both Physical & Biological Sciences Fields: SEM, EDS Contact: Tim Maugel, University of Maryland, Department of Biology, Building 144, Room 0240, College Park, Maryland, 20742, USA Tel: 301-405-6898 Fax: 301-314-9358 E-mail: tm11-at-umail.umd.edu WWW: http://www.life.umd.edu/pasem
If some of your spectra are only available in LINK AN/10000 format, then you could convert them to simple text files with a little program I wrote about a year ago. It's a 32 bit Windows program. You can select as many LINK spectrum files (in their original, not converted format) as you need. All the selected files will be converted in one run. I have tried it with more than 1700 spectra. The result of the conversion is a text file with the same name but with different file extension. An example from a converted spectrum file (from a series): -------------------------------LAK11S103.SP--------------------------- ak11s** 3
preset live time: 40 live time: 40 real time: 49 20.000000 eV/channel
Energy[keV], counts -0.200000, 0 -0.180000, 0 -0.160000, 0 -0.140000, 0 -0.120000, 3 -0.100000, 16 -0.080000, 42 -0.060000, 98 .. ---------------------------------------------------------------------- If you are still interested in a program like this, drop me a line and I'll send it to you.
With the best regards: Laszlo Varga
On 20 Apr 00, at 10:21, Filion, Christian 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 } -----------------------------------------------------------------------. } } } Good morning, } } I have both a Spectra file in Link format and txt format saved as a MSA } type file. Is there a software or an excel macro that could read these } spectra ? } } Thank you } } Christian Filion } Superviseur Laboratoires } Aciers Inoxydables Atlas } 1640 Marie-Victorin } Tracy, Québec } J3R 5R5 } Tel : 450-746-5243 } fax : 450-746-5241 } }
For those of you interested the Microscopy and Microanalysis 2000 Meeting Search Engine is now on line.
Using the Search engine you can find out dates/times/locations of any author, paper, subject or session at the upcoming meeting this August in Philadelphia.
We are looking for vendors of Peltier cooling stages for our Hitachi 2460N VPSEM. If you know of a company, please drop me a line. Also, if you have opinions on what features you would look for in such an item, I'd like to hear it. Thanks in advance, Randy -- Randy Nessler Views expressed are my own.
Never replaced a Pyramitome belt - but LKB microtome belts can be replaced by "regular" belt material - just match the width and springiness (did you keep any of the old belt fragments?). If your institution doesn't have a machine shop (those guys usually have extra belts floating around), try your local hardware store/superstore. I've had good luck replacing standard parts this way, and it is probably going to be light years cheaper than an "official" replacement part, if such a thing is available.
} From experience, giant rubber bands do not work very well.........
Tamara Howard CSHL
On Tue, 2 May 2000 mganger-at-aol.com-at-sparc5.microscopy.com 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 } -----------------------------------------------------------------------. } } } Date: Tue, 2 May 2000 08:08:07 EDT } Subject: Drive Belt for Pyramitome } To: Microscopy-at-sparc5.microscopy.com } MIME-Version: 1.0 } Content-Type: text/plain; charset="US-ASCII" } Content-Transfer-Encoding: 7bit } X-Mailer: AOL 5.0 for Windows sub 104 } } Greetings, } } I would like to know if anyone out there knows of a repair place that sells } parts for an LKB 11800 Pyramitome. The drive belt for the specimen arm has } literally fallen apart and I need to replace it. Any suggestions on where I } could get one would be greatly appreciated. } } Thanks in advance. } } Mike Ganger } Montclair State University } Montclair, New Jersey } mganger-at-aol.com } } } }
} I would like to know if anyone out there knows of a repair place that sells } parts for an LKB 11800 Pyramitome. The drive belt for the specimen arm has } literally fallen apart and I need to replace it. Any suggestions on where I } could get one would be greatly appreciated.
Be sure to look at vacuum cleaner drive belts. They come in many sizes and I've used them as drive belts for a lot of devices. They look like a giant O-ring, up to 1/4 inch thick and in many lengths.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Ernest Orlando Berkeley National Laboratory One Cyclotron Road, Berkeley, California 94720 FOR IMMEDIATE RELEASE
Two places left for students to attend the NCEM Summer School on Computing in Electron Microscopy June 26-30, 2000 in Berkeley, California
(Berkeley, CA) The seventh annual Summer School on Computer-Interactive HRTEM Image Acquisition, Processing and Simulation will be held at the National Center for Electron Microscopy (NCEM), Lawrence Berkeley National Laboratory, University of California, Berkeley from June 26 through June 30, 1999.
The curriculum will focus on training participants in techniques of computer-assisted acquisition and interpretation of high-resolution electron microscope images, including HRTEM image processing and simulation, electron holography, focal-series reconstruction, and remote-control microscopy. Participants will learn general principles and apply them to specific cases. Instruction will focus on the use of computer assistance rather than microscope training, although participants will acquire images on NCEM microscopes as well as using specific application programs for image interpretation. Class size will be limited to 16. Deadline for applications, May 01, 2000, has been extended to May 04, 2000..
Participants who wish to apply newly acquired techniques to their own projects will be encouraged to extend their visit at NCEM into the next week. Please note: this type of arrangement requires advance submission of an NCEM microscope proposal (see: http://ncem.lbl.gov/frames/user.htm). Projects may involve prepared specimens for microscopy, images and diffraction patterns for processing, or crystal and defect data for simulations.
For more information and application materials, contact:
I have been reading a great deal on polymer microscopy lately and came across the statement that there was very little information to be gained by staining polymers and (as might be expected) that they did not take up stain well. Many years ago, I had heard of an application in which osmium tetroxide was used, I believe on polyethylene. Can anyone shed any light, either on the general comment or on the application using osmium tetroxide?
Many thanks, Barbara Foster,President Microscopy/Microscopy Education 125 Paridon Street Suite 102 Springfield, MA 01118-2130 PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com Website: www.MME-Microscopy.com/education
America's first national consortium of microscopy specialists offering customized on-site training in all areas of microscopy, image analysis, and sample preparation ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Does anyone have a good, safe way of mounting Codonics dye sublimation prints onto poster boards? We would like to avoid the spray on "rubber cement" types of glues due to the mess.
Thank you.
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 750 Communications Drive - MC 4402 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 ####################################################################
Double sided tape or a product that picture framers use that just the glue without the tape. It is on a tape that releases the glue. It does an excellent job of mounting. I have pictures that were mounted with the Scotch brand 20 years ago that are just as good as the day I did them. It will also stick to the plastic use in resin coated photographic paper.
Any picture framing shop should have it.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 ----- Original Message ----- } From: "John J. Bozzola" {bozzola-at-siu.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, May 02, 2000 6:58 PM
O.k., whereas not directly a microscopy question I was hoping some one would have a solution. Look for something that will function like aluminium foil but be non- reflective.
(1) shapable around objects (we're covering petri dishes), but not too massive
(2) be opaque (visble light 420-720nm)
(3) be non-reflective on at least one side. "Non-reflective": less than 15% reflectivity to visible spectrum 420-720nm.
Any suggestions? (Any good ideas for turning Al foil nice and black?)
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
11th International Congress of histochemistry and Cytochemistry (ICHC 2000)
Cell Biology Tools for the New Century
ICHC 2000 is the premier meeting this year to address the very latest developments in visualisation techniques for Cell Biology. to be held in the magical medieval city of York, England between 3-8 September 2000.
We have a fabulous speaker listof over 100 including some of the biggest names in Cell Biology and Imaging such as Roger Tsien, Richard Haugland, Fred Bosman, Alan Boyde, Stefan Hell, Alan Fine, Jim Smith, Margaret Buckingham, Nick White, Angus Lamond, David Becker, Jeniffer Lipincott-Schwartz and many, many more.
Topics include -Tracking Molecules in Cells, Fluorescent markers of Gene Expression, Live Cell Imaging in plants and Fungi, Apoptosis, Cell Signalling, Environmental toxicology, Imaging Embryonic Development, Ion Imaging in Cells, Imaging in the Neurosciences etc., etc. etc.
See the science and have a chance to talk to the best in the field.
Come for a holiday as well as the science we can offer many great excursions.
For more information and details of how to register please visit our web-site at
www.med.ic.ac.uk/external/ichc_2000
Student bursaries are still available!
DON'T MISS OUT IT WILL BE ONE OF THE CONFERENCES OF THE YEAR
Gary Coulton
Dr. Gary Coulton Molecular Pathology Division of Biomedical Sciences Imperial College School of Medicine The Sir Alexander Fleming Building South Kensington London SW7 2AZ
tel 0044 (0)171 594 3190 fax 0044 (0)171 594 3022
PLEASE NOTE AS FROM APRIL 1ST 2000 THE FOLLOWING NUMBERS WILL REPLACE THOSE ABOVE AND SHOULD BE USED.
------------------------------------- Announcing the 11th International Congress of Histochemistry and Cytochemistry (ICHC 2000)
"Understanding Biocomplexity: The Post-Genome Challenge"
September 3-8, 2000, York, United Kingdom
ICHC 2000 will comprise 27 symposia addressing the latest developments and applications of histochemistry and cytochemistry in the life sciences including medicine.
Many leading experts to speak
On-line registration now open!!!!!!!!!!
For further details of the meeting and how to pre-register please visit our web-site at http://www.med.ic.ac.uk/external/ichc_2000
} O.k., whereas not directly a microscopy question I was hoping some one would } have a solution. Look for something that will function like aluminium foil but be non- } reflective. } } (1) shapable around objects (we're covering petri dishes), but not too massive } } (2) be opaque (visble light 420-720nm) } } (3) be non-reflective on at least one side. "Non-reflective": less than 15% } reflectivity to visible spectrum 420-720nm. } } Any suggestions? (Any good ideas for turning Al foil nice and black?) } } Thanks. }
I seem to remember that studio photographers use flat black aluminum foil. Try Porter's Camera, I think they are in Iowa. They are a large mail-order firm and probably have a website. Or try Calumet camera, 800 Supreme Drive, Bensenville, IL 1-800-225-8638.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
The best stuff that I ever found for mounting pictures is 3M Positionable Mounting Adhesive. This is very good stuff and not messy at all. It comes in rolls and is a transfer type material. You roll it out, place your picture on it and cut it out with a sharp knife. You then press the adhesive with the backing sheet and peel off the sheet. The adhesive is transferred to every square inch of the print. You can position this stuff around and then you press it down hard and it sticks forever. The roll that I use is PMA 568. There is larger and smaller sizes available. I think that you have to order through a photographic supply house because I don't think any of the EM supply houses carry it.
-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)
} -----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] } Sent: Tuesday, May 02, 2000 7:59 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: dry mounting dye sub prints } } } -------------------------------------------------------------- } ---------- } 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/MicroscopyListse} rver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } Does anyone have a good, safe way of mounting Codonics dye } sublimation prints onto poster boards? We would like to avoid the } spray on "rubber cement" types of glues due to the mess. } } Thank you. } } #################################################################### } John J. Bozzola, Ph.D., Director } Micro-Imaging and Analysis Center } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } #################################################################### }
} } O.k., whereas not directly a microscopy question I was hoping some one would } have a solution. Look for something that will function like aluminium foil but be non- } reflective. } } (1) shapable around objects (we're covering petri dishes), but not too massive } } (2) be opaque (visble light 420-720nm) } } (3) be non-reflective on at least one side. "Non-reflective": less than 15% } reflectivity to visible spectrum 420-720nm. } } } Any suggestions? (Any good ideas for turning Al foil nice and black?) } } 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." } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
As to pyramitomes, if anyone has one that is not in use and would like to sell it, please contact me. I used one years ago and wouldn't mind having one around. 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
--------------------------------------
Greetings, Babara Foster asked about staining polymers. By chance, I recently looked at paper on staining polymers for electron microscopy. The paper compared osmium and ruthenium tetroxides and concluded that ruthenenium tet stained many more polymers than did the osmium. This paper has a kind of encylopedic feel to it because they checked a lot of polymers. The citation is Trent et al., 1983 Macromolecules 16: 589-598. I have no idea whether this would work with light microscopy or what info you could get from it exactly besides contrast, but perhaps you will find the paper interesting.
As ever, Tobias } } Hi, } } I have been reading a great deal on polymer microscopy lately and came } across the statement that there was very little information to be gained by } staining polymers and (as might be expected) that they did not take up } stain well. Many years ago, I had heard of an application in which osmium } tetroxide was used, I believe on polyethylene. Can anyone shed any light, } either on the general comment or on the application using osmium tetroxide? } } Many thanks, } Barbara Foster,President } Microscopy/Microscopy Education } 125 Paridon Street Suite 102 } Springfield, MA 01118-2130 } PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com } Website: www.MME-Microscopy.com/education } } America's first national consortium of microscopy specialists offering } customized on-site training in all areas of microscopy, image analysis, and } sample preparation } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Hi, We got a bolt of black cloth from a theatrical supply house. It is the kind of thing used to make curtains and other masking on the stage. I think it was called "covert black" but I could have that wrong. We were doing photobiological work and were forever having to limit the pesky wandering photons. The cloth is thick but not too thick to wrap around a dish. It is not perfectly opaque but if you need that I bet you could line the cloth with al foil, on the inside. Hope this helps, Tobias } } } O.k., whereas not directly a microscopy question I was hoping } some one would } have a solution. Look for something that will function like } aluminium foil but be non- } reflective. } } (1) shapable around objects (we're covering petri dishes), } but not too massive } } (2) be opaque (visble light 420-720nm) } } (3) be non-reflective on at least one side. } "Non-reflective": less than 15% } reflectivity to visible spectrum 420-720nm. } } } Any suggestions? (Any good ideas for turning Al foil nice and black?) } } 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."
O.k., whereas not directly a microscopy question I was hoping some one would have a solution. Look for something that will function like aluminium foil but be non- reflective.
(1) shapable around objects (we're covering petri dishes), but not too massive
(2) be opaque (visble light 420-720nm)
(3) be non-reflective on at least one side. "Non-reflective": less than 15% reflectivity to visible spectrum 420-720nm.
Any suggestions? (Any good ideas for turning Al foil nice and black?)
Dear Richard,
How about depositing carbon black onto Al foil? It's messy, but fulfills
your requirements. Evaporating carbon seems like an inferior proceedure, but
that should also work. Just holding the foil above a low-temp flame (candle or
bunsen burner with the air turned down) seems the simplest. You can then
cover the carbon layer with something if necessary. Dipping the foil into laser-
printer toner could also work, and the toner could then be heated. Putting the
foil (cut to 8.5" x 11") through the printer is asking for trouble, but that could
also work if you can find a solid black character--WordPerfect 5.1 has such
a character, which can be accessed by ctrl-v, 3,3. Good luck.
I am looking for a protocol for Immunogold labelling membrane proteins on mitochondria / peroxisomes fractions..., I am hoping to use a formaldehyde type fixation..............to LR white resin, I would appreciate any advice, references, and if possible actual relevant protocols. Thank you in advance.
Dear John, Years ago I purchased "Mounting Film" from an EM supply house. This is double-sided sticky tape, two feet wide. You cut it to any size and press on. Mine is made by Lomacoll and is obviously German. At 06:58 PM 5/2/00 -0500, you wrote: } } Does anyone have a good, safe way of mounting Codonics dye } sublimation prints onto poster boards? We would like to avoid the } spray on "rubber cement" types of glues due to the mess. } } Thank you. } Best 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
We've used DryBond Adhesive Pads for mounting dye-sublimation prints. It's manufactured by Chartpak (their part number is DBS25 for 25 11" x 17" sheets). I order it from EMS (Cat. #77612-25), described on Page 311 of EMS Catalog XIII. Basically it consists of an array of tiny (well, actually quite large to electron microscopists) dots of rubber cement like stuff that you apply by placing the print on top, covering with a protective sheet then rubbing with a rubber brayer. Peel up the print, place on mounting board, cover with sheet and rub with brayer. We've used it on color dye-subs with and without the XtraLife coating with no problems. As far as longevity, I made a poster two years ago that is still adhered tight to the board, but that was with RC B&W paper. The oldest dye-sub has only been observed for about a year now.
I have no financial interest in Chartpak or EMS, the stuff is just nice to use.
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
I use a product called Liquid Paper Dryline Permanent Adhesive made by the Gillette Company. It is supposedly acid free, glues instantly, no drying time, and comes as either permanent or temporary. I get it a office supply store.
} Does anyone have a good, safe way of mounting Codonics dye sublimation } prints onto poster boards? We would like to avoid the spray on "rubber } cement" types of glues due to the mess.
Damian Neuberger, Ph.D. Baxter Healthcare, Inc. WG3-2S P.O. Box 490 Round Lake, IL 60073 e-mail: damian_neuberger-at-baxter.co, Tel: 847-270-5888 Fax: 847-270-5897
I want to buy an inspection scope from which I can also acquire digital images.
I remembered that I had a flyer for the Kodak MDS 120 system ($1895).
I called Kodak, they still sell the same system. It seems a little "out dated" now w/ only 1.2 pixels and no USB connection.
Are there other product that offer similar simplicity and versatility (macroscopic off the scope capability), that also fall in the same price range, and have 2 million pixels w/ a USB connection?
Now that the program for the meeting is available on the website - Thanks Nestor - I would like to advertize the "LATE-BREAKING POSTER SESSION AT MICROSCOPY AND MICROANALYSIS 2000" for any remaining authors (or potential authors) who would like to contribute to the meeting.
Microscopy and Microanalysis 2000 will feature a poster session composed of presentations of newly acquired data or analyses which were unavailable for submission by the February 15 deadline. A short, half page abstract describing the studies is required. The abstract should include: Title, Authors, Authors affiliation, and a Brief Description of the studies. The description should include the Aim of the studies, a short characterization of the Methods, and a brief account of the Results and their Importance.
Abstracts should be e-mailed or faxed to the program chair, Stuart McKernan, at stuartm-at-tc.umn.edu (email) or 612-625-5368 (fax). Abstracts may be submitted immediately but must be received by June 23, 2000. Abstracts will be reviewed by members of the program committee. A limited number of poster boards are available and preference will be given to early submissions. Abstract authors will be notified of acceptance of their abstracts no later than July 1 (earlier for early submissions).
__________________ Stuart McKernan stuartm-at-tc.umn.edu Director Office:(612) 626-7594 IT Characterization Facility, University of Minnesota Desk:(612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 NEW-} Fax:(612) 625-5368
Can someone on the listserver offer any help, I am seeking the JEOL publication of the list of O Rings (part numbers, or size/type) that are used in the JEOL COATER JEE-400 and the JEE 4B/4C. If it is available could you please either email me a copy of Fax me a copy Fax Number is (in australia) +61 (2) 9385 6400. Thankyou for your help, Barry EM UNIT UNSW
My question is : I need to obtain a estimate of virus particle concentration in a sample of cell culture supernatant for an article to be submitted to a journal. I was told of a method called the 'latex particle reference method' for obtaining virus particle counts. Can anyone on the list supply information for this procedures ? Thanks for your time.
Sincerely,
Tom Doman, Project Associate Veterinary Science Department Penn State University
Dear Neal, A good fixative for immunogold work is still a combination of glut. + paraformaldehyde, but using a 'low' glutaraldehyde concentration. One you could try would be 0.25% glutaraldehyde + 4% paraformaldehyde in phosphate buffered saline pH 7.2. Formaldehyde alone results in poor structural preservation. From there on you could follow the instructions for dehydration, infiltration etc. from the LR White pamphlet which works well. For labelling: 1. Pre-treat with 0.02M glycine in PBS to block any free aldehyde groups (2x10mins), 2. Block with a suitable protein, e.g. BSA, ovalbuimin (1%- 1X20mins) 3. Place each grid on a 15-20ul drop of primary antibody, suitably diluted, and either leave overnight at 4 degrees C in fridge, or on lab bench for about 2 hours. 4. Rinse 6 times in drops of PBS with 1% protein block (BSA etc.) - (6X5mins). 5. Place each grid on a 20ul drop of Protein-A Gold complex, suitably diluted, for 1-1.5 hours. 6. Rinse in PBS (6X5mins). 7. Wash in distilled or millipore filtered water (in small beakers, 3 x several rapid dips(about 6), making sure the grids remain under the water at all times during dipping). 8. Stain with Uranyl Acetate and Lead Citrate.
** Blot (side-on) onto filter paper between steps 1+2, 2+3, 4+5, 7+8.
Good luck.
Cheers, Marilyn
Neal Leddy 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 looking for a protocol for Immunogold labelling membrane proteins on } mitochondria / peroxisomes fractions..., I am hoping to use a formaldehyde } type fixation..............to LR white resin, I would appreciate any advice, } references, and if possible actual relevant protocols. Thank you in } advance. } } Neal Leddy nleddy-at-tcd.ie
Please contact the person at the end of the post, not me, as I am posting at the request of someone without access. Thank you. ----- Original Message ----- Sent: Wednesday, May 03, 2000 8:59 PM
} JOB OPORTUNITY AT UNIVERSITY OF HOUSTON } } } Supervisor: Histology and Microscopy Laboratory; College of Optometry; } University of Houston, Texas } } Job Description: } } This position is in direct support of research and teaching faculty } where the subject of interest is typically ocular and neural tissue. The } duties range from tissue preparation, embedment, sectioning and } mounting, to operation of light and transmission electron microscopes, } and development and printing of photographic results. Knowledge and } competency in operation of ultramicrotomes and the transmission electron } microscope as well as current techniques in morphology, histology, and } immunocytochemistry is required. Experience with cryo-microscopy, in } situ hybridization, and computer image analysis is desirable. The } successful candidate also may participate as co-author of research } papers describing research performed in the laboratory. } } The supervisor is responsible for maintenance of stocks of laboratory } supplies and care of the equipment, and keeping the lab clean and } usable, including proper disposal procedures for hazardous wastes. Other } duties include instructing graduate students in common and specialized } anatomical techniques required for their various projects, and } supervision and consultation on their work as required. Some effort also } is applied to teaching of undergraduate optometry students including } preparation of teaching slides and technical instruction in anatomical } methods. } } This job is scheduled to begin on 1 May 1999. Salary will be negotiated } based upon qualifications and experience. For further information, you } may contact: } } Chris Kuether, Technical Services Manager } College Of Optometry, University Of Houston } 4901 Calhoun Blvd. Houston TX 77204-6052 } vox:(713)743-2049 fax:--2053; ckuether-at-uh.edu } } }
You can read up on Emitech cold stages on our site home} contents} K4 We are agents for Emitech for Australasia only (south of Singapore), just hope that you would find the info useful. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Tuesday, May 02, 2000 11:37 PM, nessler [SMTP:randy-nessler-at-uiowa.edu] wrote:
} We are looking for vendors of Peltier cooling stages for our Hitachi } 2460N VPSEM. If you know of a company, please drop me a line. Also, if } you have opinions on what features you would look for in such an item, } I'd like to hear it. } Thanks in advance, } Randy } -- } Randy Nessler } Views expressed are my own.
I have archived three discussions on this from the listserver. Go to:
http://www.biotech.ufl.edu/~emcl
and look in the TEM section of "Tips & Tricks" under virus. It also has a few other ideas. Good luck
At 06:48 PM 5/3/2000 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { 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 "
I think that I have used something that would work well for your application. It is an anodized Al foil that we use for laser safety purposes. The foil is very flat black and the black does not rub off easily. However, the foil is significantly stiffer than household Al foil. The brand we have used is BlackwrapTM and it can be purchased from The Great American Market, Hollywood CA 323-461-0200. They have a web site as well. I hope that this helps.
Matthew Ervin, Ph.D. U.S. Army Research Laboratory (301)394-0017 MErvin-at-ARL.mil
"Richard E. Edelmann" {edelmare-at-casmail.muohio.edu} on 05/03/2000 07:39:20 AM
Please respond to Edelmare-at-muohio.edu
To: microscopy-at-sparc5.microscopy.com cc:
O.k., whereas not directly a microscopy question I was hoping some one would have a solution. Look for something that will function like aluminium foil but be non- reflective.
(1) shapable around objects (we're covering petri dishes), but not too massive
(2) be opaque (visble light 420-720nm)
(3) be non-reflective on at least one side. "Non-reflective": less than 15% reflectivity to visible spectrum 420-720nm.
Any suggestions? (Any good ideas for turning Al foil nice and black?)
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
Dear Listers, Does anyone currently use the Sony DKC-5000 CatsEye Digital Camera and if so would you recommend it for a shared technology lab. Rosemary Walsh, EM Facility for the Life Sciences Penn State University
Barbara, We use both osmium and ruthenium tetroxide to stain various polymers for EM observation. Ruthenium is less selective in its staining, and therefore does stain many more polymers. Both compounds stain unsaturated polymers pretty well. In olefins, like polyethylene, RuO4 is good for staining amorphous domains within the crystalline or semi-crystalline polymer fine structure. This is typically a TEM issue. With some rather coarse polymer blends (e.g. rubber inclusions in another polymer matrix) RuO4 or OsO4 can be used with SEM and FESEM imaging. I would expect to see very little staining contrast by LM due to the polymer domain sizes of typical co-polymers, polymer blends, or crystalline/amorphous polymer structure. Again the best reference I have is the book by L. Sawyer, Polymer Microscopy.
Brad Huggins BP Amoco, Naperville, IL
} ---------- } From: Barbara Foster[SMTP:mme-at-map.com] } Sent: Tuesday, May 02, 2000 4:57 PM } To: Confocal Microscopy List; microscopy-at-sparc5.microscopy.com } Subject: LM: Stains for polymers? } } ------------------------------------------------------------------------ } 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 been reading a great deal on polymer microscopy lately and came } across the statement that there was very little information to be gained } by } staining polymers and (as might be expected) that they did not take up } stain well. Many years ago, I had heard of an application in which osmium } tetroxide was used, I believe on polyethylene. Can anyone shed any } light, } either on the general comment or on the application using osmium } tetroxide? } } Many thanks, } Barbara Foster,President } Microscopy/Microscopy Education } 125 Paridon Street Suite 102 } Springfield, MA 01118-2130 } PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com } Website: www.MME-Microscopy.com/education } } America's first national consortium of microscopy specialists offering } customized on-site training in all areas of microscopy, image analysis, } and } sample preparation } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } ^^ }
We use routinely this method in our laboratory. Here is the used protocol of quantitation of virus particles by negative stain electron microscopy:
A selected volume (100 uL) of supernatant containing virus particles is mixed with a selected volume (100 uL) of polystyrene latex beads of known concentration (about 10e8 beads by mL) and a size between 100 - 200 nm in diameter. The mixture is placed in an Beckman Airfuge 240 µL-tube. A Formvar and carbon-coated grid is inserted into the bottom of the microtubes.* The tubes are placed in a Airfuge A-100 fixed angle rotor (30¡) and centrifuge at 20 psi (120 000 g) for 5 minutes. The grids are recovered with fine self-closing tweezers, dried with bibulous paper, stained 1 minute with phosphotungstic acid (PTA 3%, pH6.0) and dried again with bibulous paper. Samples are visualized under a transmission electron microscope with an approppriate magnification. On two different grids, 200-500 particles (latex beads or virus particles) are counted from at least five different areas on each grid. That way, from the ratio of the two types of particles in the suspension, the ratio of the volumes added and the known concentration of latex particles, the concentration of viral particles can be calculated.
Virus particle concentration (particles/mL) = (virus count / latex beads count) X (latex beads concentration) X (1/test article dilution)
The level of sensitivity of this procedure is between 10e6 and 10e10 particles per mL. Less than 10e6, there is not enough virus to get a realistic count and more than 10e10, there are too much virus to well differentiate them. This procedure is used principally to quantify Retrovirus type-A and -C particles in cells supernatant, but can be used for any virus particles.
*R. Alain et al, J.Virol. Meths, 16 (1987), 209-216 *Alain, R. Microscopy Today, May 1997, issue#97-4, D. Grimes Ed, p. 20
If you need more explanation you can contact us. If you are not the equipment to do this technique, we can offer this service at low price.
Robert Alain
********************************************************** Robert Alain, M.Sc. Microscopie Žlectronique
INRS-Institut Armand-Frappier Centre de Microbiologie et Biotechnologie 531 boul. des Prairies Laval, QuŽbec CANADA H7V 4B7 Tel: (450)687-5010 ext#4388 Fax: (450)686-5626 e-mail: Robert.alain-at-INRS-iaf.uquebec.ca or Robert_alain-at-hotmail.com Http://www.iaf.uquebec.ca/iaf/recherche/viro/me.html **********************************************************
----- Original Message ----- } From: Tom Doman {jtd1-at-psu.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, May 03, 2000 7:48 PM
{html} You may want to look at the Olympus DP-11 digital camera. You can find it on their web page at: {a href="http:///"} http://www.olympusamerica.com/product.asp?c=21&s=11&p=18&product=612 {/a} {br} It has very high resolution, live video output (which is quite useful at low magnification as a focusing and framing aid) and features a c-mount so that it can be used on any microscope, or with a macro lens. It uses SmartMedia cards as the recording media, and combined with a USB SmartMedia card reader, download to the computer is fast and easy. {br} {br} Configured with an AC adapter, 64MB SmartMedia card, USB card reader and a 9" Sony video monitor, it is priced at $4,349.00. This is more than you indicated you would like to spend, but I think that you will find that its features, resolution and ease of use may justify the additional expense. {br} {br} At 03:23 PM 5/3/00 -0400, you wrote: {br} >------------------------------------------------------------------------ {br} >The Microscopy ListServer -- Sponsor: The Microscopy Society of America {br}
Thanks to everyone who responded - I was thinking anodizing myself but Matt Ervin actually had an Off-the-Shelf source.
===============
Its exam time here and I wanted to share this answer from my EM Theory Final:
Ques: What does "EELS" stand for? What does it detect (Be specific!)?
Ansr: Energized Eucaliptic Leaf Shooter. If properly used it can be used to lure the better part of the world's Koala Bear population in a general area to get a more precise population density reading.
[The student recieved 1 point for creativity]
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
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Hello listers- I am hoping that someone might have a current address or phone for "Cambride Image Technologies Ltd." or "Nuclide" A colleague of mine is looking to purchase a cold cathode luminoscope and was given these two names. Or if someone knows of another supplier any information would be appreciated. Thank you, Sarah
Sarah A.W. Lundberg Lab (EPMA) (702) 895-2660 or Electron Microanalysis and (SEM) (702) 895-2462 Imaging Laboratory Office (702) 895-1134 Department of Geoscience, UNLV Fax (702) 895-4064 4505 S. Maryland Parkway Box 454010 Las Vegas, NV 89154-4010 Dept Office (702) 895-3262
I believe that what Geoff was referring to is called "Black Wrap." It is a heavy foil with a matte black coating. It is used by theater lighting people worldwide to create custom masks and light shields. It is made by several companies. My local lighting house sells black wrap made by Great American ... in 12 inch by 50 foot or 24 inch by 25 foot rolls for US$27.50. Since you're at a university you could probably go over to the theater department and get a little piece to try.
Larry
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
At 05:59 AM 5/4/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
After some time of getting used to it, the 5000 is OK. It uses a small CCD so the image files are small and not really high resolution. But for relatively modest final image output size, it is a good camera. It provides real time focusing via a separate RGB+sync color monitor. However, adjusting the monitor to match the captured image's exposure is tricky.
If you are going to be moving the camera around, remember that it uses a SCSI interface to download captured images. You won't be able to move it very far from the SCSI port unless you use a notebook or laptop computer.
Scotch 924 Transfer Tape 1/2 inch wide is the product I mentioned earlier. It cost about 6.95 a roll for a life time supply for many of us. http://productinfo.mmm.com/us/office/products/office.jhtml?powurl=4JLD1SMBbe ZCZYKRQ9geT1T4S9TCgvPDJGVQ33gl
The way I used it to mount photos was stick a band of it around the edge of the photo as close as I could to the edge. That's all I did for 5X7's for 8X10 I put a cross diagonally from corner to corner. I didn't mount prints larger than that but If I had I would have added some tape in the open areas. Some one made a small version that had an applicator but the refills cost as much as a full sized roll and it is not much if any easier to use.
I tried a lot of things spray glue, two sided tissue and a number of others. This was the easiest and most consistent other than Seal mount tissue and a hot press.
The hardware store where I drink coffee is will sell mail order. I have not interest in the hardware store other than a bunch of us drink their coffee. Their web page address is www.studyboards.com or 405 372 2644 ask of Sandy or A.J.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
You can find information on www.bal-tec.com --} products--} vct100
Best regards, Udo Graf BAL-TEC AG +423 388 12 26 +423 388 12 60 (Fax) udo.graf-at-bal-tec.com
+ -----Ursprüngliche Nachricht----- + Von: nessler [mailto:randy-nessler-at-uiowa.edu] + Gesendet: Dienstag, 2. Mai 2000 14:37 + An: miscroscopy listserver + Betreff: SEM Peltier stage vendors? + + + We are looking for vendors of Peltier cooling stages + for our Hitachi + 2460N VPSEM. If you know of a company, please drop me a + line. Also, if + you have opinions on what features you would look for in + such an item, + I'd like to hear it. + Thanks in advance, + Randy + -- + Randy Nessler + Views expressed are my own. +
I have customer here requiring some help. They are making replicas of fossil mandibles for SEM analysis. Their problem is how to localise the areas of interest when the specimen is in the SEM. I was sent the quotation below, and we are trying to find out whether we, or anyone else, can supply the sort of material they need. My background is mainly materials, so I'm farming this out to the list to see if anyone has any suggestions, or maybe even recognises the fragment below.
"Fiberglas screening material: fiber thickness (310 microns), hole width (1, 240 microns), and length (1,340 microns), wiped with Permabond 910 adhesive and pressed onto the surface so that contact was made everywhere".
Thanks in advance
Tim
***************************************************************** Tim E. Harper Managing Director CMP Cientifica s.l. Space & NanoTechnology Division Phone +34 91 640 71 85 Fax +34 91 640 71 86 http://www.cmp-cientifica.com/ {http://www.cmp-cientifica.com/}
That sounds a lot like fiberglass window screen to me, but the holes here are probably a bit smaller. There is also a fiberglass screening used for backing plaster repairs. It might have a pitch more similar to what you described.
At 05:09 PM 5/5/2000 +0200, you wrote: } Hi, } } I have customer here requiring some help. They are making replicas of fossil } mandibles for SEM analysis. Their problem is how to localise the areas of } interest when the specimen is in the SEM. I was sent the quotation below, } and we are trying to find out whether we, or anyone else, can supply the } sort of material they need. My background is mainly materials, so I'm } farming this out to the list to see if anyone has any suggestions, or maybe } even recognises the fragment below. } } } "Fiberglas screening material: } fiber thickness (310 microns), hole width (1, } 240 microns), and length (1,340 microns), wiped with Permabond 910 adhesive } and pressed onto the surface so that contact was made everywhere". } } } Thanks in advance } } Tim } } ***************************************************************** } Tim E. Harper Managing Director } CMP Cientifica s.l. } Space & NanoTechnology Division } Phone +34 91 640 71 85 Fax +34 91 640 71 86 } http://www.cmp-cientifica.com/ {http://www.cmp-cientifica.com/}
We have had the Sony DKC-5000 for quite a few years and hve never had a problem with the hardware despite many clumsy users. We have it running on a Mac computer taking the Sony RGB signal to SVHS via a Harmonic Research CV-233P video encoder. The SVHS signal is input to a ATI Xclaim video board in the Mac. So the live window and the Sony acquire plug in run on the same monitor. The density and color balance rarely coincide between the two displays. We have had many problems with users changing settings and adding software, etc. on the Mac resulting in problems with the video display. As Dr Gaugler said, the resolution is not very high. There should be better systems now but I continue to recommend that our scientists record their images on color slide film and then scan it with a $2000 slide scanner (Nikon, Polaroid etc.) much cheaper, higher resolution and bandwidth and easy convenient storage!
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
We are still not set-up well for Cryo-SEM, and although we have made some progress and learned much about these techniques from you all on this Listserver and others, I am still looking for a "cryomicroscopy facility" who has, for example, an E-SEM with Peltier Stage/controlled cooling rate capabilities and with cryostage (capable down to temperatures as low as approximately -75C); who would be interested in working together with one of my clients, in a large Chemical/Oil Company, to investigate/observe the low-temperature structures of various Synthetic Fluids.
Anyone interested, please contact: Brad Huggins at BPAmoco, Naperville, IL hugginbj-at-bp.com 630 420-3668
Hi All, Does anyone have a good protocol for silver enhancement of immunogold labeled sections for TEM? I have a paper in front of me, but they used a kit. We do light level silver enhancement without a kit all of the time, but I don't want to chance trying to adapt that method before consulting all of you. Thanks for your help! Have a wonderful weekend, Kristen Kristen A. Lennon Cell, Molecular & Developmental Biology Group Department of Botany & Plant Sciences University of California Riverside, CA 92521 kalen-at-citrus.ucr.edu 909-787-4525
If I understand correctly how you are using the DKC-5000, then you are actually obtaining half the image resolution that the system can provide.
The DKC-5000 has a 1/2" 440,000 pixel CCD with an effective image area of 795x598 pixels (RGB). The actual captured image area is probably closer to 740x580 pixels (RGB). Thus, if my calculations are correct, you should obtain RGB TIF files which are about 1.29MB in size.
The image processor in the DKC-5000 digital processor system unit expands (interpolates?) the original image to one that is 1520x1144 pixels (RGB). This results in a TIF file that is about 5.21MB in size. However, this resolution is only obtained via the SCSI interface on the back of the system box. This is likely why you are obtaining low resolution results. This is effectively taking a consumer grade RGB video camera and performing an RS-170 frame grab. This will produce up to 800x600 pixels (RGB) maximum. This is OK, but the higher cost of the DKC-5000 was due, I think, to the ability to obtain higher quality images and to do so without any separate/extra image capture hardware. The DKC-5000 was about $14K I think versus about $800 for a really nice RGB video camera with close to 800x600 pixels resolution.
Therefore, if you have any sort of typical Mac, it should have a legacy SCSI-I connector on the rear. This is easily connected to the digital processor box. Then, load the Photoshop plug-in and acquire the higher resolution images using Photoshop and the SCSI bus. You should still be able to use the RGB outputs for framing and focusing. Just don't use the RGB outputs for image capture.
Have you tried this?
gary g.
At 09:57 AM 5/5/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings Listers, I am looking for a couple decomissioned electron microscopes to be used as a source for parts. I would greatly appreciate any information or leads. The instruments are the Hitachi 7000 TEM and the Hitachi S-510 (or 515 or 520) SEM. Thank you very much, Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB-499, 1807 West Slaughter Lane, Number 200 Austin, Texas 78748-6200 Phone: 512/282-5507 Fax: 512/280-0702 QUALITY ELECTRON MICROSCOPE REPAIR
At http://www.couger.com/gcouger/leitz/ are some pictures of a very old Leitz projection microscope. For it's age and rough construction it works extreemly well. It is of cast iorn, steel and brass construction and carries no serial number. It has two objective lenses that are about 6 and 12 x It has a simple condenser in the lamp housing and every thing is adjustable on the stand and the objectives have a heilical ajustment.
I have as several musem curators and infivigules that have a life long assoitions wiht Leitz. No one has seen anything like it. It appears to be all origianl except for the light bulb, its mounting and the cord. They look like they came from the 1920's
} From my decussions with Leitz sholers and deduction I think this was made about the same time as the electric light bulb. It would be possible that it use a gas or lime light but I don't think so. The mechenism for centering a light bulb just looks like it was designed for a bulb.
Any help is welcome. If anyone knows when Leitz started putting serial numbers on their products it would be a great help in dating it.
Thanks Gordon Couger Stillwater, OK 74075 405 624 2855 GMT - 6:00 www.couger.com/gcouger
I am sorry the last post some how escaped my spell check on my email program. It is set to check every email. But it misses some. For Microsoft an occasional miss is pretty good:)
At http://www.couger.com/gcouger/leitz/ are some pictures of a very old Leitz projection microscope. For it's age and rough construction it works extremely well. It is of cast iron, steel and brass construction and carries no serial number. It has two objective lenses that are about 6 and 12 x It has a simple condenser in the lamp housing and every thing is adjustable on the stand and the objectives have a helical adjustment.
I have as several museum curators and individuals that have a life long association with Leitz. No one has seen anything like it. It appears to be all original except for the light bulb, its mounting and the cord. They look like they came from the 1920's
} From my decisions with Leitz scholars and deduction I think this was made about the same time as the electric light bulb. It would be possible that it use a gas or lime light but I don't think so. The mechanism for centering a light bulb just looks like it was designed for a bulb.
Any help is welcome. If anyone knows when Leitz started putting serial numbers on their products it would be a great help in dating it.
Thanks Gordon Couger Stillwater, OK 74075 405 624 2855 GMT - 6:00 www.couger.com/gcouger
Sarah- I can't help you with Cambridge Image Technologies Ltd., but I do know about Nuclide. Nuclide went bankrupt several years ago and was bought up and reborn as Premier American Technologies Corp. which continued making the luminoscopes among other things. I worked for PATCO for a couple of years. I don't know the details, but PATCO has since evolved into Spectru Medix and I imagine they still sell luminoscopes as it was probably their most consistent seller in the past. You can call Spectru Medix at 814-867-8600, they are at 2124 Old Gatesburg Rd., State College, PA. I would suggest you talk to Mike Vollero as I know he still works there and will be able to put you in contact with the proper people. I hope this helps. Matt Ervin
Sarah Lundberg {lundberg-at-nevada.edu} on 05/04/2000 02:10:20 PM
To: MSA listserver {Microscopy-at-sparc5.microscopy.com} cc:
Hello listers- I am hoping that someone might have a current address or phone for "Cambride Image Technologies Ltd." or "Nuclide" A colleague of mine is looking to purchase a cold cathode luminoscope and was given these two names. Or if someone knows of another supplier any information would be appreciated. Thank you, Sarah
Sarah A.W. Lundberg Lab (EPMA) (702) 895-2660 or Electron Microanalysis and (SEM) (702) 895-2462 Imaging Laboratory Office (702) 895-1134 Department of Geoscience, UNLV Fax (702) 895-4064 4505 S. Maryland Parkway Box 454010 Las Vegas, NV 89154-4010 Dept Office (702) 895-3262
We have recently encountered some perplexing problems staining tissue embedded in Spurr's resin with uranyl acetate and calcined lead. The tissue is not taking up the stain although there is some stain precipitate on the sections. Any suggestions?
Sincerely, Harry Grier Stock Enhancement Research Facility Florida Marine Research Institute 14495 Harllee Road Palmetto, FL 34221 harry.grier-at-fwc.state.fl.us
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id NAA21976 for dist-Microscopy; Mon, 8 May 2000 13:34:05 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id NAA21973 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 8 May 2000 13:33:34 -0500 (CDT) Received: from mail.ucsf.edu (mail.ucsf.edu [128.218.95.23]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id NAA21966 for {microscopy-at-Sparc5.microscopy.com} ; Mon, 8 May 2000 13:33:23 -0500 (CDT) Received: from JanLab.ucsf.edu (u226-mac-01.ucsf.edu [128.218.106.131]) by mail.ucsf.edu (Pro-8.9.3/CDR-Pro-8.9.3) with SMTP id LAA13666 for {microscopy-at-Sparc5.microscopy.com} ; Mon, 8 May 2000 11:27:55 -0700 (PDT) Message-Id: {3.0.2.32.20000508112409.015b9168-at-itsa.ucsf.edu} X-Sender: mishot-at-itsa.ucsf.edu X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.2 (32)
Apparently I was not clear in my description of our DKC-5000 setup. The RGB signal displayed in a live ATI video window is for focusing and composing. Acqusitition of the image is via the Sony plug-in and SCSI transfer. The resulting image file is about 5MB. It does not meet my standards for a publication quality image. Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
I have a question about how best to maintain a sputter coater that is used infrequently.
We are a small college with correspondingly small EM facility. Our SEM and prep equipment may go for several weeks without being used, then someone will need it heavily for a class for three or four weeks. Invariably we find that our sputter coater seems to be the weak link in our plans, since it rarely works well when we fire it up after long periods of dormancy. I understand this is common with vacuum equipment: better to use it often rather than shut it off.
Am I correct that we should we have a plan to regularly pump down the sputter coater, and if that is good idea, how often should that be? Every day? Once a week? Once a month?
