Can anybody kindly provide me with the following info about "Carbon films on Metal grids" for HRTEM sample preparation. Thank you in advance.
1. How high temperature the carbon films can withstand in the reactors (e.g. oven) without damage? What negative effect can result from the high temperature on the C films?
2. We need to collect TEM samples directly from the high-temperature (around 1200 C) reactor onto Carbon-support films (or any other alternative) on some metal grids. Do you have any good suggestions?
Thank you very much in advance.
- juha
Electron Microscope Unit, Helsinki Univ. Tech.
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From MicroscopyL-request-at-ns.microscopy.com Thu Apr 1 09:15:15 2004
We have an archeologist here who routinely makes replicas of beads and other objects that can't be sputter coated. He uses 3m Express, vinyl polysiloxane impression material, light body, medium set, product number 7302HB and gets fine-scale impressions. Fine enough to determine the technology used the drill the hole, including if the drilling was done recently using an ancient method (commonly done to make "ancient beads" to sell to tourists). This material should do for your student's work. There have also been several articles on replicating surfaces run in Microscopy Today which might help.
Phil
} Hi: } } I need some advice so I can help a student in the lab. } } She is looking at sea shells to check for grinding marks. The shells were } used for beads and 'money' by ancient people. She wants to see if the } shells have tool marks, or were collected from the beach with holes made by } natural processes. } } She has some 'modern' shells she had prepared. These are expendable and } will be easy to cut and coat for SEM. } } She has some museum specimens that she cannot do much to, making it } difficult to do the SEM. } } I do not have access to an ESEM. I do have a Robinson BSED. I don't think } she can coat these samples. } } She might be able to make a replica of the museum samples using dental } impression material. Anybody tried this? What are the details? } } Any other ideas? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 1 10:06:03 2004
} I was supposed to receive TEM pictures of my silica powder (SiO2), but } that never happened. So, I am looking for a way to analyze the surface } and structure of my silica - the sooner, the better.
} Does Wellesley own a TEM machine or any other device that would give me } more insight into the surface structure of my powder?
We need to view silica powder (SiO2) surface structure with an electron microscope. We are in a biological facility and have no experience with this type of sample. What sort of instrument is best suited for viewing this sample? What are the sample preparation techniques? Are there labs that we can bring our sample to for, let's say, an hour of observation and a few pictures? We are in the Boston, Massachusetts area. Thank you in advance.
Vachik Hacopian
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 1 10:49:43 2004
I've actually done some backscatter SEM on uncoated shells, which worked pretty well. The resolution isn't as good as a coated specimen, of course, but would probably work for what your student needs to see. So if you have a backscatter detector on your SEM, I would try that. I can forward you some images that I have taken if you like. Let me know. Regards, Aaron Bell
} From: Gary Gaugler {gary-at-gaugler.com} } To: jmkrupp-at-cats.ucsc.edu (Jon Krupp) } CC: MSA listserver {Microscopy-at-MSA.Microscopy.Com} } Subject: [Microscopy] Re: SEM of shells } Date: Wed, 31 Mar 2004 15:07:41 -0800 } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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From MicroscopyL-request-at-ns.microscopy.com Thu Apr 1 12:47:18 2004
I would think that a Robinson BSE should give excellent image results in BSE mode. If you have the mixer option or can mix SE+BSE, that may allow optimization of the image. The problem, I would think, is that you are looking for morphology info whereas the BSE produces Z contrast info. Low KV (2-4KV) should produce excellent surface detail. At this low KV, you may avoid charging. If you mix the Robinson with SE, it may do what you need. I've done this with coral and got good results.
gary g.