Or, if we don't need to pump it down regularly, are there other things we should be doing do it the down time instead of letting it just sit there?
and finally, are some designs better able to handle long periods of disuse? Do we just have the wrong sputter coater?
Gary P. Radice gradice-at-richmond.edu Associate Professor of Biology 804 289 8107 (voice) University of Richmond 804 289 8233 (FAX) Richmond VA 23173 http://www.science.richmond.edu/~radice
Does anyone have any old parts for the See-Vac, Inc. Autoconductavac sputter coaters? Especially the power feed-through that connects through the cap to the target. Thanks.
Phil -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC Dept. of Animal Health and Biomedical Sciences University of Wisconsin 1656 Linden Drive Madison, WI 53706-1581 voice: (608) 263-4162 fax: (608) 262-7420 (dept. fax)
Microscopist: To manage light and electron microscopy facility. Minimal degree requirement BS (MS preferred). Experience with confocal microscopy, computerized image analysis and histological sample preparation essential. Experience in SEM/TEM sample preparation and video microscopy desirable. Must be an interactive person willing to facilitate microscopy experiments for faculty and students with a wide variety of interests in a University setting. Salary commensurate with experience. Full time desired but will consider part time. For further information on the department see http://www.umbc.edu/biosci UMBC is an AA/EOE.
Contact: Dr. Daphne Blumberg, Chair, Microscopist Search Committee, Dept. of Biological Sciences, University of Maryland Baltimore County (UMBC), Baltimore MD 21250.
All vacuum systems are better "stored" under vacuum, with the exception of any systems that cannot vent the rotary pump when power is off. Those systems may suck oil from the pump towards the vacuum chamber.
The more common problem for infrequently used systems is moisture in the pump oil. Particularly in moist climates and when relatively short pumping times are employed a good deal of water is absorbed in the pump. When the pump is not used for a lengthy period this may cause corrosion and certainly lowers the vapour pressure of the contaminated oil and so lower performance results.
I suggest that at the end of your spasmodic activities the pump should be run with the baffle valve partially open for at least 30 minutes. The baffle valve is usually atop the rotary pump. You could also use the sputter coater's needle valve, partially open. Under these conditions the pump will run hotter and throw-out a good deal of oil mist and water vapour. Vent the exhaust into a fume hood since oil mist is not just unpleasant. You will find that the pump performs much better after a baffle run.
SEM and TEM usually do not need this treatment, but they may, for instance if a TEM has been used to "dry" film that was not previously dried in another system. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Tuesday, May 09, 2000 5:11 AM, Gary Radice [SMTP:gradice-at-richmond.edu] 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 have a question about how best to maintain a sputter coater that is used } infrequently. } } We are a small college with correspondingly small EM facility. Our SEM and } prep equipment may go for several weeks without being used, then someone } will need it heavily for a class for three or four weeks. Invariably we } find that our sputter coater seems to be the weak link in our plans, since } it rarely works well when we fire it up after long periods of dormancy. I } understand this is common with vacuum equipment: better to use it often } rather than shut it off. } } Am I correct that we should we have a plan to regularly pump down the } sputter coater, and if that is good idea, how often should that be? Every } day? Once a week? Once a month? } } Or, if we don't need to pump it down regularly, are there other things we } should be doing do it the down time instead of letting it just sit there? } } and finally, are some designs better able to handle long periods of disuse? } Do we just have the wrong sputter coater? } } Gary P. Radice gradice-at-richmond.edu } Associate Professor of Biology 804 289 8107 (voice) } University of Richmond 804 289 8233 (FAX) } Richmond VA 23173 http://www.science.richmond.edu/~radice } }
The new BAL-TEC 'VCT 100' Vacuum-Cryo-Transfer-Equipment is a modular system consisting of:
- Shuttle (Cryo-Vacuum Conditions) - Docking station at any preparation system - Docking station at any analysis system (SEM, ESEM, Cryo-AFM) - Cryo equipment for any analysis system (SEM, ESEM)
The modules can be arranged just to meet your needs.
VCT 100 Cryo equipment has been adapted to a Philips XL30 e.g.
Additional information you will find on www.bal-tec.com --} products--} vct100
Best regards, Udo Graf BAL-TEC AG +423 388 12 26 +423 388 12 60 (Fax) udo.graf-at-bal-tec.com
-----Ursprüngliche Nachricht----- Von: Huggins, Bradley J [mailto:HUGGINBJ-at-bp.com] Gesendet: Freitag, 5. Mai 2000 19:23 An: Huggins, Brad; Microscopy listserver Betreff: low temperature microscopy of synthetic fluids
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We are still not set-up well for Cryo-SEM, and although we have made some progress and learned much about these techniques from you all on this Listserver and others, I am still looking for a "cryomicroscopy facility" who has, for example, an E-SEM with Peltier Stage/controlled cooling rate capabilities and with cryostage (capable down to temperatures as low as approximately -75C); who would be interested in working together with one of my clients, in a large Chemical/Oil Company, to investigate/observe the low-temperature structures of various Synthetic Fluids.
Anyone interested, please contact: Brad Huggins at BPAmoco, Naperville, IL hugginbj-at-bp.com 630 420-3668
Gary- I am not sure if you are saying that your sputter coater is pumping down poorly or that it is depositing poor quality films. In either case, it sounds like poor vacuum is the problem. Water vapor adsorbing to the deposition chamber walls over a period of disuse will pump off of the walls very slowly. A "wet" chamber can take 10 times as long to pump down to ultimate vacuum as a "dry" one. If this sounds like it might be causing the symptoms you are seeing, I would suggest you try one of two things:
1. Place a valve between the sputter chamber and the pump. Valve off the chamber and leave it under vacuum between uses. As long as it retains any vacuum this will help when you try to use it again. You don't want to be pumping on the chamber at the pump's ultimate vacuum for any extended period of time because that will allow oil vapor from the mechanical pump to backstream into your sputter chamber and possibly onto your specimen if present! (I am assuming that you have an oil based pump.) It is also nice to spare the pump the wear and tear of pumping continuously over periods of disuse.
OR 2. Several hours or the day before you want to use it, start pumping on the sputter chamber while admitting a small flow of argon. The argon is an important part of this procedure. First of all, the argon will prevent the backstreaming described above. Second, I have been told that the argon helps to desorb the water from the chamber surfaces. I don't know if that is an old wives tale or not, but it does seem reasonable. The argon flow may also help in purging any condensed vapors from the pump's oil. Don't use too much argon though or you may overheat your pump.
I hope that this addresses the problem you are experiencing. Matt Ervin (301)394-0017 U.S. Army Research Laboratory Adelphi MD
Gary Radice {gradice-at-richmond.edu} on 05/08/2000 03:10:46 PM
To: Microscopy-at-sparc5.microscopy.com cc:
I have a question about how best to maintain a sputter coater that is used infrequently.
We are a small college with correspondingly small EM facility. Our SEM and prep equipment may go for several weeks without being used, then someone will need it heavily for a class for three or four weeks. Invariably we find that our sputter coater seems to be the weak link in our plans, since it rarely works well when we fire it up after long periods of dormancy. I understand this is common with vacuum equipment: better to use it often rather than shut it off.
Am I correct that we should we have a plan to regularly pump down the sputter coater, and if that is good idea, how often should that be? Every day? Once a week? Once a month?
Or, if we don't need to pump it down regularly, are there other things we should be doing do it the down time instead of letting it just sit there?
and finally, are some designs better able to handle long periods of disuse? Do we just have the wrong sputter coater?
Gary P. Radice gradice-at-richmond.edu Associate Professor of Biology 804 289 8107 (voice) University of Richmond 804 289 8233 (FAX) Richmond VA 23173 http://www.science.richmond.edu/~radice
I agree. In addition... Given a choice, I would always use the process gas to purge - using the "leak valve" to feed a partial pressure to the system until the vacuum pump heats up. Opening the ballast valve will help (lowers ultimate vacuum when open) purge the pump, but IMHO, it is better to use dry gas than atmosphere.
Woody White McDermott Technology
{SNIP} I suggest that at the end of your spasmodic activities the pump should be run with the baffle valve partially open for at least 30 minutes. The baffle valve is usually atop the rotary pump. You could also use the sputter coater's needle valve, partially open.
Thanks to all who responded to my query about how best to maintain my infrequently used sputter coater. Since some have replied off list and others have asked to see all the responses I thought I would append all of the responses here, (without direct attribution, since strictly speaking I don't have all the authors permission to do this....I hope they understand).
Thanks to all who responded. Based on the comments, I think my problem was accumulation of water in the oil and vacuum surfaces, and dried out gaskets. Greasing the gaskets and running the pump longer got the pump-down time from 20 minutes to 3 minutes. Keeping the chamber under vacuum isn't practical with our coater design, but I can probably solve my problem by arranging to pump down the chamber once a week, paying attention to keeping the seals lightly greased, and changing the pump oil on a regular schedule.
******************
} Hi Gary, } } How are you? Our sputter coater has sat for over a year at a time without } use. It had no problems when it was started. However, that is not an } ideal situation. Vacuum equipment whould be regularly pumped down to } out-gas the chamber, etc. We have a Denton Desk II Sputter Coater. It is } by far the best I have used. It is now used regulary because we have a new } SEM. If I were you I would pump down the chamber on the coater at least } once a month. Change the oil in the pumps once/year. Check the seal } between the chamber and the base and the glass and the seal before the } class use starts. Use fomblin grease or some other non-hydrocarbon vacuum } grease. *************
} There really should not be any heroics needed in order to snap your figers } and have the sputter coater work well. We ship our coaters all over the } world to trade shows, open up the boxes, take them out, put them on the } table, and a few minutes later we are coating samples for prospective } customers. And I think that would probably be the case for most } commercially made coaters today that are used in the SEM market. } } But I will tell you one thing that does happen and that is that the needle } valve can develop a "set" if it is left tightened down real tight over long } periods of time. } } The when you go to use it again, because of the "set", there are problems } controlling its action and therefore the bleed rate of air or inert gas. I } know that conventional wisdom says a vacuum system should be stored "under } vacuum" but the typical coater is sufficiently leaky, that the vacuum is } going to disappear shortly anyhow. } } So you might want to try storing it no under vacuum, that is, with the need } valve open, and see if that does not make your problems disappear.
******************
} Hello Dr. Radice, } Yes, the worst thing you can do to a vacuum system is to not use it. The } least you should do is keep the bell jar under vacuum when it is not being } used. Do you vent with Dry Nitrogen or room air? All high voltage leads } and feed throughs should be kept very clean since they have a tendency to } collect Carbon and crud. It might not be a bad idea to pre-run your high } voltage up rather high (higher than you would normally use it), pror to a } run. This should stabilize things a bit.
***********
} Gary, } After many years of using sputter coaters I have found that it is best } that they are kept under vacuum all the time. Many sputter coaters do } not let you maintain a vacuum when they are off. With this type I have } found that I have to keep the glass cylinder and the metal target very } clean and allow plenty of pump down time when first using the unit after } prolonged shut down. Also any solvent based glues that may be used, } silver dag or colloidal carbon, must be dry before putting the sample in } the coater, at least 2 hours after mounting. **************
} I have a similar use pattern as yours, but I have never had any problems } with my coater. Sometimes, our coater my sit for a period of months before } a pump down is required. Back in 1993 I purchased an Emitech, the same } type that is sold through EMS today, and have had but only one failure. } That failure was due to foil on a circut board that was not heavy enough to } handle the current required by the vacuum pump. The foil had melted, but } was an easy fix for me. Otherwise I have had no failures. } } It certainly can help to pump it down once a week, just like it helps } ink-jet printers to print a test page once a week. Other than keeping } non-contaminated oil in the pump, a light coating of vacuum grease on the 0 } rings to keep them from drying out and getting attacked by ozone, I have } not had other maintenance issues. Perhaps your vacuum problems are related } more to your pump and the need for some maintenance and oil change, perhaps } your seals need replacing, or perhaps your coater is going through a period } where it requires higher than normal maintenance. I'm curious, what type } of problems are you having? } } Good luck with your facility. I always enjoy meeting others who are } running small EM labs. *******************
} All vacuum systems are better "stored" under vacuum, with the exception of } any } systems that cannot vent the rotary pump when power is off. Those systems may } suck oil from the pump towards the vacuum chamber. } } The more common problem for infrequently used systems is moisture in the pump } oil. Particularly in moist climates and when relatively short pumping } times are } employed a good deal of water is absorbed in the pump. When the pump is not } used for a lengthy period this may cause corrosion and certainly lowers the } vapour pressure of the contaminated oil and so lower performance results. } } I suggest that at the end of your spasmodic activities the pump should be run } with the baffle valve partially open for at least 30 minutes. The baffle } valve } is usually atop the rotary pump. You could also use the sputter coater's } needle } valve, partially open. Under these conditions the pump will run hotter and } throw-out a good deal of oil mist and water vapour. Vent the exhaust into a } fume hood since oil mist is not just unpleasant. You will find that the pump } performs much better after a baffle run. } } SEM and TEM usually do not need this treatment, but they may, for instance } if a } TEM has been used to "dry" film that was not previously dried in another } system. *************
} We used to leave the sputter coater sitting for weeks after pumping it down } and never had any problems. It sounds as if you may have a leak at your } sealing surface. We always had to be very careful about cleanliness of the } bell jars surface and the plate it seals on. A small grain of sand or other } material can fracture the glass so that you have leaks and requires } repolishing of the glass bell jar. } } I hope that this helps you. ***************
} Gary- } I am not sure if you are saying that your sputter coater is pumping } down poorly or that it is depositing poor quality films. In either case, } it sounds like poor vacuum is the problem. Water vapor adsorbing to the } deposition chamber walls over a period of disuse will pump off of the walls } very slowly. A "wet" chamber can take 10 times as long to pump down to } ultimate vacuum as a "dry" one. If this sounds like it might be causing } the symptoms you are seeing, I would suggest you try one of two things: } } 1. Place a valve between the sputter chamber and the pump. Valve off the } chamber and leave it under vacuum between uses. As long as it retains any } vacuum this will help when you try to use it again. You don't want to be } pumping on the chamber at the pump's ultimate vacuum for any extended } period of time because that will allow oil vapor from the mechanical pump } to backstream into your sputter chamber and possibly onto your specimen if } present! (I am assuming that you have an oil based pump.) It is also nice } to spare the pump the wear and tear of pumping continuously over periods of } disuse. } } OR } 2. Several hours or the day before you want to use it, start pumping on the } sputter chamber while admitting a small flow of argon. The argon is an } important part of this procedure. First of all, the argon will prevent the } backstreaming described above. Second, I have been told that the argon } helps to desorb the water from the chamber surfaces. I don't know if that } is an old wives tale or not, but it does seem reasonable. The argon flow } may also help in purging any condensed vapors from the pump's oil. Don't } use too much argon though or you may overheat your pump. ******************
} While I have done no experimental protocol to prove this schedule is } optimum, during the off season I try to remember to run the sputter } coaters overnight one night a week. This keeps things in good shape. I } have read that the slow pumpdown of unused vacuum systems is caused mostly } by water vapor adsorbed on interior surfaces, and by traces of moisture in } the pump oil.
Gary P. Radice gradice-at-richmond.edu Associate Professor of Biology 804 289 8107 (voice) University of Richmond 804 289 8233 (FAX) Richmond VA 23173 http://www.science.richmond.edu/~radice
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Experience in electron microscopy and a Bachelor's degree in a science or related field required. Degree in a biological science, experience in light microscopy, and familiarity with microcomputers preferred. Operates electron microscopes, prepares specimens for microscopy, produces photographic and digital micrographs, analyzes data and maintains accurate work records. Necessary training will be provided. Applicants must submit a University of Utah Application for Employment.
Erik M. Jorgensen, Ph.D. Assistant Professor Department of Biology University of Utah 257 South 1400 East Salt Lake City, UT 84112-0840 PHN: (801) 585-3517 FAX: (801) 581-4668 jorgensen-at-biology.utah.edu
Yes, we find that sputter coaters do not like being ignored for long periods. Probably, the specimen chamber and vac lines adsorb moisture and other gases from the laboratory and it takes the rotary pump considerably longer to pump down (many hours versus 15-20 minutes).
You should pump the system down at least weekly for at least an hour. After pumping for about 30 minutes, allow the Argon gas to leak through the system. This really purges the residual gases. Then, we find that if you fill the system with Argon, rather than letting it fill with room air, it will take considerably less time to get into a usable range next time.
John B.
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Dear experts,
I know I should be kept out of the lab but I had to do some special resin embedding. The problem is that I didn't read my own instructions and made up Spurr resin by adding all the ingredients together and then mixing. This means I added the DMAE before mixing the other components and have ended up with brittle blocks of inconsistant hardness.
I never thought I would be asking this but is there any way that I can recover these blocks to allow me to examine what I have embedded? 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
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id TAA25040 for dist-Microscopy; Tue, 9 May 2000 19:52:21 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id TAA25034 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 9 May 2000 19:51:50 -0500 (CDT) Received: from ulu.pbrc.hawaii.edu (ulu.pbrc.hawaii.edu [128.171.22.11]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id TAA25027 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 9 May 2000 19:51:39 -0500 (CDT) Received: from halia.pbrc.hawaii.edu (halia [128.171.22.7]) by ulu.pbrc.hawaii.edu (8.8.8+Sun/8.8.8) with ESMTP id OAA04064 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 9 May 2000 14:46:15 -1000 (HST) Received: from localhost (tina-at-localhost) by halia.pbrc.hawaii.edu (8.8.8+Sun/8.8.8) with ESMTP id OAA16907 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 9 May 2000 14:46:14 -1000 (HST) X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs
Hi, all you experts...
In spite of the digital camera on our LEO 912 EFTEM, we have a couple of users who are going through huge quantities of film. I need to set up an evacuated film dessicator (separate from the one on our older TEM), but I find the non-glass, non-clear-plastic vacuum dessicators in the catalogs at hand to be enormously expensive. Does anyone have a favorite vendor and model, or a kludge? I remember one at Berkeley that I think was made out of a pressure cooker hooked to a vacuum pump...
Mahalo! Tina
80 degrees F, sunny blue skies, everything in bloom, and promise of surf.
**************************************************************************** * 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'm looking for a working used sputter coater for a descent price or donation for a university lab. Please contact me at bcraft-at-uci.edu if you have one available.
While everyone is focussed on the ease with which viruses can be transmitted as and within attachments, maybe it's a good time to ask that postings to the list be only as text messages, not as attachments.
cheers
rtch
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
Paul, I am not writing because of your "Dear experts" address. An expert is a squirt under pressure, maybe that suited better the person who mis-mixed the Spurr's. Long time ago I read a note that brittle blocks sectioned better after soaking them overnight in ethanol. I've never tried that, but there is a possibility. Please let us know if that method is any good. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Wednesday, May 10, 2000 4:09 AM, Paul Webster [SMTP:pwebster-at-mailhouse.hei.org] wrote: } } } Dear experts, } } I know I should be kept out of the lab but I had to do some special resin } embedding. } The problem is that I didn't read my own instructions and made up Spurr resin } by adding all the ingredients together and then mixing. This means I added } the DMAE before mixing the other components and have ended up with brittle } blocks of inconsistant hardness. } } I never thought I would be asking this but is there any way that I can recover } these blocks to allow me to examine what I have embedded? } 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 }
Margaret, Your message struck a chord here, and I would like to extend my heartfelt sympathy. We've been there too. I have reached the point where all the suppliers I contact have a 15-year history of preparing quotations and tenders almost completely in vain. I don't know why the sales reps. (or I) put up with it. That they do is a triumph of hope over realistic expectation, and therefore a credit to all of them. I think there should be a club for stressed out managers of poorly- funded EM facilities. I would be one of the first to join. perhaps some day we should organise a world congress if we can find a venue large enough, thoough whether it would be advisable to have a trade exhibition is open to question. Best wishes Chris
Date sent: Thu, 11 May 2000 14:43:05 -0400 To: Microscopy-at-sparc5.microscopy.com } From: Margaret Dienelt {brannign-at-asrr.arsusda.gov}
Dear Colleagues,
I want to test a new computer program of mine that processes electron diffraction ring-patterns from polycrystalline samples. I did test it with patterns recorded on film. However, I would also like to test it on energy filtered patterns that were recorded with a CCD (or imaging plate).
Could anyone of you send me such patterns from a single phase material with random orientation? If you also characterized the same sample (especially if you proved that the sample is not textured) and you published the results, I could reference this publication of yours.
Thank you in advance.
Dr. Janos L. Labar Research Institute for Technical Physics and Materials Science H-1121 Budapest, Konkoly-Thege u. 29-33 Postal address: H-1525 Budapest-114, Po Box 49 Tel: (36)(1) 392-26-92 Fax: (36)(1) 275-49-96 Fax/phone: (36)(1) 395-92-32 home page: www.mfa.kfki.hu/~labar
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Your concerns and frustrations expressed are a recurrent thread on this site. It would be nice to be able to sit down with other facility managers and discuss common problems and possible solutions. I think our common concerns are such that they effect both materials and biological facilities. I am thinking of things such as: use guidelines, multi-user vs. service functions, formal courses and informal instruction for new users, equipment maintenance costs, justification of new equipment needs to administrators, advisor committee formats, funding sources, etc. Perhaps we could arrange such an informal session at the upcoming M&M meeting. If you would be interested in such a session, take a look at the meeting schedule and suggest a time. Then I will ask the organizing committee to designate a room. 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
--------------------------------------
Margaret, Your message struck a chord here, and I would like to extend my heartfelt sympathy. We've been there too. I have reached the point where all the suppliers I contact have a 15-year history of preparing quotations and tenders almost completely in vain. I don't know why the sales reps. (or I) put up with it. That they do is a triumph of hope over realistic expectation, and therefore a credit to all of them. I think there should be a club for stressed out managers of poorly- funded EM facilities. I would be one of the first to join. perhaps some day we should organise a world congress if we can find a venue large enough, thoough whether it would be advisable to have a trade exhibition is open to question. Best wishes Chris
Date sent: Thu, 11 May 2000 14:43:05 -0400 To: Microscopy-at-sparc5.microscopy.com } From: Margaret Dienelt {brannign-at-asrr.arsusda.gov}
recall reading of ibm's hunt for a silicon wafer cleaving tool capable of handling samples 5mm in diameter. any luck out there?
mark riggs svg lithography wilton, ct 06897 riggsm-at-svg.com
We are contrasting connective tissue filaments by negative stain technique using uranyl formate. Our protocol is to adsorb filaments onto carbon film coated grids, then to wash in two drops of water and two drops of uranyl formate, removing each drop using filter paper but not allowing the grid to dry until after the second drop of UF. We can not charge the grid surface prior to specimen adsorption or our specimens will not adsorb.
My question is to do with making uranyl formate. Our formula is to boil 5ml water, add .0375 g uranyl formate (our solid is very old...), stir for 20 minutes, then add 10 microlitter 5 M NaOH, stir for 20 minutes, then filter through a 0.1 micron before use. We are not getting a consistant staining pattern. Our wish is to see a uniform coating of stain and we seldom see even single grid squares evenly coated. We've tried different concentrations of stain and also different volumes of NaOH. Any suggestions? Does anyone know the purpose of boiling the water?
Thanks in advance,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Research Facilities 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 phone: 503-221-3434 FAX: 503-412-6894 (9-5 PST) e-mail: DRK-at-shcc.org
As a former user and current vendor of such systems as you are inquiring about I can try to provide a bit of information regarding camera improvements:
There have been a number of improvements, but I am not sure what you are comparing the latest cameras against. Cameras are now usually cooled and provide 12 bits per pixel, the number of pixels has gone up a bit (but not much in general), and cameras read out faster than they used to (up to 20 fps and more). I think all cameras now use a line transfer mechanism, which makes shutters obsolete. On the software side, real-time FFT and real-time shading correction can be done now due to faster computers without special processing boards, and there have been other software developments that make using the cameras and computers easier. Other changes that affect the usability of cameras is the use of pneumatics to insert and retract the phosphors, higher frame rates for live viewing with the camera, etc.
If you have questions, please give me a call, drop me an email, or go to our web site.
Michael
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: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Margaret Dienelt [mailto:brannign-at-asrr.arsusda.gov] Sent: Thursday, May 11, 2000 12:43 PM To: Microscopy-at-sparc5.microscopy.com
Hi,
Year after year I hopefully gather information about digital imaging systems for TEMs (ours is a JEOL 100CX) , only to learn we have no money. This year it looks like it might really happen but I have not kept up with innovations in the field and am wondering the following:
1. Anything new in the last two years -- especially in terms of cameras? I'm most familiar with the Gatan and AMT systems but their web sites don't reflect much in the way of changes over a year ago. 2. With more and more microscopists finally getting their systems -- I'd love to get feedback.
Thanks, Margaret
P.S. Would welcome contacts from vendors.
-- Margaret Dienelt
Plant Pathologist Electron Microscopy Lab
Floral and Nursery Plants Research Unit U.S. National Arboretum/Agricultural Research Service/USDA
B. 010A, Rm. 238, BARC-W 10300 Baltimore Avenue Beltsville MD. 20705 USA
Patrick Echlin from Cambridge UK noted in private message that "strong" agent who may dissolve epoxy is sodium methoxide. Thanks, Patrick.
Sergey.
} Date: Thu, 11 May 2000 16:06:39 -0700 } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: Re: Spurr resin problem } X-Sender: sryazant-at-pop.ben2.ucla.edu } To: Microscopy-at-sparc5.microscopy.com } X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
My procedure for uranyl formate is a little bit simpler than yours: 0.05-0.1 g uranyl formate (0.5-1% final) + 10 ml deinozed (cell culture quality or double distilled in the glass) water in the 15 ml plastic tube. It dissolved at the same speed (even better) as acetate salt (UA). Usually I am using rotator to shake slowly the tube with stain. It takes 0.5-1 hour to dissolve salt completely. I do not filter solution yet. The difference between formate and acetate salts of the uranium is that formate is light sensitive (UA - too, but less, less sensitive). You have to avoid direct high intensity light. Usually I wrapped tube in alumina foil and prepared the samples under diffused light moderate intensity (general illumination in the lab, no local lights). Staining procedure is exact the same as for acetate salt. The advantage of formate salt - it generates smaller granularity (and less contrast than UA), spreaded sometime better than acetate salt, and pH is higher. The disadvantage of the formate is that the water solution is not stable: I do prepare fresh solution every time I have to work with it. This is great disadvantage of the uranyl formate. I guess, you may store solution in the dark at +4oC for couple of days, but this is your own risk to experiment with that. As for staining procedure, I would avoid any washes with just water. As a biochemist I am under impression that ionic conditions is important to preserve "native" structure of the sample. Therefore I am using the same buffer as for sample to wash (usually I do not wash at all). Of coarse for any uranium salts you have to avoid any phosphates in the buffer. Any Tris, MES, HEPES buffers may be the good point to start. I don't know exactly how it works, but it seems to me, that buffer in the wash may help spread satin better (don't ask me why, I have no idea). If you have problem to dissolve uranyl formate, you probably have to replace it on the fresh one (it is cheap). Double-carbon technique may also help (you may call off line for details). Good luck and sorry for the long message.
Sergey
} Date: Fri, 12 May 2000 13:00:33 -0700 (Pacific Daylight Time) } From: Douglas Keene {DRK-at-shcc.org} } Subject: Uranyl Formate } Sender: drk-at-shcc.org } To: microscopy-listserver {Microscopy-at-sparc5.microscopy.com} } Reply-to: DRK-at-shcc.org } X-Mailer: Simeon for Win32 Version 4.1.3 Build (39) } Priority: NORMAL } X-Authentication: none } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
} From Debby Sherman: } } Your concerns and frustrations expressed are a recurrent thread on this } site. It would be nice to be able to sit down with other facility managers } and discuss common problems and possible solutions. I think our common } concerns are such that they effect both materials and biological } facilities. I am thinking of things such as: use guidelines, multi-user } vs. service functions, formal courses and informal instruction for new } users, equipment maintenance costs, justification of new equipment needs } to administrators, advisor committee formats, funding sources, etc. } Perhaps we could arrange such an informal session at the upcoming M&M } meeting. If you would be interested in such a session, take a look at the } meeting schedule and suggest a time. Then I will ask the organizing } committee to designate a room.
} Debby -
It's too late to add to the program now, but I attended a Long Beach 2001 LAC meeting last week, and there's a committee member there who wants to organize something. It's almost too late to add programming even for that one! The solution that I suggested is to start an annual breakfast or lunch for facility managers, taking care to avoid other large scheduled meetings (which aren't listed in the program summary). It might even be possible to get more time Sunday afternoon, pre-opening reception. Contact the LAC chairs for that: Stacie Kirsch for Philly & Zed Mason for Long Beach.
Caroline Schooley
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
University of Connecticut Institute for Materials Science
Postdoctoral Research Position in Electron Microscopy Studies of Fatigue Crack Initiation Sites in Ti-6-4 Alloys
The Institute for Materials Science (IMS) at UConn is an interdisciplinary center with the threefold mission of fostering education, research and outreach in all areas of the materials sciences. Applications are invited for a Postdoctoral Position to study the microcrystallography of fatigue crack initiation sites in Ti 6-4 alloys. The appointee will be involved in electron microscopy studies of failed test pieces produced at Pratt and Whitney in the previous phase of this program. It is envisaged that this work will involve extensive SEM and TEM studies in the IMS at UConn with some use of the FIB/TEM/STEM and OIM facilities in the High Temperature Materials Laboratory at Oak Ridge National Laboratory. Candidates should hold a PhD in Materials Science, Physics or a related discipline and must have extensive hands-on experience in a broad range of electron microscopy techniques. Experience in the assessment of deformation substructures would also be beneficial. The appointment is for one year in the first instance and is available from June 1st. Screening of the applications will begin immediately and will continue until the post is filled.
Applications from under-represented groups, including minorities, women and people with disabilities are encouraged.
Interested candidates should send a curriculum vitae, including publication list, and the names of at least three referees with postal addresses, telephone numbers and Email addresses to: Dr. M. Aindow, Institute for Materials Science, University of Connecticut, 97 North Eagleville Road, U-3136, Storrs, CT 06269-3136 USA. Email: maindow-at-ims.uconn.edu -- *********************************************************
Mark Aindow, Associate Professor, Department of Metallurgy and Materials Engineering Institute of Materials Science, U-136 University of Connecticut, Storrs, CT 06269-3136, USA
I have a question concerning sample preparation for brightfield and polarized particle size analysis. My customer is trying to devise an experiment to analyze fly ash collected on a filter during smokestack emissions testing. He collects ~1gr. per sample, however, more is easily possible. The sample is clumped and incongruent and he would like a simple protocol for proper sample dispersion on a slide.
Hello there everyone, I am having hard time cutting gold coated polypropylene (pp) wires. I am using a diamond knife, I have embedded the samples into epoxy and hardener, I will be doing TEM. But when I cut, I get good thickness but the slice keep rolling up. I have been trying for the past three days and do not have any luck. I have change the ratio of the epoxy to hardener from10/4 to 10/6 still no luck. Can anyone suggest some changes. Is cryogenic an option here.
Thank you all,
Briget Ngampa 28 Cedar street LOWELL,MA 01852 Tel (978) 970 0433 Fax (978) 937 2297 EMAIL: hdmhos-at-aol.com
Hi, You may want to check SELA. They have an evergrowing line of cleavers for the semiconductor industry. Contact Efrat Raz: efrat-at-sela.com
Caveat: MME has no financial interest in this product
Good hunting Barbara Foster,President Microscopy/Microscopy Education 125 Paridon Street Suite 102 Springfield, MA 01118-2130 PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com Website: www.MME-Microscopy.com/education
America's first national consortium of microscopy specialists offering customized on-site training in all areas of microscopy, image analysis, and sample preparation ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
At 03:01 PM 5/12/00 -0400, Mark Riggs wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Hildy hello, } } I could not understand, how polymerized (mean crosslinked) epoxy may be } dissolved back in PO? You have to break chemical bonds between polymer's } chains first and than it will become soluble. I believe, there are some } very strong oxidizing agents should be used in order to break chemical } bonds in the epoxies. } } Sergey. } } } } } Date: Thu, 11 May 2000 11:28:43 -0600 (MDT) } } From: HILDEGARD CROWLEY {hcrowley-at-du.edu} } } Subject: Re: Spurr resin problem } } X-Sender: hcrowley-at-odin.cair.du.edu } } To: Paul Webster {pwebster-at-mailhouse.hei.org} } } Cc: MSA listserver submission {Microscopy-at-sparc5.microscopy.com} } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } } } } } On Tue, 9 May 2000, Paul Webster 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 experts, } } } } } } I know I should be kept out of the lab but I had to do some special } resin embedding. } } } The problem is that I didn't read my own instructions and made up Spurr } resin by adding all the ingredients together and then mixing. This means I } added the DMAE before mixing the other components and have ended up with } brittle blocks of inconsistant hardness. } } } } } } I never thought I would be asking this but is there any way that I can } recover these blocks to allow me to examine what I have embedded? } } } 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 } } } } } } } } } } } Hi, } } } } First try this: Get a beaker of water, heat it to the highest temperature } } at which your blocks were polymerized, then subtract 5 deg. After temp } } has been reached, put in one block. Leave it for 15 min. Take it out, } } trim it, and section it immediately. Too brittle? Repeat the water soak } } for 15 min. Repeat again. Not much can be accomplished after one hour of } } soaking, but this may be different in your case. Sometimes in desperation } } when I wanted 4 micrometer thick sections from difficult material I have } } soaked blocks overnight with good success. } } } } One time I was given immensely valuable blocks which were so bad (kind } } expression) that they curled my hair. They could not be cut. I had to } } get the epoxy out and reembed. (The micrographs later ended up in a } } publication in the Comp. Neurol. Journal) } } } } What I did was to rotate the blocks in vials continuously in the REVERSE } } order in which they were embedded, with much elongated times. That is, } } first they went into PO and epoxy (same formulation as the bad embedment) } } 3:1, for hours, the 2:1 for the rest of the day, then 1:1 for overnight, } } then pure PO for a whole day. All the above were done with numerous } } changes. Once I was in PO, pure for a day, most of the epoxy had left the } } tissue. I then remembedded the same way I deembedded. What a pain. The } } blocks were never wonderful, but they sectioned OK and went for } } publication. It should work for Spurr's. Don't give up. When } } deembedding, I left out the accelerator, of course. It helps to have some } } very good chocolate on hand for this maneuver. } } } } Good luck, } } Hildy } } } } Hildegard H. Crowley } } University of Denver } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant } } } } Hi,
Yes, it is a mystery to me that one can get old epoxy out of tissue with PO. However, I have done it at least 3 times with enough success to collect data. The only thing I can think of is that at least 10% of monomers never bind due to the low embed temps we use for our TEM work. Those will surely dissolve out leaving holes. (I have seen this). PO is the simplest of epoxies and a very strong solvent. That is all I know. I never do the "REVERSE" embed unless forced to do it, because it is so difficult dealing with the final cutting and the staining. And then the "new" sections are unstable and uneven. I would rather clean a bathroom! Bye, Hildy
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Patrick Echlin from Cambridge UK noted in private message that "strong" } agent who may dissolve epoxy is sodium methoxide. Thanks, Patrick. } } Sergey. } } } } Date: Thu, 11 May 2000 16:06:39 -0700 } } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } } Subject: Re: Spurr resin problem } } X-Sender: sryazant-at-pop.ben2.ucla.edu } } To: Microscopy-at-sparc5.microscopy.com } } X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (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 } } -----------------------------------------------------------------------. } } } } } } Hildy hello, } } } } I could not understand, how polymerized (mean crosslinked) epoxy may be } } dissolved back in PO? You have to break chemical bonds between polymer's } } chains first and than it will become soluble. I believe, there are some } } very strong oxidizing agents should be used in order to break chemical } } bonds in the epoxies. } } } } Sergey. } } } } } } } } } Date: Thu, 11 May 2000 11:28:43 -0600 (MDT) } } } From: HILDEGARD CROWLEY {hcrowley-at-du.edu} } } } Subject: Re: Spurr resin problem } } } X-Sender: hcrowley-at-odin.cair.du.edu } } } To: Paul Webster {pwebster-at-mailhouse.hei.org} } } } Cc: MSA listserver submission {Microscopy-at-sparc5.microscopy.com} } } } } } } ------------------------------------------------------------------------ } } } 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 } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } On Tue, 9 May 2000, Paul Webster 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 experts, } } } } } } } } I know I should be kept out of the lab but I had to do some special } } resin embedding. } } } } The problem is that I didn't read my own instructions and made up Spurr } } resin by adding all the ingredients together and then mixing. This means I } } added the DMAE before mixing the other components and have ended up with } } brittle blocks of inconsistant hardness. } } } } } } } } I never thought I would be asking this but is there any way that I can } } recover these blocks to allow me to examine what I have embedded? } } } } 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 } } } } } } } } } } } } } } } Hi, } } } } } } First try this: Get a beaker of water, heat it to the highest temperature } } } at which your blocks were polymerized, then subtract 5 deg. After temp } } } has been reached, put in one block. Leave it for 15 min. Take it out, } } } trim it, and section it immediately. Too brittle? Repeat the water soak } } } for 15 min. Repeat again. Not much can be accomplished after one hour of } } } soaking, but this may be different in your case. Sometimes in desperation } } } when I wanted 4 micrometer thick sections from difficult material I have } } } soaked blocks overnight with good success. } } } } } } One time I was given immensely valuable blocks which were so bad (kind } } } expression) that they curled my hair. They could not be cut. I had to } } } get the epoxy out and reembed. (The micrographs later ended up in a } } } publication in the Comp. Neurol. Journal) } } } } } } What I did was to rotate the blocks in vials continuously in the REVERSE } } } order in which they were embedded, with much elongated times. That is, } } } first they went into PO and epoxy (same formulation as the bad embedment) } } } 3:1, for hours, the 2:1 for the rest of the day, then 1:1 for overnight, } } } then pure PO for a whole day. All the above were done with numerous } } } changes. Once I was in PO, pure for a day, most of the epoxy had left the } } } tissue. I then remembedded the same way I deembedded. What a pain. The } } } blocks were never wonderful, but they sectioned OK and went for } } } publication. It should work for Spurr's. Don't give up. When } } } deembedding, I left out the accelerator, of course. It helps to have some } } } very good chocolate on hand for this maneuver. } } } } } } Good luck, } } } Hildy } } } } } } Hildegard H. Crowley } } } University of Denver } } } } } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } http://www.bol.ucla.edu/~sryazant } } } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant } } } } Hi,
Absolutely sodium methoxide will take the epoxy out of the tissue. However, in our laboratory it caused so much tissue damage (since epoxies actually bind with proteins in the tissue and not simply throw a net through and around the tissue like the acrylics) that after reembedding it was not useful for collecting data. Somebody else might have better results than myself with that method, so we should not discard the idea.
Briget: Polypropylene has a Tg of about -19C. Whenever I microtome I always cut at least 15-20 degrees below the Tg. If you are trying to cut these samples at room temperature my guess is this is your problem. If you need to embed you can still do this when cutting at cryo temperatures by trimming as much of the epoxy away as possible. If you leave a very thin layer of epoxy around your sample you should still get good sections even though you will get some chatter in the epoxy region. Steve
Hello there everyone, I am having hard time cutting gold coated polypropylene (pp) wires. I am using a diamond knife, I have embedded the samples into epoxy and hardener, I will be doing TEM. But when I cut, I get good thickness but the slice keep rolling up. I have been trying for the past three days and do not have any luck. I have change the ratio of the epoxy to hardener from10/4 to 10/6 still no luck. Can anyone suggest some changes. Is cryogenic an option here.
Thank you all,
Briget Ngampa 28 Cedar street LOWELL,MA 01852 Tel (978) 970 0433 Fax (978) 937 2297 EMAIL: hdmhos-at-aol.com
Stephen McCartney Research Associate 2108 Hahn Hall Materials Institute Virginia Tech Blacksburg, VA 24061-0344 USA
A newly appointed researcher here has asked my advice on several pieces of TEM spec. prep equipment. I turn to you for helpful suggestions.
The research involves serial sectioning biological tissues and many grids. EM is a minor, but essential component of the project. The lab runs through dozens of grids and hundreds of pictures, but only 4 - 6 times a year. Big bursts of activity followed by long periods of analysis and investigations using other techniques. Of the hundreds of pictures taken, they may only use a few for data.
The researcher is looking for ideas on what choices are available, how useful, and approximate costs of the following:
Digital imaging to add to our existing JEOL 1200EX TEM - I have seen other threads on this topic and would welcome any new ideas. Digital imaging will save a lot of time and money since they discard so many pictuers, but the question of image quality is one I am to investigate.
Ultra microtome - We have an older A/O Ultracut (the model before it became the Reichert Ultracut E) which is OK for the sectioning we do in the general lab. But she wants a new one for her exclusive use. The question is whether a new microtome will allow folks in her lab to do serial sectioning any faster or easier, or by less skilled users, than our current system.
Staining machine - Anyone have info on staining machines or systems for lots of TEM grids. I have never had to do so many grids that this was an issue, so I have never kept up on the offerings. If you know of something and/or have experience let me know. Again, this is something she would keep in her lab.
Tissue processing machine - Same as above for me, never did so much at one time that I ever thought I needed one of these. The samples to be processed are C. elegans, anything available that could do these unattended? How about upkeep and volumes of chemicals needed. Also an item to be kept in her lab.
I will filter and pass on your comments. Anything you might offer will, as always, be received with appreciation and thanks.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Jon, I can speak to the digital camera at least. We have been using a Gatan BioScan on our microscope and it serves our needs for 95% of the images that we produce. It is a very good system, when it is working. We have had more problems with it than I would have expected. We bought the system early in the production and perhaps they have worked out the bugs by now. -------------------------------------.
} Dear List: } } A newly appointed researcher here has asked my advice on several pieces of } TEM spec. prep equipment. I turn to you for helpful suggestions. } } The research involves serial sectioning biological tissues and many grids. } EM is a minor, but essential component of the project. The lab runs through } dozens of grids and hundreds of pictures, but only 4 - 6 times a year. Big } bursts of activity followed by long periods of analysis and investigations } using other techniques. Of the hundreds of pictures taken, they may only } use a few for data. } } The researcher is looking for ideas on what choices are available, how } useful, and approximate costs of the following: } } Digital imaging to add to our existing JEOL 1200EX TEM - I have seen other } threads on this topic and would welcome any new ideas. Digital imaging will } save a lot of time and money since they discard so many pictuers, but the } question of image quality is one I am to investigate. } } Ultra microtome - We have an older A/O Ultracut (the model before it became } the Reichert Ultracut E) which is OK for the sectioning we do in the } general lab. But she wants a new one for her exclusive use. The question is } whether a new microtome will allow folks in her lab to do serial } sectioning any faster or easier, or by less skilled users, than our current } system. } } Staining machine - Anyone have info on staining machines or systems for } lots of TEM grids. I have never had to do so many grids that this was an } issue, so I have never kept up on the offerings. If you know of something } and/or have experience let me know. Again, this is something she would keep } in her lab. } } Tissue processing machine - Same as above for me, never did so much at one } time that I ever thought I needed one of these. The samples to be processed } are C. elegans, anything available that could do these unattended? How } about upkeep and volumes of chemicals needed. Also an item to be kept in } her lab. } } I will filter and pass on your comments. Anything you might offer will, as } always, be received with appreciation and thanks. } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } }
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
Hi all - a first attempt at unloading a problem onto the listserver!
I am trying to get thin sections of blue-green algae that are loaded with starch granules. The problem is that every time we do the preps, we end up with cells that lose some granules, leaving a gaping hole, or of starch granules that have holes in the centre. I'm using a standard sort of method - fix in phosphate + 2.5% glut 2 hours, osmium 1% 2 hours, dehydrate in ethanol series, finishing with prop. ox, 3:1 prop. ox/resin around 2 h, 1:1 2 or 3h then another change overnight, a couple of 100% resin changes for 1/2 to one day, then polymerization 48h/60C. The fixing and EtOH steps are done on ice, the rest at room temp, the resin infiltration is done on a rotating mixer. We've tweaked the method around but nothing seems to work. I've tried using LR White as well, but with no luck. Can anybody out there help!?
Mark
******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
"I am Mark Armogida, VP of Engineering at Ted Pella, Inc. in Redding, California, USA. We have an old (1964) TEM, Hitachi model Hu 11a, that is a decommissioned instrument which we are trying to give away for removal/shipping costs. It was operational a year ago and kept under vacuum since then. We have the instruction manual. This unit consists of the main body with diffusion pumps, a large electronics console, a large oil filled resistor and mechanical pumps. The oil was analyzed and does not contain PCB's. I estimate that this system weighs several thousand pounds. I will have it removed for salvage in a week, so please contact me if you are interested in this system."
Mark Armogida VP, Engineering & Production Ted Pella, Inc.
I am sending this for a colleague. It was a wonderful little scope while I was there. } LifeCell Corporation (The Woodlands, TX and Branchburg, NJ) is closing their facility in The Woodlands. } LifeCell has available for donation a Philips EM300 TEM. The scope was in } working order when shut down in December 1999. } If you are interested please contact: } Sy Griffey, Ph.D. } 908-947-1143 or sgriffey-at-lifecell.com }
At 01:38 PM 5/10/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It seems to me that no digital camera system would work on a SEM in place of a Polaroid or other film-based output device. Since the recording CRT in a SEM is based on a sequential line scan, one would need a camera that would capture each line as it is produced. Most digital backs are single or triple pass units of a single linear set of sensors. There are other cameras that do snapshot capture but even these would not work since the whole image is not present on the record CRT at the time of taking a picture with the digital camera. The final image is generated sequentially, line by line, on the record CRT.
If the SEM image is stored in a frame buffer, the buffer can be converted to RS-170 TV video and frame grabbed. But the best that this would typically do is 640 lines.
Its an interesting problem and dilemma about being in a situation where digital camera products simply won't work in place of film. But since the goal is to obtain a digital file, why not start with a digital interface? For example, a passive digital capture system would transfer the record CRT information to computer and directly result in a nice digital file. Alternatively, for some systems, an active system can be applied to directly control the SEM's beam. In doing so, the range of final digital image resolution is limited only by the attached hardware system.
gary g.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I am Mark Armogida, VP of Engineering at Ted Pella, Inc. in Redding, California, USA. We have an old (1964) TEM, Hitachi model Hu 11a, that is a decommissioned instrument which we are trying to give away for removal/shipping costs. It was operational a year ago and kept under vacuum since then. We have the instruction manual. This unit consists of the main body with diffusion pumps, a large electronics console, a large oil filled resistor and mechanical pumps. The oil was analyzed and does not contain PCB's. I estimate that this system weighs several thousand pounds. I will have it removed for salvage in a week, so please contact me if you are interested in this system. I can be reached by e-mail or by phone at 530-241-2200 ext 212 between the hours of 8:00am and 5:00pm pacific time zone.
Electron Microscopist / Materials Scientist Materials Science Division, Argonne National Laboratory
The Materials Science Division at Argonne National Laboratory invites applications for a Staff Scientist position in Electron Microscopy. The candidate should have a strong background in the various techniques of electron microscopy and a very strong interest in application of these techniques in materials science. The candidate should have a state-of-the-art knowledge of either analytical transmission electron microscopy including high spatial resolution x-ray and electron spectroscopy or the application of electron holography and Lorentz imaging, with a particular emphasis on field emission (S)TEM. Candidates with exceptional experience in other areas of microscopy will also be considered.
Areas of particular research interest include defects and interfaces in materials, in situ studies of critical phenomena, irradiation and ion implantation effects, and applications of electron holography and Lorentz imaging. A familiarity with research in one or more of the following areas is highly desirable: magnetic and superconducting materials, irradiation effects, ferroelectrics, nanoscale materials, diamond films, and non-crystalline materials. The successful candidate will work closely with Electron Microscopy Center personnel and other research groups within the Materials Science Division to develop strong research efforts in one or more of these areas.
Interested candidates should send a curriculum vitae, a brief statement of research interests and plans, and the names and contact information of three references to: Susan Walker, Employment and Placement Box MSD-210121 9700 S. Cass Avenue Argonne, IL 60439 TDD: 630-252-7722
Questions can be addressed to Dr. Dean J. Miller, Materials Science Division, Argonne National Laboratory, 9700 S. Cass Ave., Argonne, IL 60439, tel. 630-252-4108 (with voice mail), fax 630-252-7529, or miller-at-anl.gov.
Argonne National Laboratory is a multidisciplinary center of energy research and related scientific studies and is operated by the University of Chicago for the U.S. Department of Energy. Argonne National Laboratory is a federal contractor and complies with all federal contractor rules and regulations regarding the maintenance and implementation of our Affirmative Action Program.
--------------------------- Dean J. Miller Materials Science Division Argonne National Laboratory Argonne, IL 60439
Dr. Gary Gaugler 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 } -----------------------------------------------------------------------. } } At 01:38 PM 5/10/00, you 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 curious ... has anyone tried simply installing a digital } } 4x5 back in place of a Polaroid 4x5 back??? For example, see: } } } } http://www.phaseone.com/brochures/powerfx.html } } } } cheerios, shAf } } It seems to me that no digital camera system would work on a SEM } in place of a Polaroid or other film-based output device. Since the } recording CRT in a SEM is based on a sequential line scan, one } would need a camera that would capture each line as it is produced. } Most digital backs are single or triple pass units of a single linear } set of sensors. There are other cameras that do snapshot capture } but even these would not work since the whole image is not present } on the record CRT at the time of taking a picture with the digital } camera. The final image is generated sequentially, line by line, } on the record CRT. } } If the SEM image is stored in a frame buffer, the buffer can be } converted to RS-170 TV video and frame grabbed. But the } best that this would typically do is 640 lines. } } Its an interesting problem and dilemma about being in a situation } where digital camera products simply won't work in place of } film. But since the goal is to obtain a digital file, why not start } with a digital interface? For example, a passive digital capture } system would transfer the record CRT information to computer } and directly result in a nice digital file. Alternatively, for some } systems, an active system can be applied to directly control the } SEM's beam. In doing so, the range of final digital image } resolution is limited only by the attached hardware system. } } gary g. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Modern surfers use PC boards. You can too at } http://photoweb.net } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Shaf, Aside from the problems already mentioned, the back alone (without computer) is almost as expensive as the smallest SEMs. It also has about 4 times the resolution of the best recording systems out there (ETEC) so what you'd get is a lot of empty (information-wise) pixels. It might be adaptable to TEM, though.
For an SEM an active digital control could be set up to gather 10K x 10K images, but the stability of the column drivers becomes very important. Some could handle it fairly well while some would again be useless because their drivers have no noise immunity due to the fact that their push-pull mag drivers were designed backwards and all powere supply noise is passed directly to the scan coils.
Ken Converse Quality Images third party SEM service Delta, PA
Embedding and sectioning material containing a lot of starch is always a problem. If you do not need to look at the structure of the mature starch grains, you could try embedding the algae first thing in the morning before they have had time to produce the starch. Most plants have a low starch content after being kept in the dark for approx 12 hours. For instance, active, growing leaves are difficult to section when collected midday, but early morning collecting makes for easy sectioning.
Jan Coetzee
Mark West wrote:
} I am trying to get thin sections of blue-green algae that are loaded with } starch granules. The problem is that every time we do the preps, we end up } with cells that lose some granules, leaving a gaping hole, or of starch } granules that have holes in the centre. I'm using a standard sort of } method - fix in phosphate + 2.5% glut 2 hours, osmium 1% 2 hours, } dehydrate in ethanol series, finishing with prop. ox, 3:1 prop. ox/resin } around 2 h, 1:1 2 or 3h then another change overnight, a couple of 100% } resin changes for 1/2 to one day, then polymerization 48h/60C. The fixing } and EtOH steps are done on ice, the rest at room temp, the resin } infiltration is done on a rotating mixer. We've tweaked the method around } but nothing seems to work. I've tried using LR White as well, but with no } luck. Can anybody out there help!? }
-- Prof Jan Coetzee Lab for Microscopy and Microanalysis University of Pretoria, South Africa. Tel: 012-420-2075, Fax 012-362-5150 www.up.ac.za/science/electron/emunit1.htm
Question: Dear sir In order to test the quality of difraction limited microscope I am looking for reticles that will enable me to produce difraction patterns of about 0.2 micron source can you advice where can I find such reticles . Who can produce them for me if they are not available comercially . Best regards Moshe Marcovitch
I've seen references to using potassium ferrocyanide reduced osmium tetroxide to help preserve glycogen. Karnovsky(1971) Use of ferrocyanide-reduced OsO4 in EM. In Proc.14th Annu.Meet. Am. Soc. Cell Biol., p. 146. Abstract 284.
Tracy Gales Fox Chase Cancer Center Electron Microscopy Facility 7701 Burholme Ave. Philadelphia PA 19111 Tel. 215-728-2484 Fax 215-728-2412
At 04:02 PM 5/15/00 -0700, Dr. Gary Gaugler wrote: } It seems to me that no digital camera system would work on a SEM } in place of a Polaroid or other film-based output device. Since the } recording CRT in a SEM is based on a sequential line scan, one } would need a camera that would capture each line as it is produced.
What you're saying is there must be a system out there that digitizes that single stream of line scan intensities, then processes all that data inside the computer as an image as opposed to trying to digitize the frame buffer.
Try Klarmann Rulings, Inc., they manufacture reticles. If not a stock item they will make one to your specification.
There URL is http://www.reticles.com
Best Regards
Joseph Passero
mailto:jp-at-spacelab.net
On Tuesday the 16th of May, 2000 at 07:39:23 -0500, moshe_marc-at-gohip.com wrote and posted:
} Email: moshe_marc-at-gohip.com } Name: moshe marcovitch } } Question: Dear sir } In order to test the quality of difraction limited microscope I am looking } for reticles that will enable me to produce difraction patterns of about } 0.2 micron source can you advice where can I find such reticles . Who can } produce them for me if they are not available comercially . } Best regards } Moshe Marcovitch } } ---------------------------------------------------------------------------
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Dear colleagues there are new websites names available. It will look like: www.diptera.ws or www.????.ws more information you can find on www.coleoptera.org in section {software house} Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Collorado-Mansfeld
website: http://www.coleoptera.org listserver: coleoptera on www.egroup.com/group/coleoptera/info.html Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 email: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
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In response to the current thread on the problems of facility managers, I say count me in as being highly interested! If a discussion group does get together at M&M it would be great to see a report posted on this listserver for those of us who unfortunately can't attend the meeting. If somebody could take notes and post them, I for one would be very appreciative.
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Mark, I have been working with cyanobacteria for over 15 years. We found out in the late 80's that the only way to really hold the starch granules together is by using plunge freezing and freeze substitution techniques. I would substitute in acetone + osmium and embed in Spurr's for normal ultrastructure and use ETOH and embed in HM-20 for immuno. Check out the following references for details of method and contact me for further explanations:
Schneegart, M.A., D. M. Sherman, S. Nayar, and L. A. Sherman (1994). Oscillating Behavior of Carbohydrate Granule Formation and Dinitrogen Fixation in the Cyanobacterium Cyanothese sp. strain ATCC 51142. J. Bacteriology. 176:1586-1597.
Sherman, D. M., T. Troyan, and L. A. Sherman (1994). Localization of Membrane Proteins in the Cyanobacterium Synechococcus sp. PCC7942. Plant Physiol. 106:251-262
Plunge freezing apparatus can be made in a university shop at relatively low cost and works great for many unicellular organisms. 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
--------------------------------------
Hi all - a first attempt at unloading a problem onto the listserver!
I am trying to get thin sections of blue-green algae that are loaded with starch granules. The problem is that every time we do the preps, we end up with cells that lose some granules, leaving a gaping hole, or of starch granules that have holes in the centre. I'm using a standard sort of method - fix in phosphate + 2.5% glut 2 hours, osmium 1% 2 hours, dehydrate in ethanol series, finishing with prop. ox, 3:1 prop. ox/resin around 2 h, 1:1 2 or 3h then another change overnight, a couple of 100% resin changes for 1/2 to one day, then polymerization 48h/60C. The fixing and EtOH steps are done on ice, the rest at room temp, the resin infiltration is done on a rotating mixer. We've tweaked the method around but nothing seems to work. I've tried using LR White as well, but with no luck. Can anybody out there help!?
Mark
******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
Responding to the message of {Pine.WNT.4.10.10005151651240.-3843205-100000-at-mwest.ifisiol.unam.mx} from Mark West {mwest-at-ifcsun1.ifisiol.unam.mx} : } Mark,
If you don't really care about seeing the starch, perhaps you could "light starve" the algae, put them in the dark for some period of time to deplete or minimize granule size of the starch, without killing them of course. Then process as usual. We do this with plant samples to avoid the embedding problems and the resultant holes in the setions that you describe.
Good luck!
Gib
} Hi all - a first attempt at unloading a problem onto the listserver! } } I am trying to get thin sections of blue-green algae that are loaded with } starch granules. The problem is that every time we do the preps, we end up } with cells that lose some granules, leaving a gaping hole, or of starch } granules that have holes in the centre. I'm using a standard sort of } method - fix in phosphate + 2.5% glut 2 hours, osmium 1% 2 hours, } dehydrate in ethanol series, finishing with prop. ox, 3:1 prop. ox/resin } around 2 h, 1:1 2 or 3h then another change overnight, a couple of 100% } resin changes for 1/2 to one day, then polymerization 48h/60C. The fixing } and EtOH steps are done on ice, the rest at room temp, the resin } infiltration is done on a rotating mixer. We've tweaked the method around } but nothing seems to work. I've tried using LR White as well, but with no } luck. Can anybody out there help!? } } Mark ******************************************** } Mark West, } Unidad de Microscopia Electronica, } (Electron Microscopy Unit) } Instituto de Fisiologia Celular, } Universidad Nacional Autonoma de Mexico, } 04510 Mexico D.F. } } tel (unidad/lab) *(525) 622 5610* } (casa/home) (525) 619 3020 } Fax (525) 616 2282 } ********************************************
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.cdl.umn.edu http://biosci.umn.edu/MIC/consortium.html
Me too, though I would like to attend the meeting in Philadelphia. Elaine } } } Collegues, } } In response to the current thread on the problems of facility managers, I } say count me in as being highly interested! If a discussion group does get } together at M&M it would be great to see a report posted on this listserver } for those of us who unfortunately can't attend the meeting. If somebody } could take notes and post them, I for one would be very appreciative. } } Thanks! } Dee
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
In my experience, two weeks of downtime a year is not bad at all, for any EM. Especially when you are dealing with many users with varying levels of expertise and a heavy usage schedule. EM's are maintenance-intensive instruments (especially TEM's), and even performing preventive maintenance routines can easily eat up a week a year.
In my opinion, you're doing great.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: "HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com [mailto:"HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com] Sent: Tuesday, May 23, 2000 5:02 AM To: Microscopy-at-sparc5.microscopy.com
A how long is a piece of string ? type question
I would be pleased if anyone could broaden my views on equipment reliability . Basically we have a 3yr old FESEM which I consider to be fairly reliable in that it is on call 24hrs/day , has numerous ( non dedicated users ) and apart from downtime for filament change and the odd wear and tear type problems answers our needs . As we do not have a back up instrument and when we do experience problems it's always at the worse time certain personnel have the impression that it is unreliable .
What do other sem users expect in terms of reliability , apart from my ' subjective ' comments is it quantifiable , would approx 2wks /year downtime including planned maintenance be considered excessive ?
Two weeks a year (avg) seems like a lot of downtime for a FE-SEM to me. Our Hitachi S-800 SEM, for example, is 13 years old, just recently had only it's second emitter replacement, and, not counting building air, water and power problems, has certainly had only about 4 weeks of instrument-related downtime in that 13 year period, including annual PM services.
I guess it's tough to break an anvil... ;-).
Larry PS it's a heavily used multi-user instrument...
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Gib, Have you thought about microwave fixation in the pressence of aldehyes (para/GA) followed by microwave fixation in osmium. each takes seconds and fixations are as good or better (for rapid fixation) than standard fixation. The key is to keep heat off the sample (use water baths and/or ice bath). The theory is that the microwave pulsations increase the penetration speed of the fixatives. Call Ted Pella's tech divison for more info or protocols.
Dear Woody, I had a problem once that really strained the ultimate Z number sensitivity of my BSE detector. This was a case of a small amount of a marker chemical, I think Strontium, being fed to growing fish and trying to detect it in a scale of the fish. I was definintely able to detect the faint, brighter band on my GW BSE detector (solid state), but not on my Robinson (scinntilator) on the other SEM. I had to use more beam current for the GW, but it saw the contrast when the Robinson didn't. More sensitivity does not mean better Z resolution. At 01:36 PM 5/22/00 -0500, you wrote: } } Hello Gary, } } I have never used a sintillator type BSE detector, but the major differences } are } two fold. } } Typically (though diodes are getting better all the time) the Robinson has a } better low energy BSE detection effiency. It follows that for the same } noise } level electronics, it would exhibit better sensitivity. The dynamic range } problem will still exist for very different Zs in the field of view. In the } middle and upper portions of the sensitivity range (low and lower), I would } not } expect much difference between them. ...Any Comments from users of both???? } } I like the 4 quad diodes since I can go differential mode for macro } topography } (like fracture surfaces) and show the gross features while suppressing the } fine } detail. } } The Pt coating can have a profound negative effect on sensitivity if not } extremely thin. If given a choice, I would always use carbon for the best } BSE } sensitivity. ...If not hazardous, I would use Be, BUT!!!! For elements in } the } range of Al & Si, I prefer to use carbon and lower beam voltages to minimize } penetration, especially if they are films or very small features. } } I once presented some data illustrating the BSE signal attenuation as a } function } of sputtered Au thickness. But that data would be hard to retrieve now. } } Woody White } McDermott Technology
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
Dear Listers, I now know that the size of the gold used in the label was 0.8 nm and the DNA in another exp. will be ssDNA (7249 bases, circular). We're prepared to try 1.4 nm and 3.0 nm and darkfield before shadowing. I have copied the repsonses below and will post the results of our efforts. Once again, I am most grateful for your help. Rosemary
I don't think the PT shadowing would obscuring the gold labelling. I did some rotary shadowing of myosin molecules with antibody attached. You couldn't see the actual Y structure but you could see arrowheads. If you can see isolated IgG molecules you should be able to pick up the gold particles. Patty Jansma
Do you mean you want to see a gold particle in a preparation that is shadowed with Pt after the gold labelling step has take place? If so, the answer is probably "yes." The Pt gives such a high-contrast shadow that it may be difficult to pick out the small gold probe against the contrasty background. Carol Heckman
NO, it should enhance the whole image so that you can more readily see exactly where the gold is labelling. Cheers,Marilyn Henderson
I would suggest to use dark field imaging of your labeled DNA-protein complexes in TEM. By using this technique, you do not need to use Pt- shadowing of your samples. Best regards from Prague. O. Benada
Rosemary, The typical "grain" size using Pt/C is on the order of 0.8 nm. So the answer to your question would be dependent on the size of the gold probe you are using. I would think that you would be OK with a gold probe larger than 3 nm, but this is speculation on my part, I have not actually done that exact thing with my own two hands. Chuck (Charles Garber)
Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
Sure it is possible to melt samples by sputter coating but it is usually when you are using the old fashioned "brute force" systems that use a variac to adjust the operating voltage.
A big problem with sputter coating is that many of the solutions to one problem are the reasons for others!
1. To help prevent melting increase the working distance (} 5cms) and cut down the current (~10mA) - problem this cuts down the coating thickness so I need to coat for longer!
2. To help prevent melting use a number of short coating periods with a cooling down period between - problem multi coats build structure on the specimen and from tests I have conducted the first 10 seconds of the plasma are the hottest! We actually use the multi coat method to make test specimens ( 5 x 1 minute coats at 20mA 5cms working distance with 1 minute between coats).
3. During the early days of SEM sputter coating I would place the sputter head in a refrigerator for an hour prior to use, part of my method in order to tray and track down the heat problems. There was considerably less heating under these circumstances but we were very very careful with condensation on the then "high voltage" connection. This test led us to talk very seriously about water cooling the sputter head, a route that was taken by Baltzers at one stage.
4. My route for a specimen that melts would be to cut down the current, give a longer coating time and provided you were not working above 5,000X have a few minutes "cooling time" between no more than three coating sessions.
Good luck
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
I have been involved with clients around the world since FEG systems first became available. I have found this style of equipment to be very reliable with if anything less down time than the conventional W hairpin instruments.
I must say that I have seen a considerable difference in the performance of these instruments with a certain manufacturer's range of FEG microscopes being far less of a problem in the production of very high quality results than any of the others.
This said if you are only averaging two weeks per year down time there should be no one within your organisation who should complain.
As a consultant and ex service engineer I can only suggest that people look at other instruments within your establishment of equal complexity and compare the up time performances.
If we can help a phone call consultation costs nothing?
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
And another - same as Larry really, we have a four year-old Hitachi 4500 that has had no emitter deterioration and probably 5 days total downtime counting bakeouts. Touch wood. Also in a multiuser facility. But this FESEM series are proverbially reliable and in general two weeks average per year doesnt seem excessive, particularly if the FESEM is only three years old, and some teething problems with any EM column would not be unusual in the first couple of years. Depends on the context what is acceptable I guess, but if absolute reliability is a requirement, 24 hr access by "non-dedicated" users might be the first point to examine? good luck, Sally Stowe
} } } Larry Allard {l2a-at-ornl.gov} 05/24/00 12:21am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Just another opinion...
Two weeks a year (avg) seems like a lot of downtime for a FE-SEM to me. Our Hitachi S-800 SEM, for example, is 13 years old, just recently had only it's second emitter replacement, and, not counting building air, water and power problems, has certainly had only about 4 weeks of instrument-related downtime in that 13 year period, including annual PM services.
I guess it's tough to break an anvil... ;-).
Larry PS it's a heavily used multi-user instrument...
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
I hope that someone can make a suggestion to me. I am looking for a sample to use as a standard for low level nanoindentation.
What would be ideal is a gold film of perhaps 1 micron or more on perhaps polished silicon wafer. This needs to be as smooth as possible, rms roughness of less than a nanometer. I know that this is quite possible, I have samples that are just like this but they are too thin (15nm).
We have a JOEL 5200 SEM with a Link Analytical QX 2000 EDS detector. The problem is that we always get an aluminum peak in every EDS sprectrum regardless of sample & scan size. Does anyone have any suggestions on why this is?
Thanks, Hanson Fong
Univ. of Washington Materials Science & Engineering Department Box 352120 Seattle, WA 98195 USA
This may be off-topic for this listserver but I'll give it a shot for those who are into materials science.
Suppose that you have an etching system based on a small plasma chamber using CF4 and O. Is there some simple/convenient way to measure species during the etching process to indicate that some end point has been reached? i.e., a major etching area has been etched and the nature of the species has dramatically or noticeably changed?
I can think of many x-ray methods, but what I am looking for is basically a sensor of some sort at a port in the etching chamber system. There is no SEM beam--and presumedly, no direct or indirect x-rays.
Are you sure it's Al and not Br? If your accelerating voltage is not high enough to stimulate the emission of Br K radiation, it can be easy to mistake the Br L lines for the K lines of Al which are slightly narrower but at the same peak position. Like the other halogens, Br can initiate corrosion of metals and end up in the corrosion products. And if this metal is part of your detector window.....
John Twilley Art Conservation Scientist
H. Fong 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 have a JOEL 5200 SEM with a Link Analytical QX 2000 EDS detector. The } problem is that we always get an aluminum peak in every EDS sprectrum } regardless of sample & scan size. Does anyone have any suggestions on why } this is? } } Thanks, } Hanson Fong } } Univ. of Washington } Materials Science & Engineering Department } Box 352120 } Seattle, WA 98195 } USA
A change in the absorption spectrum of light could to be used as a cut off. The photon penetration is of the order of the first few monolayers. The difficulty is using sources and filters of the right frequency range that is compatible with the etchants and the material you want to detect (when it becomes exposed).
You would need to find out the optical reflectivity/absorption of the materials involved. Is this feasible? ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 Phone: Microscope room +46 18 471 6365 http://www.angstrom.uu.se/analytical/home.html ********************************************************
Hi Hanson Fong, A possible explanation is that you have a collimator made of aluminium. Either the collimator is not properly aligned, or, the carbon paint inside the collimator has broken. Another explanation can be high energy x-rays hitting the collimator.
Vennlig Hilsen dr.ing Gunnar Kopstad overingeni¿r Avd f Patologi, Rit
Dr. Gary Gaugler 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 } -----------------------------------------------------------------------. } } This may be off-topic for this listserver but I'll give it } a shot for those who are into materials science. } } Suppose that you have an etching system based } on a small plasma chamber using CF4 and O. } Is there some simple/convenient way to measure } species during the etching process to indicate that some } end point has been reached? i.e., a major etching } area has been etched and the nature of the species } has dramatically or noticeably changed? } } I can think of many x-ray methods, but what I am } looking for is basically a sensor of some sort at a } port in the etching chamber system. There is no } SEM beam--and presumedly, no direct or indirect } x-rays. } } Any ideas? } } gary g.
Gary, There are some small, inexpensive mass-specs available that can operate at pressures up to 25 mT. I believe Ferranti in New Mexico is one that I've seen. I don't know what pressure your plasma system operates at, but if it's too high, you'd only need a roughing pump to operate this system.
When I looked into it a few years ago, it was about $3000 plus a computer to plug it into.
H. Fong 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 have a JOEL 5200 SEM with a Link Analytical QX 2000 EDS detector. The } problem is that we always get an aluminum peak in every EDS sprectrum } regardless of sample & scan size. Does anyone have any suggestions on why } this is? } } Thanks, } Hanson Fong } } Univ. of Washington } Materials Science & Engineering Department } Box 352120 } Seattle, WA 98195 } USA
Hanson, I'm not familiar with the 5200 chamber geometry, but I'm sure there is aluminum in there. The question is: can the detector see the aluminum and can the aluminum be excited by either backscattered electrons or x-rays generated by from the specimen? The detector doesn't care or know WHERE the x-rays come from, only that they are being generated and are within line of sight of the detector. Often, moving the detector closer to the specimen and making sure that the collimator is properly placed on the detector nose will help narrow the field of view.
I would check your electron trap and make sure that it is installed correctly, your detector window is almost certainly coated with Aluminum to keep light out, electrons striking the window can produce the effect you are observing. If this is the case you should also be seeing a large hump in the high end of your continuum. Good luck and let the list know what you discover. Scott
} } We have a JOEL 5200 SEM with a Link Analytical QX 2000 EDS detector. The } problem is that we always get an aluminum peak in every EDS sprectrum } regardless of sample & scan size. Does anyone have any suggestions on why } this is? } } Thanks, } Hanson Fong } } Univ. of Washington } Materials Science & Engineering Department } Box 352120 } Seattle, WA 98195 } USA
------------------- note: new mailing address ------------------------ Scott Wight fax: 301-417-1321 NIST 222/A113 W voice: 301-975-3949 100 Bureau Dr STOP 8371 | email: scott.wight-at-nist.gov Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
If you use a tride sputter coater, you should not experience any significant heat build-up during a normal coating process, because the geometry of the sputter target, employing a central magnet element to create a shaped magnetic field, directs the electrons in the plasma away from the sample, thus reducing the "I-square-R" heating. If you are using an older diode sputter coater, it is a simple matter to use a pulsed coating process to reduce heating effects. We used this process many years ago before the triode sputter coaters were introduced. It turns out that a cycle of 1 sec ON, 2 sec OFF (or maybe it was 2 ON, 1 OFF) ended up generating an increase in temperature on an insulating sample only to about 40¡C, or about body temperature (directly measured using thin-wire thermocouples). We found we could coat a piece of styrofoam cup with 200 of gold using this process in a diode sputterer, and see no evidence of melting of the structure.
This is basically the suggestion Steve Chapman makes, to use several brief coating times. The more regulated, very short heating times with some cooling time in between seemed to do the trick for us. Try it, you'll like it... :-).
Larry PS I think we published this somewhere...if I find it, I'll post.
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
How big is the Al peak compared to other peaks? Does it vary according to the sample composition? Do you get it even if there is no sample in place? Are you sure you have the sample height correct (the "nominal" height is often not right - you need to check the X-ray count rate vs. sample height)? Are you certain it is Al (the position is accurate, the peak width right)? Do you always use the same voltage, or is the peak there at different beam voltages? Do you have a thin window or Be detector? Is the detector working normally in all other respects? Has this spurious response always been present (i.e. since installation in the '80's) or have you only recently observed it?
Sorry to ask these questions, but they are relevant.
If there really is an Al peak, it means that within the detector's field of view is something made primarily of Al which is being irradiated either with x-rays or electrons. Assuming the working distence (sample height) is correct, this implies some problem with the collimator, because it's purpose is to eliminate exactly this type of spurious x-ray signal. You do have your original collimator and electron trap, do you? Has it been damaged or moved in some accident with the sample stage?
If the Al signal varies strongly with sample composition (for example, much weaker with a carbon sample than with a tungsten sample) then it could well be related to backscattered electrons or secondary x-ray flourescence.
If, on the other hand, the signal varies strongly with operating voltage (especially if the peak moves) then it probably isn't Al at all, but a spurious response related either to electrons getting through the window or to electron noise.
No solutions here (and some of my thoughts are included for completeness as this is a positing going out for everyone to read), but I hope my musings help.
Tony Garratt-Reed.
At 05:48 PM 05/23/2000 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6479 **
1) You should be able to get some spectral data from the plasma itself. Hook up a spectrometer and you should be able to see the components in the spectrum. You may have to excite the plasma with some light.
2) Hook up a mass spectrometer (quadrupole). That should be able to give you masses of the components.
Don't people do that on a regular basis? You may check the web or literature for "residual gas analysis" or similar.
Michael
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: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Dr. Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Tuesday, May 23, 2000 9:27 PM To: MSA listserver Cc: just_in_case_I_bounce
This may be off-topic for this listserver but I'll give it a shot for those who are into materials science.
Suppose that you have an etching system based on a small plasma chamber using CF4 and O. Is there some simple/convenient way to measure species during the etching process to indicate that some end point has been reached? i.e., a major etching area has been etched and the nature of the species has dramatically or noticeably changed?
I can think of many x-ray methods, but what I am looking for is basically a sensor of some sort at a port in the etching chamber system. There is no SEM beam--and presumedly, no direct or indirect x-rays.
This is the same thing as I am trying to do. In my case, the passivation will be either sinox or PSG. At present, I remove them using two methods. One is to do a short soak in BOE, rinse & dry. Then plasma etch with CF4 at about 80 mTorr. Power and gas flow have dramatic effects on etch rate. Same for dilution of BOE. The challenge is to nicely get through the passivation and stop at the uppermost SiO2 dry ox layer.
The physical size of my specimens is small. A 3" diameter cylindrical chamber would be fine. I use a sputter coater now in etch mode (quartz chamber of course). It seems to me that there would not be much of a change from a detection/measuring system after the process finishes off the passivation and reaches the SiO2. Maybe not true.
There have been several good suggestions on the list so far. One I will check out right away is the residual gas analyzer. I also may need some different type of etching unit rather than the coater operating in etch mode. Since I am interested in FA too, the specimens are too small to justify the cost of impressive huge etching units.
gary
At 06:27 AM 5/24/00, you wrote: } Hello Gary, } I am also interested in this. We make PLD and I work in the FA group. Being } this as it may I am new to this field and would wish to find a technique to } get through the passivation and intrametal layers. } Thank you for any help. } } Sincerely, } Robb Westby } Associate Reliability Engineer } Lattice Semiconductor } } "Dr. Gary Gaugler" wrote: } } } This may be off-topic for this listserver but I'll give it } } a shot for those who are into materials science. } } } } Suppose that you have an etching system based } } on a small plasma chamber using CF4 and O. } } Is there some simple/convenient way to measure } } species during the etching process to indicate that some } } end point has been reached? i.e., a major etching } } area has been etched and the nature of the species } } has dramatically or noticeably changed? } } } } I can think of many x-ray methods, but what I am } } looking for is basically a sensor of some sort at a } } port in the etching chamber system. There is no } } SEM beam--and presumedly, no direct or indirect } } x-rays. } } } } Any ideas? } } } } gary g.
The weekend workshop on Chemical Microscopy, sponsered by the New York Microscopical Society, originally sheduled for the weekend of May 20 has been rescheduled for the weekend of June 17 & 18.
This is an opportunty to learn some of the fundamentals of Chemical Microscopy from Skip Panenik of Trace Analysis.
The course will be held in West Paterson, NJ.
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I am looking at this same problem for my own Plasma cleaning process. I just purchased a small, fiber optic, emission spectrometer (USB 2000) from OCEAN OPTICS to examine the light coming from my cleaning plasma. I just setup the software yesterday and will be trying it today. If I get some end point results I will repost to this thread in a few days. There is plasma etch literature that suggests that end points can be observed in the plasma emission lines.
Ronald Vane XEI Scientific (650) 369-0133
-----Original Message----- } From: Dr. Gary Gaugler {gary-at-gaugler.com} To: MSA listserver {Microscopy-at-sparc5.microscopy.com} Cc: just_in_case_I_bounce {zaluzec-at-sparc5.microscopy.com}
Al is present everywhere in the SEM. X-rays are produced by emission (electrons hitting the material) and Fluorescence (X-rays hitting the material). Collimator fluorescence is a major design problem for EDS system designers. High Z materials for stopping X-rays fluoresce strongly, and Low Z materials have no stopping power. Collimators often have high Z material lined with Aluminum to stop X-rays and then filter out the Fluorescence. If some of the AL is displaced it can be excited by either stray electrons or x-rays and cause the Al peak you see.
Ronald Vane XEI Scientific
-----Original Message----- } From: Gunnar Kopstad {gunnar.kopstad-at-medisin.ntnu.no} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
On many scopes I've found this was an artifact of an aluminum sample holder. Painting the surface of the sample holder with colloidal graphite or using a carbon planchet usually eliminates the problem.
Phil Swab Engineering Development Deposition Sciences Inc. Santa Rosa, CA 707-566-3718 phil.swab-at-depsci.com
-----Original Message----- } From: H. Fong [SMTP:hfong-at-u.washington.edu] Sent: Tuesday, May 23, 2000 5:49 PM To: microscopy-at-sparc5.microscopy.com
We have a JOEL 5200 SEM with a Link Analytical QX 2000 EDS detector. The problem is that we always get an aluminum peak in every EDS sprectrum regardless of sample & scan size. Does anyone have any suggestions on why this is?
Thanks, Hanson Fong
Univ. of Washington Materials Science & Engineering Department Box 352120 Seattle, WA 98195 USA
Dear Hanson, I solved that problem by cutting a circle of thin lead (Pb) foil to line the inside of my Al collimator. Poke a hole in the foil for the hole in the collimator. At 05:48 PM 5/23/00 -0700, you wrote:
} } } } We have a JOEL 5200 SEM with a Link Analytical QX 2000 EDS detector. The } problem is that we always get an aluminum peak in every EDS sprectrum } regardless of sample & scan size. Does anyone have any suggestions on why } this is? } } Thanks, } Hanson Fong } 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
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Hello Friends, Does anyone know if one needs to make any adjustments/compromises to Koehler illumination when using a digital camera? We are trying out a few demo's and get circular patterns or edge unevenness at optimum Koehler. If the condenser is defocused things are better...but...isn't the entire purpose of Koehler to get the most out of Abbe's equation that we can?? How much of a loss of resolution can be expected without optimum condenser settings. My sense is that one should always go with great scope alignment. I am just checking to see if any of us old microscopists can be taught a thing or two regarding digital camera set up. Thanks, Linda Fox lfox1-at-wpo.it.lumc.edu
Hi We have a client on an older Leo S200 with the standard solid state BSD fitted who has to look at slag off their stainless steel plant. In this slag there are slight compositional differences which they can only define should they run their filament on first peak. Strange but true! At first peak the slightest difference can be seen very easily, at saturation not a chance. Now I remember that Steve Chapman did give us an explanation for this the first time we mentioned it but, alas age catches up on me too and I have forgotten what it was.
Point is, try this on your system. It works for them. Better resolution on the BSD at first peak than at saturation.
Cheers Luc Harmsen Anaspec, South Africa Technical support on microscopy. www.anaspec.co.za
Remember, ICEM 15 will be in 2002, Durban, South Africa. www.icem15.com
-----Original Message----- } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: Tuesday, May 23, 2000 6:17 PM To: White, Woody N Cc: Microscopy-at-sparc5.microscopy.com
Dear Woody, I had a problem once that really strained the ultimate Z number sensitivity of my BSE detector. This was a case of a small amount of a marker chemical, I think Strontium, being fed to growing fish and trying to detect it in a scale of the fish. I was definintely able to detect the faint, brighter band on my GW BSE detector (solid state), but not on my Robinson (scinntilator) on the other SEM. I had to use more beam current for the GW, but it saw the contrast when the Robinson didn't. More sensitivity does not mean better Z resolution. At 01:36 PM 5/22/00 -0500, you wrote: } } Hello Gary, } } I have never used a sintillator type BSE detector, but the major differences } are } two fold. } } Typically (though diodes are getting better all the time) the Robinson has a } better low energy BSE detection effiency. It follows that for the same } noise } level electronics, it would exhibit better sensitivity. The dynamic range } problem will still exist for very different Zs in the field of view. In the } middle and upper portions of the sensitivity range (low and lower), I would } not } expect much difference between them. ...Any Comments from users of both???? } } I like the 4 quad diodes since I can go differential mode for macro } topography } (like fracture surfaces) and show the gross features while suppressing the } fine } detail. } } The Pt coating can have a profound negative effect on sensitivity if not } extremely thin. If given a choice, I would always use carbon for the best } BSE } sensitivity. ...If not hazardous, I would use Be, BUT!!!! For elements in } the } range of Al & Si, I prefer to use carbon and lower beam voltages to minimize } penetration, especially if they are films or very small features. } } I once presented some data illustrating the BSE signal attenuation as a } function } of sputtered Au thickness. But that data would be hard to retrieve now. } } Woody White } McDermott Technology
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
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Our SEM room is a little too noise for high res SEM work, so the service engineering recommended to dampen the noise. I know there is panels I can put up on the wall to do this. What is the cheapest solution for doing this?
I have worked on Noran Systems for 10 years. I have seen this problem several times and always on a JEOL SEM. Most times it was not the detector. It usually was a problem with alignment of the beam. The obvious point here is that detectors do not produce x-rays, they detect them.
} Hello Friends, } Does anyone know if one needs to make any adjustments/compromises to Koehler illumination when using a digital camera? We are trying out a few demo's and get circular patterns or edge unevenness at optimum Koehler. If the condenser is defocused things are better...but...isn't the entire purpose of Koehler to get the most out of Abbe's equation that we can?? How much of a loss of resolution can be expected without optimum condenser settings. } My sense is that one should always go with great scope alignment. I am just checking to see if any of us old microscopists can be taught a thing or two regarding digital camera set up. .} } } } } } } } } } } } } } } } } } .
You are giving up a lot of resolution with a digital camera due to the pixel spacing so the loss of resolution due to non optimal illumination have little effect on the image quality.
You can also solve the problem by increasing the distance from the CCD to the eyepeice so the ragged edge doesn't fall on the the CCD. This would also reduce the loss of resolution due to the spacing of the pixels. Of course is aslo decreases the coverage of the image.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
My Amray 1910 FESEM is only used by me. It is up as long as I say so (don't shut it down, put it in failsafe mode, etc.). PM takes about one day and this is done per contract twice a year. The only real bummer is when all 3 apertures have gone bad, one-by-one over time. Vent, pull the holder, change out the apertures and evacuate. This takes me about 45 minutes to accomplish. Each aperture typically lasts about 2 months. Since Amray gold flashes them, they cannot be flamed. Guess that is why they call them "consumables."
The only major down time I experienced was with my 1830 load lock system. The Balzers 240 turbo was going out (high frequency oscillation). That took about 3 days to fix for a total pump exchange. Other than this, both systems are very easy to keep running.
If they were in a mixed user environment, I'd opt for the FESEM over the LaB6. The FESEM is rather tough to screw up....unless of course, someone really worked at it.
gary g.
At 04:37 PM 5/23/00, you wrote: } [snip] } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } } } A how long is a piece of string ? type question } } } } } } I would be pleased if anyone could broaden my views on equipment } } reliability . } } Basically we have a 3yr old FESEM which I consider to be fairly } } reliable in that it is on call 24hrs/day , has numerous ( non } } dedicated users ) and apart from downtime for filament change and } the } } odd wear and tear type problems answers our needs . } } As we do not have a back up instrument and when we do experience } } problems it's always at the worse time certain personnel have the } } impression that it is unreliable . } } } } What do other sem users expect in terms of reliability , apart from } my } } ' subjective ' comments is it quantifiable , would approx 2wks } /year } } downtime including planned maintenance be considered excessive ? } } } } Regards } } Martyn Harris } } harrism-at-esm-semi.co.uk } } Dr. Lawrence F. Allard } Senior Research Staff Member } High Temperature Materials Laboratory } Oak Ridge National Laboratory } 1 Bethel Valley Road } Bldg. 4515, MS 6064 } PO Box 2008 } Oak Ridge, TN 37831-6064 } } 865-574-4981 } 865-576-5413 Fax } allardlfjr-at-ornl.gov
Assuming that the noise is being generated from equipment in the room then curtains will reduce the noise level quite effectively.
Ron
On Wed, 24 May 2000 19:22:19 -0700 (PDT) Ben Craft {bcraft-at-uci.edu} 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 } -----------------------------------------------------------------------. } } } Our SEM room is a little too noise for high res SEM work, so the service } engineering recommended to dampen the noise. I know there is panels I can } put up on the wall to do this. What is the cheapest solution for doing } this? } } } } ####### } #####\_O -Ben Craft- } ####/\/} } #### /" } ### \ } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
} ------------------------------------------------------------------------ } 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 Sergey and others, } } I want to add my 2 cents as I like the topic. } Most of this is a result of my experience with our TEM 2Kx2K CCD. } We are working in bright field - so I cannot comment on dark field } performance } Several points: } } 1. The CCD is much more convenient - you get your pictures instantly. } } 2. CCD is linear and has larger dynamic range than the film. } } 3. The bad thing about the CCD is resolution - about 4 times lower than that } of the film (in our camera the pixel size is 30 microns).
If You would choose a CCD with smaller pixel size You would get a better resolution.
} So if you want to } work in minimum dose you will do better with film.
Never, a good CCD is much more sensitive than a film.
} As I know the CCD is not } performing well in terms of signal to noise at low doses (and if you have to } work at 4 times higher magnification because of the resolution the things } become much worse).
see above , a smaller pixel gives a better sensitivity and a better resolution.
} } } 4. The CCD has smaller observation area - again loss of information.
use the so called Image mounting, than You get very large images with much more image information due to the larger dynamic range and better sensitivity.
} } } 5. I don't know about the detection efficiency compared to the film - it } depends on the thickness of the phosphorous and the accelerating voltage. If } a photon reaches the CCD chip it will be detected ... the problems are in } the conversion electron-photon.
The currently leading CCD systems reach single electron sensitivity at thin phosphor screens and good resolution.
} There are two sides - if you make the } phosphorous thicker you will get higher detection efficiency but the point } spread also increases so always a compromise is made between detection } efficiency and resolution. When I say detection efficiency this is not only } related to the detection of single electrons (as it detects single } electrons) but more to the actual signal detected on the background of the } noise. Apart from the shot noise additional noise is added due to the } scintillator driven detection and thermal noise in the CCD chip.
In good, highly sensitive CCD systems the Poisson noise of the incoming signal is dominating, not the CCD noise.
} } } The CCDs are now very popular in diffraction work because of the dynamic } range and linearity. } } Here is one reference where a nice comparison between 2Kx2K CCD and film has } been made: } } Downing, K., Hendrickson, F., Ultramicroscopy, 75, (1999), 215-233
Thanks for that.
} } } Best regards, } } Rado } } --------------------------------------------------------------------- } Radostin Danev } Laboratory of Ultrastructure Research } National Institute for Physiological Sciences } Myodaiji-cho, Okazaki 444-8585, JAPAN } e-mail: rado-at-nips.ac.jp } ---------------------------------------------------------------------
-- Best regards / Mit freundlichen Gruessen Dr. Frank Jenichen Proscan elektronische Systeme GmbH Tel.: +49 8195 999 -511 Fax: -512 mailto:Jenichen-at-proscan.de ------------------------------------------------------------ More information concerning our products and services can be found on our website http://www.proscan.de
I have a question about the study of for instance the translocation of a fluorescent labeled cellular component from the cytoplasm to the nucleus. At first it seems that labeling the component you want to study and do a counterstain for the nucleus would be sufficient to give an idea of the migration of a componenent from the cytoplasm to the nucleus or not.