At 09:01 AM 4/1/2004, you wrote:
} I've actually done some backscatter SEM on uncoated shells, which worked } pretty well. The resolution isn't as good as a coated specimen, of } course, but would probably work for what your student needs to see. So if } you have a backscatter detector on your SEM, I would try that. I can } forward you some images that I have taken if you like. Let me know. } Regards, } Aaron Bell } } } } } } At 02:54 PM 3/31/2004, you wrote: } } } } } } } Hi: } } } } } } I need some advice so I can help a student in the lab. } } } } } } She is looking at sea shells to check for grinding marks. The shells were } } } used for beads and 'money' by ancient people. She wants to see if the } } } shells have tool marks, or were collected from the beach with holes made by } } } natural processes. } } } } } } She has some 'modern' shells she had prepared. These are expendable and } } } will be easy to cut and coat for SEM. } } } } } } She has some museum specimens that she cannot do much to, making it } } } difficult to do the SEM. } } } } } } I do not have access to an ESEM. I do have a Robinson BSED. I don't think } } } she can coat these samples. } } } } } } She might be able to make a replica of the museum samples using dental } } } impression material. Anybody tried this? What are the details? } } } } } } Any other ideas? } } } } } } Thanks } } } } } } Jonathan Krupp } } } Microscopy & Imaging Lab } } } University of California } } } Santa Cruz, CA 95064 } } } (831) 459-2477 } } } jmkrupp-at-cats.ucsc.edu } } } } _________________________________________________________________ } Get tax tips, tools and access to IRS forms all in one place at MSN } Money! http://moneycentral.msn.com/tax/home.asp }
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 1 14:23:28 2004
} Can anybody kindly provide me with the following info } about "Carbon films on Metal grids" for HRTEM sample } preparation. Thank you in advance. } } 1. How high temperature the carbon films can withstand } in the reactors (e.g. oven) without damage? What } negative effect can result from the high temperature } on the C films? } } 2. We need to collect TEM samples directly from the } high-temperature (around 1200 C) reactor onto } Carbon-support films (or any other alternative) on } some metal grids. Do you have any good suggestions?
Dear Juha, For depositing samples at high temperature, you want the support film to have the same coefficient of expansion as the grid material, and the easiest way to do that is to make the film from the same material as the grid. You also want a low-z material that is strong, so that a thin film will not interfere with microscopy of the sample. If the pieces of the sample are larger than 2 um, you can use a holey film and examine the areas of the specimen that lie over the holes. Cu grids themselves will not work, since Cu melts at 1083 C. Ti grids will be useable at 1200 C, and either Ti or Ti-Si (88%Ti + 12% Si) should make a suitable film. A lower-z alternative is Be, but it is very toxic. Mo grids have a similar coefficient of expansion to C at LN2 temp, so these grids have been used with cryo specimens to eliminate crinkling, but I don't know what will happen to this combination at high temps. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 1 14:46:11 2004
Dear Juha, I think that the carbon film on your TEM grids would burn away if you put it in a furnace at 1200 degrees C, unless you could exclude all oxygen or oxidizing sources. If it is a carbon-coated formvar or collodion (plastic) film it will be even more heat-sensitive. You may find a SiO film is more oxidation-resistant. It evaporates much like carbon in an evaporator, but I have not tried to heat it. When we did heat-treat studies of an Al alloy by sandwiching the TEM foil in a Cu folding grid and putting it back and forth from the TEM to the furnace, we found the Cu grid oxidized too much, and so we switched to Ni folding grids, and they worked fine. Can you sample the outlet of the reactor, where the gas has cooled a bit? Regards, 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-interchange.ubc.ca ----- Original Message ----- } From: "Juha Dapper" {juha_dapper-at-yahoo.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, April 01, 2004 7:19 AM
A not unknown fenomena: in our lab there are too some people who have problems with it, although not that strong, when moving away from the microscope for a short time (a minute or so), they feel already better. And I must admit, I also suffer a very little bit from it, and (but this is feed for psychologists I think), more when working on a Leica then on a Zeiss... No offence Leica! ;-) And more on high then on low magnification... A solution:, easy in a way, but possibly not immediate applicable: project the image on a screen. You also have those small screens attached on an ocular which project the image of the ocular onto this 'personal', small screen, so a camera is not needed. Also, move away often and let the eyes relax. Good luck,
Sven Terclavers
-----Original Message----- } From: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Friday, March 05, 2004 16:17 PM To: Microscopy-at-MSA.Microscopy.com
My undergrad histology class has given me a new and novel problem. One of the students is experiencing severe motion sickness while she views specimens on a standard binocular compound microscope. I have had students bothered when viewing specimens on the 6-headed scope and someone else is moving the specimen field but this is a first. Any one have an easy suggestion?
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pwebster-at-hei.org) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, April 1, 2004 at 11:12:05 ---------------------------------------------------------------------------
Question: I have been watching this disccusions with some interest. However, the replies have confused me. It seems that high resolution sputter coating requires a clean, high vacuum - yet most sputter coaters use an intert gas to create an inonizable environment and operate at a pre-set, low vacuum. What is the need for high vacuum?
There also seems to be a need for magnetron cooling, but other messages suggest that magnetrons do not warm up when being used. Which is correct?