By doing the experiment this way however you do not have a clue about the total cell content, because the cytoplasm is not counterstained with a background stain to show the cell outlines to give an idea of the actual cell extent. Without a cytoplasm stain, you have no idea of the actual size of the cell in which the label for the cellular componenent resides. For an "absolute" idea of the migration I think a background cellular counterstain is necessary ?
Also the nuceus is thicker than the cytoplasm, so for a given focuslevel inside the nucleus there is more light falling in the lens form above and below than in the cytoplasm, which probably will give a non-linear response curve for the quantification of the translocation ?
A clear case for doing the experiment with a fluorescent probe with a confocal or multiphoton microscope! Either way, out-of-focus contributions to intensity are not important. Your confocal image is also a sample from a well-defined volume of cell or tissue. Therefore, provided you can regard each compartment as homogeneously labelled the total compartment (e.g. cytoplasm, nucleus) volume does not need to be determined.
} From: "Van Osta, Peter [JanBe]" {PVOSTA-at-janbe.jnj.com} To: Microscopy-at-sparc5.microscopy.com
McMaster-Carr has some of the best selection for sound deadening material and generally a better price than specialty dealers or other distributors. Their Web site is very functional http://www.mcmaster.com Delivery has always been more than prompt and they have more indespensible items for any microscope lab. No lab should go without one of their catalogs.
Sound control products are in my catalog on page 2777-2779 products ranging from flat foam to sculptured foam to "sono-tech" foam to acoustical quilts to acoustical cylinders. Looking through this stuff isn't cheap but of the products I have seen offered other places the prices here are competitive.
Of course you could always head to a carpet store and dig through their dumpsters for throw-away remnants and hang them on the walls in the scope room. Two or three layers might work well enough - not sure about the smell though. . .
Good luck Geoff
Ben Craft wrote:
} Our SEM room is a little too noise for high res SEM work, so the service } engineering recommended to dampen the noise. I know there is panels I can } put up on the wall to do this. What is the cheapest solution for doing } this? } } ####### } #####\_O -Ben Craft- } ####/\/} } #### /" } ### \
-- Geoff Williams,
Electron Microscope Facility Supervisor Biology Department Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
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Linda, Look for a digital camera that allows you to capture an image of the illumination pattern with your specimen slide removed from the beam path. This "background" image should then be automatically subtracted from your final image. This will eliminate not only uneven illumination but also small light distortions due to dirt on lenses (that you cannot get off by cleaning external surfaces), etc. Using this feature permits good Koehler illumination and very even illumination on your final image file. As an example, the SPOT RT software has a feature called "Flatscreen". I capture images from each objective after checking for proper Koehler illumination. These are stored and easily called up as needed. However, the microscope alignment should still be rechecked prior to capturing images. This is a very important feature if you do Nomarski/DIC imaging. You can easily smooth out the very directional illumination pattern by capturing the lighting pattern without the sample and then subtracting it automatically when capturing the sample image.
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
--------------------------------------
Hello Friends, Does anyone know if one needs to make any adjustments/compromises to Koehler illumination when using a digital camera? We are trying out a few demo's and get circular patterns or edge unevenness at optimum Koehler. If the condenser is defocused things are better...but...isn't the entire purpose of Koehler to get the most out of Abbe's equation that we can?? How much of a loss of resolution can be expected without optimum condenser settings. My sense is that one should always go with great scope alignment. I am just checking to see if any of us old microscopists can be taught a thing or two regarding digital camera set up. Thanks, Linda Fox lfox1-at-wpo.it.lumc.edu
In a message dated 5/25/00 10:26:43 AM, sherman-at-btny.purdue.edu writes:
} Look for a digital camera that allows you to capture an image of the } illumination pattern with your specimen slide removed from the beam path. } This "background" image should then be automatically subtracted from your } final image. This will eliminate not only uneven illumination but also } small light distortions due to dirt on lenses (that you cannot get off } by cleaning external surfaces), etc. Using this feature permits good Koehler } illumination and very even illumination on your final image file.
One additional note. Using a background image captured with a log-response camera (e.g., a Vidicon) does call for subtraction. For a linear response camera (most CCDs unless you are using some built-in gamma circuitry) you want to divide by the background (ratio of signal to background). Also, the problem with this method is that it uses some of your dynamic range, so you effectively cannot handle as great a range from bright to dark.
Ben:Ê we had the same problem in in our SEM/FIB rooms.Ê We got large sheets of egg-crate foam and glued them to the walls.Ê It gives the place sort of a "rubber room" appearance, but it works really well.
Ben Craft wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } ToÊ Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Our SEM room is a little too noise for high res SEM work, so the service } engineering recommended to dampen the noise.Ê I know there is panels I can } put up on the wall to do this. What is the cheapest solution for doing } this? } } ####### } #####\_OÊÊÊÊÊÊÊÊÊÊÊ -Ben Craft- } ####/\/} } #### /" } ###Ê \
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky HoldfordÊ (r-holdford-at-ti.com) 972-598-1291 (pager) KFAB Physical Analysis Labs--SEM/FIB/FA Kilby Center West Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Ê
I received an AO fluorescence microscope from a kind colleague but need to find an instruction manual for it. The microscope is an AO model 10 or 20 and it has a vertical fluorescence illuminator (model 2071) attached. Also, I'm looking for a trinocular head for the microscope to do photography. Any assistance in locating these would be greatly appreciated.
David A. Doe -- Dr. David A. Doe Biology Department Westfield State College Westfield, MA 01086 413/572-5291 fax: 413-562-3613
The amount of backscatter generated from a specimen depends on the kV, the probe size and the composition of the material under investigation. If the level of backscatter is insufficient under "normal" saturation conditions i.e. the gun is correctly saturated, then by de-saturating the larger source will result in a larger probe dimension on the specimen; increasing the probe diameter increases the volume of material involved in the production of BSE.
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
I have been following off and on this discussion on CCD cameras for EM. Much of this discussion has been concerned with "sensitivity". I am being naive here, but when we talk about "sensitivity" of CCDs, isn't this the same thing as the QE of camera/chip? I don't think I have ever seen a QE for em CCD's for various accelerating voltages/wavelenghts. Does the electron beam directly hit the silicon photodyodes or it there an interface that converts the incoming electrons to different (longer?) wavelengts? I know em films are sensitive to specific acc voltages. Are CCD cameras for em the same. For long exposures it may not matter, but for short exposures or low level intensity does the QE of the camera come into play?
} ===== Original Message From Jenichen {Jenichen-at-proscan.de} ===== } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hank Adams Manager Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx 77025
Microscopy Experts, We have recently inherited a Varian 936-40 Porta-Test leak detector. It came with out any documentation. Big surprise, I know. I called Varian and they offered to sell me an operators manual for $185.00. This seems just a bit excessive. If anyone has one, I would be happy to pay a Xeroxing and shipping fee. TIA Kim DeRuyter Electron Microscopy Technician 308 Natural Science Facility P.O. Box 755780 University of Alaska Fairbanks, AK 99775-5780 907-474-5452 907-474-5163 fax
} } I have worked on Noran Systems for 10 years. I have seen this problem several } times and always on a JEOL SEM. Most times it was not the detector. It } usually was a problem with alignment of the beam. The obvious point here is } that detectors do not produce x-rays, they detect them. } } Regards, } } Craig Theberge }
Did you ever figure out what sort of alignment problem it was, and from exactly what the Al X-rays were being produced?
cheers
rtch
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
Looking for spare parts for an ISI-40 SEM. Particularly, column pieces, apertures, filament assembly and wehnelt cylinders. Any suggestions would be welcome.
Kim DeRuyter Electron Microscopy Technician Room 308 Natural Sciences Facility P.O. Box 755780 University of Alaska Fairbanks, AK 99775-5780 907-474-5452 907-474-5163 fax
Hi friends I agree Steve, sometimes (I seem it depends on electron gun design and distance between wenelt and anode, wenelt and filament) the first peak on saturation curve is more than saturation level even in secondary emission signal. Victor Sidorenko, ANTRON Co.Ltd., Moscow, Russia
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Has anyone knowledge of a method to rapidly fix adherent cell cultures so as to prevent the loss of small hydrophobic molecules? The problem involves subsequent diffusion of the antibody marker into the cytoplasm. Material is examined using fluorescence/confocal microscopy The organelles of interest in this case are mitochondia but general recommendations would be most appreciated too.
Regards
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
At 05:07 PM 25-05-2000 +0100, you wrote: Steve Chapman wrote,
} Hi } } Yes Luc is right I did give him an explanation. } } The amount of backscatter generated from a specimen depends on the kV, the } probe size and the composition of the material under investigation. If the } level of backscatter is insufficient under "normal" saturation conditions } i.e. the gun is correctly saturated, then by de-saturating the larger source } will result in a larger probe dimension on the specimen; increasing the } probe diameter increases the volume of material involved in the production } of BSE. } But how does this lead to an increase in the BSE differentiation (implying more signal and hence less noise). Surely just defocussing would do the same thing. Defocussing as such should not effect the BSE coeffecient, unless you had sub surface charging or some other artefact.
Very interesting! Ken.
Moran Scientific Pty Ltd P.O. Box 651 Goulburn NSW 2580 Australia Tel 02 4844 4234 (International, 61 2 48444234) Fax 02 4844 4291 (International, 61 2 48444291) Email {kmoran-at-goulburn.net.au} Web Page http://www.goulburn.net.au/~kmoran/ "Patience accomplishes its object, while hurry speeds to its ruin. Gulistan 1258"
} ------------------------------------------------------------------------ } 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 have been following off and on this discussion on CCD cameras for EM. } Much of this discussion has been concerned with "sensitivity". I am being } naive here, but when we talk about "sensitivity" of CCDs, isn't this the same } thing as the QE of camera/chip?
No, sensitivity means how many electrons You need for a digital response from the CCD.
} I don't think I have ever seen a QE for em } CCD's for various accelerating voltages/wavelenghts. Does the electron beam } directly hit the silicon photodyodes or it there an interface that converts } the incoming electrons to different (longer?) wavelengts?
For TEM investigations the high energy electrons (80 - 400 keV) hit a scintillator (YAG- or Phosphor-screen). These screen emits visible photons (energy in the region 2eV) which are detected by the CCD. If we would use the high energy electrons directly onto the CCD the CCD would be damaged.
} I know em films are } sensitive to specific acc voltages. Are CCD cameras for em the same.
The response for a phosphor scintillator rises linear with the energy, but reaches saturation for high energies (higher than 200keV). This response depends also on the material You use and on the thickness of the screen.
} For long } exposures it may not matter, but for short exposures or low level intensity } does the QE of the camera come into play?
If You want to get a good statistics of Your signal a response of one digital count for one electron would be very good. If the application gives You only a small amount of electrons to detect (Filter applications, low contrast applications, biological application and other) the sensitivity (reponse) of the camera should be higher to avoid long exposure times to overcome problems with drift an sample damage. So the best cameras optimized for high sensitivity (the screen is directly coupled with a fibreoptic to a cooled slow-scan CCD with up to 16bit digitization) reach more than one digital count per incident electron to have a good compromise between sensitivity and good statistics. Standard system with optical coupling do not reach this sensitivity and can be used only for applications with high beam density.
} } } } ===== Original Message From Jenichen {Jenichen-at-proscan.de} ===== } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } } } } } Radostin Danev 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 } } } -----------------------------------------------------------------------. } } } } } } Dear Sergey and others, } } } } } } I want to add my 2 cents as I like the topic. } } } Most of this is a result of my experience with our TEM 2Kx2K CCD. } } } We are working in bright field - so I cannot comment on dark field } } } performance } } } Several points: } } } } } } 1. The CCD is much more convenient - you get your pictures instantly. } } } } } } 2. CCD is linear and has larger dynamic range than the film. } } } } } } 3. The bad thing about the CCD is resolution - about 4 times lower than } that } } } of the film (in our camera the pixel size is 30 microns). } } } } If You would choose a CCD with smaller pixel size You would get a better } } resolution. } } } } } So if you want to } } } work in minimum dose you will do better with film. } } } } Never, a good CCD is much more sensitive than a film. } } } } } As I know the CCD is not } } } performing well in terms of signal to noise at low doses (and if you have } to } } } work at 4 times higher magnification because of the resolution the things } } } become much worse). } } } } see above , a smaller pixel gives a better sensitivity and a better } resolution. } } } } } } } } } } } 4. The CCD has smaller observation area - again loss of information. } } } } use the so called Image mounting, than You get very large images with much } more } } image information due to the larger dynamic range and better sensitivity. } } } } } } } } } } } 5. I don't know about the detection efficiency compared to the film - it } } } depends on the thickness of the phosphorous and the accelerating voltage. } If } } } a photon reaches the CCD chip it will be detected ... the problems are in } } } the conversion electron-photon. } } } } The currently leading CCD systems reach single electron sensitivity at thin } } phosphor screens and good resolution. } } } } } There are two sides - if you make the } } } phosphorous thicker you will get higher detection efficiency but the point } } } spread also increases so always a compromise is made between detection } } } efficiency and resolution. When I say detection efficiency this is not only } } } related to the detection of single electrons (as it detects single } } } electrons) but more to the actual signal detected on the background of the } } } noise. Apart from the shot noise additional noise is added due to the } } } scintillator driven detection and thermal noise in the CCD chip. } } } } In good, highly sensitive CCD systems the Poisson noise of the incoming } signal } } is dominating, not the CCD noise. } } } } } } } } } } } The CCDs are now very popular in diffraction work because of the dynamic } } } range and linearity. } } } } } } Here is one reference where a nice comparison between 2Kx2K CCD and film } has } } } been made: } } } } } } Downing, K., Hendrickson, F., Ultramicroscopy, 75, (1999), 215-233 } } } } Thanks for that. } } } } } } } } } } } Best regards, } } } } } } Rado } } } } } } --------------------------------------------------------------------- } } } Radostin Danev } } } Laboratory of Ultrastructure Research } } } National Institute for Physiological Sciences } } } Myodaiji-cho, Okazaki 444-8585, JAPAN } } } e-mail: rado-at-nips.ac.jp } } } --------------------------------------------------------------------- } } } } -- } } Best regards / Mit freundlichen Gruessen } } Dr. Frank Jenichen } } Proscan elektronische Systeme GmbH } } Tel.: +49 8195 999 -511 Fax: -512 } } mailto:Jenichen-at-proscan.de } } ------------------------------------------------------------ } } More information concerning our products } } and services can be found on our website } } http://www.proscan.de } } Hank Adams } Manager } Integrated Microscopy Core } Molecular and Cellular Biology } Baylor College of Medicine } Houston, Tx 77025
-- Best regards / Mit freundlichen Gruessen Dr. Frank Jenichen Proscan elektronische Systeme GmbH Tel.: +49 8195 999 -511 Fax: -512 mailto:Jenichen-at-proscan.de ------------------------------------------------------------ More information concerning our products and services can be found on our website http://www.proscan.de
I suppose in principle a line-scanner type of CCD could be made to do this, but the practical difficulties in setting it up would be enormous - the scan system would have to be synchronously stepping with the vertical progression of the SEM scan, and the alignment and line geometry of the system would also have to be exactly right. Not at all easy to achieve. Also, it is fairly well understood that the resolution of the record screen underrepresents the resolution of the raw signal fed to it (partly to ensure that lines are not visible in the image). Therefore this just seems to be the wrong approach. There are plenty of low-cost (~10% of the cost of this camera) image grabbers for SEM that digitise the stream of analogue data fed to the record tube.
It would be more interesting to consider whether this type of camera can contribute to high quality TEM imaging. I am not clear what is limiting the resolution of current TEM digital cameras - could a high-resolution CCD camera approach the resolution of TEM film more closely than the current generation of these, or is the phosphor/YAG not good enough to make it worthwhile.
Chris
Date sent: Tue, 16 May 2000 09:06:16 -0500 To: {Microscopy-at-sparc5.microscopy.com} } From: John Foust {jfoust-at-threedee.com}
Dear List Members,
Does anyone know of a source for glass coverslips which have been treated with something to optimize cell growth? A colleague of mine is looking for some, particularly round ones.
Thanks.
Matthew J. Schibler Ph.D. UCLA Brain Research Institute 1524A Gonda (Goldschmied) Center for Neuroscience and Genetics Los Angeles, CA 90095-1761
On Mon, 15 May 2000 11:19:12 -0700, Jon Krupp 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 List: } } A newly appointed researcher here has asked my advice on several pieces of } TEM spec. prep equipment. I turn to you for helpful suggestions. } } The research involves serial sectioning biological tissues and many grids. } EM is a minor, but essential component of the project. The lab runs through } dozens of grids and hundreds of pictures, but only 4 - 6 times a year. Big } bursts of activity followed by long periods of analysis and investigations } using other techniques. Of the hundreds of pictures taken, they may only } use a few for data. } } The researcher is looking for ideas on what choices are available, how } useful, and approximate costs of the following: } } Digital imaging to add to our existing JEOL 1200EX TEM - I have seen other } threads on this topic and would welcome any new ideas. Digital imaging will } save a lot of time and money since they discard so many pictuers, but the } question of image quality is one I am to investigate. } } Ultra microtome - We have an older A/O Ultracut (the model before it became } the Reichert Ultracut E) which is OK for the sectioning we do in the } general lab. But she wants a new one for her exclusive use. The question is } whether a new microtome will allow folks in her lab to do serial } sectioning any faster or easier, or by less skilled users, than our current } system. } } Staining machine - Anyone have info on staining machines or systems for } lots of TEM grids. I have never had to do so many grids that this was an } issue, so I have never kept up on the offerings. If you know of something } and/or have experience let me know. Again, this is something she would keep } in her lab. } } Tissue processing machine - Same as above for me, never did so much at one } time that I ever thought I needed one of these. The samples to be processed } are C. elegans, anything available that could do these unattended? How } about upkeep and volumes of chemicals needed. Also an item to be kept in } her lab. } } I will filter and pass on your comments. Anything you might offer will, as } always, be received with appreciation and thanks. } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } Jon:
A few comments about the equipment. I have done literally thousands of serial sections, so I think I can offer some first hand comments.
First: My microtome of choice for serial sectioning has been the old Reichert OMU-3--the predecessor of the Ultracut E. I have to admit that part of the reason was not wanting to take the time for a learning curve. The other reason was that I found the illumination system on the old OMU-3 to be outstanding (I did repetitive serial thins and thicks over several hundred microns to about a millimeter).
Second:Staining machine--I use/have used the LKB/Leice Ultrastainer. The original LKB unit was a marvel--we did over 300 grids at one point, losing only one (stuck the forceps through the formvar!!!), and no precipitates. Can't say the same for the Leice unit, although it should be pretty much the same. The stains are the biggest variable, as is a really rigorous cleaning regime. Check it out.
Third: Processor--I use/love/hate the Lynx unit (currently available through EM Sciences). I had an early unit (from Australia)--no problems over 3 or 4 years. Have a Leica unit here--it took 4 or 5 years to get it to work consistently, but now seems ok, and it gets substantial use in bursts. Check out the RMC/Ventana unit--it is the progeny of the old LKB unit, and is probably a worthy competitor. (I just can't get one for demo--they've been promising for nearly 4 years, but I think they gave up after last year.) Maintenance on both is fairly routine, and consumables are not prohibitive. Both use very small volumes, and can be used osmium through pure epoxy (if your protocol is limited to 20 or so steps). Personally wouldn't be without one.
Hope this helps.
Roger C Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals
Disclaimer: I have no financial interest in any of the products or suppliers. Just a long time user.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freelane.excite.com/freeisp
Do the cells need some sort of support matrix (e.g., collagen) or are they just not fond of glass? We do our own treating....collagen, poly-lysine, there is a mussel protein (not a spelling error - I think it is the byssal thread stuff from Mytilus sp.) that is sticky (CellTak?)...you can also dissolve PS culture dishes in solvent and coat glass coverslips with the resulting goo.
Tamara Howard CSHL
On Tue, 16 May 2000, Schibler, Matthew 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 List Members, } } Does anyone know of a source for glass coverslips which have been treated } with something to optimize cell growth? A colleague of mine is looking for } some, particularly round ones. } } Thanks. } } } Matthew J. Schibler Ph.D. } UCLA Brain Research Institute } 1524A Gonda (Goldschmied) Center } for Neuroscience and Genetics } Los Angeles, CA 90095-1761 } } (310) 825-9783 } FAX (310) 206-5855 } E-mail: mschibler-at-mednet.ucla.edu } } }
I just got off the phone with Stacie Kirsch, and discovered that I was the 3rd caller today asking about 'something in Philaldelphia'! Debby Sherman was first and is to talk to the Meeting Arrangements person, or some such, but I (and probably some accompanying colleagues) would be happy to participate if something informal were organized for the Sunday afternoon. Our Group at this Canadian federal lab:
- has 8 different beam instruments in 7 distinct 'labs' (and formally accesses another 2 at an on-site private sector service provider), - is manned by 18 scientists and technologists, the majority of which are permanent, - does a ~50-100 project/yr mix of contract work and core research projects for and with internal programs and external clients (academics, industrial, other federal), - has a significant number (too many!) of internal and external operators, and - has a reasonable operating budget (but currently no capital), so - we have likely encountered some variation of almost every conceivable problem (and solved only a fraction).
If something comes to pass, now or next year, I would gladly share experiences with others, especially as this is an aspect of delivering science that is commonly overlooked. A couple of thoughts:
- this could easily turn into a 'gripe-athon'. Someone should be prepared to lead it and direct it towards problem-solving if this occurs. - Ron Anderson and I have made sporadic attempts over the years to lead something called " Factors Influencing the Establishment of a New TEM Facility" at numerous workshops, often with great success. This also touched upon many of the real-world issues of an EM lab, though from the slightly more positive viewpoint of actually some hard cash in hand. -
---------- From: Elaine Humphrey [SMTP:ech-at-unixg.ubc.ca] Sent: May 16, 2000 11:39 AM To: Microscopy-at-sparc5.microscopy.com Subject: Re: SEM facility managers
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Me too, though I would like to attend the meeting in Philadelphia. Elaine } } } Collegues, } } In response to the current thread on the problems of facility managers, I } say count me in as being highly interested! If a discussion group does get } together at M&M it would be great to see a report posted on this listserver } for those of us who unfortunately can't attend the meeting. If somebody } could take notes and post them, I for one would be very appreciative. } } Thanks! } Dee
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
Guess it depends on what you mean by optimize. The intestinal cell lines i work with prefer bare glass (i wash in acetone, then ethanol, then lots of dH2O, then boil in dH2O, then place each one on a piece of filter paper so they aren't overlapping and then autoclave - a pain in the neck but I think it really matters). i seed the cells in serum containing medium without any other pre-treatement
I have coated with collagen, fibronectin, etc by placing coverslips in 24 well trays and adding the matrix material. this makes no difference or reduces differentiation of my cells.
many labs report cell lines that differentiate better on permeable filters so nutrients have access to the basolateral membrane. } } Dear List Members, } } Does anyone know of a source for glass coverslips which have been treated } with something to optimize cell growth? A colleague of mine is looking for } some, particularly round ones. } } Thanks. } } } Matthew J. Schibler Ph.D. } UCLA Brain Research Institute } 1524A Gonda (Goldschmied) Center } for Neuroscience and Genetics } Los Angeles, CA 90095-1761 } } (310) 825-9783 } FAX (310) 206-5855 } E-mail: mschibler-at-mednet.ucla.edu
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I very much agree with others in this issue. Unfortunately, I won't be able to attend this year's meeting, but I will love to find out what others had to say about managing a multi-user EM facility. Please post the highlights of that meeting if it takes place in Philadelphia.
Thanks in advance,
Soumitra
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
Hi all, I believe to be on track with mounting clay rich rocks (that disagregate readily in perturbed water (i.e. shaken or sonicated), but not in ethylene glycol or ethanol (at rest). My goal is to mount the samples (whole, not seperates) for analysis in TEM. I have been soaking them in LR-White at 60C, where fixation occurs without accelerator in less than 24 hours. I am planning to (but have not yet) microtome the samples, or potentially ion-mill. Thoughts for the future are to use a set of glassware with a Millipore filter and vacuum to pull ethanol(cleanser and dilator), ethylene glycol(for swelling clay component), and an LR White "chaser" to view "saturated" textures, as opposed to compacted. My question is this: Can anyone point out pitfalls with this approach that I am not seeing, again I haven't tried the whole thing yet, but am in a position to start prepping the samples. In particular, is microtoming preferred to ion-milling for weak, soft samples that rely on epoxy for reinforcement? Does LR-white readily pull through a sample given a weak vacuum and a porous plate (its viscous, but not as viscous as water, for example)? Does swelling the clays with ethylene glycol and the like, and then directly mounting, introduce volatiles into the column under 120-200 kV? Any recommendations are welcome, the literature helps a bit, but is usually sketchy about these fine details of preparation procedure. thanks in advance, N
_____________________________ Nicholas W. Hayman \ Dept. of Geological Sci., U.W.\ http://www.geology.washington.edu/~hayman Box 351310, Seattle WA 98195 \_________________________________________
Dear John, That is exactly what passive image capture systems like Quartz PCI do. At 09:06 AM 5/16/00 -0500, you wrote:
} At 04:02 PM 5/15/00 -0700, Dr. Gary Gaugler wrote: } } It seems to me that no digital camera system would work on a SEM } } in place of a Polaroid or other film-based output device. Since the } } recording CRT in a SEM is based on a sequential line scan, one } } would need a camera that would capture each line as it is produced. } } What you're saying is there must be a system out there that } digitizes that single stream of line scan intensities, then } processes all that data inside the computer as an image as } opposed to trying to digitize the frame buffer. } } - John 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
some of my users think it is tome to retire our ancient but still functional cryostat. Could anyone who has bought one in the recent past give me some info about who is making them these days as well as any pros and cons of these new-fangled models.
} From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } } } I suppose in principle a line-scanner type of CCD could be made to } do this, but the practical difficulties in setting it up would be } enormous - the scan system would have to be synchronously } stepping with the vertical progression of the SEM scan, and the } alignment and line geometry of the system would also have to be } exactly right. Not at all easy to achieve. Also, it is fairly well } understood that the resolution of the record screen underrepresents } the resolution of the raw signal fed to it (partly to ensure that lines } are not visible in the image). } Therefore this just seems to be the wrong approach. There are } plenty of low-cost (~10% of the cost of this camera) image } grabbers for SEM that digitise the stream of analogue data fed to } the record tube. } } It would be more interesting to consider whether this type of } camera can contribute to high quality TEM imaging. I am not clear } what is limiting the resolution of current TEM digital cameras - } could a high-resolution CCD camera approach the resolution of } TEM film more closely than the current generation of these, or is } the phosphor/YAG not good enough to make it worthwhile. }
If you control the scan you only need a single light sensitive element. You step the beam digitize the light, step the beam, wait for the last spot to go out and repeat. You don't need a camera. You do need a very fast responding system for changing electron beams to light.
This system has the resolution of the scan beam. It would not be particualy fast but you could increase the number of bits resolution to as large a number as you wanted.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
At 10:08 AM 5/16/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This is true except that the record CRT has a fixed resolution. These are typically 2000 horizontal lines. Different scan rates change the pixel dwell time. But the final image is still not much more than 2000 lines. This is fine for a Polaroid print. But if using real film, it is less than ideal or optimum. I find that film has much more resolution than a Polaroid print. A good alternative is the Polaroid PN (positive/negative). But one would still have to scan the negative to get a digital file. This is another topic, all together.
} It would be more interesting to consider whether this type of } camera can contribute to high quality TEM imaging. I am not clear } what is limiting the resolution of current TEM digital cameras - } could a high-resolution CCD camera approach the resolution of } TEM film more closely than the current generation of these, or is } the phosphor/YAG not good enough to make it worthwhile.
Not having TEM experience, I cannot comment on this type of application. But I am experienced with SEM usage. It would seem to me at first blush that TEM images are continuous whereas the SEM images are discrete. This would mean that CCD imaging devices would work for TEM applications but not for SEM. The common denominator remains the Polaroid and silver emulsion film.
gg
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
It's called a "passive" or "listening" digital image acquisition system, such as our ADDA II or similar devices from other manufacturers. Benefits: fairly easy to set up and use. Disadvantages (as opposed to an "active" or "talking" system): You're still limited to what the microscope can provide in terms of resolution, dwell time, etc.
Michael
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: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: John Foust [mailto:jfoust-at-threedee.com] Sent: Tuesday, May 16, 2000 8:06 AM To: Microscopy-at-sparc5.microscopy.com
At 04:02 PM 5/15/00 -0700, Dr. Gary Gaugler wrote: } It seems to me that no digital camera system would work on a SEM } in place of a Polaroid or other film-based output device. Since the } recording CRT in a SEM is based on a sequential line scan, one } would need a camera that would capture each line as it is produced.
What you're saying is there must be a system out there that digitizes that single stream of line scan intensities, then processes all that data inside the computer as an image as opposed to trying to digitize the frame buffer.
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Thanks to all who indicated an interest in meeting to discuss some common problems associated with managing microscopy facilities at the M&M meeting. I am in the process of arranging this and will send details once we are a bit further along. 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
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Like Gary and Ken have pointed out, there are several reasons why 4x5 backs won't work. Even if they did, there is another issue. A 4x5 back would incorporate the photo CRT with its phosphor into the loop. That means the electronic signal from the PMT or BSE detector has to be converted back into an analog brightness via the photo CRT, then the digital 4x5 back would be used to digitize the signal, and that in a most unwieldly manner.
As a rule, if you don't have to convert signals back and forth or pass them through extra stages of processing, don't do it. I understand photo CRTs are quite good, but there are extra focus, noise, and calibration factors involved passing the signal through the CRT. It is far better to take the signal straight over to digital using a good, single channel A/D converter for the video (plus one for X and Y position rather than using the millions of A/D converters in a CCD. Makes for a lot cheaper system, too.
Warren S.
At 04:02 PM 5/15/2000 -0700, Gary Gaugler wrote: } At 01:38 PM 5/10/00, you wrote: } } } } I'm curious ... has anyone tried simply installing a digital } } 4x5 back in place of a Polaroid 4x5 back??? For example, see: } } } } http://www.phaseone.com/brochures/powerfx.html } } } } cheerios, shAf } } It seems to me that no digital camera system would work on a SEM } in place of a Polaroid or other film-based output device. Since the } recording CRT in a SEM is based on a sequential line scan, one } would need a camera that would capture each line as it is produced. } Most digital backs are single or triple pass units of a single linear } set of sensors. There are other cameras that do snapshot capture } but even these would not work since the whole image is not present } on the record CRT at the time of taking a picture with the digital } camera. The final image is generated sequentially, line by line, } on the record CRT. } } If the SEM image is stored in a frame buffer, the buffer can be } converted to RS-170 TV video and frame grabbed. But the } best that this would typically do is 640 lines. } } Its an interesting problem and dilemma about being in a situation } where digital camera products simply won't work in place of } film. But since the goal is to obtain a digital file, why not start } with a digital interface? For example, a passive digital capture } system would transfer the record CRT information to computer } and directly result in a nice digital file. Alternatively, for some } systems, an active system can be applied to directly control the } SEM's beam. In doing so, the range of final digital image } resolution is limited only by the attached hardware system. } } gary g.
} Date: Tue, 16 May 2000 14:27:27 -0700 } To: John Foust {jfoust-at-threedee.com} } From: "Dr. Gary Gaugler" {gary-at-gaugler.com} } Subject: Re: SEM: digital 4x5 backs } } No. What I was saying was that there is not a digital } camera system out there that will capture the line-by-line } recording CRT output--as far as I see it at present. } } To get a digital file from the SEM, the method needs to } be digital but either passively attached to the record CRT } or actively connected as a replacement for the SEM's } internal scan generator. } } gary } } } } At 07:06 AM 5/16/00, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
ELECTRON MICROSCOPE TECHNICIAN Naval Research Laboratory
Announcement Number 74-0448-00 http://amp.nrl.navy.mil/code1800/74-0448.htm Job Title: Physical Scientist, NP-1301-II, $22,563* to $49,794* (*Includes locality pay)
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All candidates will be rated on the following factors: 1) Knowledge in the preparation of transmission electron microscope (TEM) thin specimens (preferably from fine-grained sediment/soil specimens and /or other geological samples). 2) Knowledge in the operation of TEMâs and scanning electron microscopes (SEMâs) and their analytical detectors. 3) Knowledge in the basic interpretation of TEM data. 4) Ability to communicate technical concepts orally and in writing.
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-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
} If you control the scan you only need a single light sensitive } element. You step the beam digitize the light, step the beam, } wait for the last spot to go out and repeat. You don't need } a camera. You do need a very fast responding system for } changing electron beams to light. } } This system has the resolution of the scan beam. It would not } be particualy fast but you could increase the number of bits } resolution to as large a number as you wanted. } } Gordon } } Gordon Couger gcouger-at-couger.com } } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00
The light sensitive element is already in-place. It is the Everhart-Thornley scintillator detector, or any other SE or BSE detector on a SEM. Just passively tap into the record CRT signals, digitize the detected video (SE/BSE), and save the resulting image as a digital file. Changing scan rates will change pixel dwell time. But the final image ought to still be a digital representation of what was intended to go to the film/Polaroid camera.
Again, the other option is active control of the SEM's beam. This just swaps the internal scan generator with an external one. The video system and digitization is the same as for a passive system. A robust active system will be capable of producing higher resolution digital images than a passive system. This is because a good active system can go up to 4096 horizontal pixels (12-bit D/A converter) and the pixel dwell time is usually adjustable. The only down side is the total frame time based on pixel dimensions and dwell time. At extremes, one could be waiting a very long time for a single image. If the beam is not stable, and as pointed out in an earlier posting, the drive circuits are not stable, there would be some upper limit on pixel dimensions and dwell time. Beyond this limit, the image would appear to shift as the beam shifted during the capture.
Several of the current generation SEMs have incorporated digital control and digital image capture. But these SEMs are of course at today's prices. Its rather easy to breathe new life into an older SEM by adding active or passive third party digital control/capture systems. One gets essentially a "new" modern SEM at a fraction of the cost of buying a new one.
We are exploring the possibility of purchasing a freeze fracture system to do FFTEM of emulsions/microemulsions. Does anyone have any recommendations? Does anyone have a used system available?
Hello Nicholas, I've looked at several soils similar to your easily disaggregated clay rich rocks in the TEM and find that the best procedure is to imbed aggregates in 2% agar to hold the aggregates together. The aggregates are then cut out as agar-soil cubes and chemical processing (fixation, Os-fix, buffer rinse, dehydration, resin-solvent washes, resin infiltration) done directly onto the agar cubes. The cubes are placed into molds and after curing, microtomed to 40-60nm sections. The sections come out nicely, but if you have a high primary mineral content (quartz, feldspars } 15%) then you'll have a lot of torn sections. Use of a diamond knife rather than glass gives you significantly better results.
My methods used in my thesis are on the web at: http://wilfred.berkeley.edu/~gordon/PHD Look particularly at chapter four which concentrates on TEM methods and results. If it would be of use, I'll be glad to send a cdrom copy of my thesis to you.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Tue, 16 May 2000, N. Hayman 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 all, } I believe to be on track with mounting clay rich rocks (that } disagregate readily in perturbed water (i.e. shaken or sonicated), but not } in ethylene glycol or ethanol (at rest). My goal is to mount the samples } (whole, not seperates) for analysis in TEM. I have been soaking them in } LR-White at 60C, where fixation occurs without accelerator in less than 24 } hours. I am planning to (but have not yet) microtome the samples, or } potentially ion-mill. } Thoughts for the future are to use a set of glassware with a } Millipore filter and vacuum to pull ethanol(cleanser and dilator), } ethylene glycol(for swelling clay component), and an LR White "chaser" to } view "saturated" textures, as opposed to compacted. } My question is this: Can anyone point out pitfalls with this } approach that I am not seeing, again I haven't tried the whole thing yet, } but am in a position to start prepping the samples. In particular, is } microtoming preferred to ion-milling for weak, soft samples that rely on } epoxy for reinforcement? Does LR-white readily pull through a sample given } a weak vacuum and a porous plate (its viscous, but not as viscous as } water, for example)? Does swelling the clays with ethylene glycol and the } like, and then directly mounting, introduce volatiles into the column } under 120-200 kV? Any recommendations are welcome, the literature helps a } bit, but is usually sketchy about these fine details of preparation } procedure. } thanks in advance, } N } } _____________________________ } Nicholas W. Hayman \ } Dept. of Geological Sci., U.W.\ http://www.geology.washington.edu/~hayman } Box 351310, Seattle WA 98195 \_________________________________________ } } } }
Dear All, Does anyone know of an EDS mineral spectrum database or mineral identification program? Is there such a thing available? Possibly a program that would allow the input of an image file of the unknown mineral spectrum thereby generating a list of possible 'best-fit' candidates. Or maybe it would work by predicting what the spectrum of a certain mineral should look like for a particular beam voltage etc. Regards Martin Roe
Martin J. Roe Macaulay Land Use Research Institute Craigiebuckler Aberdeen Scotland UK Phone 01224 318611 e-mail m.roe-at-mluri.sari.ac.uk
I will wait for info about sensitivity. I think, for EM in particular, sensitivity is very important parameter of the system. I am surprised that manufacturers don't have data on this matter. If sensitivity, say 10 times higher than SO-163 film - it may be a great reason to switch from film to the CCD. The major disadvantage of the CCD cameras for TEM is their price in my point of view. Thanks for your respond.
Sergey
} Date: Wed, 17 May 2000 11:34:05 -0600 } From: Michael Bode {mb-at-Soft-Imaging.com} } Subject: RE: new developments in imaging systems? } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} } X-Mailer: Internet Mail Service (5.0.1457.3) } } Sergey, } } I will try to find out what we have regarding the relative } sensitivities. One thing that I can say now is, that we acquire images } with a 50 or 100 msec exposure of the digital camera when the film } requires an exposure of about 2 seconds. This would mean a 20 to 40 } times better sensitivity of the CCD camera. But to answer your question } in more detail would require to compare also the resolution of film and } camera and that is where it becomes very difficult, as it is not easy to } determine the resolution of film in terms of spatial resolution and } dynamic range, as both are interwoven. I will try to find some answers } for you. } } Regarding the linearity: As the CCDs simply count Photons, they have } almost perfect linearity over their complete dynamic range. Even more so } for TEMs, where all Photons have the same energy and the quantum } efficiency does not change from photon to photon. The Phosphor is a part } of a system that could theoretically introduce some non-linearities. } However, I did measure the linearity of some TEM camera systems a few } years back and did not find any significant deviations from linearity. } } Michael } } 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: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } -----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] } Sent: Wednesday, May 17, 2000 1:29 AM } To: Michael Bode } Subject: RE: new developments in imaging systems? } } } Mickhael hello } } I have question for you. I am thinking about adding CCD camera to my } JEM1200EX. The information I gathered from Internet is not so } optimistic. } The standard CCD resolution is about 1-1.5 million pixels, 2 million is } a } max as I understand. The price for cheaper camera is about 20-30 K$ - } much } more that I expect to spend for "film" process. There are two things } may } attract me to the modern CCD camera: dynamic range and sensitivity. I } am } pretty sure that dynamic range for CCD itself is a few orders better } than } any film available. But what about phosphorus screen? Does it reduce } dynamic range for the EM images? How dramatic? This is my first } question. } The next question is: could you tell me something about sensitivity of } the } modern CCD cameras used in EM? I am using dark field imaging at x80K } magnification and the exposure time for SO-163 (non deluded D-19, 7 min, } 20oC) is about 2 sec. I called GATAN, but they did not say anything } useful. } Could you provide some comparison of your side-mount camera with } sensitivity of the SO-163 film at condition I mentioned? I will greatly } appreciate any information in this matter. Thanks. Sergey. } } } } Date: Fri, 12 May 2000 13:46:01 -0600 } } From: Michael Bode {mb-at-Soft-Imaging.com} } } Subject: RE: new developments in imaging systems? } } To: "'Microscopy-at-MSA.Microscopy.Com'" } {Microscopy-at-sparc5.microscopy.com} } } Cc: 'Margaret Dienelt' {brannign-at-asrr.arsusda.gov} } } X-Mailer: Internet Mail Service (5.0.1457.3) } } } } ----------------------------------------------------------------------- } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America }
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I'm no digital imaging guru, but I have been following this thread with interest. This whole discussion has been of interest to me because of the idea of the passive 4x5 (camera back) "detector" that could provide an extremely simple user venue for converting those countless analog SEMs into a very usable digital output format.
We have a 10 year old JEOL 840 microanalysis system that we would like to see in operation for another 10 years, and I think it may be possible. However, digital imaging, although "doable", is not "convenient" for us, at this point for this instrument. Too much time and effort is required in digital acquisition, maintenance of instrument condition info with the image, labeling the image, and archiving. Then on top of it all, the digital image is often not quite as good as the analog Polaroid shot that also has the Mag, Bar Scale, KeV, WD, and other info permanently incorporated (We use Type 53 for much of our work.) Don't get me wrong, we do some digital imaging with it, but "it ain't the same" as using a newer digital scope with integrated digital control and digital interface. If someone put such a passive digital detector into a 4x5 camera back-type mount, and it was capable of passively detecting at least 2500 horizontal lines and an approximately equivalent vertical resolution, I believe it would be a huge success with the analog SEMs. The 840 and many other instruments like it are great conventional SEMs, and such a simple interface (if affordable) would greatly extend their value. Many of these scanners have exceptional imaging capability, but often the easiest/best way to record that impressive image is by Polaroid film.
Is it doable? Brad Huggins
} ---------- } From: Michael Bode[SMTP:mb-at-Soft-Imaging.com] } Sent: Wednesday, May 17, 2000 1:36 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Cc: 'John Foust' } Subject: RE: SEM: digital 4x5 backs } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Yes, there is. } } It's called a "passive" or "listening" digital image acquisition system, } such as our ADDA II or similar devices from other manufacturers. } Benefits: fairly easy to set up and use. Disadvantages (as opposed to an } "active" or "talking" system): You're still limited to what the } microscope can provide in terms of resolution, dwell time, etc. } } Michael } } 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: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } -----Original Message----- } } From: John Foust [mailto:jfoust-at-threedee.com] } Sent: Tuesday, May 16, 2000 8:06 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: SEM: digital 4x5 backs } } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } At 04:02 PM 5/15/00 -0700, Dr. Gary Gaugler wrote: } } It seems to me that no digital camera system would work on a SEM } } in place of a Polaroid or other film-based output device. Since the } } recording CRT in a SEM is based on a sequential line scan, one } } would need a camera that would capture each line as it is produced. } } What you're saying is there must be a system out there that } digitizes that single stream of line scan intensities, then } processes all that data inside the computer as an image as } opposed to trying to digitize the frame buffer. } } - John } }
} Dear All, } Does anyone know of an EDS mineral spectrum database or } mineral identification program? Is there such a thing available? } Possibly a program that would allow the input of an image file of } the unknown mineral spectrum thereby generating a list of possible } 'best-fit' candidates. Or maybe it would work by predicting what the } spectrum of a certain mineral should look like for a particular beam } voltage etc. } Regards
} Martin Roe
Martin,
I know of no publicly available database of mineral EDX spectra or identification software. However, I believe several commercial EDX systems have a facility for generating your own database by collecting spectra from standard mineral samples. The software can then compare a spectrum from an unknown with those in the database.
One must be very careful of comparing like with like. All instrument parameters such as kV, tilt, etc, etc must be equivalent for a reasonable chance of a correct match.
I use the Desk Top Spectrum Analyser (DTSA) program from NIST to simulate EDX spectra from minerals, ceramics etc from the known composition. It requires a fair bit of effort to master the program but I feel it is worth the effort. Of course it does a lot more than just simulate spectra, including analysis of real measured spectra with a huge amount of flexibility in choosing analysis parameters.