} From the literature on metal coating of freeze-fractured specimens, it is clear that the best (fine grain) coats are obtained when the specimen (not the source) is cooled. From what I understand of this literature it seems that warm metal grains can actually move around on the specimen before settling in place. It is during this migration that they aggregate to form the grains we see. If specimen cooling is important, why don't any of the commercial sputter coaters cool the specimens?
No-one has mentioned the possibility of using non-coating methods that employ multiple exposures to osmium tetroxide so that metal is placed in the specimen and not over it. Are there so many disadvantages to this approach for it to be disregarded so easily?
} From my experience in coating very complex specimens it is clear that the metal coat really has no chance of covering large parts of the specimen, yet even small coverings of platinum over only small areas seem to improve the ability of the SEM to image the specimen. Given this proactical observation, do we really need to make sure the specimen is properly grounded before examining it?
Regards,
Paul Webster.
Hosue Ear Institute 2100 W 3rd St. Los Angeles, CA 90057.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (basgen-at-umn.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, April 1, 2004 at 11:44:59 ---------------------------------------------------------------------------
Question: The 8th American Stereology Course sponsored by the International Society for Stereology will be held September 26-October 1, 2004 in Minneapolis, Minnesota, USA.
The course is an intense 6-day learning experience, which includes lectures and hands on exercises each day. In the evenings, participants have the opportunity to discuss their individual projects with the instructors. Topics covered include: 1) The measurement of volumes, surfaces (area), lengths, number, and connectivity in 3-dimensional space using 2-dimensional images; 2) The measurement of these structural parameters, in homogeneous and non-homogeneous tissues; 3) The design of efficient, unbiased sampling strategies; 4) How to determine the optimal number of animals per experimental group and the optimal number of measurements per animal.
Instructors: Hans J¯rgen G. Gundersen, Stereological Research Laboratory, University of Aarhus, Aarhus, Denmark Jens R. Nyengaard, Stereological and Electron Microscopy Laboratory, University of Aarhus, Aarhus, Denmark, Bente Pakkenberg, Research Laboratory for Stereology and Neuroscience, Bispebjerg University Hospital, Copenhagen, Denmark, Karl-Anton Dorph-Petersen, Department of Neuroscience, University of Pittsburgh School of Medicine, Pittsburgh, PA
The course is intended for persons interested in obtaining morphometric data from biological specimens using unbiased and efficient sampling designs. No prior stereological experience is necessary. The course will be limited to 25 participants.
Osmium tetroxide has been discussed and is available for coating specimens. It is just too costly to be viable...IMO. SPI offers this unit. Check it out.
What is interesting is to actually define what "high resolution" really means. Let's assume that it is fine grain...whatever that means. Au is definitely out in this regard. I have had better fine grain results with Au/Pd and Pt. The key is to keep a high vacuum ( {=100mT) and low current ( {=20mA). Keep these constant and use time as the final determinator of end point thickness.
Modern (and most older) coaters use magnetron heads and do not require water cooling. They work very well. The differences are in how they allow different variables to be introduced (time, pressure, current) and the thickness of the original target material.
At {= 75KX, Au is OK. At or above that, Au/Pd is better. Ultimately, Pt is best for non-complex sputter coating. It is fine grain and is not seen below 200KX. Au is definitely seen as a honeycomb web. Au/Pd is more difficult to image.
As far as grains moving, they probably do. However, the issue is "how much?" I do not see them move at all at normal temperatures. At elevated temperatures, they do. but this is a separate and more complex issue.
If the coating is done right, the whole specimen will be conductive and will not charge. But one must keep in mind the 3-D nature of specimens. i.e., if a high dimension specimen is coated without rotating and/or tilting, it may not have a conductive path to the specimen stub. Depending on what you are trying to see, some coloidal silver dag will help form a conductive path from the specimen to the stub and to ground. This is the way it is for large and irregular specimens.
gary g.
At 04:47 PM 4/1/2004, you wrote:
} Email: pwebster-at-hei.org } Name: Paul Webster } } Organization: House Ear Institute } } Title-Subject: [Microscopy] [Filtered] MListserver: HR Sputter Coater } } Question: I have been watching this disccusions with some interest. } However, the replies have confused me. It seems that high resolution } sputter coating requires a clean, high vacuum - yet most sputter coaters } use an intert gas to create an inonizable environment and operate at a } pre-set, low vacuum. What is the need for high vacuum? } } There also seems to be a need for magnetron cooling, but other messages } suggest that magnetrons do not warm up when being used. Which is correct? } } } From the literature on metal coating of freeze-fractured specimens, it } is clear that the best (fine grain) coats are obtained when the specimen } (not the source) is cooled. From what I understand of this literature it } seems that warm metal grains can actually move around on the specimen } before settling in place. It is during this migration that they aggregate } to form the grains we see. If specimen cooling is important, why don't } any of the commercial sputter coaters cool the specimens? } } No-one has mentioned the possibility of using non-coating methods that } employ multiple exposures to osmium tetroxide so that metal is placed in } the specimen and not over it. Are there so many disadvantages to this } approach for it to be disregarded so easily? } } } From my experience in coating very complex specimens it is clear that } the metal coat really has no chance of covering large parts of the } specimen, yet even small coverings of platinum over only small areas seem } to improve the ability of the SEM to image the specimen. Given this } proactical observation, do we really need to make sure the specimen is } properly grounded before examining it? } } Regards, } } Paul Webster. } } Hosue Ear Institute } 2100 W 3rd St. } Los Angeles, CA 90057. } } } } } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 2 01:19:40 2004
Why high vaccum before sputtering ? To remove most of the water vapor from the vaccum, before introducing the (dry) inert gas. One reason for grain formation, typicaly with gold, is the formation of gold clusters arround water molecules from the residual vaccum. For the same reason one will prefere high (dry) gas flow, to diluate and evacuate the water desorbing from the walls of the chamber. It-s the same goal when one purge the chamber with argon, before sputtering, on coater which have only a roughing pump. The effet is only not suffisent to avoid grain formation.
Cooling the head or not ? Depends on the sputter time and current. For 30" at 20 mA, no matter in cooling. For 5' at 80 mA, depending how the head is built, but I would cool it... On production magnetrons, the head is always cooled.
A last word about the grain formation . We are always comparing coating materials, without any consideration of the substrate. In a good clean vaccum, saying UHV (!), the grain or film formation depends on the pair substrate/adsorbate. Of coarse, on a clean substrate, without water and organics on it, what's not the case with SEM samples ! An for example, on glass or in general on oxydes, gold will give in most cases grains. Ir not. But gold on mica or MoS2 gives a layer. So there are general rules and particuar cases ! In a good sputter coater, Ir, Pt or Cr (apart the oxydation pb) seems to give acpetable results for HR SEM, what not the case with Au. The goal is not to build a conducting film, but to have an minimum, homogeneous and stable charging of the sample. Carbon and low voltage work too.
Hope it helps;
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (staylor-at-fit.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, April 1, 2004 at 11:02:08 ---------------------------------------------------------------------------
Email: staylor-at-fit.edu Name: Scott Taylor
Organization: Florida Institute of Technology
Education: Graduate College
Location: Melbourne, FL 32901
Question: We are considering purchasing a used Sorvall MT-5000 ultramicrotome, and would like some information about this particular model. It appears that the older MT-2 models are more widely used, and we would like to know if the MT-5000 is an instrument of as high as, or better quality than the MT-1 and 2 models. Can you also suggest one or more companies that can service either of these pieces of equipment in the state of Florida? Thank you very much for your assistance.
Has anybody tried direct scanning of glass microscope slides using one of the Nikon Coolscan series scanners? I would appreciate any comments or experiences. Feel free to contact me directly.
Thank you!
George
George Laing National Graphic Supply ph 800.223.7130 x3109 f 800.832.2205 email scisales-at-ngscorp.com
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 2 12:57:45 2004
I worked in the area of precipitated silica powders and other submicron particles and aggregates for many years. My TEM lab was closed down in an ongoing restructuring and I am currently available for free consulting while I find a new job in electron microscopy somewhere in the USA.
I probably ran 4,000+ samples of silica in the past. I assume you are not talking about quartz particles but a higher tech material like a manufactured silica. First of all, what is the primary particle size of the silica that you are trying to view or disperse? Is it a precipitated silica, a colloidal silica, a clustered nanoparticle powder, a CMP silica, or what? Is it agglomerated or aggregated? If it's aggregated or agglomerated, it has pores and intergrowth can come into play with the surface area and structural interpretation from supporting techniques outside microscopy. Is the sample hydrophobic or hydrophilic? The sample prep can depend on the degree of dispersion you want and if the sample is hydrophilic or not.
Start with alcohol and water but you might have to switch to pure alcohol for some level of dispersion. Some people use acidic water but that can change the aggregate size. Stick to pure water, if you feel you must use water with hydrophilic silicas. I suggest 100KV for submicron primary particle sized silicas.