DTSA is available free from the NIST web site:
http://www.cstl.nist.gov/div837/837.02/dtsa.html
Hope this helps,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
For what it's worth - if anything, there is a company out here in California called Silicon Film Technologies that has developed a technology that converts a traditional 35mm SLR camera into a digital camera. You essentially just replace the film with something they call (e)film. I don't know if they have any interest in developing something for this application, but it may be worth a look. It's a pretty nifty concept for people who want to do both traditional film photography and also digital photography with the same equipment.
I have copied a blurb from their website to give you an overview of what they do:
"Our EFS product suite is a digital photo system comprised of an electronic film cartridge and a carrier/adapter. The user simply inserts the cartridge, called (e)film, into the film cavity in the back of a conventional 35mm SLR camera. After recording up to 24 images, the cartridge is placed into a carrier/adapter, called (e)port, which may then be connected directly to a personal computer, enabling images to be downloaded quickly into the computer and then printed, sent via email, or modified into a photo end product using photo management software such as Adobe PhotoShop LE, which comes bundled with the EFS system. We will also market a digital photo storage module, called (e)box, which can store and transport hundreds of digital images in the field when the photographer does not have access to a computer.
The EFS system transforms conventional camera equipment into a digital image capture system. It is the only system currently available that provides the convenience and flexibility of choosing between conventional and electronic film formats with the same camera body. The product is aimed toward the large, installed base of 35mm camera owners who would like to enjoy the benefits of digital imaging while not giving up the cameras, lenses, and photo accessories with which they are familiar."
You can check out their website for more information at www.siliconfilm.com.
DISCLAIMER: I do own a small amount of stock in their parent company, Irvine Sensors Corporation. Of course, even if all of you bought a dozen of their neat little cameras, I'd still have to keep my day job, but I thought I should disclose it.
David Henriks Vice President TEL: 800-728-2233 (toll free in the USA) South Bay Technology, Inc. +1-949-492-2600 1120 Via Callejon FAX: +1-949-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by "Huggins, Bradley J" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm no digital imaging guru, but I have been following this thread with interest. This whole discussion has been of interest to me because of the idea of the passive 4x5 (camera back) "detector" that could provide an extremely simple user venue for converting those countless analog SEMs into a very usable digital output format.
We have a 10 year old JEOL 840 microanalysis system that we would like to see in operation for another 10 years, and I think it may be possible. However, digital imaging, although "doable", is not "convenient" for us, at this point for this instrument. Too much time and effort is required in digital acquisition, maintenance of instrument condition info with the image, labeling the image, and archiving. Then on top of it all, the digital image is often not quite as good as the analog Polaroid shot that also has the Mag, Bar Scale, KeV, WD, and other info permanently incorporated (We use Type 53 for much of our work.) Don't get me wrong, we do some digital imaging with it, but "it ain't the same" as using a newer digital scope with integrated digital control and digital interface. If someone put such a passive digital detector into a 4x5 camera back-type mount, and it was capable of passively detecting at least 2500 horizontal lines and an approximately equivalent vertical resolution, I believe it would be a huge success with the analog SEMs. The 840 and many other instruments like it are great conventional SEMs, and such a simple interface (if affordable) would greatly extend their value. Many of these scanners have exceptional imaging capability, but often the easiest/best way to record that impressive image is by Polaroid film.
Dear Martin: An interesting, but difficult idea. Besides the problem of instrument parameters, and settings, there is the problem of mineral compositions being in many cases very similar, but having different structures. For example, there are several iron oxide/hydroxide phases that would be difficult to tell apart with an EDS analysis. More complex are the silicates which have a number of different major structural families, many having the same or similar chemical compositions. There are sites on the web that would allow you to input an element list and it will output all the minerals with those elements and their formulas. It won't identify the minerals, but it would give you a start. To identify them you need to do optical or x-ray diffraction or some other more appropriate technique. Michael L. Boucher Sr. mboucher-at-isd.net http://www.isd.net/mboucher -----Original Message----- } From: Martin J. Roe {m.roe-at-mluri.sari.ac.uk} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
If sensitivity was ten times higher, one would in some cases save some beam damage and in most cases suffer excessive electron noise. As a rough guide, electron noise becomes apparent at 30x enlargement, rarely a problem. Magnification is a linear function and electron density relates to an area, but clearly very short exposure images would be much noisier and could not be enlarged nearly as much. When not enough electrons form an image it appears grainy, which makes it unsuitable for further enlarging. Photo enlarging utilises higher depths-of-field and without this, very high power TEM is much, much harder. By nature, slower emulsions have finer grain and higher resolution and contrast. It is fortuitous that these desirable characteristics run in tandem with relatively long exposures, so the image is formed by more electrons. TEM films rate about 6 ISO, so X-ray film at about ISI 3000 is about a 10x faster than TEM films. Unfortunately its rather grainy, but if the exposure was well adjusted, chances are that electron noise would be more bothersome than the film's grain. Digital is inevitable and already fairly common, but some aces remain with conventional film. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, May 18, 2000 5:06 AM, Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] wrote: } } } Thank you Mickhael. } } I will wait for info about sensitivity. I think, for EM in particular, } sensitivity is very important parameter of the system. I am surprised that } manufacturers don't have data on this matter. If sensitivity, say 10 times } higher than SO-163 film - it may be a great reason to switch from film to } the CCD. The major disadvantage of the CCD cameras for TEM is their price } in my point of view. } Thanks for your respond. } } Sergey } } } } Date: Wed, 17 May 2000 11:34:05 -0600 } } From: Michael Bode {mb-at-Soft-Imaging.com} } } Subject: RE: new developments in imaging systems? } } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} } } X-Mailer: Internet Mail Service (5.0.1457.3) } } } } Sergey, } } } } I will try to find out what we have regarding the relative } } sensitivities. One thing that I can say now is, that we acquire images } } with a 50 or 100 msec exposure of the digital camera when the film } } requires an exposure of about 2 seconds. This would mean a 20 to 40 } } times better sensitivity of the CCD camera. But to answer your question } } in more detail would require to compare also the resolution of film and } } camera and that is where it becomes very difficult, as it is not easy to } } determine the resolution of film in terms of spatial resolution and } } dynamic range, as both are interwoven. I will try to find some answers } } for you. } } } } Regarding the linearity: As the CCDs simply count Photons, they have } } almost perfect linearity over their complete dynamic range. Even more so } } for TEMs, where all Photons have the same energy and the quantum } } efficiency does not change from photon to photon. The Phosphor is a part } } of a system that could theoretically introduce some non-linearities. } } However, I did measure the linearity of some TEM camera systems a few } } years back and did not find any significant deviations from linearity. } } } } Michael } } } } 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: mailto:info-at-soft-imaging.com } } web: http://www.soft-imaging.com } } =================================== } } } } } } } } -----Original Message----- } } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] } } Sent: Wednesday, May 17, 2000 1:29 AM } } To: Michael Bode } } Subject: RE: new developments in imaging systems? } } } } } } Mickhael hello } } } } I have question for you. I am thinking about adding CCD camera to my } } JEM1200EX. The information I gathered from Internet is not so } } optimistic. } } The standard CCD resolution is about 1-1.5 million pixels, 2 million is } } a } } max as I understand. The price for cheaper camera is about 20-30 K$ - } } much } } more that I expect to spend for "film" process. There are two things } } may } } attract me to the modern CCD camera: dynamic range and sensitivity. I } } am } } pretty sure that dynamic range for CCD itself is a few orders better } } than } } any film available. But what about phosphorus screen? Does it reduce } } dynamic range for the EM images? How dramatic? This is my first } } question. } } The next question is: could you tell me something about sensitivity of } } the } } modern CCD cameras used in EM? I am using dark field imaging at x80K } } magnification and the exposure time for SO-163 (non deluded D-19, 7 min, } } 20oC) is about 2 sec. I called GATAN, but they did not say anything } } useful. } } Could you provide some comparison of your side-mount camera with } } sensitivity of the SO-163 film at condition I mentioned? I will greatly } } appreciate any information in this matter. Thanks. Sergey. } } } } } } } Date: Fri, 12 May 2000 13:46:01 -0600 } } } From: Michael Bode {mb-at-Soft-Imaging.com} } } } Subject: RE: new developments in imaging systems? } } } To: "'Microscopy-at-MSA.Microscopy.Com'" } } {Microscopy-at-sparc5.microscopy.com} } } } Cc: 'Margaret Dienelt' {brannign-at-asrr.arsusda.gov} } } } X-Mailer: Internet Mail Service (5.0.1457.3) } } } } } } ----------------------------------------------------------------------- } } - } } } 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 } } } ----------------------------------------------------------------------- } } . } } } } } } } } } Margaret, } } } } } } As a former user and current vendor of such systems as you are } } inquiring } } } about I can try to provide a bit of information regarding camera } } } improvements: } } } } } } There have been a number of improvements, but I am not sure what you } } are } } } comparing the latest cameras against. Cameras are now usually cooled } } and } } } provide 12 bits per pixel, the number of pixels has gone up a bit (but } } } not much in general), and cameras read out faster than they used to (up } } } to 20 fps and more). I think all cameras now use a line transfer } } } mechanism, which makes shutters obsolete. } } } On the software side, real-time FFT and real-time shading correction } } can } } } be done now due to faster computers without special processing boards, } } } and there have been other software developments that make using the } } } cameras and computers easier. } } } Other changes that affect the usability of cameras is the use of } } } pneumatics to insert and retract the phosphors, higher frame rates for } } } live viewing with the camera, etc. } } } } } } If you have questions, please give me a call, drop me an email, or go } } to } } } our web site. } } } } } } Michael } } } } } } } } } 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: mailto:info-at-soft-imaging.com } } } web: http://www.soft-imaging.com } } } =================================== } } } } } } } } } } } } -----Original Message----- } } } } From: Margaret Dienelt [mailto:brannign-at-asrr.arsusda.gov] } } } Sent: Thursday, May 11, 2000 12:43 PM } } } To: Microscopy-at-sparc5.microscopy.com } } } Subject: TEM: new developments in imaging systems? } } } } } } } } } ----------------------------------------------------------------------- } } - } } } 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, } } } } } } Year after year I hopefully gather information about digital imaging } } } systems for TEMs (ours is a JEOL 100CX) , only to learn we have no } } } money. This year it looks like it might really happen but I have not } } } kept up with innovations in the field and am wondering the following: } } } } } } 1. Anything new in the last two years -- especially in terms of } } } cameras? I'm most familiar with the Gatan and AMT systems but their } } } web sites don't reflect much in the way of changes over a year ago. } } } 2. With more and more microscopists finally getting their systems -- } } } I'd love to get feedback. } } } } } } Thanks, } } } Margaret } } } } } } P.S. Would welcome contacts from vendors. } } } } } } -- } } } Margaret Dienelt } } } } } } Plant Pathologist } } } Electron Microscopy Lab } } } } } } Floral and Nursery Plants Research Unit } } } U.S. National Arboretum/Agricultural Research Service/USDA } } } } } } B. 010A, Rm. 238, BARC-W } } } 10300 Baltimore Avenue } } } Beltsville MD. 20705 USA } } } } } } (301) 504-6097 } } } Fax: (301) 504-5096 } } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } http://www.bol.ucla.edu/~sryazant } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant } }
I can identify with your situation. While I do not know the intricacies of the JEOL instruments, some SEMs are made to accept external drive for x-ray analysis. This same input scheme works perfectly for active control of the SEM and direct digital capture of images. At the worst, you can replicate the resolution of your record CRT using a passive mode. How easy either of these modes are depends greatly on how the SEM system was designed.
gary g.
At 01:44 PM 5/17/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Dear Martin: } An interesting, but difficult idea. Besides the problem of instrument } parameters, and settings, there is the problem of mineral compositions being } in many cases very similar, but having different structures.
And then there is also the converse: the problem of mineral compositions being in many cases very different, but having the same structures. One of a huge number of examples would be the feldspar series where Na and Ca can substitute for each other continuously from Na to Ca endmembers.
Various other bits of crystallographic information, some intangibles such as crystal shape and growth habit, associations with other minerals and so on, would have to be taken into account in such software for it to really zero in on an unambiguous ID for you.
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
Hello all ..... we have an old ETEC Omniscan SEM, and we need to contact with any people that can supply manuals ....( fundamentally electric and electronic schematic )
any help is welcome
best regards
=================================================== Fernando D. Balducci Laboratorio de Microscopia Electr—nica Facultad de Ingenier’a - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
At 07:02 PM 5/17/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think that their aim is a bit off the target. Its nice to not have to give up the hardware but their offering makes the user give up the benefits of 35mm film--frame size and selection of speed. And, a field of view factor of 2.8 is absurd.
I beta tested what must have been an initial offering of this product early last year. It was by a different company. Same idea though. Same deficiencies. The sensor is 1.3M pixels and has an odd aspect ratio of 1.25 (35mm frame is 1.5). Also, and most importantly, the product does not image the entire 35mm frame, only a small central portion. 1.3M pixel point and shoot cameras are a lot cheaper than this silicon film thing. A really dedicated user would have to buy more than one silicon film insert. At $600 each, that would buy a lot of P&S cameras. But there is no need to do that since one just pops out a SmartMedia or Compact Flash module ($150 or so each).
I still say wait and see how the Fuji Finepix S1 Pro performs. If it lives up to spec, I'd say that it will be a raging success and a major turning point in digital cameras which are based on the 35mm format.
gary g.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Dear Friends, Can anyone give me advice regarding the dismantling and the packing for shipment of a Philips 201 TEM? I need to pay particular attention to not disturbing the alignment. I plan to move the TEM from Univ. of Delaware to Naples, Florida in an enclosed U-Haul Trailer. Any help or suggestions will be most welcome. If anyone has hands-on experience in doing this sort of thing and is willing to give me a hand at U-Del, please contact me and name your price. I'll gladly pay for the assistance. Best regards, Mike Urbanik www.crystalguru.com
I will reiterate my point and that of others. An SEM technically has video only coming from one point in the image at a time. Sure, you could pay lots of $$ for a big CCD with the resolution you desire, or you could simply digitize the one (or more) video signals as the beam scans the screen. You have the choice of either asserting the x-y positions through active digital control or you can read them off passively.
From a hardware perspective, it is a much easier (and cheaper) task to build an active or passive system like those on the market than to build a 4x5 camera back detector. For both systems you would still need the software and computer
At 10:58 PM 5/17/2000 -0700, Gary Gaugler wrote:
} I can identify with your situation. While I do not know the intricacies } of the JEOL instruments, some SEMs are made to accept external } drive for x-ray analysis. This same input scheme works perfectly } for active control of the SEM and direct digital capture of images. } At the worst, you can replicate the resolution of your record CRT } using a passive mode. How easy either of these modes are depends } greatly on how the SEM system was designed. } } gary g. } } At 01:44 PM 5/17/00, you wrote: } } } } I'm no digital imaging guru, but I have been following this thread with } } interest. This whole discussion has been of interest to me because of the } } idea of the passive 4x5 (camera back) "detector" that could provide an } } extremely simple user venue for converting those countless analog SEMs into } } a very usable digital output format. } } } } We have a 10 year old JEOL 840 microanalysis system that we would like to } } see in operation for another 10 years, and I think it may be possible. } } However, digital imaging, although "doable", is not "convenient" for us, at } } this point for this instrument. Too much time and effort is required in } } digital acquisition, maintenance of instrument condition info with the } } image, labeling the image, and archiving. Then on top of it all, the } } digital image is often not quite as good as the analog Polaroid shot that } } also has the Mag, Bar Scale, KeV, WD, and other info permanently } } incorporated (We use Type 53 for much of our work.) Don't get me wrong, we } } do some digital imaging with it, but "it ain't the same" as using a newer } } digital scope with integrated digital control and digital interface. If } } someone put such a passive digital detector into a 4x5 camera back-type } } mount, and it was capable of passively detecting at least 2500 horizontal } } lines and an approximately equivalent vertical resolution, I believe it } } would be a huge success with the analog SEMs. The 840 and many other } } instruments like it are great conventional SEMs, and such a simple interface } } (if affordable) would greatly extend their value. Many of these scanners } } have exceptional imaging capability, but often the easiest/best way to } } record that impressive image is by Polaroid film. } } } } Is it doable? } } Brad Huggins
I fuss with mineral identification via EDS spectra every day. I keep the program "Mineral" running at all times. It is a Windows Filemaker Pro database which includes the chemical formula, physical properties, type locale and 7 to 10 x-ray diffraction lines for 4500 accredited mineral species and many un-named ones. Searches can be constructed from any single or combination of fields. The software is available from Aleph Enterprises in Livermore, CA (510 443 7319) The price was around $550 a few years ago. Less expensive DOS based, but similar software is available from the Fersmann Institute.
More complete chemistry (but no spectra) is offered by Alexander Holzel's MDAT program. It includes a database of chemical analyses of many minerals and can perform a search based on weight per cents of elements present as well the unknown's physical properties and x-ray diffraction data. This software is $1000 and up depending upon options. Dr. Holzel's e-mail address was Compuserve100333,2771 as of last August. He is in Ober-Olm, Germany.
None of the software programs work directly from spectra and only MDAT contains sample analyses so you will likely be estimating peak heights from chemical formula. For more definitive results you will need to start building your own spectrum library from your own reference materials.
Even with a high elemental analysis correlation, one will almost always need supplemental methods for a definitive ID. This is usually x-ray diffraction or optical microscopy.
Some day affordable EBSP retrofitted into SEMS will allow the analyst to distinguish, in situ in polished section between orthorhombic FeS2 (marcasite) and cubic FeS2 (pyrite). Currently available systems are $90,000+, I think.
If you need help obtaining grains of some of the less common minerals, I can help. I supply a catalog which includes hundreds of well identified reference quality mineral grains in addition to synthetic probe standards.
Bart Cannon Cannon Microprobe 1041 NE 100th Street Seattle, WA 98125 206 522 9233 (3947 fax)
In evaluating some electron beam damage data, I am trying to determine the electron dose to the sample using a small screen current reading from a JEOL 2010 TEM.
The screen current density is given in pA/cm^2 on the microscope terminal, and can be converted to dose (electrons/cm^2*s) in the sample. I have spoken to JEOL, and they indicated that the actual current density on the screen is the reading multiplied by a factor of 10.
I am unable to use a borrowed stage with a Farraday cup for calibration on this microscope because it is suspect for internal radioactive contamination. Thus, I would be grateful for insights from other users of 2010's who may have checked the accuracy of the reading (whether or not the factor of 10 is right, etc.) on their instruments.
Many Thanks, Wharton -- ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724
Dear Martin, As the other responders have stated, minerals can be difficult to identify with EDS only. There is no substitute for experience (and some diffraction data). My rule of thumb is to group minerals as do most mineral classification schemes; is it a silicate, oxide, carbonate, sulfide, phosphate, salt, organic, or metalic phase? Sometimes it's hard to decide even at this level (i.e., If you don't have a light element detector, you can't differentiate between oxides and carbonates, P and Si substitute for each other). Most times you can. Since I often work with silicates, my second observation is to look at Si/Al. This will help limit the choices greatly, but is not usually diagnostic by itself. There are relatively few minerals with Al} Si. Knowledge of the approximate Si/Al (the most common tetrahedral cations) combined with the ratio of Si and Al with other elements like Mg, Ti, Cr, Mn and Fe (sometimes Ca) will give you an idea of the tetrahedral to octahedral ratio. The alkali elements Na and K, and also Ca in significant abundances are very important for identifying feldspars and sheet silicates, but again, there are no hard and fast rules that I've come up with. The best thing is to know your mineral compositions (and their structures) or hire a mineralogist!;-) I also agree that even the mineralogist can find the DTSA program very useful and worth investing in a MAC. Hope this helps. Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets Johns Hopkins University Baltimore, Maryland 21218 USA Phone: (410) 516-8342 Fax: (410) 516-7933 e-mail: klivi-at-jhu.edu
Due to the price of today's microscopes I am in need of advice concerning the quality of Russian microscopes. I had one in Spain and it was excellent. Please, advice. Thanks, Jose
Hello Folks, In reading the third edition of Hayat's Principles and Techniques of Electron Microscopy, there was mention of a 'Safety Chart, Chemicals in Electron Microscopy', for free distribution, by EMscope Laboratories Ltd (Kingsnorth Industrial Estate, Ashford, Kent). Does anyone out there have it? Or can anyone tell me how to get it or any wall chart that lists chemicals/hazards specific to EM? Thanks, Winnie
Lou, Very true. However, not one instrument can unambigously identify ALL minerals! In the case of my interests, PLM is not very useful since most of the crystals in shale are less than 2 um. Martin's original question was regarding how to identify EDS patterns, not what is the best way to identify a mineral. Ciao for now, Ken
} Colleagues; } } My question is why have you forgotten that polarized light microscopes were } devised for a need to identify and classify minerals? It almost seems } intentional to ignore it. Perhaps I am beginning to sound like Dr. McCrone } as I get older, but you dont have to throw a million dollar instrument at a } problem to solve it. If people have a problem with a $20k light microscope } doing a job better, that is there problem not mine. Just dont neglect the } fact that PLM is still a powerful technique in the hands of a competent } microscopist. PLM is still the standard for characterizing new crystal } compounds at the be.....visit any pharmaceutical companys R&D department and } you will see that it is so. } } Lou Solebello } } } ----- Original Message ----- } From: Kenneth JT Livi {klivi-at-jhu.edu} } To: {microscopy-at-sparc5.microscopy.com} } Cc: {m.roe-at-mluri.sari.ac.uk} } Sent: Thursday, May 18, 2000 8:15 AM } Subject: Re: EDS mineral identification and database } } } } ------------------------------------------------------------------------ } } 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 Martin, } } As the other responders have stated, minerals can be difficult to identify } } with EDS only. There is no substitute for experience (and some diffraction } } data). My rule of thumb is to group minerals as do most mineral } } classification schemes; is it a silicate, oxide, carbonate, sulfide, } } phosphate, salt, organic, or metalic phase? Sometimes it's hard to decide } } even at this level (i.e., If you don't have a light element detector, you } } can't differentiate between oxides and carbonates, P and Si substitute for } } each other). Most times you can. Since I often work with silicates, my } } second observation is to look at Si/Al. This will help limit the choices } } greatly, but is not usually diagnostic by itself. There are relatively few } } minerals with Al} Si. Knowledge of the approximate Si/Al (the most common } } tetrahedral cations) combined with the ratio of Si and Al with other } } elements like Mg, Ti, Cr, Mn and Fe (sometimes Ca) will give you an idea } of } } the tetrahedral to octahedral ratio. The alkali elements Na and K, and } also } } Ca in significant abundances are very important for identifying feldspars } } and sheet silicates, but again, there are no hard and fast rules that I've } } come up with. The best thing is to know your mineral compositions (and } } their structures) or hire a mineralogist!;-) I also agree that even the } } mineralogist can find the DTSA program very useful and worth investing in } a } } MAC. Hope this helps. } } Ciao for now, } } Ken } } } } Kenneth JT Livi } } Department of Earth and Planetary Sciences } } 34th and Charles Streets } } Johns Hopkins University } } Baltimore, Maryland 21218 USA } } Phone: (410) 516-8342 } } Fax: (410) 516-7933 } } e-mail: klivi-at-jhu.edu } } } } } }
Yes, we are aware of the problems in reaching our web site in the last few days. We have posted our new web site of microscopy supplies at a new URL and it is now up and running. Please change your bookmarks to:
http://www.laddresearch.com
Thank you,
John Arnott --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net
Warren's point is right on. The JEOL 840 or any other SEM that has a record CRT for 4x5 film or Polaroid is the signal source of passive information. It is not a big technical deal to do this. Depending on how readily available the CRT signals are (blank, frame, etc.), it may not be convenient or really simple. Some systems use BNC connectors to snake the signals throughout the system. T-ing off of these makes digitizing the SEM quite easy. If the signals are hardwired, it is more work to install the capture system. But in either case, it is a one time effort.
A passive system will record all legends just like the Polaroid does. But it will do it a lot better. The reason is that the tonal range and exposure latitude of a Polaroid is rather poor compared to real film. Digital capture systems are typically 10-bits; or 12-bits for ones with really exceptional dynamic range. Either of these are vastly superior to Polaroid prints.
The number of horizontal lines that a passive system will capture is the same as a Polaroid. This is because the record CRT circuitry fixes this dimension. However, the scan rate alters the pixel dwell time. Slower scan rates produce images that have less noise--be it Polaroid or digital capture. If the Polaroid print works OK for you, I would suggest that a digital capture system would be even better.
Compared to the cost of a modern computerized SEM, a digitizing attachment can be the key factor in keeping a good old SEM.
gg
At 06:42 AM 5/18/00, you wrote:
} I will reiterate my point and that of others. An SEM technically has video } only coming from one point in the image at a time. Sure, you could pay } lots of $$ for a big CCD with the resolution you desire, or you could } simply digitize the one (or more) video signals as the beam scans the } screen. You have the choice of either asserting the x-y positions through } active digital control or you can read them off passively. } } From a hardware perspective, it is a much easier (and cheaper) task to } build an active or passive system like those on the market than to build } a 4x5 camera back detector. For both systems you would still need the } software and computer } } At 10:58 PM 5/17/2000 -0700, Gary Gaugler wrote: } } } I can identify with your situation. While I do not know the intricacies } } of the JEOL instruments, some SEMs are made to accept external } } drive for x-ray analysis. This same input scheme works perfectly } } for active control of the SEM and direct digital capture of images. } } At the worst, you can replicate the resolution of your record CRT } } using a passive mode. How easy either of these modes are depends } } greatly on how the SEM system was designed. } } } } gary g. } } } } At 01:44 PM 5/17/00, you wrote: } } } } } } I'm no digital imaging guru, but I have been following this thread with } } } interest. This whole discussion has been of interest to me because of the } } } idea of the passive 4x5 (camera back) "detector" that could provide an } } } extremely simple user venue for converting those countless analog SEMs into } } } a very usable digital output format. } } } } } } We have a 10 year old JEOL 840 microanalysis system that we would like to } } } see in operation for another 10 years, and I think it may be possible. } } } However, digital imaging, although "doable", is not "convenient" for us, at } } } this point for this instrument. Too much time and effort is required in } } } digital acquisition, maintenance of instrument condition info with the } } } image, labeling the image, and archiving. Then on top of it all, the } } } digital image is often not quite as good as the analog Polaroid shot that } } } also has the Mag, Bar Scale, KeV, WD, and other info permanently } } } incorporated (We use Type 53 for much of our work.) Don't get me wrong, we } } } do some digital imaging with it, but "it ain't the same" as using a newer } } } digital scope with integrated digital control and digital interface. If } } } someone put such a passive digital detector into a 4x5 camera back-type } } } mount, and it was capable of passively detecting at least 2500 horizontal } } } lines and an approximately equivalent vertical resolution, I believe it } } } would be a huge success with the analog SEMs. The 840 and many other } } } instruments like it are great conventional SEMs, and such a simple interface } } } (if affordable) would greatly extend their value. Many of these scanners } } } have exceptional imaging capability, but often the easiest/best way to } } } record that impressive image is by Polaroid film. } } } } } } Is it doable? } } } Brad Huggins }
I am in need of a way to modify my current method of preparing TEM cross-sections as to introduce no heating. Specifically, I need to find new adhesive materials. We currently use thermally cured M-bond, Gatan G1 epoxy and crystal bond wax - all of which require heat.
If anyone has information on an adhesive that ion mills at a rate to similar Si, is stable under the electron beam, will cure at room temperature within about 24 hours, and once cured is impervious to solvents such as acetone, please respond. It would be an additional plus if the adhesive has low viscosity, so it can be used to secure TEM grids to the specimens.
Also I am looking for a replacement for the wax that we currently use to affix the samples to the polishing studs. This should cure quickly, bond strongly enough to endure the mechanical stresses of grinding, and be readily soluble in a solvent other than water so the samples can be removed from the studs.
Any help would be greatly appreciated.
Brenda
Brenda I. Prenitzer, Ph.D. Member of Technical Staff Cirent Semiconductor (Lucent Technologies) 9333 S. John Young Parkway 6D-Lab Orlando, FL 32819-8612
Our management has finally agreed to connect my electron microscopes to a recirculating water system.
I need some ideas what type of systems are on the market and the Australian agent for these units. We need to attach 2 SEMs, 1 TEM plus 1 coating unit to the new reciculating line.
Thank you
Hans Brinkies Senior Lecturer Swinburne, University of Technology School of Engineering and Science P.O.Box 218 - Hawthorn - Vic -3122 - Australia Phone: +61 3 9214 8657 Fax: +61 3 9214 8264 Email: Hbrinkies-at-swin.edu.au
1) Sensitivity: The CCD cameras are sensitive enough to see single electrons striking the phosphor. You can't get much more sensitive than that.
2) Price: I had a discussion about this with George McAuliffe in this forum about that theme a while ago. That thread was also printed in Microscopy Today. You may want to check the archives of this list server for "digital archiving/cost". I think George would agree with me, that the calculations of cost can swing in one or the other direction, depending on how you define cost, what you need to include and how many images you take (remember, a negative is on the order of $1 for the material alone, not labor time or anything else).
Michael
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: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Wednesday, May 17, 2000 1:06 PM To: Microscopy-at-sparc5.microscopy.com
Thank you Mickhael.
I will wait for info about sensitivity. I think, for EM in particular, sensitivity is very important parameter of the system. I am surprised that manufacturers don't have data on this matter. If sensitivity, say 10 times higher than SO-163 film - it may be a great reason to switch from film to the CCD. The major disadvantage of the CCD cameras for TEM is their price in my point of view. Thanks for your respond.
Sergey
} Date: Wed, 17 May 2000 11:34:05 -0600 } From: Michael Bode {mb-at-Soft-Imaging.com} } Subject: RE: new developments in imaging systems? } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} } X-Mailer: Internet Mail Service (5.0.1457.3) } } Sergey, } } I will try to find out what we have regarding the relative } sensitivities. One thing that I can say now is, that we acquire images } with a 50 or 100 msec exposure of the digital camera when the film } requires an exposure of about 2 seconds. This would mean a 20 to 40 } times better sensitivity of the CCD camera. But to answer your question } in more detail would require to compare also the resolution of film and } camera and that is where it becomes very difficult, as it is not easy to } determine the resolution of film in terms of spatial resolution and } dynamic range, as both are interwoven. I will try to find some answers } for you. } } Regarding the linearity: As the CCDs simply count Photons, they have } almost perfect linearity over their complete dynamic range. Even more so } for TEMs, where all Photons have the same energy and the quantum } efficiency does not change from photon to photon. The Phosphor is a part } of a system that could theoretically introduce some non-linearities. } However, I did measure the linearity of some TEM camera systems a few } years back and did not find any significant deviations from linearity. } } Michael } } 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: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } -----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] } Sent: Wednesday, May 17, 2000 1:29 AM } To: Michael Bode } Subject: RE: new developments in imaging systems? } } } Mickhael hello } } I have question for you. I am thinking about adding CCD camera to my } JEM1200EX. The information I gathered from Internet is not so } optimistic. } The standard CCD resolution is about 1-1.5 million pixels, 2 million is } a } max as I understand. The price for cheaper camera is about 20-30 K$ - } much } more that I expect to spend for "film" process. There are two things } may } attract me to the modern CCD camera: dynamic range and sensitivity. I } am } pretty sure that dynamic range for CCD itself is a few orders better } than } any film available. But what about phosphorus screen? Does it reduce } dynamic range for the EM images? How dramatic? This is my first } question. } The next question is: could you tell me something about sensitivity of } the } modern CCD cameras used in EM? I am using dark field imaging at x80K } magnification and the exposure time for SO-163 (non deluded D-19, 7 min, } 20oC) is about 2 sec. I called GATAN, but they did not say anything } useful. } Could you provide some comparison of your side-mount camera with } sensitivity of the SO-163 film at condition I mentioned? I will greatly } appreciate any information in this matter. Thanks. Sergey. } } } } Date: Fri, 12 May 2000 13:46:01 -0600 } } From: Michael Bode {mb-at-Soft-Imaging.com} } } Subject: RE: new developments in imaging systems? } } To: "'Microscopy-at-MSA.Microscopy.Com'" } {Microscopy-at-sparc5.microscopy.com} } } Cc: 'Margaret Dienelt' {brannign-at-asrr.arsusda.gov} } } X-Mailer: Internet Mail Service (5.0.1457.3) } } } } ---------------------------------------------------------------------- - } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America }
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
A few months ago, I asked the same question and received the answers I copied below. By the way, I would like to thank again to all who respond my question. I do appreciate it. I still use these procedures and am planning to prepare a study comparing these techniques. All of them work very well.
Hope these helps. Good luck.
Ranan Gulhan AKTAS, M.D. Trakya University, Faculty of Medicine Pathology Department Edirne 22030 TURKEY
Dear Dr. Ranan Gulhan Aktas Protocol as follows: De-wax in 100% xylene - small pieces of tissue - 1mm3 (3X) Re-hydrate to water xylene/ethanol 100% each 100% ethanol 96% ethanol 70% ethanol H2O GA in buffer} Normal EM processing from here on Buffer }
Contact me if you have any further queries
Kind regards John
Mr John F. Putterill Electron Microscopy Unit Tel: (Int) 27-12-529-9174 Pathology Section Fax: (Int) 27-12-529-9165 Onderstepoort Veterinary Institute E-mail: john-at-moon.ovi.ac.za Private Bag X05 http://www.ovi.ac.za Onderstepoort 0110 South Africa
The basic procedure for re-embedding paraffin embedded is relatively simple. We had a Lynx Tissue Processor set up to do it automatically. Cut the paraffin tissue into 1-2 mm cubes with a warm scalpel. Trim as much excess paraffin away as possible. De-paraffinize with 6 alternately warm/room temp xylene washes with agitation for 30 minutes each. Reverse the dehydration process from 100% EtOH to your normal EM buffer. Refix and postfix with appropriate EM fixatives, dehydrate, and embed. The smaller the paraffin pieces, typically the better the deparaffinization. Your tissue will never look great though. The initial fixation for LM is a poor TEM fixative. good Luck.
Chuck Butterick Engineered Carbons, Inc.
Ranan,
Kai Chein did some nice work in this in the 1970's. He dissolved osmium in xylene(1 percent)and then deparaffinized in that solution. This took care of osmicating and depariffinization in one step (assuming you want to use osmium). After several changes in this solution you can go right into resin.. I've done this many times and it works very well and saves a lot of time.
Tim Morken, B.S., EMT(MSA), HTL(ASCP) Infectious Disease Pathology Centers for Disease Control MS-G32 1600 Clifton Rd. Atlanta, GA 30333 USA
email: tim9-at-cdc.gov timcdc-at-hotmail.com
FAX: (404)639-3043
MICROSCOPY AND IMAGING SERVICE CENTER CELL BIOLOGY AND NEUROSCIENCES
PROCEDURE FOR PRELIMINARY PREPARATION OF PARAFFIN EMBEDDED TISSUE FOR ELECTRON MICROSCOPY:
1. PLACE 0.5-1.0 mm CUBES OF TISSUE FROM PARAFFIN BLOCK IN XYLENE FOR 3 CHANGES OF 30 MINUTES EACH.
2. PUT IN 100% ETHANOL FOR 2 CHANGES OF 15 MINUTES WITH AGITATION.
3. PUT IN 95% ETHANOL FOR 2 CHANGES FOR 15 MINUTES WITH AGITATION.
4. PUT IN 70% ETHANOL FOR 2 CHANGES FOR 15 MINUTES WITH AGITATION.
5. PUT IN 50% ETHANOL FOR 2 CHANGES FOR 15 MINUTES WITH AGITATION.
6. RINSE IN PBS FOR 2 CHANGES OF 5 MINUTES EACH WITH AGITATION.
7. FOLLOW ROUTINE PREPARATION OF TISSUE FOR ELECTRON MICROSCOPY.
REFERENCE:
PIERCE, A., 1972. "A MANUAL FOR HISTOLOGICAL TECHNICIANS", LITTLE AND BROWN, BOSTON.
Your morphology will not be great since the tissue was probably fixed in formalin and not glut. I've done this many times and have even had publications with this method. Make sure you get all of the paraffin out.
George Lawton Chief Electron Microscopist Microscopy and Imaging Service Center UT Southwestern Medical Center at Dallas Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
Hi: This is Bob Mixon ( I work with Bob Kayton on the PNEMS society and am on the OHSU campus) . I have re-embedded tissues from paraffin many times and many ways. I would recommend cutting out the piece of tissue from the paraffin block and putting through at least two changes of xylene for about 30 minutes each. Then you could put the tissue through either absolute alcohol or acetone to remove the rest of the paraffin. Probably 30 minutes and two changes. Some folks continue to run the tissue through a series of alcohol and try to refix in EM fix and post-fix in osmium. THIS IS FUTILE! I have never found any enhancement by doing this. You can simply dissolve osmium in the acetone and try fixing in this (two percent). and then running through by your routine into plastic (through alcohol, acetone, or propylene oxide etc). Thanks Bob Mixon
Hello Ranan.
Here is the method used by the pathology people around here.
- Remove tissue of interest from parafin embedded specimen, cut in 1-2 mm cubes. - Leave the specimens in fresh 1% OsO4 in XYLENE for 1,5 - 2 hrs. - Wash in xylene 2 times 10 min - One part resin(we use Epon-Araldite) + two parts xylene for 1 hr. - 1 part resin + 1 part xylene for 1 hr. - Resin for 30 min to 1 hour without a lid on the glass
All this steps on a carousel.
Embedd as usual, but keep the specimens in the resin over night before polymerization.
Good luck!
Best regards Randi Olsen
Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso MH-Breivika N-9037 TROMSO NORWAY
The following procedure may sound a little obvious, but I imagine you may be facing a rather limited access to all those books and journals, and I would feel really happy if this could help. I do wish you all the best in your EM lab-raising mission!
First, you will have to select smaller, "EM-size", portions of your paraffin-embedded specimens, cut them out and thoroughly deparaffinate. I would recommend at least 2 hours in xylene with frequent changes of xylene and some agitation; you should be able to see when it's all gone. Then several changes of 100% (anhydrous) ethanol, at least 1-1.5 h total time, and then gradually down the ethanol concentrations, like 95-85-70-50, 30 min or more at each step. Rinse 3x5 min in your choice of phosphate or cacodylate buffer, and then osmicate for 1-2 h, 1% OsO4 in the same buffer. Then dehydrate and embed for EM as you normally would.
Needless to say, even with the best original fixation the material will look pityful, but most of those diagnostically significant cell-to-cell junctions, filaments, etc. must still be there.
Best of luck! Sincerely, Vlad.
Vladislav V. Speransky Postdoctoral Research Associate School of Marine Sciences University of Maine 5722 Deering Hall Orono ME 04469-5722 Ph: 207 581 2998 FAX: 207 581 2969 Email: vladis-at-maine.maine.edu
Hi Keith,
At least for the specimens I happened to deal with so far (vertebrate tissues, cell culture, bacteria, microalgae), the material normally will not really blacken until you wash and start dehydrating it AFTER osmication in traditional aqueous OsO4 solutions. While in aqueous OsO4, it will remain brownish. (Unless, of course, you are using that simultaneous, single solution glutaraldehyde/OsO4 fixation, or a reducing buffer like PIPES, and have passed the time when it all turns black...)
The explanation used to be that, at the stage of osmication, while some of the OsO4, indeed, oxidizes nonsaturated lipids, etc., a lot of it simply dissolves in the lipids of the specimen as nonreduced OsO4. Then, say, the ethanol reduces that specifically accumulated OsO4, to form the so called "osmium black".
As for making FRESH solutions of OsO4 in xylene (or CCl4, etc.), I would just never have a spare OsO4 ampule (and enough paraffine blocks to EM-reevaluate) to try that! :-)
Sincerely, Vlad.
Vladislav V. Speransky Postdoctoral Research Associate School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Ph: 207 581 2998 FAX: 207 581 2969 Email: vladis-at-maine.maine.edu
} I was interested in the use of osmium in xylene that you described. Does the tissue blacken as } in a water-solution or does it simply become yellowish/brown? I am curious because the latter is } what happens in the absence of water during freeze-substitution using e.g. 1% in acetone at -80 } *C, although the colour change probably happens when warming up from low temperature.
Hello Keith.
According to the people here that most often works with this (Irene Lund) the blocks are not as dark as with standard methods, but light brown. We haven't done freeze-substitution with osmium, so it's not easy to compare directly. For this purpose we buy ampoulas with 0,1 gram OsO4.
Best regards Randi Olsen Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso MH-Breivika N-9037 TROMSO NORWAY
At least for the specimens I happened to deal with so far (vertebrate tissues, cell culture, bacteria, microalgae), the material normally will not really blacken until you wash and start dehydrating it AFTER osmication in traditional aqueous OsO4 solutions. While in aqueous OsO4, it will remain brownish. (Unless, of course, you are using that simultaneous, single solution glutaraldehyde/OsO4 fixation, or a reducing buffer like PIPES, and have passed the time when it all turns black...)
The explanation used to be that, at the stage of osmication, while some of the OsO4, indeed, oxidizes nonsaturated lipids, etc., a lot of it simply dissolves in the lipids of the specimen as nonreduced OsO4. Then, say, the ethanol reduces that specifically accumulated OsO4, to form the so called "osmium black".
As for making FRESH solutions of OsO4 in xylene (or CCl4, etc.), I would just never have a spare OsO4 ampule (and enough paraffine blocks to EM-reevaluate) to try that! :-)
Sincerely, Vlad.
Vladislav V. Speransky Postdoctoral Research Associate School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Ph: 207 581 2998 FAX: 207 581 2969 Email: vladis-at-maine.maine.edu
Dear HistoNetters,
Dr. Aktas asked about the re-embedding of paraffin embedded tissues in resin for observation by electron microscopy. There are two papers at our web site which describe this "Pop-Off" technique. Go to the URL provided in my signature file and follow the links to the JB-4 microtomy system, then to "Technical Information"
If anyone needs reprints of these articles and has no WWW access, send a note to Sonja White in my office (sales-at-ebsciences.com) {mailto:sales-at-ebsciences.com)} , and she'll s-mail you the dead tree version.
Best regards, Steven E. Slap, Vice-President
******************************************* Energy Beam Sciences, Inc. The Laboratory Microwave Company Adding Brilliance to Your Vision http://www.ebsciences.com {http://www.ebsciences.com} Ranan,
Kai Chein did some nice work in this in the 1970's. He dissolved osmium in xylene(1 percent)and then deparaffinized in that solution. This took care of osmicating and depariffinization in one step (assuming you want to use osmium). After several changes in this solution you can go right into resin.. I've done this many times and it works very well and saves a lot of time.
Tim Morken, B.S., EMT(MSA), HTL(ASCP) Infectious Disease Pathology Centers for Disease Control MS-G32 1600 Clifton Rd. Atlanta, GA 30333 USA
email: tim9-at-cdc.gov timcdc-at-hotmail.com
FAX: (404)639-3043 The basic procedure for re-embedding paraffin embedded is relatively simple. We had a Lynx Tissue Processor set up to do it automatically. Cut the paraffin tissue into 1-2 mm cubes with a warm scalpel. Trim as much excess paraffin away as possible. De-paraffinize with 6 alternately warm/room temp xylene washes with agitation for 30 minutes each. Reverse the dehydration process from 100% EtOH to your normal EM buffer. Refix and postfix with appropriate EM fixatives, dehydrate, and embed. The smaller the paraffin pieces, typically the better the deparaffinization. Your tissue will never look great though. The initial fixation for LM is a poor TEM fixative. good Luck.
Chuck Butterick Engineered Carbons,
Inc."Michelle.Taurino-at-aventis.com"-at-sparc5.microscopy.com wrote: ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Any suggestions on re-embedding paraffin sections in epon? Has anyone worked with the "pop out" method using BEEM capsules?
Any suggestions would be helpful...
Michelle Taurino Aventis Pharmaceuticals Bioimaging and Molecular Histology Michelle.Taurino-at-aventis.com 908-231-3357 "Michelle.Taurino-at-aventis.com"-at-sparc5.microscopy.com wrote: ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Any suggestions on re-embedding paraffin sections in epon? Has anyone worked with the "pop out" method using BEEM capsules?
Any suggestions would be helpful...
Michelle Taurino Aventis Pharmaceuticals Bioimaging and Molecular Histology Michelle.Taurino-at-aventis.com 908-231-3357
____________________________________________________________________ Get free email and a permanent address at http://www.netaddress.com/?N=1
I was talking about very special EM case: dark-field TEM. At this point we are talking about a few electrons per square angstrom, even less. In high resolution EM crystallography people have deal with 0.6 e/A2. At such conditions, high sensitivity and linearity of the modern digital cameras may be a plus. Talking about long exposures --what about drift? I never had good pictures at x100K with exposure longer that a few seconds. I don't understand your point about noise. In case of digital camera the noise is a function of camera. Current cameras has a very low level of noise (they uses cooling, etc) and we have to pay for that astronomical price. This is a life. I am not friendly with TEM cameras, but I do know that in the light microscopy people count individual photons using CCD cameras. In TEM camera we have phosphorous screen as a source of image and my questions to Mickhael were addressed mostly to the problem how effectively (and correctly) information is traveled thought that funny screen. The Mickhael's answer is that screen does not affect dynamic range and sensitivity is much higher than on convention films. Am I correct, Mickhaels?
Sergey
} Date: Thu, 18 May 2000 14:15:02 +1000 } From: jim {jim-at-proscitech.com.au} } Subject: RE: new developments in imaging systems? } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} , } "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-sparc5.microscopy.com} } Reply-to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } Organization: ProSciTech } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Hi We have a JEOL 733 with a two-sector PN type BS detector. Anyone out there have any idea on the resolution of these things in terms of mean atomic number. What I want to know basically is the sensitivity of these things to compositional variation. Cheers, Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: malc-at-rock.ru.ac.za SOUTH AFRICA
Thanks for all the responses to this posting both on and off the list. I routinely analyse minerals found in soils and reservoir sandstones and can easily recognise most of the common minerals by their EDS spectra. The problem for me recently has been looking at samples containing some of the more 'exotic' minerals, or put simply minerals I'm not used to looking at, e.g. sphene and clinpyroxenes. This has been more of a problem because we had no supporting XRD data for these particular analyses. What I was asking for, perhaps naively, was a program that would help identify an unknown mineral from its spectrum and give a list of possible alternatives. This is evidently not available. What is available: Thereis a free Mac program DTSA that can simulate spectra of a mineral for a particular detector and kV etc. Scott Wight/Mark Blackford). This can be downloaded from http://www.cstl.nist.gov/div837/837.02/dtsa.html Thereis a EDS spectrum database with search capabilities (based on archived spectra) written for the US FBI which may be commercially available soon. I will certainly follow up this up soon (thanks Dennis Ward and Nicholas Ritchie.) Thereseems to be various databases including MDAT that allow a search by chemical elements but not actual analyses. (thanks Bart Cannon) About a year ago, I started building a spectrum library collected on our system recording the various beam operating parameters, preparation (rough or polished, C or Au coated) etc. I agree with Bart Cannon that this is probably the best way forward. Just one thing - my database needs many more spectra added to it, especially different variations of many of the solid solution minerals. I suppose the idea of the database is that it should be used as a reference guide only. Although some minerals could never be identified by their EDS spectrum alone this is where an extensive reference library of my own would come in so useful and help answer the question: is this spectrum a Ca feldspar or Ca-rich zeolite? Although the best answer to this specific question would probably be the unknown spectrum indicates it may be a Zeolite because it is more similar to the zeolite reference we collected under similar operating parameters. Of course there would be no substitute if there were XRD data telling you in the same sample of zeolite and no Ca-feldspar. Thanks again to all of you who responded. Best regards Martin Roe
Martin J. Roe Macaulay Land Use Research Institute Craigiebuckler Aberdeen Scotland UK Phone 01224 318611 e-mail m.roe-at-mluri.sari.ac.uk
---------- } From: Hans Brinkies {HBrinkies-at-groupwise.swin.edu.au} } To: microscopy-at-sparc5.microscopy.com } Subject: Our management has finally agreed to connect my electronmicroscopes to a recirculating water system } Date: May 18, 2000 9:49 PM } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Our management has finally agreed to connect my electron microscopes to } a recirculating water system. } } I need some ideas what type of systems are on the market and the } Australian agent for these units. We need to attach 2 SEMs, 1 TEM plus 1 } coating unit to the new reciculating line. } } Thank you } } Hans Brinkies } Senior Lecturer } Swinburne, University of Technology } School of Engineering and Science } P.O.Box 218 - Hawthorn - Vic -3122 - Australia } Phone: +61 3 9214 8657 } Fax: +61 3 9214 8264 } Email: Hbrinkies-at-swin.edu.au } } I don't really know what companies distribute such systems in Australia, but one thing occurs to me....if you set up all your instruments on one circulator, it's nice and cost-effective, but when the pump crashes or motor burns out, all your toys are down until the one circulation system is repaired. Separate systems for each 'scope would be better, (that'll probably give your managers a heart attack) or at least try and retain the capability of switching back to the old constant-loss system during emergencies. Our own ESEM was recently down for nearly three weeks while I was trying to replace the circulator pump motor. Turned out to be kind of a hard one to find...
Frank Thomas MicroAnalysis Facility Geological Survey of Canada Atlantic Bedford Institute of Oceanography Dartmouth, Nova Scotia
For the past 20 years I have used the Philips TEM heating holder furnace and heater elements to build hotstages for a range of microscopes. Unfortunately, Philips have told me that they can no longer supply the Pt furnace bodies and I cannot get information on possible replacements.
If anyone has a spare furnace (or more?) - Philips part number 5322 265 70028 - that they wish to part with I am willing to pay a `Philips' price. I am also willing to consider lightly used furnaces or complete holders that are now surplus to requirements and will pay a price depending on condition and history.
If anyone has any information on replacements to the previous style or on present day heaters I would be most grateful.
Thanks, Ron
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Atomic number contrast sensitivity using BSE is dependent on several variables in addition to the equipment. I may not cover everything, but what quickly comes to mind....
Specimen surface condition - If a specimen is well polished, compo imaging is best. Topography of rough specimens (like a fracture) will generate feature contrast, "diluting" the compo contrast.
Beam current and potential affect sensitivity.
Conductive films will reduce sensitivity. Use carbon rather than sputtered Au, Pd etc. to minimize the effect if the specimen must be coated.
Depending on the detector geometry, working distance can affect the solid angle of collection thus changing BSE collection effiency.
The absloute atomic numbers present in the specimen will affect sensitivity in two ways.
Sensitivity will be different for a composition of lighter elements compared to a specimen composed of higher atomic numbers.
Also, if it is desired to NOT saturate the video, the range of atomic numbers can limit sensitivity. This is a typically a result of limited dynamic range in the image capture device/photo. For example, if C and W are present as pure inclusions in a brass, it will be difficult (to say the least) to contrast the difference between Cu and Zn phases whild not loosing C & W contrast variability.
Considering the above, any single statement of sensitivity is difficult, but if I had to generalize... My 4 diode GW Electronics BSE system will resolve 0.1 atomic number differencees under "good" conditions.
Woody White McDermott Technology
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Hi We have a JEOL 733 with a two-sector PN type BS detector. Anyone out there have any idea on the resolution of these things in terms of mean atomic number. What I want to know basically is the sensitivity of these things to compositional variation. Cheers, Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: malc-at-rock.ru.ac.za SOUTH AFRICA
I want to add my 2 cents as I like the topic. Most of this is a result of my experience with our TEM 2Kx2K CCD. We are working in bright field - so I cannot comment on dark field performance Several points:
1. The CCD is much more convenient - you get your pictures instantly.
2. CCD is linear and has larger dynamic range than the film.
3. The bad thing about the CCD is resolution - about 4 times lower than that of the film (in our camera the pixel size is 30 microns). So if you want to work in minimum dose you will do better with film. As I know the CCD is not performing well in terms of signal to noise at low doses (and if you have to work at 4 times higher magnification because of the resolution the things become much worse).
4. The CCD has smaller observation area - again loss of information.
5. I don't know about the detection efficiency compared to the film - it depends on the thickness of the phosphorous and the accelerating voltage. If a photon reaches the CCD chip it will be detected ... the problems are in the conversion electron-photon. There are two sides - if you make the phosphorous thicker you will get higher detection efficiency but the point spread also increases so always a compromise is made between detection efficiency and resolution. When I say detection efficiency this is not only related to the detection of single electrons (as it detects single electrons) but more to the actual signal detected on the background of the noise. Apart from the shot noise additional noise is added due to the scintillator driven detection and thermal noise in the CCD chip.
The CCDs are now very popular in diffraction work because of the dynamic range and linearity.
Here is one reference where a nice comparison between 2Kx2K CCD and film has been made:
Downing, K., Hendrickson, F., Ultramicroscopy, 75, (1999), 215-233
Best regards,
Rado
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp --------------------------------------------------------------------- ----- Original Message ----- } From: Sergey Ryazantsev {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, May 19, 2000 2:03 PM
If your samples are not site specific, then I would recommend a modified version of the small angle cleavage technique. You can get a detailed description of this technique in the MRS TEM Sample Prep IV book, vol 480.
Instead of the low temperature melting wax to hold the samples down, you use superglue. It takes longer to soak the samples off in acetone, but there is no heat. Then instead of the silver epoxy that is normally used, you need to use a slow curing viscous epoxy to mount the samples on the copper grid. There may be a temperature spike in the curing, but you would have to experiment and find out yourself. The amount of epoxy that is needed is very small and I doubt that it would raise the temperature an appreciable amount. The down side is that it will take a day to fully cure the epoxy. The net result is that the only heating the sample really sees is the heat generated during grinding the back side of the samples down.
-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)
} -----Original Message----- } From: prenitzer,brenda s [mailto:prenitzer-at-lucent.com] } Sent: Thursday, May 18, 2000 7:06 PM } To: 'List Server' } Subject: Room Temperature TEM 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/MicroscopyListse} rver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } Hello all, } } I am in need of a way to modify my current method of preparing TEM } cross-sections as to introduce no heating. Specifically, I } need to find new } adhesive materials. We currently use thermally cured M-bond, } Gatan G1 epoxy } and crystal bond wax - all of which require heat. } } If anyone has information on an adhesive that ion mills at a } rate to similar } Si, is stable under the electron beam, will cure at room } temperature within } about 24 hours, and once cured is impervious to solvents such } as acetone, } please respond. It would be an additional plus if the } adhesive has low } viscosity, so it can be used to secure TEM grids to the specimens. } } } Also I am looking for a replacement for the wax that we } currently use to } affix the samples to the polishing studs. This should cure } quickly, bond } strongly enough to endure the mechanical stresses of grinding, and be } readily soluble in a solvent other than water so the samples } can be removed } from the studs. } } Any help would be greatly appreciated. } } Brenda } } Brenda I. Prenitzer, Ph.D. } Member of Technical Staff } Cirent Semiconductor (Lucent Technologies) } 9333 S. John Young Parkway } 6D-Lab } Orlando, FL 32819-8612 } } Phone: 407 371 7108 } Fax: 407 371 6999 } } prenitzer-at-lucent.com } bsp101-at-worldnet.att.net } }
as I said in my response, a direct comparison between the sensitivities of film and a CCD camera are very complicated. The film has a non-linear response curve, it reacts to electrons directly, there is grain size to take into account, etc. All I wanted to say is the following: For a film camera setting of 2 seconds, we acquire an image in approx. 50 to 100 msec with similar contrasts to that of film.
Regarding the linearity of the phosphor-CCD system: I have measured the linearity by measuring the beam current and the image intensity and I found it to be linear over the entire range that I measured. The deviations were insignificant and probably due to noise (I would give you the numbers, but this was in a different life and I don't have them). There may be nonlinearities that show up if you reach extreme illumination levels of the phosphor, but I did not measure them and it will probably depend on the phosphor itself.
Michael
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: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Thursday, May 18, 2000 11:04 PM To: Microscopy-at-sparc5.microscopy.com
Jim hello
I was talking about very special EM case: dark-field TEM. At this point we are talking about a few electrons per square angstrom, even less. In high resolution EM crystallography people have deal with 0.6 e/A2. At such conditions, high sensitivity and linearity of the modern digital cameras may be a plus. Talking about long exposures --what about drift? I never had good pictures at x100K with exposure longer that a few seconds. I don't understand your point about noise. In case of digital camera the noise is a function of camera. Current cameras has a very low level of noise (they uses cooling, etc) and we have to pay for that astronomical price. This is a life. I am not friendly with TEM cameras, but I do know that in the light microscopy people count individual photons using CCD cameras. In TEM camera we have phosphorous screen as a source of image and my questions to Mickhael were addressed mostly to the problem how effectively (and correctly) information is traveled thought that funny screen. The Mickhael's answer is that screen does not affect dynamic range and sensitivity is much higher than on convention films. Am I correct, Mickhaels?
Sergey
} Date: Thu, 18 May 2000 14:15:02 +1000 } From: jim {jim-at-proscitech.com.au} } Subject: RE: new developments in imaging systems? } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} , } "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-sparc5.microscopy.com} } Reply-to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } Organization: ProSciTech } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hello,
I am new to this list. I would like to know if anyone has information I could use to write an article on the analytical chemistry techniques used to authenticate art objects using light microscopes or any other type of microscope. Also, if anyone has information on any of the other techniques used to authenticate art objects such as x-ray diffraction, x-ray fluorescence, accelerator mass spectrometry, that would be helpful too.
Sincerely,
Margaret M. Mitchell Assistant Editor AOAC INTERNATIONAL 301-924-7077 (tel) 301-924-7089 (fax) mmitchell-at-aoac.org
I'm looking for information regarding final polishing solutions for preparing metallic cross-section samples for TEM analysis.
I've looked into colloidal silica suspensions such as Ludox and Syton, and am concerned that their alkilinity may chemically etch my samples. I'm analyzing different compositions of NiFe on copper substrates, so the specimens are prone to preferential etching of different phases.
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With that agreement you also have a few new problems to consider. I suggest that: In Australia you could not buy a large system like that as a supplied item. Several smaller plants would be much too expensive and cumbersome. Any imported system will result in problems with fittings and parts
I expect that it would be best (been there, done that a couple of times) to design your own large system with a bit of spare capacity.
Beer chillers as used in hotels are most suitable. You want at least two large chillers, so if one fails you still could run more than half of the equipment). Calculate total heater element wattage with some spare capacity.
Diffusion pumps are designed to work best at 16 degrees. At that temperature the TEM column will condense water. Depending on ambient temperature, it might be sufficient to use the dif pump return line to cool the column and so avoid that problem. The chillers should be installed under an open shelter as they generate a lot of heat. Chiller temperature cut-in and out requires a thermostat each.
I suggest that a centrifugal pump of 1hp capacity will give enough pressure at the relative small flow rates. Chose a very common pump make so you can install a spare quickly without replacing fittings. Other pump types give more pressure, but they are more expensive and generally less reliable.
12mm reinforced garden hoses with hose clamps to and from the instruments make installation easy, pretty secure and cause minimal pressure drop.
Required is a SS tank (insulated) or a fiberglass lined tank to act as a heat sink. 100 liters would be a reasonable minimum capacity.
A good inline filter is required and a bypass, so the system can run during filter change.
If a mains water line is maintained, it is very important to have the returning water going to a drain whenever mains water is in use. I have lived through some horrid floods, which resulted from people opening mains but not the bypass to the drain.
If you have the components any plumber can install the system in under a day. The plumber would supply gate valves, temperature and pressure gauges etc.
Hans, you may fax your rough design to me, I'll be happy to make any suggestions. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, May 19, 2000 10:50 AM, Hans Brinkies [SMTP:HBrinkies-at-groupwise.swin.edu.au] wrote: } } Our management has finally agreed to connect my electron microscopes to } a recirculating water system. } } I need some ideas what type of systems are on the market and the } Australian agent for these units. We need to attach 2 SEMs, 1 TEM plus 1 } coating unit to the new reciculating line. } } Thank you } } Hans Brinkies } Senior Lecturer } Swinburne, University of Technology } School of Engineering and Science } P.O.Box 218 - Hawthorn - Vic -3122 - Australia } Phone: +61 3 9214 8657 } Fax: +61 3 9214 8264 } Email: Hbrinkies-at-swin.edu.au }
John, The solutions that the colloidal silica are in have a ph of 8.5 to 9.8 depending on the formulation. I suggest you try our 0.05 micron water based polycrystalline diamond suspension. Allied High Tech http://www.alliedhightech.com sells this product. I would also suggest our Final A cloth for this polishing step. If you would like samples of either of these products please let me know and I will be happy to send it to you.
If you need further technical assistance please contact me off-line and I will be happy to help or you may contact our main office at (800)675-1118 located in CA.
I hope this helps. Ed Please note, I have a financial interest in providing you with these products and other sample preparation equipment and consumable items.
************************************************* Edward A. Hirsch Product Application Specialist Allied High Tech Products 2376 East Pacifica Place Rancho Dominguez, CA 90220 ph: (919) 846-9628 vm:(800)675-1118 x245 fx: (310)762-6808 http://www.alliedhightech.com
Equipment and Consumables for Metallurgical Sample Preparation *************************************************
-----Original Message----- } From: john david whitaker [mailto:jwhitake-at-u.washington.edu] Sent: Friday, May 19, 2000 4:01 PM To: Microscopy Lister Server
I'm looking for information regarding final polishing solutions for preparing metallic cross-section samples for TEM analysis.
I've looked into colloidal silica suspensions such as Ludox and Syton, and am concerned that their alkilinity may chemically etch my samples. I'm analyzing different compositions of NiFe on copper substrates, so the specimens are prone to preferential etching of different phases.
Dear Friends, Can anyone give me advice regarding the dismantling and the packing for shipment of a Philips 201 TEM? I need to pay particular attention to not disturbing the alignment. I plan to move the TEM from Univ. of Delaware to Naples, Florida in an enclosed U-Haul Trailer. Any help or suggestions will be most welcome. If anyone has hands-on experience in doing this sort of thing and is willing to give me a hand at U-Del, please contact me and name your price. I'll gladly pay for the assistance. Best regards, Mike Urbanik www.crystalguru.com
I find what you are trying to do interesting, but there are things that may never work and this may be one.
We had it confirmed by Michael Bode that good digital system "see "single photons - so that is not likely to improve much. Film too registers single electrons. We were told that in digital 1 electron can expose one pixel. On film one electron initiates the exposure of a silver halide and subsequent electrons in a near identical location increase the size of the grain.
Low noise (cooled cameras) hopefully do not add their own noise, but they cannot make up for lack of information in the image.
My case (below) was that in high resolution TEM at least, less exposed images will be inferior because there are simply not enough electrons to produce a good image. A more sensitive digital system uses fewer electrons and so makes matters worse.
In dark field EM we are using the beam indirectly and such images too suffer particularly from electron noise. Here too the use of a digital system would only make this worse. Furthermore, the slower 4489 film would be better than SO-163.
If you don't like long exposures (4 to 8 seconds I found a practical limit), you cannot ignore the reality of electron noise and hope to correct that with yet fewer electrons, albeit processed in a "noise-free" digital system. I think that the answer is more electrons, i.e. a field emission source or a slower digital camera. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, May 19, 2000 3:04 PM, Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] wrote: } } } Jim hello } } I was talking about very special EM case: dark-field TEM. At this point we } are talking about a few electrons per square angstrom, even less. In high } resolution EM crystallography people have deal with 0.6 e/A2. At such } conditions, high sensitivity and linearity of the modern digital cameras } may be a plus. Talking about long exposures --what about drift? I never } had good pictures at x100K with exposure longer that a few seconds. I } don't understand your point about noise. In case of digital camera the } noise is a function of camera. Current cameras has a very low level of } noise (they uses cooling, etc) and we have to pay for that astronomical } price. This is a life. I am not friendly with TEM cameras, but I do know } that in the light microscopy people count individual photons using CCD } cameras. In TEM camera we have phosphorous screen as a source of image and } my questions to Mickhael were addressed mostly to the problem how } effectively (and correctly) information is traveled thought that funny } screen. The Mickhael's answer is that screen does not affect dynamic range } and sensitivity is much higher than on convention films. Am I correct, } Mickhaels? } } Sergey } } } Date: Thu, 18 May 2000 14:15:02 +1000 } } From: jim {jim-at-proscitech.com.au} } } Subject: RE: new developments in imaging systems? } } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} , } } "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-sparc5.microscopy.com} } } Reply-to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } } Organization: ProSciTech } } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } If sensitivity was ten times higher, one would in some cases save some beam } } damage and in most cases suffer excessive electron noise. } } As a rough guide, electron noise becomes apparent at 30x enlargement, } rarely a } } problem. Magnification is a linear function and electron density relates } to an } } area, but clearly very short exposure images would be much noisier and } could } } not be enlarged nearly as much. When not enough electrons form an image it } } appears grainy, which makes it unsuitable for further enlarging. Photo } } enlarging utilises higher depths-of-field and without this, very high } power TEM } } is much, much harder. } } By nature, slower emulsions have finer grain and higher resolution and } } contrast. It is fortuitous that these desirable characteristics run in } tandem } } with relatively long exposures, so the image is formed by more electrons. } } TEM films rate about 6 ISO, so X-ray film at about ISI 3000 is about a 10x } } faster than TEM films. Unfortunately its rather grainy, but if the } exposure was } } well adjusted, chances are that electron noise would be more bothersome than } } } } the film's grain. } } Digital is inevitable and already fairly common, but some aces remain with } } conventional film. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } www.proscitech.com } } } } On Thursday, May 18, 2000 5:06 AM, Sergey Ryazantsev } [SMTP:sryazant-at-ucla.edu] } } wrote: } } } } } } } } } Thank you Mickhael. } } } } } } I will wait for info about sensitivity. I think, for EM in particular, } } } sensitivity is very important parameter of the system. I am surprised } } } that } } } manufacturers don't have data on this matter. If sensitivity, say 10 } } } times } } } higher than SO-163 film - it may be a great reason to switch from film to } } } the CCD. The major disadvantage of the CCD cameras for TEM is their price } } } in my point of view. } } } Thanks for your respond. } } } } } } Sergey } } } } } } } } } } Date: Wed, 17 May 2000 11:34:05 -0600 } } } } From: Michael Bode {mb-at-Soft-Imaging.com} } } } } Subject: RE: new developments in imaging systems? } } } } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} } } } } X-Mailer: Internet Mail Service (5.0.1457.3) } } } } } } } } Sergey, } } } } } } } } I will try to find out what we have regarding the relative } } } } sensitivities. One thing that I can say now is, that we acquire images } } } } with a 50 or 100 msec exposure of the digital camera when the film } } } } requires an exposure of about 2 seconds. This would mean a 20 to 40 } } } } times better sensitivity of the CCD camera. But to answer your question } } } } in more detail would require to compare also the resolution of film and } } } } camera and that is where it becomes very difficult, as it is not easy to } } } } determine the resolution of film in terms of spatial resolution and } } } } dynamic range, as both are interwoven. I will try to find some answers } } } } for you. } } } } } } } } Regarding the linearity: As the CCDs simply count Photons, they have } } } } almost perfect linearity over their complete dynamic range. Even more so } } } } for TEMs, where all Photons have the same energy and the quantum } } } } efficiency does not change from photon to photon. The Phosphor is a part } } } } of a system that could theoretically introduce some non-linearities. } } } } However, I did measure the linearity of some TEM camera systems a few } } } } years back and did not find any significant deviations from linearity. } } } } } } } } Michael } } } } } } } } 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: mailto:info-at-soft-imaging.com } } } } web: http://www.soft-imaging.com } } } } =================================== } } } } } } } } } } } } } } } } -----Original Message----- } } } } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] } } } } Sent: Wednesday, May 17, 2000 1:29 AM } } } } To: Michael Bode } } } } Subject: RE: new developments in imaging systems? } } } } } } } } } } } } Mickhael hello } } } } } } } } I have question for you. I am thinking about adding CCD camera to my } } } } JEM1200EX. The information I gathered from Internet is not so } } } } optimistic. } } } } The standard CCD resolution is about 1-1.5 million pixels, 2 million is } } } } a } } } } max as I understand. The price for cheaper camera is about 20-30 K$ - } } } } much } } } } more that I expect to spend for "film" process. There are two things } } } } may } } } } attract me to the modern CCD camera: dynamic range and sensitivity. I } } } } am } } } } pretty sure that dynamic range for CCD itself is a few orders better } } } } than } } } } any film available. But what about phosphorus screen? Does it reduce } } } } dynamic range for the EM images? How dramatic? This is my first } } } } question. } } } } The next question is: could you tell me something about sensitivity of } } } } the } } } } modern CCD cameras used in EM? I am using dark field imaging at x80K } } } } magnification and the exposure time for SO-163 (non deluded D-19, 7 min, } } } } 20oC) is about 2 sec. I called GATAN, but they did not say anything } } } } useful. } } } } Could you provide some comparison of your side-mount camera with } } } } sensitivity of the SO-163 film at condition I mentioned? I will greatly } } } } appreciate any information in this matter. Thanks. Sergey. } } } } } } } } } } } } } Date: Fri, 12 May 2000 13:46:01 -0600 } } } } } From: Michael Bode {mb-at-Soft-Imaging.com} } } } } } Subject: RE: new developments in imaging systems? } } } } } To: "'Microscopy-at-MSA.Microscopy.Com'" } } } } {Microscopy-at-sparc5.microscopy.com} } } } } } Cc: 'Margaret Dienelt' {brannign-at-asrr.arsusda.gov} } } } } } X-Mailer: Internet Mail Service (5.0.1457.3) } } } } } } } } } } ----------------------------------------------------------------------- } } } } - } } } } } 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 } } } } } ----------------------------------------------------------------------- } } } } . } } } } } } } } } } } } } } } Margaret, } } } } } } } } } } As a former user and current vendor of such systems as you are } } } } inquiring } } } } } about I can try to provide a bit of information regarding camera } } } } } improvements: } } } } } } } } } } There have been a number of improvements, but I am not sure what you } } } } are } } } } } comparing the latest cameras against. Cameras are now usually cooled } } } } and } } } } } provide 12 bits per pixel, the number of pixels has gone up a bit (but } } } } } not much in general), and cameras read out faster than they used to (up } } } } } to 20 fps and more). I think all cameras now use a line transfer } } } } } mechanism, which makes shutters obsolete. } } } } } On the software side, real-time FFT and real-time shading correction } } } } can } } } } } be done now due to faster computers without special processing boards, } } } } } and there have been other software developments that make using the } } } } } cameras and computers easier. } } } } } Other changes that affect the usability of cameras is the use of } } } } } pneumatics to insert and retract the phosphors, higher frame rates for } } } } } live viewing with the camera, etc. } } } } } } } } } } If you have questions, please give me a call, drop me an email, or go } } } } to } } } } } our web site. } } } } } } } } } } Michael } } } } } } } } } } } } } } } 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: mailto:info-at-soft-imaging.com } } } } } web: http://www.soft-imaging.com } } } } } =================================== } } } } } } } } } } } } } } } } } } } } -----Original Message----- } } } } } } From: Margaret Dienelt [mailto:brannign-at-asrr.arsusda.gov] } } } } } Sent: Thursday, May 11, 2000 12:43 PM } } } } } To: Microscopy-at-sparc5.microscopy.com } } } } } Subject: TEM: new developments in imaging systems? } } } } } } } } } } } } } } } ----------------------------------------------------------------------- } } } } - } } } } } 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, } } } } } } } } } } Year after year I hopefully gather information about digital imaging } } } } } systems for TEMs (ours is a JEOL 100CX) , only to learn we have no } } } } } money. This year it looks like it might really happen but I have not } } } } } kept up with innovations in the field and am wondering the following: } } } } } } } } } } 1. Anything new in the last two years -- especially in terms of } } } } } cameras? I'm most familiar with the Gatan and AMT systems but their } } } } } web sites don't reflect much in the way of changes over a year ago. } } } } } 2. With more and more microscopists finally getting their systems -- } } } } } I'd love to get feedback. } } } } } } } } } } Thanks, } } } } } Margaret } } } } } } } } } } P.S. Would welcome contacts from vendors. } } } } } } } } } } -- } } } } } Margaret Dienelt } } } } } } } } } } Plant Pathologist } } } } } Electron Microscopy Lab } } } } } } } } } } Floral and Nursery Plants Research Unit } } } } } U.S. National Arboretum/Agricultural Research Service/USDA } } } } } } } } } } B. 010A, Rm. 238, BARC-W } } } } } 10300 Baltimore Avenue } } } } } Beltsville MD. 20705 USA } } } } } } } } } } (301) 504-6097 } } } } } Fax: (301) 504-5096 } } } } } } } } } } } } } } _____________________________________ } } } } } } } } Sergey Ryazantsev Ph. D. } } } } Electron Microscopy } } } } UCLA School of Medicine } } } } Department of Biological Chemistry } } } } Box 951737 } } } } Los Angeles, CA 90095-1737 } } } } } } } } Phone: (310) 825-1144 } } } } Pager: (310) 845-0248 } } } } FAX (departmental): (310) 206-5272 } } } } mailto:sryazant-at-ucla.edu } } } } http://www.bol.ucla.edu/~sryazant } } } } } } } } } } } _____________________________________ } } } } } } Sergey Ryazantsev Ph. D. } } } Electron Microscopy } } } UCLA School of Medicine } } } Department of Biological Chemistry } } } Box 951737 } } } Los Angeles, CA 90095-1737 } } } } } } Phone: (310) 825-1144 } } } Pager: (310) 845-0248 } } } FAX (departmental): (310) 206-5272 } } } mailto:sryazant-at-ucla.edu } } } http://www.bol.ucla.edu/~sryazant } } } } } } } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant } }
Dear Malc! Some years ago I got the resolution 0.1 in the middle of periodic table (around Z=28) on standard 733 BSE. But for this purpose it is necessary to balance additionally the preamplifier to remove completely the rest of topo signal, which becomes comparable by value with a very low signal of differential compo. There are no means in the preamplifier for this additional balancing, therefore I have soldered the additional variable resistor in one of shoulders of a differential amplifier in the preamplifier. Frankly speaking I have forgotten the details already, but I can restore them, if you wait about two weeks. But basically it is very simple. Regards. Victor Sidorenko, ANTRON Co. Ltd., scientific service, Moscow, Russia.
-----é–›”Š’”Œ –””‡Œ’Œ----- ë˜: Dr Malcolm Roberts {malc-at-rock.ru.ac.za} 攓—: Microprobe discussion group {Microscopy-at-sparc5.microscopy.com} €ˆ˜ˆ: 19 “ˆØ 2000 ‹. 16:13 ñŒ“ˆ: BSE resolution
} --------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Polishing suspensions that does not interact chemically with your samples include diamond suspensions (available down to 0.1µm at least) and alumina suspensions (down to 0.05µm). Many vendors have information available on the World-Wide-Web.
Cheers, Paul =================== Paul Baggethun Alcoa Technical Center Alcoa Center, PA 15069 USA =================== + 724 - 337-1760 (tel) + 724 - 337-2044 (fax) ===================
} ---------- } From: john david whitaker[SMTP:jwhitake-at-u.washington.edu] } Sent: Friday, May 19, 2000 5:01 PM } To: Microscopy Lister Server } Subject: final polishing for metals (Cu, Ni, Fe, NiFe) } } ------------------------------------------------------------------------ } 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 looking for information regarding final polishing } solutions for preparing metallic cross-section samples } for TEM analysis. } } I've looked into colloidal silica } suspensions such as Ludox and Syton, and am concerned } that their alkilinity may chemically etch my samples. } I'm analyzing different compositions of NiFe } on copper substrates, so the specimens are prone to } preferential etching of different phases. } } Does anyone have any experience with this? } } Any input would be appreciated. } } John } }
On Thu, 18 May 2000 ComCryLab1-at-aol.com-at-sparc5.microscopy.com wrote:
} Dear Friends, } Can anyone give me advice regarding the dismantling and } the packing for shipment of a Philips 201 TEM? I need to } pay particular attention to not disturbing the alignment. } I plan to move the TEM from Univ. of Delaware to Naples, Florida } in an enclosed U-Haul Trailer. Any help or suggestions } will be most welcome.
I will make sure Ron Veil, a very experienced independent EM service tech, sees this and has a chance to reply. It was with his advice that we (my hubby and I and its new owner) packed up and shipped a Philips 201. Like you, we wanted to ship it intact, column on and all. We built a heavy duty skid and added a strong base to which we bolted the instrument. Then we wrapped it with whatever that plastic packing tape that is like Saran Wrap is called, making sure to secure any part of the column we didn't want to move, but avoiding putting any pressure on things, such as the aperture drives. Wrap the bottom, wrap the column, wrap the column to the bottom and back again, etc. Build a crate up around the instrument. I think I remember putting a wood insert with a crescent cut out near the top of, but not touching the column, but hubby thinks not. The idea is NOT to let any shock to the crate get transferred to the column, so perhaps we didn't. Add some packing material, such as old egg crate foam from your bed (it needed replacing anyway) and whatever. Then, and this was the fun part, buy that stuff that when you mix it with a catalyst, produces huge volumes of foam that hardens in a few minutes. I think you can also buy spray cans of similar stuff, but since hubby had this on hand for other things, we had the bulk stuff. Fill the voids in the crate with it. It will easily chip off later. Add a top, and away you go. The scope made it in great shape.
I think we packed the rotary pump and a couple of other things separately. Get it into the truck and TIE IT DOWN. I say this because an SEM we once packed for shipping with the base and saran, but no closed crate, made it from Hawaii to Ohio, then slid in the shippers truck in the last mile and slammed into the driver's seat. The driver was OK, the ion getter pump was bent, and the column needed a bit of work, but it could have been worse. Duh.
I've also packed up a couple of Denton vcuum evaporators and a couple of ultramicrotomes. The saran stuff and blow foam are a good way to stabilize the instruments.
Good luck!
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* ****************************************************************************
} From: "Tina Carvalho" {tina-at-pbrc.hawaii.edu} } On Thu, 18 May 2000 ComCryLab1-at-aol.com-at-sparc5.microscopy.com wrote: } } } Dear Friends, } } Can anyone give me advice regarding the dismantling and } } the packing for shipment of a Philips 201 TEM? I need to } } pay particular attention to not disturbing the alignment. } } I plan to move the TEM from Univ. of Delaware to Naples, Florida } } in an enclosed U-Haul Trailer. Any help or suggestions } } will be most welcome. } } I will make sure Ron Veil, a very experienced independent EM service tech, } sees this and has a chance to reply. It was with his advice that we (my } hubby and I and its new owner) packed up and shipped a Philips 201. Like } you, we wanted to ship it intact, column on and all. We built a heavy } duty skid and added a strong base to which we bolted the instrument. Then } we wrapped it with whatever that plastic packing tape that is like Saran } Wrap is called, making sure to secure any part of the column we didn't } want to move, but avoiding putting any pressure on things, such as the } aperture drives. Wrap the bottom, wrap the column, wrap the column to the } bottom and back again, etc. Build a crate up around the instrument. } I think I remember putting a wood insert with a crescent cut out near the } top of, but not touching the column, but hubby thinks not. The idea is } NOT to let any shock to the crate get transferred to the column, so } perhaps we didn't. Add some packing material, such as old egg crate foam } from your bed (it needed replacing anyway) and whatever. Then, and this } was the fun part, buy that stuff that when you mix it with a catalyst, } produces huge volumes of foam that hardens in a few minutes. I think you } can also buy spray cans of similar stuff, but since hubby had this on hand } for other things, we had the bulk stuff. Fill the voids in the crate with } it. It will easily chip off later. Add a top, and away you go. The } scope made it in great shape. } } I think we packed the rotary pump and a couple of other things } separately. Get it into the truck and TIE IT DOWN. I say this because an } SEM we once packed for shipping with the base and saran, but no closed } crate, made it from Hawaii to Ohio, then slid in the shippers truck in the } last mile and slammed into the driver's seat. The driver was OK, the ion } getter pump was bent, and the column needed a bit of work, but it could } have been worse. Duh. } } I've also packed up a couple of Denton vcuum evaporators and a couple of } ultramicrotomes. The saran stuff and blow foam are a good way to } stabilize the instruments. } Since you are using a U haul you don't have to worry about sides and a top on the crate. Make sure it is tied down real well. Dry wall screws through the crate and into the floor work well for locking it down but make sure you have it braced with timbers to the front and sides of the trailer in case you make a panic stop.
Also make sure that the weight is centers in front of the trailer axle if you use a trailer. Negitive weight on the trailer tounge at the very least makes driving very interesting. You have no Idea how fast a trailer can pass you while it is still tied on to the truck. I have had the privilege of experiencing this and I can promise you that you won't enjoy it:).
The foam in place stuff is great. Just cover the instrument in plastic and foam away. If you can get foam between the instrument and the Support the foam will dissipate most of those little shocks that get things out of line.
Last of all make sure you understand what you insurance covers and what it doesn't on moving equipment. You should be fine on liability but the value of the contents is probably not covered. Rental truck companies have contents insurance as an extra.
Good luck Gordon
Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
I would like to announce an exciting job opportunity to the Microscopy community.
JOB ANNOUNCEMENT
POSITION TITLE: SR. SCIENTIST/ENGINEER LIGHT MICROSCOPY NIKON INC. MELVILLE, NY Posted: May 18, 2000
As seen in the 19 May issue of Science: http://recruit.sciencemag.org/cgi/show/5469/5469x03207
Sr. Applications Scientist/Engineer Bring Your Light Microscopy Experience To The BioScience Division Of Nikon Inc.
Our world renowned company has an excellent opportunity for a professional with extensive experience in light microscopy, specifically in advanced BioScience technologies in our USA headquarters located in Melville, Long Island , NY. Today, scientists at world-renowned biological institutions are making tremendous advances in Cancer, AIDS, Alzheimer's, in-vitro fertilization and other leading-edge research. Nikon is proud to be playing a role in this enormously important work. Our ongoing commitment to optical excellence and technological advancement is allowing researchers to view objects never before seen by the human eye. We would like to invite you to investigate the new opportunities and technologies available at Nikon Inc. BioSciences.
JOB SUMMARY
Provide the principal conduit for technical information and feedback between the end user/sales network and Nikon Factory engineers on: product applications, product acceptance, competitive changes, required product improvements, new technologies and methodologies that impact Nikon's present and future business. Provide the sales force with technical assistance to successfully satisfy customer's needs and complete the sale. Create and conduct training curriculum on advanced microscopy and new technologies to Nikon distribution and end users. Attend trade shows, workshops and national dealer / sales meetings. Write technical and applications bulletins to end users and distribution channels as well articles for publication in Bio journals highlighting Nikon technology or applications.
Selected candidate will act as liaison for technical information and feedback between the end user/sales network and Nikon factory engineers on all matters pertaining to our BioScience technical product line. Applicants must have a Ph.D. in BioSciences, BioEngineering or Physics with a specialty in optics, or equivalent experience. Knowledge of advanced microscopy techniques and optical principals including fluorescence applications, imaging and confocal applications required. Excellent communications skills in English a must, along with good writing and public speaking abilities. Excellent benefits and compensation provided.
Mail or fax your resume, which MUST include salary requirements, to: HR Department, Nikon, 1300 Walt Whitman Road, Melville, NY 11747. Fax: 631-547-4025.
Or send a digital cover letter and resume / CV in (plain text, MS Word, or .pdf file) to biosales-at-nikonincmail.com
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Also See add display in NY Times: http://search.nytimes.com/classified/display/adverts/524986401/
Nikon is an Equal Opportunity Employer =============================================================
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I've had to coat some PLGA polymer samples and we think the coater may be melting them. The samples have a rolled or beaded edge and large cracks. I have jpeg images if anyone wants me to send them for a look. Are we correct in our assessment or should we look elsewhere for the source of cracks, shrinkage, beading around edges. These artifacts, BTW do not appear when the uncoated samples are checked under a light microscope.
Dear friends, I have just finished writing the programs for TEMAlert system(http://www6.ewebcity.com/temalert), which serves as a pre-print announcement system. Currently it serves only TEM-related region. Do you think this is very useful for all TEM users? The system was designed to be totally self-maintaining and everyone can post messages to announce his/her newly published (accepted) paper there.
I hope that TEMAlert will become a widespread blackboard in TEM region and tighten the relationship between electron microscopists allover the world.
I am sorry that the server is somewhat slow - because it is a free internet host. I will be very grateful if anyone of you can supply me a space (should support ASP).
With best regards, Shu-You Li ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Please visit my new homepage - a personal website on transmission electron microscopy (TEM) - at http://syli.homepage.com at your convenience! It contains my current research work, my resume TEM-related journals, instruction for authors link to on-line EELS database, periodic table, physical constants JOB list and RESUMEs (only for TEM region) TEMAlert - a self-maintaining preprint announcement system ************************************************** Shu-You Li, Dr. Institut fuer Physikalische Chemie Johannes Guttenberg Universitaet Jakob-Welder-Weg 11 D-55099 Mainz, Germany
} In Australia you could not buy a large system like that as a supplied item.
Don't know about that...We've had excellent results from a locally made system we've been using here now on the SEMs for several years. Service and parts are readily available and it has heaps of spare capacity. Contact Mark Blackford (mgb-at-postoffice.ansto.gov.au) for details on who to contact about this system because they probably have a branch in Melbourne.
} } Beer chillers as used in hotels are most suitable.
Certainly a mass produced item in Australia.
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
I have a batch of images (~300) that I would like to present as a QuickTime movie. Does anybody have experience how to do this? I am using a Macintosh. Comments welcome. Thanks,
Edgar
________________ Dr. Edgar Voelkl ORNL Bldg 4515 1 Bethel Valley Road P.O. Box 2008 Oak Ridge, TN 37831-6064
As a metallographic equipment and consumables manufacturer, BUEHLER¨ offers a number of products which would work for your application. However, I would suggest our MASTERPREP* (Part No. 40-6377-032) product. This is a 0.05 micron alumina suspension. What makes it unique is the fact that it is produced through the seeded gel process instead of by calcining. In the seeded gel process, the alumina is precipitated from a liquid phase. This results in a better controlled particulate size distribution, higher particulate density, and more consistent particle geometry. We've found this product to be superior to all of the other 0.05 micron alumina products that we sell for preparation of ductile metals.
I hope this helps. If you need further information, you can email me privately, call me at the numbers listed below, or contact our sales department for pricing.
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I am also trying to figure out how to make a QuickTime video from a series of images. I am using a PC, and the images are from a Zeiss 510 Confocal, in LSM format, which have been converted to Tiff. The number I am dealing with is 64 images. Any help would be greatly appreciated Linda Barthel Research Associate II Department of Cell and Developmental Biology University of Michigan barthel-at-umich.edu
We have a client, here in South Africa, who has recently purchased a Philips XL30 ESEM LaB6 with a cryo system, hot stage and CL detectors. We would like to now run a workshop, here on this system in South Africa, on the applications of ESEM. This system is a national facility and would therefore like to introduce the local EM users to the exciting world of ESEM.
We realise that there are a few friends of ours in Australia who would be ideal, but then we have various contacts in England and the USA too. In this way we feel we should get a chance at the best choice of getting a really exciting workshop set up or possibly a series of workshops. We expect to keep the delegates riveted for at least 4 days of workshop. The idea of the workshop is to show as many applications for ESEM as possible. Then to specialise in some of the biological areas, as this would be some of the more regular users for this system. Those who could assist should please contact us and we will pass your information on to the client. Please indicate your availability, field of interest and the, always important, costs involved.
Thanks for your time.
Luc Harmsen Anaspec, South Africa Technical support on microscopy. Tel + 27 (0) 11 476 3455 Fax + 27 (0) 11 476 7290 anaspec-at-icon.co.za www.anaspec.co.za
Remember, ICEM 15 will be in 2002, Durban, South Africa. www.icem15.com
We have a sample of 6 micrometers Ni-Zn-P electroless plated onto 1 mm thick Al and we would like to prepare cross sections for TEM examination. It looks very difficult to use the tripod polishing technique for metals.So,we are thinking at an electropolishing procedure with our double-jet Tenupol apparatus. This would be done on slices cutted from a sample initially thicked up to 3mm by electroplating.But how to avoid differential material removal problems? Which material for plating?Which electrolyte for electropolishing?Other method? Does anyone have suggestions for us?
Thanks in advance for your help.