You didn't state the level of structure you are interested in. For example, there are three basic types of silanols on the surfaces of things like precipitated silica. These molecular structures and the near molecular pores detected by nitrogen BET are not observable in a TEM. Above those levels, the structures of silicas are directly observable in a TEM. BET is made up of at least four different factors that are summed together when used to characterize molecular pores and molecular roughness in materials like ppt'd silica powders. BET can tell you about surface area but determining the structural differences from that parameter requires other techniques like TEM, for example. Argon, nitrogen and CTAB surface areas could give totally different answers for surface area on the same sample. TEM relates well to the mean surface volume diameter that you can calculate from a CTAB surface area value or the MSVD that can be determined in a TEM by image analysis and post analysis.
As for getting the samples run fast, have you tired to search the membership list to see who might be located close to you? Most professional microscopists would be willing to help you out at least once. I would think JEOL in Boston could give you a complimentary real world sample 'demo'. Call their salesman. Maybe you will buy a microscope within the next few years and that kind of PR help should give JEOL the inside track on a future purchase. Somebody in Boston will help you run the sample(s). They/you can call me free for any help you might need with the sample prep.
Sincerely,
Paul Beauregard Senior Research Associate Microscopy, Microtomy and Image Analysis (724) 834-2247 (home)
} } We need to view silica powder (SiO2) surface structure with an electron } microscope. We are in a biological facility and have no experience with } this type of sample. What sort of instrument is best suited for viewing } this sample? What are the sample preparation techniques? Are there labs } that we can bring our sample to for, let's say, an hour of observation and } a few pictures? We are in the Boston, Massachusetts area. Thank you in } advance. } } Vachik Hacopian } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 2 13:01:27 2004
After much discussion last year (6/03) on this list about problems with sticky tabs, I found another source to replace Pella's tabs. The earlier ones worked great but when I re-ordered, the new Pella tabs were unacceptable. They were stiff, not sticky and not conductive. I wound up using EMS/Nissin.
I received a nice telecon from Ted Pella last week asking if I would try their new tabs. Of course I would like to. He sent them and here are the results:
A B C Original Pella 25K not meas. thin sticky Re-order Pella open open stiff not very sticky Newest Pella 10K 55K-200K thin sticky EMS/Nissin 45K 112K-1.5M thin sticky Fullam 20K 27K-500K thin sticky
A=initial resistance (Ohms) B=overnight resistance; compressed (first value)-uncompressed C=consistency of material
Tests were conducted using a pair of Pella 16242 pin stub mounts with a sample sticky tab sandwiched in between. Resistance measurements were taken using a Simpson #467 DVM. The initial resistance was taken after pressing the stubs together. The overnight resistance was taken the next day and was recorded under two-hand compression and then after release of compression. Resistance decreases under compression.
Another test comparison was done in the sputter coater. I had noticed that different tabs were differently affected by the coating experience. The original Pella tabs would show small linear cracks in the surface. The re-ordered tabs did not show any evidence of surface effects but were unusable due to non-conductance. The EMS/Nissin tabs show circular and concentric circular deformations at numerous locations on the tab. The Fullam tab shows similar deformations. The new Pella tabs appear to be affected in the same manner. I don't think that any of these effects are killers. It is just interesting and useful to note for comparison.
Pella's new 16084-1 replacement tab is definitely a viable replacement. Thanks Ted.
Looking at the data I've taken, I conclude that I'd rate Pella #1, Fullam #2, EMS/Nissin #3. All are thin and sticky but the differentiation is initial and final resistance (lower the better). That said, it likely makes little difference in operation since the specimen currents are so low. At a high specimen current of say 10nA, using the EMS/Nissin tab, the specimen could be about 15 mV above ground. Not a big deal, IMO.
Therefore, the new Pella, EMS/Nissin and Fullan tabs are all good tabs. The only remaining discriminator is cost. The new Pella tabs are $14.50/100.
Note that all three flavors of the Pella tabs have the same 16084-1 part number. If you have any of the old Pella tabs around (esp the hard ones), be sure your techs check new tabs to be sure they are thin. If you order new Pella tabs, there could be some confusion as to which is which until all the old ones are used or trashed.
gary g.
cc: Ted Pella, Tom Pella
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 2 15:28:52 2004
Recently I tried examining some thin sections of fossil bone (hydroxylapatite, R.I. around 1.6) and siderite (extreme birefringence, R.I. from around 1.9 to about 1.6) using immersion oils that matched the R.I. of the target mineral in the thin sections. This brought out lots of detail missing with normal mounting media (nominally about R.I. 1.5). Out of curiosity, does anyone know of some permanent mounting media that has an index in the 1.6 - 1.8 range? I know high R.I. melts were used at one time, but I seem to remember they were formulated with things like arsenic and antimony (?).