Jean Dille
Materials Science and Electrochemistry Free University of Brussels CP 194/3 Avenue F.Roosevelt 50 1050 BRUSSELS BELGIUM tel:32-2-6502723 fax:32-2-6502786 e-mail: jdille-at-ulb.ac.be
The results are determined by the largest source of noise, be it the initial electron statistics or subsequent noise introduced by conversions or recording techniques. In this case (dark field imaging), the largest source for noise is probably the statistical nature of the electron beam. The lower you go in exposure (i.e., the fewer electrons you record per pixel), the higher the relative noise. And there is no way to get around this either by film or digital camera. The only way to improve the noise is to go to more electrons, i.e., longer exposure or brighter beam.
If the sample is so sensitive that even a dark field exposure damages the sample, there is probably not much you can do.
If the problem is drift, a digital system can help you: Instead of 1 exposure at, for example, 30 seconds, acquire 3 exposures at 10 seconds. Each one of them will be very noisy, but one can add them to get the same noise figure as a 30 sec exposure. And the 10 sec exposure of each one cuts down on drift. All you need to do is to align the 3 images.
I used to take dark field images with a digital camera and (of course) I loved it. It allowed me to acquire the images and immediately see them. I did not have to wait hours to develop the negatives and then find out the exposure was not long enough (or too long).
Michael
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: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: jim [mailto:jim-at-proscitech.com.au] Sent: Sunday, May 21, 2000 2:38 AM To: 'Sergey Ryazantsev'; Microscopy-at-sparc5.microscopy.com
I find what you are trying to do interesting, but there are things that may never work and this may be one.
We had it confirmed by Michael Bode that good digital system "see "single photons - so that is not likely to improve much. Film too registers single electrons. We were told that in digital 1 electron can expose one pixel. On film one electron initiates the exposure of a silver halide and subsequent electrons in a near identical location increase the size of the grain.
Low noise (cooled cameras) hopefully do not add their own noise, but they cannot make up for lack of information in the image.
My case (below) was that in high resolution TEM at least, less exposed images will be inferior because there are simply not enough electrons to produce a good image. A more sensitive digital system uses fewer electrons and so makes matters worse.
In dark field EM we are using the beam indirectly and such images too suffer particularly from electron noise. Here too the use of a digital system would only make this worse. Furthermore, the slower 4489 film would be better than SO-163.
If you don't like long exposures (4 to 8 seconds I found a practical limit), you cannot ignore the reality of electron noise and hope to correct that with yet fewer electrons, albeit processed in a "noise-free" digital system. I think that the answer is more electrons, i.e. a field emission source or a slower digital camera. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, May 19, 2000 3:04 PM, Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] wrote: } } } Jim hello } } I was talking about very special EM case: dark-field TEM. At this point we } are talking about a few electrons per square angstrom, even less. In high } resolution EM crystallography people have deal with 0.6 e/A2. At such } conditions, high sensitivity and linearity of the modern digital cameras } may be a plus. Talking about long exposures --what about drift? I never } had good pictures at x100K with exposure longer that a few seconds. I } don't understand your point about noise. In case of digital camera the } noise is a function of camera. Current cameras has a very low level of } noise (they uses cooling, etc) and we have to pay for that astronomical } price. This is a life. I am not friendly with TEM cameras, but I do know } that in the light microscopy people count individual photons using CCD } cameras. In TEM camera we have phosphorous screen as a source of image and } my questions to Mickhael were addressed mostly to the problem how } effectively (and correctly) information is traveled thought that funny } screen. The Mickhael's answer is that screen does not affect dynamic range } and sensitivity is much higher than on convention films. Am I correct, } Mickhaels? } } Sergey } } } Date: Thu, 18 May 2000 14:15:02 +1000 } } From: jim {jim-at-proscitech.com.au} } } Subject: RE: new developments in imaging systems? } } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} , } } "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-sparc5.microscopy.com} } } Reply-to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } } Organization: ProSciTech } } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } } } ----------------------------------------------------------------------- - } } 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 } } -----------------------------------------------------------------------
My thanks to all those who responded to my question about treating coverslips to make cells stick to them better. I have used one or two of these in the past, but my colleague wanted to try some different approaches. Thanks again.
Matthew J. Schibler Ph.D. UCLA Brain Research Institute 1524A Gonda (Goldschmied) Center for Neuroscience and Genetics Los Angeles, CA 90095-1761
Linda, if you get quicktime pro (its an upgrade to regular quicktime, costs about $30 from Apple) you can import a numbered series of tiffs directly into quicktime. which then can be made into a movie Simon
-----Original Message----- } From: Linda Barthel [mailto:barthel-at-umich.edu] Sent: Monday, May 22, 2000 10:44 AM To: Microscopy listserver
I am also trying to figure out how to make a QuickTime video from a series of images. I am using a PC, and the images are from a Zeiss 510 Confocal, in LSM format, which have been converted to Tiff. The number I am dealing with is 64 images. Any help would be greatly appreciated Linda Barthel Research Associate II Department of Cell and Developmental Biology University of Michigan barthel-at-umich.edu
I have a functional, but old, Balzers Freeze Fracture machine. It is headed for the scrap pile unless someone out there would like to have it. I will explore the possibilities if anyone shows an interest.
Greg Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
Thanks for the input. I was hesitant to use diamond abrasives on my samples due to previous advice. I was told that due to the hardness of diamond, it tends to embed itself in soft metals such as copper, which results in "smearing" more than polishing. Do you have any input regarding the smearing issue?
John
On Fri, 19 May 2000, Edward Hirsch wrote:
} John, } The solutions that the colloidal silica are in have a ph of 8.5 to 9.8 } depending on the formulation. I suggest you try our 0.05 micron water based } polycrystalline diamond suspension. Allied High Tech } http://www.alliedhightech.com sells this product. I would also suggest our } Final A cloth for this polishing step. If you would like samples of either } of these products please let me know and I will be happy to send it to you. } } If you need further technical assistance please contact me off-line and I } will be happy to help or you may contact our main office at (800)675-1118 } located in CA. } } I hope this helps. } Ed } Please note, I have a financial interest in providing you with these } products and other sample preparation equipment and consumable items. } } ************************************************* } Edward A. Hirsch } Product Application Specialist } Allied High Tech Products } 2376 East Pacifica Place } Rancho Dominguez, CA 90220 } ph: (919) 846-9628 } vm:(800)675-1118 x245 } fx: (310)762-6808 } http://www.alliedhightech.com } } Equipment and Consumables for Metallurgical Sample Preparation } ************************************************* } } -----Original Message----- } From: john david whitaker [mailto:jwhitake-at-u.washington.edu] } Sent: Friday, May 19, 2000 4:01 PM } To: Microscopy Lister Server } Subject: final polishing for metals (Cu, Ni, Fe, NiFe) } } } ------------------------------------------------------------------------ } 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 looking for information regarding final polishing } solutions for preparing metallic cross-section samples } for TEM analysis. } } I've looked into colloidal silica } suspensions such as Ludox and Syton, and am concerned } that their alkilinity may chemically etch my samples. } I'm analyzing different compositions of NiFe } on copper substrates, so the specimens are prone to } preferential etching of different phases. } } Does anyone have any experience with this? } } Any input would be appreciated. } } John } } } }
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Our software, 4D turnaround will make quicktime movies out of image stacks that are in biorad format, pics, or tiff. Please see our website for more details and to download: http://www.loci.wisc.edu/4d/native/4d.html
Currently we have only a mac version available, however we will be releasing a java version next month that works cross platform.
Best regards, kevin
Kevin W. Eliceiri Project Director Laboratory for Optical and Computational Instrumentation http://www.loci.wisc.edu 159 Animal Sciences 1675 Observatory Dr. Madison, WI 53706 608-263-6288 voice 608-265-4076 fax
A view days ago I found your message in my mailbox. Regarding your interest about digital imaging systems I can provide you some information on our imaging plate system for TEM.
DIBIS Imaging Plate Technology is adapted for TEMs from, FEI/Philips, LEO/Zeiss, JEOL and HITACHI. The Imaging Plate (81 x 100 mm) is inserted into the sheet film cameras of the various TEMs and is directly exposed by electrons. After that the imaging plate reader creates digital images directly from the plates without involving chemical processing thus providing extraordinary image quality with 3600 x 3200 pixel at 25 µm and 16 or 20 bit dynamic range with true linearity. The high pixel count supports printouts in real photographic quality, also on larger formats as you are used to from photographic film. One Instrument in your lab will serve all your TEMs with highest quality digital imaging technology, no matter what type and manufacturer. The high performance, resolution, sensitivity and dynamic range makes Imaging Plate Technology first choice for life science and material sciences imaging, low dose applications and high dynamic diffraction patterns.
So, DIBIS introduces MICRON Digital Imaging Plate Technology as the alternative to overcome the limits of CCD technology for TEM.
For more details visit our homepage.
R. Kuenzler ---------------------------------- DIBIS Digital Biomedical Imaging Systems AG Gewerbestra§e 11; D-75217 Birkenfeld Tel.: +49 (0)7082 940639 Fax : +49 (0)7082 940076 E-Mail: contact-at-dibis.de E-Mail: kuenzler-at-dibis.de Internet: http://www.dibis.de
Margaret Dienelt schrieb:
Hi,
} } Year after year I hopefully gather information about digital imaging } systems for TEMs (ours is a JEOL 100CX) , only to learn we have no } money. This year it looks like it might really happen but I have not } kept up with innovations in the field and am wondering the following: } } 1. Anything new in the last two years -- especially in terms of } cameras? I'm most familiar with the Gatan and AMT systems but their } web sites don't reflect much in the way of changes over a year ago. } 2. With more and more microscopists finally getting their systems -- } I'd love to get feedback. } } Thanks, } Margaret } } P.S. Would welcome contacts from vendors. } } -- } Margaret Dienelt } } Plant Pathologist } Electron Microscopy Lab } } Floral and Nursery Plants Research Unit } U.S. National Arboretum/Agricultural Research Service/USDA } } B. 010A, Rm. 238, BARC-W } 10300 Baltimore Avenue } Beltsville MD. 20705 USA } } (301) 504-6097 } Fax: (301) 504-5096
GraphicConverter will convert a folder of numbered tiffs into QT. Just be aware that after choosing the folder with your tiffs, and clicking on Convert, the next window has an easily overlooked choice at its top to save the files as ONE movie. Forget this selection and you will have converted your 300 tiffs into 300 moov files. GraphicConverter has many options for compression and output size.
NIH Image can open a folder of numbered tiffs with the 'Open All' option. Use the 'Windows to Stack' command then save as QT. Image will place your tiffs into the stack in the order in which they were opened. So if your filenames don't end with an incrementing number you could open them manually in the desired order.
Regards, 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
********************************************************************* C:} The box said "Requires Windows95 or better". So I bought a Macintosh. *********************************************************************
I am unsure what versions are available for the Mac, but I have made videos using ULEAD video editing software for the PC. TIFF images can be imported and displayed for "N" number of frames. Special effects (like a wipe or fade) can be added between sets fo (still) frame sets.
Do beware this process can tax the resources of most PCs. If the video is uncompressed, the data rate can be 10+ MB/sec. I use a Matrox video card which has MJPEG hardware compression. At full VHS resolution and 15 frames/sec (not 29.9) the data rate is below 2 MB/sec. With MJPEG, I can get about 10 minutes of "fair" quality video on one CD-R (650 MB). Mpeg compression can make a smaller file, but I havent tried a compression program I like - Lousy quality - Jumps, etc. I haven't tried the MPEG program from Xing Tech. which is rated better thatn the sharewares I have tried.
Another issue is the conversion time using software mpeg convertors is that 10 minutes of MJPEG video to mpeg on my PC (350 MHz/256 Meg Ram) takes well over an hour.
E.A. Fischione Instruments, Inc., a rapidly growing company specializing in the development and manufacture of TEM specimen preparation instrumentation, is seeking a highly motivated individual for the position of a Application Scientist. The candidate's responsibilities will include:
* Obtain and prepare TEM specimens of customer's material for sales purposes. * Analyze customers' specimens in Fischione's TEM. * Library research to support design and application work activities. * Answer customers' questions regarding use of Fischione products. * Instrument training (when required). * Provide technical support at major tradeshows. * Give product demonstrations. * Collaborate with potential customers on experiments. * Write applications notes. * Give scientific presentations. * Conduct short courses and workshops on specimen preparation and other Fischione related technology. * Write refereed journal articles for publication. * Generate information for Website. * Revise instrument instruction manuals. * Evaluate emerging microscopy related technologies and market opportunities. * Convey market opportunities to Fischione management. * Obtain input from customers on possible new products and on improvements to existing products. * Work with the product design team on new product developments. * Provide design support from the microscopist's standpoint. * Prototype and test both existing product improvements and new products. * Procure needed instrumentation to get specimen preparation facility running appropriately. * Optimize Fischione's TEM laboratory. * Work with architects to design new EM facility when building expansion occurs. * Travel approximately 10%-20%.
The candidate should have a Ph.D in Material Science, Engineering, or Physics with a specialization in Electron Microscopy.
Salary is commensurate with experience.
E.A. Fischione Instruments, Inc. is an equal opportunity employer. Please send your resume and salary requirements to:
Human Resources Director E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 Phone (724) 325-5444 FAX (724) 325-5443 E-mail: info-at-fischione.com
I have never used a sintillator type BSE detector, but the major differences are two fold.
Typically (though diodes are getting better all the time) the Robinson has a better low energy BSE detection effiency. It follows that for the same noise level electronics, it would exhibit better sensitivity. The dynamic range problem will still exist for very different Zs in the field of view. In the middle and upper portions of the sensitivity range (low and lower), I would not expect much difference between them. ...Any Comments from users of both????
I like the 4 quad diodes since I can go differential mode for macro topography (like fracture surfaces) and show the gross features while suppressing the fine detail.
The Pt coating can have a profound negative effect on sensitivity if not extremely thin. If given a choice, I would always use carbon for the best BSE sensitivity. ...If not hazardous, I would use Be, BUT!!!! For elements in the range of Al & Si, I prefer to use carbon and lower beam voltages to minimize penetration, especially if they are films or very small features.
I once presented some data illustrating the BSE signal attenuation as a function of sputtered Au thickness. But that data would be hard to retrieve now.
Woody White McDermott Technology
=================================================== I use a Robinson Model 6 which is specified at a Z contrast of 0.003 at } = 2KV. From your work with a 0.1Z detector, what would you say is the qualitative effect that a higher Z contrast detector would offer? I am especially interested in imaging Al/Si alloys and I typically sputter coat them with Pt. I may try carbon one of these days to see what the difference might be.
Yes, heating in the sputter coater is likely the cause.
Confirm this by checking the coated samples under the optical microscope before putting them in the SEM. This will eliminate from consideration any artifacts caused by higher SEM vacuum and electron beam damage.
At 2:56 AM -0400 5/22/00, lherault wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
====================================== Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. 425 E. University Ave. Ann Arbor, MI 48109-1063 USA (734) 936-1550 FAX (734) 763-4690 ======================================
Are you using a magnetron sputter coater? If so, then you should not be getting melting, etc. But ...
For how long did you coat your samples? You might try coating for short periods, with some rest in between -- say 30 sec, turn off (maybe add some argon to help carry away any heat), coat, rest, etc. If your samples are very sensitive, use 10 sec. coat times. Rest for 30 sec more or less (empirically). I've done Teflon fabrics this way, using 90 sec. coat periods without problems, but if your samples are thick, this would increase the problems.
Phil
} I've had to coat some PLGA polymer samples and we think the coater may be } melting them. The samples have a rolled or beaded edge and large cracks. I } have jpeg images if anyone wants me to send them for a look. Are we } correct in our assessment or should we look elsewhere for the source of } cracks, shrinkage, beading around edges. These artifacts, BTW do not appear } when the uncoated samples are checked under a light microscope. } } Thanks in advance. } } Ron L } lherault-at-bu.edu
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC Dept. of Animal Health and Biomedical Sciences University of Wisconsin 1656 Linden Drive Madison, WI 53706-1581 voice: (608) 263-4162 fax: (608) 262-7420 (dept. fax)
How long of running time, approximately? Do you want dissolve from image to image or simple image swap? Any audio?
gary g.
At 07:44 AM 5/22/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If your samples are particularly heat sensitive, you should probably look into an Ion Beam Sputtering System. The heating effects in such a system are negligible. Of course, you have the added advantages of thinner, more uniform coatings etc. also. We do make the IBS/e Ion Beam Sputter Deposition and Etching System. If this is an infrequent application for you, perhaps I can put you in touch with one of our local users who could help you out.
David Henriks Vice President TEL: 800-728-2233 (toll free in the USA) South Bay Technology, Inc. +1-949-492-2600 1120 Via Callejon FAX: +1-949-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy. Message text written by "lherault" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I've had to coat some PLGA polymer samples and we think the coater may be melting them. The samples have a rolled or beaded edge and large cracks. I have jpeg images if anyone wants me to send them for a look. Are we correct in our assessment or should we look elsewhere for the source of cracks, shrinkage, beading around edges. These artifacts, BTW do not appear when the uncoated samples are checked under a light microscope.
We are moving to a new building and need to get rid of some old equipment. The following are available in New Jersey, USA. All items were fully functional when last used, most have all accessories and owners manuals. Any reasonable offer considered.
For sale:
-GW Electronics attachments to go on Hitachi HS-510: Dual magnification unit, Graphics generator, and Homomorphic Processor
-LKB 8800 Ultrotome III (thermal advance, includes ALL accessories)
-Denton Critcal Point Dryer (CPD-1) With extra baskets, seals, and coupling for CO2 tank
-Ames Lab-Tek cryostat (slow leak in refrigerant line)
-Zeiss microspectrophotometer (MPM microscope photometer for cytospectrophotometry) for Zeiss Photo-microscope, Ultraphot, Standard Universal microscopes (includes light chopper, power supply, monochromator, photometer head, photomultiplier housing, indicator unit, and coupling for microscope head)
-Printz automatic print dryer model JET 260158 ferrotyper drum and canvas belt in like-new condition)
-Kodak Ektamatic Model 214-K automatic print processor -Accessories for AO/Reichert Microstar compound scope (available with or without microscope): AO Expostar photomicrographic system (lens and shutter, control unit model 1190, polaroid and 35 mm film backs), AO verical illuminator for incident light fluorescence microscopy (includes mercury lamp model 2054A, filter housing, AND microscope), Camera lucida attanchment AO #1030
- 2 ea Omni-mixer homogenizer with stand model # 17105, 16,000 rpm, no impellers -Lightnin Mixer Model F (no impellor)
-B&L Dynazoom microscope with integrated 4x5 (polaroid) and 35 mm camera backs
**Items looking for a good home (shipping cost plus a little extra for us to be able to say that we actually sold them):
-B&L # 42-63-89 projecting compound scope (used to do camera lucida)
-stainless steel frames for holding individual sheets/plates of 3 1/4 x 4 EM film
--Arthur Thomas Co., paraffin for 50-52oc (33 lib pkg of 1/4 lb bars)
Dealers are welcomed to contact us. Perhaps we could give you some items plus a cash allowance to purchase some used items that you may have that we want.
Don ______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
You can get NIH Image to do this and a great deal more. For PC's it's called Scion Image, and it's available free from www.scioncorp.com. Hope this helps.
Lesley Weston.
On Mon, 22 May 2000, Kevin W. Eliceiri 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 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 also trying to figure out how to make a QuickTime video from a series } } of images. I am using a PC, and the images are from a Zeiss 510 Confocal, } } in LSM format, which have been converted to Tiff. The number I am dealing } } with is 64 images. Any help would be greatly appreciated } } Linda Barthel } } Research Associate II } } Department of Cell and Developmental Biology } } University of Michigan } } barthel-at-umich.edu } } Our software, 4D turnaround will make quicktime movies out of image } stacks that are in biorad format, pics, or tiff. Please see our } website for more details and to download: } http://www.loci.wisc.edu/4d/native/4d.html } } Currently we have only a mac version available, however we will be } releasing a java version next month that works cross platform. } } Best regards, } kevin } } Kevin W. Eliceiri } Project Director } Laboratory for Optical and Computational Instrumentation } http://www.loci.wisc.edu } 159 Animal Sciences } 1675 Observatory Dr. } Madison, WI 53706 } 608-263-6288 voice } 608-265-4076 fax } } }
: Frieda Lim } Barbaric Yawp } 224 west 13th Street #5r } NYC, NY 10011-7775 } 212.206.0683 } barbaricyawp-at-att.net } } Dear MSA Members } } It was suggested to me that this may be the perfect spot to list my query. } I would appreciate your perusal of the below queries and am anxious to hear from you. } } Sincerely, } Frieda Lim } } } } 1st query: } } Dear MSA Members : } } I invite you to take a close look at our query. We are a film production } company seeking existing moving microscopic organism film or video footage. } We will utilize this footage for our single purpose of making a feature } film. The film we are creating is a microscopic fantasia. Zooming in on } your microscopic world, we will spin a tale through imagery and music. } } We would appreciate your help with obtaining some footage directly or any } referring leads. Please contact us at your convenience to discuss further } details. } } Sincerely, } Frieda Lim } } } } } Elaborating 2nd Query: } } Dear MSA Members: } } I thank you for your prompt response to my query for microscopic } microorganism footage. Our intended use of your existing footage is to edit } & generate a full feature film comprised solely of microscopic imagery and } music with which we hope, eventually, will be distributed to the general } public at large--large in fact by mesmerizing all ages, the young & the } young of heart. Our microscopic fantasia will bring to life a single } narrative, the classic myth of Cupid & Psyche. } } In our original query, we had cast a wide net being intentionally } non-specific with regard to the kind of microlife we are seeking. At this } stage, we have no restrictions as to what microworld would be best suited } for our story. However, our intent is to portray our story within a } scientific veritable reality. We don't want micro species colliding that } would never interact in truth. Right now we are in our initial phase of } investigative hunting & gathering-a casting call for microscopic actors. We } actually want to cast organisms as actual characters and to use others as } metaphorical imagery. My suspicion is that you have already created footage } for your scientific research and discovered many a talented organism } awaiting their big break. We further suspect that we will need to pool } microscopy resources and see what footage offered is most varied in look & } movements that will best fill the wide assortment of roles required. Since } we are approaching your world as layman, we would appreciate your expertise } in advising what microorganism microcosm might be most readily available. } } To note, this is a low budget project with big hopes and dreams. We } believe our film can achieve success similar to that of "Microcosmos"-the ' } 96 Cannes Film Festival jury prize winner. That movie was testament that } science has mass marketable enthusiasm in the entertainment world. } Comparatively, we hope to explode your frontier world onto the big screen } and excite a wide audience. } } So, if you know of some microorganisms wanting to make it big, we are } anxious to hear from you. Thank you. } } Sincerely, } Frieda Lim
Firstly a confession, that my taxonomy is badly in need of repairing - the alga we have been working with is not a blue-green algae at all - it's actually Polytomella, a eukaryote, and I think is a relative of Chlamydomonas, but doesn't have a cell wall or photosynthetic apparatus.
Anyway, embedding in Spurr's seems to have done the trick as far as stopping the starch granules dropping out. I've just been looking at grid-fulls of cells full of starch granules, and they're all there! Many thanks for the suggestions. I'll be back with more problems soon!
Mark
******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
We've applied microscopic techniques (e.g., polarized light microscopy, fluorescence microscopy, FTIR microspectroscopy, and SEM-EDS) and non-microscopic techniques to more than 700 projects involving historic and artistic works, from Egyptian antiquity to contemporary. Material evidence of authenticity -- or more likely, inauthenticity -- has been or become an objective of many of these studies.
Scientific investigation plays an important role in the authentication process, often providing indirect or direct evidence of date, and an objective basis by which to compare materials and techniques to works of unquestioned authenticity. Given sampling limitations and the layered structure of many art objects and decorative finishes, microscopy is ideally and elegantly suited to their study of the former - that is, structure and composition. We use a new infinity-corrected optical/FTIR microscopy system, that we had custom-fitted for polarized light microscopy, fluorescence microscopy, and FTIR microspectroscopy (narrow-band MCTA and wide-band MCTB detectors).
Your article sounds very interesting -- please feel free to call.
Jamie
James Martin Orion Analytical, LLC Post Office Box 550 Williamstown, MA 01267 phone: 413-458-0233 e-mail: martin-at-orionanalytical.com website: www.orionanalytical.com
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Hello,
I am new to this list. I would like to know if anyone has information I could use to write an article on the analytical chemistry techniques used to authenticate art objects using light microscopes or any other type of microscope. Also, if anyone has information on any of the other techniques used to authenticate art objects such as x-ray diffraction, x-ray fluorescence, accelerator mass spectrometry, that would be helpful too.
Sincerely,
Margaret M. Mitchell Assistant Editor AOAC INTERNATIONAL 301-924-7077 (tel) 301-924-7089 (fax) mmitchell-at-aoac.org
Jean Dille wrote: ===================================================== We have a sample of 6 micrometers Ni-Zn-P electroless plated onto 1 mm thick Al and we would like to prepare cross sections for TEM examination. It looks very difficult to use the tripod polishing technique for metals.So, we are thinking at an electropolishing procedure with our double-jet Tenupol apparatus.
This would be done on slices cutted from a sample initially thicked up to 3mm by electroplating.But how to avoid differential material removal problems? Which material for plating?Which electrolyte for electropolishing?Other method?
Does anyone have suggestions for us?
Thanks in advance for your help. ==================================================== If the aluminum substrate can be thinned down a bit more, then a coating of this thickness should be able to be thin sectioned using diamond knife ultramicrotomy. It is hard to predict whether better results will be obtained embedded or unembedded (there is a tendency for the coating to separate from the substrate). We usually find that such separation is less likely to occur if the sample is embedded. However, structure within the coating is less well preserved when embedded. Naturally our experience has been with our own SPI-Pon™ 812 epoxy embedding resin and SPI diamond knives, but I would expect that at least most of the other "Epon substitutes" (and knives) would work just as well. With regard to the substrate separation, the tendency for this to happen can be reduced by using a knife included angle that is smaller rather than larger (we were successful with 45 deg.). The use of larger included angles, at least in our experience, seemed to produce a level of compression artifacts in the sections that we found unacceptable.
In any case, we have found the diamond knife thin sectioning approach to often times offer certain advantages over the alternatives for sample preparation.
Disclaimer: SPI Supplies offers materials science diamond knives and also our preferred embedding resin, SPI-Pon 812 Embedding System. Our Structure Probe™ laboratory services division performs this kind of diamond knife thin sectioning as a service for commercial clients.
Chuck
PS: Remember that we are trying to become 100% paperless and the only way that we can manage this kind of correspondence, in a paperless environment, is for our correspondents to always reply by way of "reply" on their software so that the entire string of correspondence on that topic gets returned to us, and the entire correspondence history can be kept in one place.
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Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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Ronald J. L'Herault wrote: ==================================================================== I've had to coat some PLGA polymer samples and we think the coater may be melting them. The samples have a rolled or beaded edge and large cracks. I have jpeg images if anyone wants me to send them for a look. Are we correct in our assessment or should we look elsewhere for the source of cracks, shrinkage, beading around edges. These artifacts, BTW do not appear when the uncoated samples are checked under a light microscope. ==================================================================== If by PLGA you mean polylactic glycolic acid, which in certain forms could be a (dissolvable) surgical suture material, then the polymer could be quite hygroscopic and if it has had time to absorb enough moisture, it could be evolving moisture, and that could be the reason for your problems.
Some sputter coaters have a "test mode" which enables the user to momentarily expose in a gentle way the polymer surface to the glow of the plasma. If moisture is evolving, then there will be an immediate diminution of the quality of the vacuum. When confronted with that kind of situation, we go through the cycle of "test mode", then pump down, test mode, etc, until hitting the test mode button results in no more deflection of the vacuum. Ten or more cycles might be needed but in the end, the surface moisture is removed.
Then you are ready to coat.
If you are not talking about polylactic glycolic acid, I apologize for taking up the extra bandwidth.
I realize that not all sputter coaters have such a test mode, but that is the perfect kind of example where such a "test mode" has its greatest value.
Disclaimer: SPI Supplies manufactures sputter coaters with "test modes".
Chuck
PS: Remember that we are trying to become 100% paperless and the only way that we can manage this kind of correspondence, in a paperless environment, is for our correspondents to always reply by way of "reply" on their software so that the entire string of correspondence on that topic gets returned to us, and the entire correspondence history can be kept in one place.
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
I've applied microscopic techniques (e.g., polarized light microscopy, fluorescence microscopy, FTIR microspectroscopy, and SEM-EDS) and non-microscopic techniques to more than 700 projects involving historic and artistic works, from Egyptian antiquity to contemporary. Material evidence of authenticity -- or more likely, inauthenticity -- has been or become an objective of many of these studies.
Scientific investigation plays an important role in the authentication process, often providing indirect or direct evidence of date, and an objective basis by which to compare materials and techniques to works of unquestioned authenticity. Given sampling limitations and the layered structure of many art objects and decorative finishes, microscopy is ideally and elegantly suited to their study of the former - that is, structure and composition. We use a new infinity-corrected optical/FTIR microscopy system, that we had custom-fitted for polarized light microscopy, fluorescence microscopy, and FTIR microspectroscopy (narrow-band MCTA and wide-band MCTB detectors).
Your article sounds very interesting -- please feel free to call.
Jamie
James Martin Orion Analytical, LLC Post Office Box 550 Williamstown, MA 01267 phone: 413-458-0233 e-mail: martin-at-orionanalytical.com website: www.orionanalytical.com
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Hello,
I am new to this list. I would like to know if anyone has information I could use to write an article on the analytical chemistry techniques used to authenticate art objects using light microscopes or any other type of microscope. Also, if anyone has information on any of the other techniques used to authenticate art objects such as x-ray diffraction, x-ray fluorescence, accelerator mass spectrometry, that would be helpful too.
Sincerely,
Margaret M. Mitchell Assistant Editor AOAC INTERNATIONAL 301-924-7077 (tel) 301-924-7089 (fax) mmitchell-at-aoac.org
When I purchased a copy of Adobe Photoshop 5.5, it came bundled with their "ImageReady 2.0" This package can be used to create animated GIF images from layered images, or the animation exported in QuickTime format.
When I purchased a copy of Adobe Photoshop 5.5, it came bundled with their "ImageReady 2.0" This package can be used to create animated GIF images from layered images, or the animation exported in QuickTime format.
What I ended up doing in your situation was downloading Confocal Assistant, and reconstructing the TIFF series using that. You first need to add a BioRad header... the software and tricks can be found at:
http://www.cs.ubc.ca/spider/ladic/source.html
You can export the animation from CA as an .avi file. Then if you have Image Ready (Photoshop 5.5), you can convert the .avi to .mov. (I think Graphics Converter will also work) A roundabout way to get the job done, but it works.
All the best,
Angela
} I am also trying to figure out how to make a QuickTime video from a series } of images. I am using a PC, and the images are from a Zeiss 510 Confocal, } in LSM format, which have been converted to Tiff. The number I am dealing } with is 64 images. Any help would be greatly appreciated } Linda Barthel } Research Associate II } Department of Cell and Developmental Biology } University of Michigan } barthel-at-umich.edu } } } } } } --------------------------------------------- Angela V. Klaus
Laboratory Manager, Interdepartmental Laboratories American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
I would be pleased if anyone could broaden my views on equipment reliability . Basically we have a 3yr old FESEM which I consider to be fairly reliable in that it is on call 24hrs/day , has numerous ( non dedicated users ) and apart from downtime for filament change and the odd wear and tear type problems answers our needs . As we do not have a back up instrument and when we do experience problems it's always at the worse time certain personnel have the impression that it is unreliable .
What do other sem users expect in terms of reliability , apart from my ' subjective ' comments is it quantifiable , would approx 2wks /year downtime including planned maintenance be considered excessive ?
I have done this procedure many times using many different programs. Most often I was using TIFF files in sequence. I can't remember all the names of the programs that I tried, most were freeware or shareware including NIH image and the results were never to my satisfaction, I wanted control over Frames per second, compression, output format, light levels, you know complete control. The program that I wound up using was Adobe After Effects. An incredible program, capabilities way beyond what was needed for creating simple sequence movies from 3D volumes captured in a LSCM and rendered on a SGI. The benefit is unlimited file type importation (well nearly unlimited - hey it supports SGI .rgb files!) and complete control over the creation process. I could make an animated gif with any frame rate I wanted or a full compression-less Quicktime (.mov) or Microsoft Video for Windows format (.avi) with sound and special effects.
Most of you have used Adobe photoshop at one time or another, Adobe After Effects has the same intuitive interface and many of the same filters and image adjusting features. The educational discount for the lab brought the price down to less the $300! An incredible value for anyone doing lots of routine slice parade movies or animations for presentations.
If you have the ability to record sound on your computer you can even record a whole seminar presentation. Imagine showing up at a meeting plugging in your laptop computer (with sound) to the projection system and clicking play. You can sit down and listen to your own presentation, and then answer questions at the end. No more forgetting to mention a point, you can make sure that you are making sense. Of course this all takes away from the art of the presentation. I have never done this mind you all but the idea is intriguing isn't it?
Adobe also has a software program called Premier that is used in the film industry to do even more digital effects and post production (I think the last guess I heard was that about 75% of all rolling credits and film intros are done with this program and many of the same features are in After Effects).
-Geoff
-- Geoff Williams, M.S.
Electron Microscope Facility Supervisor Biology Department Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
Gordon Couger 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 } -----------------------------------------------------------------------. } } } From: "Tina Carvalho" {tina-at-pbrc.hawaii.edu} } } On Thu, 18 May 2000 ComCryLab1-at-aol.com-at-sparc5.microscopy.com wrote: } } } } } Dear Friends, } } } Can anyone give me advice regarding the dismantling and } } } the packing for shipment of a Philips 201 TEM? I need to } } } pay particular attention to not disturbing the alignment. } } } I plan to move the TEM from Univ. of Delaware to Naples, Florida } } } in an enclosed U-Haul Trailer. Any help or suggestions } } } will be most welcome. } } } } I will make sure Ron Veil, a very experienced independent EM service tech, } } sees this and has a chance to reply. It was with his advice that we (my } } hubby and I and its new owner) packed up and shipped a Philips 201. Like } } you, we wanted to ship it intact, column on and all. We built a heavy } } duty skid and added a strong base to which we bolted the instrument. Then } } we wrapped it with whatever that plastic packing tape that is like Saran } } Wrap is called, making sure to secure any part of the column we didn't } } want to move, but avoiding putting any pressure on things, such as the } } aperture drives. Wrap the bottom, wrap the column, wrap the column to the } } bottom and back again, etc. Build a crate up around the instrument. } } I think I remember putting a wood insert with a crescent cut out near the } } top of, but not touching the column, but hubby thinks not. The idea is } } NOT to let any shock to the crate get transferred to the column, so } } perhaps we didn't. Add some packing material, such as old egg crate foam } } from your bed (it needed replacing anyway) and whatever. Then, and this } } was the fun part, buy that stuff that when you mix it with a catalyst, } } produces huge volumes of foam that hardens in a few minutes. I think you } } can also buy spray cans of similar stuff, but since hubby had this on hand } } for other things, we had the bulk stuff. Fill the voids in the crate with } } it. It will easily chip off later. Add a top, and away you go. The } } scope made it in great shape. } } } } I think we packed the rotary pump and a couple of other things } } separately. Get it into the truck and TIE IT DOWN. I say this because an } } SEM we once packed for shipping with the base and saran, but no closed } } crate, made it from Hawaii to Ohio, then slid in the shippers truck in the } } last mile and slammed into the driver's seat. The driver was OK, the ion } } getter pump was bent, and the column needed a bit of work, but it could } } have been worse. Duh. } } } } I've also packed up a couple of Denton vcuum evaporators and a couple of } } ultramicrotomes. The saran stuff and blow foam are a good way to } } stabilize the instruments. } } } Since you are using a U haul you don't have to worry about sides } and a top on the crate. Make sure it is tied down real well. Dry } wall screws through the crate and into the floor work well for } locking it down but make sure you have it braced with timbers } to the front and sides of the trailer in case you make a panic } stop. } } Also make sure that the weight is centers in front of the trailer } axle if you use a trailer. Negitive weight on the trailer tounge } at the very least makes driving very interesting. You have no } Idea how fast a trailer can pass you while it is still tied on to the } truck. I have had the privilege of experiencing this and I can promise } you that you won't enjoy it:). } } The foam in place stuff is great. Just cover the instrument in plastic } and foam away. If you can get foam between the instrument and the } Support the foam will dissipate most of those little shocks that get } things out of line. } } Last of all make sure you understand what you insurance covers and } what it doesn't on moving equipment. You should be fine on liability but } the value of the contents is probably not covered. Rental truck companies } have contents insurance as an extra. } } Good luck } Gordon } } Gordon Couger gcouger-at-couger.com } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00
Gordon, Now I guess I know who to blame when I get U-Haul trailers that are in p----poor shape to move SEMs. Where do you get off running screws through the floor of a trailer you don't own?
You could probably expose your CCD chip directly to the electron bean -- and buy a new chip every few hundred exposures or so! Electrons have mass and carry momentum, which gets transformed into force as they are stopped in the crystal, especially at higher keV. This can lead to damage. Photons only heat up the target.
As far as I know ALL TEM cameras use a medium to convert electrons into light, and this light is then detected. The medium is either a phosphor screen or a very thin YAG crystal. From there on it is different. Some cameras use a fiber-optic to guide the photons to the chip, others use mirrors and lenses. So, it is not simply a matter of comparing the QE of the chip, the entire system has to taken into account. Also, each electron creates many photons in the phosphor or YAG, and in principle only one photon is enough to create a signal in the CCD. In other words, a CCD could theoretically detect "fractions" of an electron, whereas the film is directly exposed to the electron. Of course the electron can "rattle" around in the film and activate several grains, but as you can see, the question of comparing sensitivities now becomes one of comparing two different physical processes. It can definitely be done, but it's not easy.
Michael
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: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: hpadams [mailto:hpadams-at-bcm.tmc.edu] Sent: Thursday, May 25, 2000 12:30 PM To: microscopy-at-sparc5.microscopy.com
I have been following off and on this discussion on CCD cameras for EM. Much of this discussion has been concerned with "sensitivity". I am being naive here, but when we talk about "sensitivity" of CCDs, isn't this the same thing as the QE of camera/chip? I don't think I have ever seen a QE for em CCD's for various accelerating voltages/wavelenghts. Does the electron beam directly hit the silicon photodyodes or it there an interface that converts the incoming electrons to different (longer?) wavelengts? I know em films are sensitive to specific acc voltages. Are CCD cameras for em the same. For long exposures it may not matter, but for short exposures or low level intensity does the QE of the camera come into play?
} ===== Original Message From Jenichen {Jenichen-at-proscan.de} ===== } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id JAA13333 for dist-Microscopy; Fri, 26 May 2000 09:46:30 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id JAA13329 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 26 May 2000 09:46:00 -0500 (CDT) Received: from mailhub.iastate.edu (mailhub.iastate.edu [129.186.1.102]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id JAA13322 for {microscopy-at-sparc5.microscopy.com} ; Fri, 26 May 2000 09:45:48 -0500 (CDT) Received: from pc12223.bot.iastate.edu (pc12223.botany.iastate.edu [129.186.12.223]) by mailhub.iastate.edu (8.9.3/8.9.3) with ESMTP id JAA06942 for {microscopy-at-sparc5.microscopy.com} ; Fri, 26 May 2000 09:40:17 -0500 Message-Id: {4.2.0.58.20000526092941.00948c60-at-pop-1.iastate.edu} X-Sender: tpepper-at-pop-1.iastate.edu (Unverified) X-Mailer: QUALCOMM Windows Eudora Pro Version 4.2.0.58
Hello all, I have a researcher that is interested in utilizing florescent probes to label material with the desire to visualize with a 50 angstrom resolution. Are there any service facilities out there that have a cathodoluminescence detector on their SEM or STEM that would be able to assist him (via contract)? Thank you!
Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
} You could probably expose your CCD chip directly to the electron bean -- } and buy a new chip every few hundred exposures or so! Electrons have
There is an even worse problem with exposing the CCD directly the electron beam. The p-n junctions in the CCD chip have a certain "well capacity" - a number of electron/hole pairs they can hold before they saturate. Fast (keV) electrons are much more efficient at producing electron/hole pairs than photons - so much so that a pixel on the CCD would saturate after about 15 fast electrons hit it. Just the square root N shot noise at that level is about 25% - larger than typical TEM micrograph contrast of 10-20%. That's why a phosphor or scintillator is necessary to transform the fast electrons into photons.
In case anyone is interested, you can learn more than you ever possibly want to know about CCD cameras in:
"Applications of slow-scan CCD cameras in transmission electron microscopy" O. L. Krivanek and P. E. Mooney, Ultramicroscopy V. 49 p. 95-108 (1993). First description of Gatan CCD cameras, including some design issues, and measurements of the linearity, modulation transfer function (MTF), and detector quantum efficiency (DQE).
"Methods to measure the properties of slow-scan CCD cameras for electron detection" W. J. de Ruijter and J. K. Weiss, Rev. Sci. Instrum. V. 63, p. 4314 - 4321 (1992). Another early paper, includes comparisons to photographic plates. Uses a slightly different definition of the MTF from everyone else.
J. M. Zuo "Electron detection characteristics of slow-scan CCD camera", Ultramicroscopy V. 66 p. 21-33, (1996). Describes gain, MTF, and DQE measurements, measurement techniques based on stochastic noise (blank beam) images, and theory. This is a good place to start.
"Quantitative characterization of point spread function and detection quantum efficiency for a YAG scintillator slow scan CCD camera" A. L. Weickenmeier, W. Nuchter, and J. Mayer, Optik, V. 99, p. 147-154, (1995). Line-scan method of measuring the MTF.
Happy reading, Paul
Paul Voyles, voyles-at-research.nj.nec.com voice: (609) 951-2627, fax: (609) 951-2496 NEC Research Institute 4 Independence Way Princeton, NJ 08540 USA {http://www.neci.nj.nec.com/homepages/voyles/fluct.html}
On which systems have you observed higher focused beam current from the false (first) saturation peak rather than "true" saturation? I have not observed this in 18 years with an Etec nor on the new Hitachi.
Woody White
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Hi friends I agree Steve, sometimes (I seem it depends on electron gun design and distance between wenelt and anode, wenelt and filament) the first peak on saturation curve is more than saturation level even in secondary emission signal. Victor Sidorenko, ANTRON Co.Ltd., Moscow, Russia
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I would also be interested in learning how/why. I understand that if the larger spot size delivers more incident beam current, signal to noise is improved, but this does not seem to be the parameter to which Steve is referring.
Woody White McDermott Technology
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At 05:07 PM 25-05-2000 +0100, you wrote: Steve Chapman wrote, SNIP , then by de-saturating the larger source } will result in a larger probe dimension on the specimen; increasing the } probe diameter increases the volume of material involved in the production } of BSE. } Ken Wrote: But how does this lead to an increase in the BSE differentiation (implying more signal and hence less noise). Surely just defocussing would do the same thing. Defocussing as such should not effect the BSE coeffecient, unless you had sub surface charging or some other artefact.
Dear Walter, yes we have started one. We continue to gather as time goes on many of them are stored at the moment as the library section that will house them will not be finished until November. we are planning to offer some of them online in pdf format when we have permission from the various companies on the really old ones that they seem not to care about.
We need anything we can get our hands on and also include in this pursuit their older equipment brochures and catalogs. Any assistance of originals or good Xerox copies is great!
thanks Ed Sharpe archivist for SMECC
{ { Subj: Service Manuals Date: 5/25/00 10:54:00 PM US Mountain Standard Time From: Corvos-at-aol.com-at-sparc5.microscopy.com To: microscopy-at-sparc5.microscopy.com
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All,
I would like to know if any has started an extensive collection of Microscopy Service Manuals?
I am an author and platform presenter at the M&M 2000 conference at Philadelphia in August,2000. I am a scientist at the National Geophysical Research Insitute , India. I am looking for living accomodation during my 5 day stay at Philadelphia. I was wondering if any of you will be able to provide me with free accomodation or would be aware of any other resources who may be able to help me in this regard. Due to the limited fiancial resources avaliable to me from my lab, I am looking forward to help from my fellow researchers. Hope you can help me in this regard. I am eagerly looking forward to contributing to the M&M 2000 conference.
With kind regards R. Natarajan Scientist, National Geophysical Research Institute, Hyderabad, India e-mail: rnataraj-at-hd2.dot.net.in Phone: 91-40-7170141 X 2430 (W) Fax: 91-40-7170564
A hearty thank you to everyone who replied to my request for help. I had sources for everything I needed before the evening was out. It is the generosity of the subscribers that makes this list-serve a great resource. Thanks again.