Henry Barwood Associate Professor of Science, Earth Science Department of Math and Physics MSCX 312G Troy State University Troy, Alabama 36082 hbarwood-at-troyst.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 2 18:48:13 2004
There are number of old mounting media - I have a few -
Styrax Hyrax Styrene dissolved in methyl iodide and the highest I know of - arsenic tribromide from Cargill - R.I. 2.31
Cargill makes (or made) Carmount in a range of R.I.s - I have three different viscosities of some 1.65
Bill Miller
At 04:45 PM 4/2/2004, Henry Barwood wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sat Apr 3 01:05:17 2004
You can find such book in Earth Sciences Library, Geocentrum, next block of your department or even at Sverige Geological Survey, SGU. For a mutimedia of SEM&EDS , Oxford instrument has a CDROM set.
Hopefully you'll find it.
Paiboon
_______________________________________________ Paiboon Nuannin Department of Physics Facylty of Science Prince of Songkla University Hatyai Thailand. now on leave to Deapartment of Earth Sciences Geocentrum Uppsala University Villavagen 16 Uppsala, Sweden _________________________________________________________
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } The book below is one of the first to have a section on low } vacuum/ESEM. } } Dave } } 3rd Edition (2003) } Scanning electron microscopy and X-ray microanalysis a text for } biologists, materials scientists, and geologists Joseph I. } Goldstein } ... [et al] } } } } On Tue, 9 Mar 2004 11:28:33 +0100 Stefan Gunnarsson } {Stefan.Gunnarsson-at-ebc.uu.se} wrote: } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------- } } } } Hi all! } } } } What textbooks and multimedia teaching aids for EM in general, SEM and } } EDS in particular, would you recommend? I am especially looking for } } those with (at least some) emphasis on low voltage FEG-SEM and low } } vacuum techinques and theory. } } } } tia, } } Stefan } } } } +++++++++++++++++++++++++++++++++++++++++++++++++++++++ } } Dr Stefan Gunnarsson } } Evolutionsbiologiskt Centrum Evolutionary Biology Centre } } Enheten för biologisk strukturanalys Microscopy and Imaging Unit } } Norbyvägen 18A } } SE75236 Uppsala, Sweden Tel & Fax: +46 - 18 471 2638 } } +++++++++++++++++++++++++++++++++++++++++++++++++++++++ } } }
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From MicroscopyL-request-at-ns.microscopy.com Sat Apr 3 09:39:10 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kuwahara-at-ag.arizona.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, April 2, 2004 at 12:06:47 ---------------------------------------------------------------------------
Email: kuwahara-at-ag.arizona.edu Name: Sara Kuwahara
I'm looking for information about protocals and methods that would allow me to affix antibodies to a glass or silicon surface, but would leave the antibody freee to bind to its associated antigen. If you could direct me to any articles, procedues or texts it would be greatly appreciated.
*****note***** I am not a memeber of this Listsever, so please respond to me directly.
Thank you
Sara Kuwahara
Dept of Ag and Biosystems Engineering University of Arizona
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mingram-at-rohmhaas.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, April 2, 2004 at 14:16:11 ---------------------------------------------------------------------------
Check out SELA. They have been doing this sort of thing for a long time.
Caveat: MME has no financial interest in this product.
Best regards, Barbara Foster Microscopy/Microscopy Education, Inc. (MME, Inc.) 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Optimizing Light Microscopy is still available through MME. For information on ordering single or small quantities, visit our website. For discounts on classroom-sized, call us today. ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&
At 10:27 AM 4/4/04 -0500, mingram-at-rohmhaas.com wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sun Apr 4 15:11:47 2004
Can you elaborate a bit about what you are trying to do? I'm having trouble figuring out why anyone would want to cross section a whole wafer of any size. Typically, the individual die are sliced out of the wafer and then cross sectioned as a die. If the wafer is 8", then this strongly suggests that the feature size is { 0.35u and would be a likely candidate for FIB rather than mechanical or fracture cross sectioning.
What are you trying to do?
gary g.
At 07:28 AM 4/4/2004, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Apr 5 06:07:34 2004
someone (ican't remember who) told me to use iridium...
Does anyone have informations about iridium (Ir) metallization coating for FE-SEM observations of VLSI/ULSI microsectionned Si or AsGa dies, instead of Au/Pd?
If so, can you detail the main differences in terms of performance : granulometry, deposition process, conditions of the process (time vaccum, which gas used...), cost... ?
d'avance merci!
Sylvain MAURY, de FRANCE
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 5 10:39:32 2004
At South Bay Technology we can provide equipment and process information to handle cleaving 8" wafers to preparing SEM and TEM cross-sections. If you can supply me with more details on your application, I would be pleased to send you information on the most appropriate solution.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
by way of MicroscopyListserver wrote:
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-- David Henriks Vice President
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
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email: henriks-at-southbaytech.com
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Please visit us online at www.southbaytech.com.
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From MicroscopyL-request-at-ns.microscopy.com Mon Apr 5 13:55:09 2004
Last year another research lab and I looked into an iridium high resolution coater. I suggested Ir and someone else did the 'leg work' evaluating the contender companies. Some manufacturers said they had Ir but the only one that DELIVERED one to that lab last November was Emitech.
My retired old boss and I had a VCR Group micro sputter system that was one of the original systems sold by Vince Carlino. Later, this technology was sold and merged into South Bay and Vince worked for them. (My take on the situation.) He retired and you should send him an email because he contacted me about the Ir unit we were considering. He is very very very knowledgeable and helpful. Vince Carlino - Sales Manager 415-566-5774 Evactron.Com
Cressington says they have an Ir unit but I didn't see one on their web page. Contact them too. A lady in Europe wrote me an email and said she was getting one from Cressington shortly.
So you have the following four choices: South Bay EmiTech Talking to Vince Carlino at Evactron.Com and Cressington.
Vince can tell you where the original Ir idea came from in the US and how the various metal high resoution coatings perform; Cr, Pt, and Ir. He's a great guy to get in touch with about these coatings and knows the strength and weakness of each of them.
Hope this helps,
Paul Beauregard Senior Research Associate Microscopy and Image Analysis
Disclaimer: I have no interest in any of these companies mentioned.
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 5 17:59:50 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (muchphoto-at-earthlink.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, April 5, 2004 at 10:23:03 ---------------------------------------------------------------------------
Email: muchphoto-at-earthlink.net Name: Michael Reese Much
Organization: Michael Reese Much Photographic Illustration
Education: Undergraduate College
Location: Bethlehem, PA
Question: I have recently purchased a microscope and have adapted it for digital photography. Could you recommend a good textbook about the preparation of specimen slides (staining, preparing protozoa for mounting and staining, etc.). I am also intereted in an atlas of protozoans.
This may be of NO use to you whatsoever, but when I heard that Ir gave a smaller grain size, I bought some wire and wadded it up and put it in a drilled-out carbon rod and deposited it with our (mostly unused) Balzers 400T freeze-fracture device (electron gun deposition). Worked great! More complicated than a sputter coater, but we needed the resolution. I believe Kent MacDonald's EM lab at Berkeley has a Balzers 300 that they have used to coat for high-res SEM, using Pt. Again, kind of a pain, but if one of these machines, used and about to be discarded, crosses your path... They generally have a thickness monitor as well.
Aloha, Tina
**************************************************************************** * 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 MicroscopyL-request-at-ns.microscopy.com Mon Apr 5 23:51:49 2004
muchphoto-at-earthlink.net (by way of Ask-A-Microscopist) wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (muchphoto-at-earthlink.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, April 5, 2004 at 10:23:03 } --------------------------------------------------------------------------- } } Email: muchphoto-at-earthlink.net } Name: Michael Reese Much } } Organization: Michael Reese Much Photographic Illustration } } Education: Undergraduate College } } Location: Bethlehem, PA } } Question: I have recently purchased a microscope and have adapted it for digital photography. Could you recommend a good textbook about the preparation of specimen slides (staining, preparing protozoa for mounting and staining, etc.). I am also intereted in an atlas of protozoans. } } Hello Michael - I am not familiar with a textbook about specimen slide preparation, especially at it concerns protozoa. However, I did extensive work with ciliates while I was pursuing my M.A. degree. I used basic microscopy techniques--that is, take a sample containing protozoa, put it on a slide, line the edges of the coverslip with vaseline (to act as a humid chamber), and then put it in a light microscope. About an atlas..the Society of Protozoologists (http://www.protozoa.uga.edu) (pretty sure) sells a book on protozoa called the Illustrated Guide To The Protozoa. I am unsure how the public can buy it, but perhaps if you visited that web site, you may be able to find some helpful information. Good luck, Nelson Conti
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From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 01:15:26 2004
We at South Bay Technology can help you with any of your cross-sectioning requirements from cleaving an 8" wafer to making SEM and TEM cross-sections. If you can narrow down your requirements a little further, I'll be happy to direct you to our most appropriate solution.