Kim DeRuyter Electron Microscopy Technician 308 Natural Science Facility P.O. Box 755780 University of Alaska Fairbanks, AK 99775-5780 907-474-5452 907-474-5163 fax
I am an author and platform presenter at the M&M 2000 conference at Philadelphia in August,2000. I am a male scientist at the National Geophysical Research Insitute , India. I am looking for living accomodation during my 5 day stay at Philadelphia. I was wondering if any of you will be able to provide me with free accomodation or would be aware of any other resources who may be able to help me in this regard. Due to the limited fiancial resources avaliable to me from my lab, I am looking forward to help from my fellow researchers. Hope you can help me in this regard. I am eagerly looking forward to contributing to the M&M 2000 conference.
With kind regards Mr. R. Natarajan Scientist, National Geophysical Research Institute, Hyderabad, India e-mail: rnataraj-at-hd2.dot.net.in Phone: 91-40-7170141 X 2430 (W) Fax: 91-40-7170564
Dear Fellow Microscopists - Someone in the lab in which I work wants to do SEM of yeast colonies. I was able to locate some references and a general protocol, which seems rather straight forward. However, the authors two of the papers refer to an "Edwards Pearce Tissue Drier, EPD3". I contacted Edwards to find out what this tissue drier is, however they said they could not give me any info because it is no longer manufactured. I am thinking that this tissue drier is simply a freeze drier, but I am not sure. Does anyone out there have any insight into what this thing is or what it does? Thanks for your help!!
Tony
Tony Kowal Research Assistant for Dr. Susan Lindquist
Howard Hughes Medical Institute The University of Chicago 5841 S. Maryland MC 1028, Room N339 Chicago, IL 60637
A student has asked me if I can find a way to tell whether small plant seeds have been charred or not. She is examining seeds from an anthropological dig on a site probably inhabited both in historic times (170 years ago) and prehistoric (1000+ years ago). It seems that for seeds to date from the earlier inhabitation, they would need to be charred, which apparently preserves them.
I looked at some (uncoated) in the SEM, along with some 'fresh' seeds of same species as controls. The seeds of unknown age showed a lot more charging than the fresh ones. Could this be evidence that they are uncharred (i.e., would charring make them more conductive/better secondary electron producers?) Does anyone know of a method of differentiating between charred & uncharred seeds?
Thank you thank you thank you :0)
Paul Grover Chief Microscopist and Bottle Washer Microvista Laboratory Lafayette, IN
The Edwards Pearce Tissue drier was a freeze drier with a small peltier cooled specimen stage. The peltier device was a water- cooled 3-stage stack, achieving about -60oC on a stage about the size of a 35mm negative. There was provision for a small tray of phosphorus pentoxide, and the specimen chamber was a simple Pyrex glass bell, pumped by a rotary pump.
Date sent: Sat, 27 May 2000 12:38:36 -0500 To: Microscopy-at-sparc5.microscopy.com } From: Tony Kowal {askowal-at-midway.uchicago.edu}
How about using the low angle cleaving method of Rafferty? Polish with the SiC close to say the {110} planes (e.g. ten degrees) and then cleave along both the scratches and the {110} plane to get a thin acute wedge. It will take some practice, but should work generally with most Perovskites.
references: 1/Thin Solid Films Vol308-309 (1997) pp399-405 S.D.Walck & J.P.McCaffrey: The small angle cleavage technique applied to coatings and thin films
2/Thin Solid Films Vol304 (1997) pp157-159 Suli Suder, C.A.Faunce & S.E.Donelly: Thin solid film preparation by a small-angle cleavage for transmission electron microscopy
-I hope thses can provide some help. By the way this is not a definitive list, but I am sure Scott Walck can give you a far greater insight to this method.
Regards, Jon
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 Phone: Microscope room +46 18 471 6365 http://www.angstrom.uu.se/analytical/home.html ********************************************************
Chuck CPD certainly did become the most popular method by a country mile for SEM specimen drying, but freeze-drying still has advantages over it in some situations. These include, for example, specimens where lipid content or lipid structures must be retained (e.g. plant and insect epicuticles), where the specimen is an aggregate of objects loosely bound by a fluid or a mucilaginous matrix (e.g. it could be an advantage in Tony Kowal's yeast and bacterial colonies, soils and clays), or where the specimen is mechanically fragile and the components would be dispersed on submersion in baths of liquid during fixation and solvent drying and CPD (soils & clays, fungal sporangia, yeast and bacterial colonies).
The down side of freeze-drying in most of these contexts is that some shrinkage and distortion almost always results. Consequently, for almost all the situations listed above, and a host of others as well, Low-temperature SEM became the method of choice. In LTSEM the specimen can be viewed fully frozen- hydrated, but most commercially-available LTSEM systems have specimen temperature control, and full or partial freeze-drying can be undertaken either on the SEM specimen stage or in the cryo- preparation unit if required.
Anyone seeking a freeze-drier unit for EM specimens should contact Emitech who make a peltier-cooled unit (K750) which operates around -60oC (and is not unlike the Edwards-Pearce tissue drier) and a turbo molecular pumped Liquid nitrogen cooled low-temperature freeze drier (K775) which operates {-80oC.
http://www.Emitech.co.uk/
I have no financial interest in this company
Chris
} Hi Chris, } } Am I not remembering correctly, or was this earlier approach sort of a } precursor to the critical point drying technique? I mean, did not people use } this earlier technique because that was all they knew and then when the } option of CPD came along, everyone switched over to it? } } I myself was reluctant to say that because that was a very long time ago and } the memory does dull at the edges. } } Chuck } -------- REPLY, Original message follows -------- } } } Date: Sunday, 28-May-00 08:57 PM } } } } From: Chris Jeffree \ Internet: (cjeffree-at-srv0.bio.ed.ac.uk) } } To: Tony Kowal \ Internet: (askowal-at-midway.uchicago.edu } ) } } cc: MICROSCOPY BB \ Internet: } } (microscopy-at-sparc5.microscopy.com) } } } } Subject: Re: Tissue Drier } } } } ------------------------------------------------------------------------ } } 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 Edwards Pearce Tissue drier was a freeze drier with a small peltier } cooled } } specimen stage. The peltier device was a water- cooled 3-stage stack, } achieving } } about -60oC on a stage about the size of a 35mm negative. There was } provision } } for a small tray of phosphorus pentoxide, and the specimen chamber was a } } simple Pyrex glass bell, pumped by a rotary pump. } } } } Date sent: Sat, 27 May 2000 12:38:36 -0500 } } To: Microscopy-at-sparc5.microscopy.com } } } From: Tony Kowal {askowal-at-midway.uchicago.edu} } } Subject: Tissue Drier } } } } } ------------------------------------------------------------------------ } } } 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 - } } } Someone in the lab in which I work wants to do SEM of yeast colonies. I } } } was able to locate some references and a general protocol, which seems } } } rather straight forward. However, the authors two of the papers refer } to } } } an "Edwards Pearce Tissue Drier, EPD3". I contacted Edwards to find out } } } what this tissue drier is, however they said they could not give me any } } } info because it is no longer manufactured. } } } I am thinking that this tissue drier is simply a freeze drier, but I am } not } } } sure. Does anyone out there have any insight into what this thing is or } } } what it does? Thanks for your help!! } } } } } } Tony } } } } } } } } } Tony Kowal } } } Research Assistant } } } for } } } Dr. Susan Lindquist } } } } } } Howard Hughes Medical Institute } } } The University of Chicago } } } 5841 S. Maryland } } } MC 1028, Room N339 } } } Chicago, IL 60637 } } } } } } Phone: (773) 702-8795 } } } Fax: (773)702-7254 } } } e-mail: askowal-at-midway.uchicago.edu } } } Pager: on campus - 188 - 9668 (YNOT) } } } off campus - (773)753-1880 - 9668 } } } } } } } } } } } } } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } Dr Chris Jeffree } } University of Edinburgh } } Biological Sciences EM Facility } } Daniel Rutherford Building } } King's Buildings EDINBURGH EH9 3JH } } Tel: +44 (0) 131 650 5345 } } FAX: +44 (0) 131 650 6563 } } } } Inveresk Cottage, 26 Carberry Road, } } Inveresk, Musselburgh, Midlothian EH21 8PR, UK } } Tel. +44 (0) 131 665 6062 / Mobile 0410 585 401 } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } } -------- REPLY, End of original message -------- } }
===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
When using published methodology, some things matter, others do not. How to know which and what? I don't know, other then a good understanding of the field. Anyway, I have been amused over time with researchers insisting on outdated or cumbersome methods, because "it was published". In this particular case I would like to note that the tissue dryer is a freeze dryer designed for microscopy samples. Several other such instruments would do equally well and I expect that rather more researchers have prepared yeast by the critical point method. Freeze drying or critical point drying both work well for numerous samples. There will be a few specimens that are better prepared by one means or the other, but I wonder how frequently the claim "this gives better preservation" should have the disclaimer "in my hands" added.
Point is that you want to dry the yeast for SEM and you may be wasting your time chasing a particular instrument, when another, perhaps already in the department would do equally well. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Sunday, May 28, 2000 3:39 AM, Tony Kowal [SMTP:askowal-at-midway.uchicago.edu] 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 - } Someone in the lab in which I work wants to do SEM of yeast colonies. I } was able to locate some references and a general protocol, which seems } rather straight forward. However, the authors two of the papers refer to } an "Edwards Pearce Tissue Drier, EPD3". I contacted Edwards to find out } what this tissue drier is, however they said they could not give me any } info because it is no longer manufactured. } I am thinking that this tissue drier is simply a freeze drier, but I am not } sure. Does anyone out there have any insight into what this thing is or } what it does? Thanks for your help!! } } Tony } } } Tony Kowal } Research Assistant } for } Dr. Susan Lindquist } } Howard Hughes Medical Institute } The University of Chicago } 5841 S. Maryland } MC 1028, Room N339 } Chicago, IL 60637 } } Phone: (773) 702-8795 } Fax: (773)702-7254 } e-mail: askowal-at-midway.uchicago.edu } Pager: on campus - 188 - 9668 (YNOT) } off campus - (773)753-1880 - 9668 } }
Has she looked at them with conventional light microscopy, either stereo or compound? Following Oxam's Razor, this approach seems to be the simplest and should be tried first.
Best regards Barbara Foster,President Microscopy/Microscopy Education 125 Paridon Street Suite 102 Springfield, MA 01118-2130 PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com Website: www.MME-Microscopy.com/education
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At 04:19 PM 5/27/00 EDT, Pbgrover-at-aol.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It's been awhile since I was involved in this kind of research (i.e., graduate work in palaeoethnobotany, but I do not recall that seeds need to be "charred", per se, to be preserved over large periods of time. I believe that seeds and other plant remains can also become carbonized without fire, as a result of oxidation, etc., in the depositional environment.
To detect charring in the SEM might be a real problem, especially if trying to differentiate it from carbonization by means other than fire. I suppose that "charred" seeds might show physical damage more than other seeds, but even that might not always be true. The charging effects you observed, on the other hand, might possibly be the effects of mineralization of the seeds as the original seed components become replaced with non-conductive soil constituents over time. If so, this might indicate that these seeds are indeed quite old.
I guess if I was faced with this problem as an archaeologist I would try to identify other indications of fire in the context in which the seeds were found, such as wood charcoal, hearths, etc. On the EM side, if I had access to WDS, I might try comparing amounts of carbon in the old seeds, versus fresh ones.
Finally, the simplest thing to try might just be to toast some fresh seeds and compare them to non-toasted ones of the same type. Try a couple different charring methods, like throwing some in a campfire and collecting them later, and toasting them on a frying pan. A reference you might look at is "Palaeoethnobotany", a book by Dr. Deborah Pearsall published by Academic Press. I think it just came out in a new edition.
Good luck.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: "Pbgrover-at-aol.com"-at-sparc5.microscopy.com [mailto:"Pbgrover-at-aol.com"-at-sparc5.microscopy.com] Sent: Saturday, May 27, 2000 3:19 PM To: Microscopy-at-sparc5.microscopy.com
Esteemed colleagues,
A student has asked me if I can find a way to tell whether small plant seeds
have been charred or not. She is examining seeds from an anthropological dig on a site probably inhabited both in historic times (170 years ago) and prehistoric (1000+ years ago). It seems that for seeds to date from the earlier inhabitation, they would need to be charred, which apparently preserves them.
I looked at some (uncoated) in the SEM, along with some 'fresh' seeds of same species as controls. The seeds of unknown age showed a lot more charging than the fresh ones. Could this be evidence that they are uncharred (i.e., would charring make them more conductive/better secondary electron producers?) Does anyone know of a method of differentiating between charred
& uncharred seeds?
Thank you thank you thank you :0)
Paul Grover Chief Microscopist and Bottle Washer Microvista Laboratory Lafayette, IN
The continuing discussion on whether film is better than digital, and whether we can directly compare their resolving ability is very interesting and throwing up some really useful technical stuff. But aren't we missing the point a bit? Surely the image quality is user defined - if you or your customer, are satisfied with the end product then the equipment has done its job. How often does one need to examine a micrograph to assess its limits? If are getting that close to a picture then you may be taking things out of context a bit and losing the whole concept.
The future is with digital image capture, let's hope the price of the equipment comes down to something more attainable!
Pete
Peter Bond Plymouth Electron Microscopy Unit University of Plymouth Drake Circus Plymouth Devon UK PL4 8AA Tel/Fax: 01752 233092 email: pbond-at-plymouth.ac.uk
A discussion is developing from my comment that a de-saturated W hairpin source provides a better BSE image. This "problem" first came to light when a South African client commented that they have far better atomic number contrast images if they run on "first peak" rather than gun saturation.
I based my explanation on some work by C-R Peters, he relates BSE and SE signals to probe size. The bigger the probe the bigger the reaction volume that is the source of the BSE contribution to an image.
In the de-saturated state a normal W hairpin system has a very much bigger source which reflects in a very much larger probe being placed on the specimen than would be obtained under a "normal saturation" situation and therefore provides the operator with more BSE.
Some instruments do give a higher signal at their false peak than at what we would know as saturation. I have seen this on Cambridge (Leica, Leo) instruments and no matter how hard you try to "correct" what you feel is a mis alignment you just do not win. On some occasions I have also seen this on a Philips but it is much more rare.
I put this down to gun design as you do not see this in a Japanese designed instrument. In their case a higher false peak indicates a definite mis alignment!
I base my comments on running courses with a different SEM in a different laboratory almost every week, not on what happens with one laboratory on one instrument.
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
Laboratories who have to do pathology work and research also, have particular problems in that they are faced with a wide variety of specimens not all of which are suitable for a single epoxy formulation mixture during embedment.
Many pathology laboratories in the US use the medium hard formulation by Luft. This is indeed a good, nearly all purpose formulation, however, in laboratories that have to use glass knives for thick sectioning the NMA contained in this formulation deterioates the edge of the glass knife very quickly. If glass knives have to be used for thin sectioning, then this formulation cannot be used at all. Otherwise with diamond knives it cuts very well, assuming of course, that the embedding has been done well. The original formulation by Luft required two mixtures to be made up seperately. Years ago I combined this into a single mixture which can be easily frozen (without accelerator!) and kept for months at -80 or -20.
Eponate 12 148 ml DDSA 100 ml NMA 76 ml
This makes 324 ml. Accelerate it with 1.5% DMP-30 before use. If you are interested in this formulation, please print it out now, because this is the last time I will address it.
Sometimes laboratories are faced with processing floating cells. These ar e best enrobed in agarose, hopefully after osmication. In order to avoid enormous cell loss during spinning down repeatedly into embedding medium, it is fortuitous to draw off all buffer after osmium, and spin the cells into a mixture of 3% Ultra-low temperature agarose, Type IX (Sigma). This agarose type gells at 15 deg C, and it has the property of not being so dense and fibrous that embedding medium cannot penetrate it. After the gel is cooled (in the refrigerator, or on ice), the blocks are cut of desired size. It is important to note that one is no longer dealing with cells, but actual blocks and the protocol for dehydration and embedding must fit the requirements of the block size. The resulting blocks also are very well embedded in the above formulation.
The above formulation is good for skin and muscle, but the protocol must be adjusted to allow adequate penetration. We do 2 hours for dehydration, use propylene oxide for an intermediate, and start infiltrating with mixtures of PO and Epoxy, 2:1, 1:1, 1:3, pure, pure, pure, etc. When the tissue goes into the newly accelerated pure mixtures, I put an ordinary 60W bulb in the vicinity so that the mixture can heat to about 37 deg. At this point the formulation becomes very liquid and infiltration is enhanced. After 1.5 hours the lamp is turned off as minor polymerization begins about 40 deg C, which is undesirable during infiltration.
A problem arises when laboratories have to produce a large number of thick sections with glass knives, or they have to also use glass knives for thin sectioning. Then the above formulation cannot be used. A new embedding medium containing no NMA is needed. Many years ago, Mollenhauer invented a "Mixed Embedding" system which contained Araldite 502, Epon, DDSA, and dibutyl pthalate. This is an extremely useful system - a joy to section with glass knives. The downside is that it is more viscous than media without Araldite, and it takes longer to infiltrate. Here is the formulation, again reconfigured by me years ago into a single mixture.
Araldite 502 30ml Eponate 12 50 ml DDSA 55 ml Dibutyl (EMS) 0.75% (mix in with the 3 items above DMP-30 1.5% (add at time of use)
Freeze mixture without the accelerator. Will keep many months at -80. Can also keep at -20. For thick sectioning I polymerize just 24 hours. If blocks are to be thin sectioned, then I put them back into the oven for another 24 or 48 hours. This formulation has the capability of being soft and stiff, as you wish. If only thick sections are to be done, use 1% dibutyl. If blocks are too soft for your liking, just put them into a 60 deg C oven for several days, or use 95 deg C for an hour or two. One time I forgot some blocks in the 95 deg oven for a week. They still sectioned well.
Again, anyone interested please print this out now. I will not be answering these questions again, nor write these formulations again.
This is fun! Keep accurate records of what you do, particularly the infiltration and dehydration times. Finally you will have every block a joy to handle. No frustrations!
Bye, Hildy Crowley Sr. Electron Microscopy Specialist University of Denver Denver, CO
P.S. It is assumed that all processing is done with vials in constant motion! Once infiltration starts, vials must be on a rotator, not on a rocker or a shaker, as the monomers of the formulations will seperate and poor embedding will result.
} The continuing discussion on whether film is better than digital, and } whether we can directly compare their resolving ability is very interesting } and throwing up some really useful technical stuff. But aren't we missing } the point a bit? Surely the image quality is user defined - if you or your } customer, are satisfied with the end product then the equipment has done } its job. How often does one need to examine a micrograph to assess its } limits? If are getting that close to a picture then you may be taking } things out of context a bit and losing the whole concept. } } The future is with digital image capture, let's hope the price of the } equipment comes down to something more attainable! }
Dear Pete, If one is doing quantitative image processing, the better resolution available from film can be relevant. For example, if one wants to do corellation averaging, one needs as many objects in the picture as possible, and also as good resolution as needed--e.g., for 1 nm resolution of the reconstruction, a pixel size of 0.25 nm times the magnification is necessary, so the recording medium must have resolution equal to or better than that. The pixel size of the scanner must also be this small--obviously irrelevant for digital recording. A 5 micrometer scanning-pixel size at 20kx mag gives a pixel resolution of 0.25 nm, and for 6.5 by 9 cm film, one will have about 12,000 by 16,000 (taking the header into account) pixels of useful information. This will give a broader area than that available at equal resolution from any CCD chip now out there. I can assure you that I have often had to determine the information limits in a particular micrograph. Yours, Bill Tivol
Has anyone ever cured Epon-Araldite with UV? I was given a cobbled-together protocol by a non-microscopist post-doc; the protocol calls for a dry-ice, UV cure of Epon-Araldite. It says it can take from 3-7 days. I'm wondering if this is either a resin mix-up *or* the resin just set up all by itself (over that time frame I've seen it happen), just because it was a thin layer and left alone - nothing to do with the UV exposure. She has no ref. for this technique and isn't sure now where she got it. Sigh. I've only been able to find heat-cure protocols...anyone have any leads or thoughts on this UV thing?
(Sorry about the cross-post for those of you on both of these servers)
Thanks!
Tamara (Planning to use the oven.....) Howard CSHL
} ... } } I based my explanation on some work by C-R Peters, he } relates BSE and SE signals to probe size. } The bigger the probe the bigger the reaction volume } that is the source of the BSE contribution to an image. } } In the de-saturated state a normal W hairpin system has a } very much bigger source which reflects in a very much } larger probe being placed on the specimen than would be } obtained under a "normal saturation" situation and } therefore provides the operator with more BSE.
I have seen the 1st peak provide more beam current, but I fail to see how a larger probe diameter at similar beam currents can provide better z contrast. Do you know what the beam current is ... "1st peak" vs "saturated"??
I might also suggest the 1st peak may provide emission from several areas of the filament instead of just from the tip. This would manifest as "ghost" images or double images ... leastwise, I hope this type of phenomenon couldn't be confused with "better z contrast" :o)
Does anyone know of anybody who sells motors with suitable mechanical interfaces to drive X and Y of a JEOL 840A stage?
Replies from suppliers very welcome.
thanks
rtch
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
This is my last posting, I promise, looking for a Probe Current Detector (PCD) for a JEOL 840. The pneumatically-powered Faraday Cup which shoots into the beam just below the objective aperture, samples the beam, and retracts.
Does anyone have one which is redundant or otherwise spare?
Alternatively, does anyone know of a third-party manufacturer of this sort of thing?
cheers
rtch
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
At 12:17 PM 5/30/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The best CCD imagers are typically 6u in size. Complicating this is that they may be square or rectangular. Fuji's new hex-shaped pixels may be a major improvement. We'll see.
Our civil engineering department is trying to look at the bonding between bitumen and aggregate with the aid of a SEM. However, we do not have ESEM facilities (as I suggested). Now I was ask to see if anyone in the EM fraternity has any experience in using standard SEMs for examining bitumen containing specimens (I have not!!; and I don't want to ruin my microscopes).The supplied samples have been cut into slices of approximately 4mm thickness.
Any ideas out there? Thanks
Hans Brinkies Senior Lecturer Swinburne, University of Technology School of Engineering and Science P.O.Box 218 - Hawthorn - Vic -3122 - Australia Phone: +61 3 9214 8657 Fax: +61 3 9214 8264 Email: Hbrinkies-at-swin.edu.au
Peter Bond is right in saying "The future is with digital image capture" if we were at some future date to count heads of digital versus film TEM users. That change will be for reasons of convenience and to save labour. These are powerful and valid reasons.
Bill Tivol has given one set of applications where digital currently does not always measure up to film. Here are another couple of such applications. 1 When great enlargements are required film is superior. This is because of film's greater resolution, but more importantly, because its much, much easier to take images at moderate powers and highly enlarge. That way we take advantage of the TEM's greater depths of focus at low powers and of film's higher resolution/ image detail. 2 Whenever a TEM image is taken at low brightness (to avoid beam damage or at very high powers or in dark-field) relatively few electrons form the image and make that image grainy. Film is very slow and requires then a longer exposure, thus boosting the quantity of electrons used and improving the image. Digital is much more sensitive and so the exposure must be shortened. As a result the best digital camera will record, quiet faithfully the grainy, unsharp image.
Many labs rarely or never use such applications and for them the reasons to change to digital now may be overwhelming. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Wednesday, May 31, 2000 5:18 AM, William Tivol [SMTP:tivol-at-wadsworth.org] 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 } -----------------------------------------------------------------------. } } } Peter Bond wrote: } } } The continuing discussion on whether film is better than digital, and } } whether we can directly compare their resolving ability is very interesting } } and throwing up some really useful technical stuff. But aren't we missing } } the point a bit? Surely the image quality is user defined - if you or your } } customer, are satisfied with the end product then the equipment has done } } its job. How often does one need to examine a micrograph to assess its } } limits? If are getting that close to a picture then you may be taking } } things out of context a bit and losing the whole concept. } } } } The future is with digital image capture, let's hope the price of the } } equipment comes down to something more attainable! } } } } Dear Pete, } If one is doing quantitative image processing, the better } resolution } available from film can be relevant. For example, if one wants to do } corellation averaging, one needs as many objects in the picture as possible, } and also as good resolution as needed--e.g., for 1 nm resolution of the } reconstruction, a pixel size of 0.25 nm times the magnification is necessary, } so the recording medium must have resolution equal to or better than that. } The pixel size of the scanner must also be this small--obviously irrelevant } for digital recording. A 5 micrometer scanning-pixel size at 20kx mag } gives a pixel resolution of 0.25 nm, and for 6.5 by 9 cm film, one will have } about 12,000 by 16,000 (taking the header into account) pixels of useful } information. This will give a broader area than that available at equal } resolution from any CCD chip now out there. I can assure you that I have } often had to determine the information limits in a particular micrograph. } Yours, } Bill Tivol } }
I have a question about the structure of 2H-SiC (x-ray card No.29-1130), which belong to No. 186 space group (P63mc). (a=0.3076 nm, c=0.5048 nm, alfa=beta=120 degrees, gama=90 degrees)
My question is as below:
(A) Based on the symmetry of space group No. 186, the coordinates of the atoms in a unit cell should be
Si 2a : (0,0,0), and (0,0,0.5)
C 2b : (2/3, 1/3, 0.125) and (1/3,2/3,0.625)
(B) However, acoording to the real stacking sequence, the coordinates of the Si and C atoms in a unit cell are
Si sites: (0, 0, 0) and (2/3, 1/3, 0.5) C sites: (2/3, 1/3, 0.125) and (0, 0, 0.625)
It seems that there are some conflict between the two sets of coordinates, could someone help me to solve such a problem.
} Does anyone know of anybody who sells motors with suitable mechanical } interfaces to drive X and Y of a JEOL 840A stage? } } Replies from suppliers very welcome.
Ritchie,
Deben UK Ltd, 11-15 High St, Stowmarket, Suffolk UK IP14 6QL make very nice motors and controllers for X, Y and Z control of SEM stages. You can also contact them at : info-at-deben.co.uk or via their webpage : http://www.deben.co.uk
Cheers,
David Vowles Electron Microscopy Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 1223 334325 Fax: +44 1223 334567 Email: djv23-at-msm.cam.ac.uk
Dear Gao, we have used simulation software for CBED for both ZnO and GaN which are isomorphic with 2H-SiC (same space group:P6_3mc). For our software we use the following atom locations (I will use Si & C instead):
Si positions: (1/3, 2/3, 0) and (2/3, 1/3, 1/2)
C positions: (1/3, 2/3, u) and (2/3, 1/3, u + 1/2)
The Si-C bond length is parameterized by the quantity u. For the ideal (perfect stacking) arrangement u=3/8. However I am reliably informed that for SiC the u parameter is a great mystery at the present. However using u=3/8 should prove to be a good starting point.
Please note that to place Si at (0,0,0) you will have to remove (1/3,2/3,0) from each coordinate above which gives the following locations:
Si: (0,0,0) and (1/3, 2/3, 1/2)
C: (0,0,u) and (1/3, 2/3, u + 1/2)
Does this make sense?
Regards, Jonathan ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 Phone: Microscope room +46 18 471 6365 http://www.angstrom.uu.se/analytical/home.html ********************************************************
Dear List, I'm looking for methods for measuring the roughness of a sheet of polyethylene. The film consists of grains (80 to 150 microns in diameter) which are loosely packed. Large area analysis is prefered since the surface is very irregular locally.
Hello everybody: I will like to do cryomicrotomy of PP wires. My question is : can anyone suggest what kind of epoxy is appropriate for cryomicrotomy? I used Epoxy Mount and it was chattered during cryo process. The manufacturer confirm to me that epoxy mount is not appropriate for cryomicrotomy. Can anynone help me?
HI Hildy, You are always a great source of information....thanks.
Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
2 Whenever a TEM image is taken at low brightness (to avoid beam damage or at very high powers or in dark-field) relatively few electrons form the image and make that image grainy. Film is very slow and requires then a longer exposure, thus boosting the quantity of electrons used and improving the image. Digital is much more sensitive and so the exposure must be shortened. As a result the best digital camera will record, quiet faithfully the grainy, unsharp image.
Dear Jim,
Your points are good; however, exposure times are not, by
themselves, relevant. An exception to this is if the damage depends
on the dose rate; i.e., if there are mechanisms to dissipate the absorbed
beam energy which limit damage at low illumination levels. The
image quality depends on how much information is carried by each elec-
tron (basically, the number of photons produced in the scintillator times
the quantum efficiency of the system for these photons for digital, and the
probability of a darkened film grain for film or image plates) times the
number of electrons. The damage--except for dose-rate-dependence--
is linear with the number of electrons, so it is the efficiency of the pro-
cess of obtaining the information from each electron which determines
the ultimate resolution limit. Also, the use of image averaging can make
one clear image from many grainy ones in the case that one has a large
Randy, Don't forget about other analytical techniques, such as XPS. Some well engineered surface analysis may reveal significant chemical differences between "charred", new, and aged or mineralized specimens. In addition to Randy's good advice on the microscopy, I would also utilize the power of both TEM and EELS in comparing morphology and carbon and oxygen chemistry. Electron energy loss spectroscopy combined with EDS is one of my favorites for carbon micro-analyses. Looking at both the low loss and the core loss structure of EELS spectra, there is tremendous detail and differentiation to be obtained. If TEM and "PEELS" are available, and you don't mind sacrificing a bit of the artifact, a series of control specimens and analysis by "AEM" (TEM, EDS, and PEELS) would be worth a shot. Sounds like fun, Good Luck, Brad Huggins BP Amoco, Analytical Naperville IL
} ---------- } From: Tindall, Randy D.[SMTP:TindallR-at-missouri.edu] } Sent: Tuesday, May 30, 2000 9:13 AM } To: '"Pbgrover-at-aol.com"-at-sparc5.microscopy.com' } Cc: 'microscopy-at-sparc5.microscopy.com' } Subject: RE: charred plant seeds } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Paul, } } It's been awhile since I was involved in this kind of research (i.e., } graduate work in palaeoethnobotany, but I do not recall that seeds need to } be "charred", per se, to be preserved over large periods of time. I } believe } that seeds and other plant remains can also become carbonized without } fire, } as a result of oxidation, etc., in the depositional environment. } } To detect charring in the SEM might be a real problem, especially if } trying } to differentiate it from carbonization by means other than fire. I } suppose } that "charred" seeds might show physical damage more than other seeds, but } even that might not always be true. The charging effects you observed, on } the other hand, might possibly be the effects of mineralization of the } seeds } as the original seed components become replaced with non-conductive soil } constituents over time. If so, this might indicate that these seeds are } indeed quite old. } } I guess if I was faced with this problem as an archaeologist I would try } to } identify other indications of fire in the context in which the seeds were } found, such as wood charcoal, hearths, etc. On the EM side, if I had } access } to WDS, I might try comparing amounts of carbon in the old seeds, versus } fresh ones. } } Finally, the simplest thing to try might just be to toast some fresh seeds } and compare them to non-toasted ones of the same type. Try a couple } different charring methods, like throwing some in a campfire and } collecting } them later, and toasting them on a frying pan. A reference you might look } at is "Palaeoethnobotany", a book by Dr. Deborah Pearsall published by } Academic Press. I think it just came out in a new edition. } } Good luck. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } } } -----Original Message----- } } From: "Pbgrover-at-aol.com"-at-sparc5.microscopy.com } [mailto:"Pbgrover-at-aol.com"-at-sparc5.microscopy.com] } Sent: Saturday, May 27, 2000 3:19 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: charred plant seeds } } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Esteemed colleagues, } } A student has asked me if I can find a way to tell whether small plant } seeds } } have been charred or not. She is examining seeds from an anthropological } dig } on a site probably inhabited both in historic times (170 years ago) and } prehistoric (1000+ years ago). It seems that for seeds to date from the } earlier inhabitation, they would need to be charred, which apparently } preserves them. } } I looked at some (uncoated) in the SEM, along with some 'fresh' seeds of } same } species as controls. The seeds of unknown age showed a lot more charging } } than the fresh ones. Could this be evidence that they are uncharred } (i.e., } would charring make them more conductive/better secondary electron } producers?) Does anyone know of a method of differentiating between } charred } } & uncharred seeds? } } Thank you thank you thank you :0) } } Paul Grover } Chief Microscopist and Bottle Washer } Microvista Laboratory } Lafayette, IN }
Could it be that moisture escaping from the fresh seeds somehow helps to dissipate the charge so that new seeds do not charge like the old ones?
I would also be interested in the x-ray spectra of the two seeds. Perhaps there is partial mineralization as Randy Tindall suggested. A light element detector should also reveal a difference in O/C ratio due to differences in moisture content, or perhaps due to charring.
Warren S.
At 09:13 AM 5/30/2000 -0500, you wrote: } Esteemed colleagues, } } A student has asked me if I can find a way to tell whether small plant seeds } } have been charred or not. She is examining seeds from an anthropological } dig } on a site probably inhabited both in historic times (170 years ago) and } prehistoric (1000+ years ago). It seems that for seeds to date from the } earlier inhabitation, they would need to be charred, which apparently } preserves them. } } I looked at some (uncoated) in the SEM, along with some 'fresh' seeds of } same } species as controls. The seeds of unknown age showed a lot more charging } than the fresh ones. Could this be evidence that they are uncharred (i.e., } would charring make them more conductive/better secondary electron } producers?) Does anyone know of a method of differentiating between charred } } & uncharred seeds? } } Thank you thank you thank you :0) } } Paul Grover } Chief Microscopist and Bottle Washer } Microvista Laboratory } Lafayette, IN
Of course there are situations where one or the other Technique may be better because of the limitations of film or digital imaging. And especially for a task like the one Bill talks about below, where field of view AND resolution are important, images taken on film may be superior to images from a digital camera. On the other hand, digital imaging may have something to offer in those cases as well:
Instead of taking one image and do the corellation averaging, why not have the computer do it? I could probably set up a system that acquires the image at a high enough resolution, extract all the necessary data, then move the stage to an adjacent area and continue the measurements there. This technique would even have an advantage over film: I can set the lower limits for the accuracy before taking the images and the system continues to measure until these limits are satisfied. This could be done without user intervention. So, instead of taking one image, scanning it in with a scanner, and then being "limited" by the field of view to a certain measurement accuracy, one could start the measurement with a predetermined accuracy, go drink a coffee, work on that paper, do the travel expense report, then go back to the microscope and collect the spreadsheet with the measurement data. Of course this requires the setup of a fairly complex system, but those are technical problems, not fundamental ones.
Just a thought ....
Michael
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: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: William Tivol [mailto:tivol-at-wadsworth.org] Sent: Tuesday, May 30, 2000 1:18 PM To: microscopy-at-sparc5.microscopy.com
Peter Bond wrote:
} The continuing discussion on whether film is better than digital, and } whether we can directly compare their resolving ability is very interesting } and throwing up some really useful technical stuff. But aren't we missing } the point a bit? Surely the image quality is user defined - if you or your } customer, are satisfied with the end product then the equipment has done } its job. How often does one need to examine a micrograph to assess its } limits? If are getting that close to a picture then you may be taking } things out of context a bit and losing the whole concept. } } The future is with digital image capture, let's hope the price of the } equipment comes down to something more attainable! }
Dear Pete, If one is doing quantitative image processing, the better resolution available from film can be relevant. For example, if one wants to do corellation averaging, one needs as many objects in the picture as possible, and also as good resolution as needed--e.g., for 1 nm resolution of the reconstruction, a pixel size of 0.25 nm times the magnification is necessary, so the recording medium must have resolution equal to or better than that. The pixel size of the scanner must also be this small--obviously irrelevant for digital recording. A 5 micrometer scanning-pixel size at 20kx mag gives a pixel resolution of 0.25 nm, and for 6.5 by 9 cm film, one will have about 12,000 by 16,000 (taking the header into account) pixels of useful information. This will give a broader area than that available at equal resolution from any CCD chip now out there. I can assure you that I have often had to determine the information limits in a particular micrograph. Yours, Bill Tivol
your remark 1) What you can do of course with digital imaging (provided you have the necessary hardware), is to automatically collect larger areas by taking several images that overlap, reducing the advantage that film has in this area and perhaps offer other possibilities as I just explained in another posting as a response to Bill Tivol's posting.
your remark 2) I am not sure you are not comparing apples and oranges. What you are saying is, that because of the "slowness" of film you need longer exposures, thereby averaging out the statistical noise of the electron, which is not the case for CCD cameras at short exposures, hence they appear more noisy. In essence what you are doing is to compare a short exposure image to a long exposure image. What you can do with a CCD camera is the following: You can get (perhaps) real-time dark field images and position your sample and/or decide if you want to actually take the image. Then you take a SERIES of images, let's say 10, each at an exposure time of 1/10 of the film exposure. This can be done automatically, of course. Finally, you add or average all of these images using a pattern recognition to align them first. The result: A dark field image that should be as noisy as the film image, with much less problems due to drift during long exposures, a higher dynamic range and visible immediately on the viewing screen.
Michael
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: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: jim [mailto:jim-at-proscitech.com.au] Sent: Tuesday, May 30, 2000 7:13 PM To: 'William Tivol' Cc: microscopy-at-sparc5.microscopy.com
Peter Bond is right in saying "The future is with digital image capture" if we were at some future date to count heads of digital versus film TEM users. That change will be for reasons of convenience and to save labour. These are powerful and valid reasons.
Bill Tivol has given one set of applications where digital currently does not always measure up to film. Here are another couple of such applications. 1 When great enlargements are required film is superior. This is because of film's greater resolution, but more importantly, because its much, much easier to take images at moderate powers and highly enlarge. That way we take advantage of the TEM's greater depths of focus at low powers and of film's higher resolution/ image detail. 2 Whenever a TEM image is taken at low brightness (to avoid beam damage or at very high powers or in dark-field) relatively few electrons form the image and make that image grainy. Film is very slow and requires then a longer exposure, thus boosting the quantity of electrons used and improving the image. Digital is much more sensitive and so the exposure must be shortened. As a result the best digital camera will record, quiet faithfully the grainy, unsharp image.
Many labs rarely or never use such applications and for them the reasons to change to digital now may be overwhelming. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Wednesday, May 31, 2000 5:18 AM, William Tivol [SMTP:tivol-at-wadsworth.org] 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 } -----------------------------------------------------------------------. } } } Peter Bond wrote: } } } The continuing discussion on whether film is better than digital, and } } whether we can directly compare their resolving ability is very interesting } } and throwing up some really useful technical stuff. But aren't we missing } } the point a bit? Surely the image quality is user defined - if you or your } } customer, are satisfied with the end product then the equipment has done } } its job. How often does one need to examine a micrograph to assess its } } limits? If are getting that close to a picture then you may be taking } } things out of context a bit and losing the whole concept. } } } } The future is with digital image capture, let's hope the price of the } } equipment comes down to something more attainable! } } } } Dear Pete, } If one is doing quantitative image processing, the better } resolution } available from film can be relevant. For example, if one wants to do } corellation averaging, one needs as many objects in the picture as possible, } and also as good resolution as needed--e.g., for 1 nm resolution of the } reconstruction, a pixel size of 0.25 nm times the magnification is necessary, } so the recording medium must have resolution equal to or better than that. } The pixel size of the scanner must also be this small--obviously irrelevant } for digital recording. A 5 micrometer scanning-pixel size at 20kx mag } gives a pixel resolution of 0.25 nm, and for 6.5 by 9 cm film, one will have } about 12,000 by 16,000 (taking the header into account) pixels of useful } information. This will give a broader area than that available at equal } resolution from any CCD chip now out there. I can assure you that I have } often had to determine the information limits in a particular micrograph. } Yours, } Bill Tivol } }
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} Instead of taking one image and do the corellation averaging, why not } have the computer do it? I could probably set up a system that acquires } the image at a high enough resolution, extract all the necessary data, } then move the stage to an adjacent area and continue the measurements } there. This technique would even have an advantage over film: I can set } the lower limits for the accuracy before taking the images and the } system continues to measure until these limits are satisfied. This could } be done without user intervention. So, instead of taking one image, } scanning it in with a scanner, and then being "limited" by the field of } view to a certain measurement accuracy, one could start the measurement } with a predetermined accuracy, go drink a coffee, work on that paper, do } the travel expense report, then go back to the microscope and collect } the spreadsheet with the measurement data. } Of course this requires the setup of a fairly complex system, but those } are technical problems, not fundamental ones. }
Dear Mike, That is an excellent suggestion. Of course, the beam should be
restricted to the area of the detector to avoid damaging the specimen except where inevitable. You are correct that setting the accuracy limits prior to taking the image avoids mixing the required data with those from excess exposure, when the specimen has undergone some radiation damage--this could perhaps be done with film, but it is simpler with electronic data collection. There one can even make sure that those pixels which are most important are the ones optimized, and, with a small enough beam, like that used in spot-scan imaging, one could expose different parts of the image for different times, so that as much as possible of the image is optimally collected. This also avoids damaging the area of the specimen not yet under investigation.
Anyone aware of software capable of generating depth profiles from digital SEM stereo pair images? I am aware of the Oxford ISIS system/program, but wondered if other companies offer similar software that would run 'stand-alone' in a PC and work with any digital files.
Any help or suggestions along these lines would be most appreciated.
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Greetings! } } Has anyone ever cured Epon-Araldite with UV? I was given a } cobbled-together protocol by a non-microscopist post-doc; the protocol } calls for a dry-ice, UV cure of Epon-Araldite. It says it can take from } 3-7 days. I'm wondering if this is either a resin mix-up *or* the resin } just set up all by itself (over that time frame I've seen it happen), just } because it was a thin layer and left alone - nothing to do with the UV } exposure. She has no ref. for this technique and isn't sure now where she } got it. Sigh. I've only been able to find heat-cure protocols...anyone } have any leads or thoughts on this UV thing? } } (Sorry about the cross-post for those of you on both of these servers) } } Thanks! } } Tamara (Planning to use the oven.....) Howard } CSHL } } } Hi,
Try it, but not on anything valuable! Resins set up by themselves at rt. I don't remember ever reading anything about UV in the Handbook of Epoxy Resins. Why do you want to do it? That is the question. What would be the advantage of that major fiddle?
When I worked for Zeiss, we did a lot of coal analysis using microspectrophotometry. I've also done some polarized light work on a sample or two for customers. Is the bonding you are looking for on the level where light microscopy might work? Just for your information, I also recently took a look at the "tie" layers between polymer films in ketsup bottles, perhaps a distant but related application. Polarized light and DIC did a great job in that instance.
.. just a thought, but hopefully helpful.
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At 12:13 PM 5/31/00 +1000, Hans Brinkies wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Have you considered structured lighting? A flat beam of light is illuminates the subject from a 45 degree angle. A camera is views it at 90 degrees. The height and size of the roughness can be reconstructed from the shadows the light makes.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 ----- Original Message ----- } From: "DAVID I SAXON" {DISAXON-at-prodigy.net} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, May 31, 2000 7:19 AM
In a message dated 5/31/00 2:52:24 PM, ferreira1-at-llnl.gov writes:
} Anyone aware of software capable of generating depth profiles from digital } SEM stereo pair images? I am aware of the Oxford ISIS system/program, } but wondered if other companies offer similar software that would run 'stand-alone' } in a PC and work with any digital files.
One of the (many) functions in Fovea Pro (http://members.aol.com/FoveaPro), a set of Photoshop-compatible plug-ins intended for the analysis of images including those from microscopes such as SEMs, is a routine that fuses stereo pair images to obtain the elevation of points on the surface. This can then be used to measure elevation profiles, or to reconstruct 3D surface images. Examples of both are included in the tutorial and are illustrated on the web site. It isn't stand-alone (you need Photoshop or a compatible program), but it will do what you ask.
The tissue drier you referred to had a "cold finger" which acted as a vapor trap to remove the water vapor produced from the sublimation of the specimen ice. This is a very important feature for achieving "distortion-free drying" which common type of freeze driers lack I believe. Check out Pearse's book Histochemistry Theoretical and Applied, vol.1. It contains an excellent chapter on this technique and others. However, I would try CPD, also.
Hank Adams Manager Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx 77030
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