Best regards-
David
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- David Henriks Vice President
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com.
The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 01:31:17 2004
Paul Beauregard was correct in that Vince Carlino did some of the early work on ion beam sputtered Ir films . In fact, I have a copy of a paper by Cardinale, Carlino and Howitt titled "Comparison of Ion Beam Sputtered Chromium and Iridium Films for Electron Microscopy Applications" which I would be pleased to share with you.
Basically, the paper indicates that Ir produces very thin, extremely fine grain, conductive coatings without the negative effects caused by oxidation of chromium films. It is also of sufficiently high atomic number to enhance electron scattering. Ir is routinely sputtered with our IBS/e system and, in fact, is one of the standard targets supplied with the system.
If you send me your maling address, I'll be pleased to send you a copy of the paper. Paul Beauregard was correct in that the original sputtering system (IBS/TM200) used in this analysis was acquired by South Bay Technology back in 1999 when they purchased VCR Group. South Bay Technology does continue to support The IBS/TM200 although a newer generation Ion Beam Sputter Deposition and Etching System (IBS/e) has been developed as well.
I have a fairly substantial library of technical papers covering ion beam deposition as well as ion beam sources that I would be pleased to share with anyone interested. Send me an email with your mailing address and I'll send out the relevent papers.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
-- David Henriks Vice President
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com.
The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.
sylvain maury wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- David Henriks Vice President
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com.
The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 07:49:36 2004
Director of Microscopy and Imaging University of Richmond
The Department of Biology at this highly selective, private, primarily undergraduate university invites applications for a Director of Microscopy and Imaging. Duties including supervising operation of a new biological imaging facility, including operation and routine maintenance of a TEM, SEM, and laser scanning confocal microscope and associated equipment. Teaching responsibilities include training and supervising faculty and student users in research and classroom activities and courses based on interests and department needs; assisting faculty and students in experimental design, specimen preparation, digital photography, and analysis of results.
Qualifications: MS or PhD degree and experience with light and electron microscopy and digital imaging, strong interpersonal skills and desire and ability to work well in a team environment. This is a continuing appointment that is not eligible for tenure but may include faculty status.
Applications including a letter of interest, CV, evidence of expertise in microscopy, and three letters of recommendation should be sent to Dr. Gary Radice, Department of Biology, University of Richmond, Richmond VA 23173 (gradice-at-richmond.edu). Review of applications will begin on April 23, 2004. The appointment is expected to begin June 1, 2004.
The University of Richmond is committed to increasing the diversity of our faculty and strongly encourages applications from women and minorities. For more information on the department, resources, and teaching assignment, see (http://biology.richmond.edu/)
Gary P. Radice gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond 804-289-8233 (FAX) Richmond VA 23173 http://www.richmond.edu/~gradice
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 08:51:01 2004
A rather strange story read in the BBC online this morning. A new chemical to become regulated in EM labs in the US (after uranyl acetate)? Let's hope not! { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 10:02:08 2004
I did a search on Amazon for "Illustrated Guide To The Protozoa" and came up with a book by John Lee, which is out of print but they say they have limited availability. Found several others from that search as well including: Marine Interstitial Ciliates: An Illustrated Key (Chapman & Hall Identification Guide, No 2) by Philip G. Carey (Hardcover - July 1992) (though it's a bit pricey)
Bettye
On Apr 5, 2004, at 10:06 PM, Nelson Conti wrote:
} Hello Michael - } I am not familiar with a textbook about specimen slide preparation, } especially at it concerns protozoa. However, I did extensive work with } ciliates while I was pursuing my M.A. degree. I used basic microscopy } techniques--that is, take a sample containing protozoa, put it on a } slide, line the edges of the coverslip with vaseline (to act as a } humid chamber), and then put it in a light microscope. About an } atlas..the Society of Protozoologists (http://www.protozoa.uga.edu) } (pretty sure) sells a book on protozoa called the Illustrated Guide To } The Protozoa. I am unsure how the public can buy it, but perhaps if } you visited that web site, you may be able to find some helpful } information. } Good luck, } Nelson Conti } } } -- } 164 Ferne Court } Palo Alto, CA 94306 } Home: (650) 494-0472 } Cell: (650) 906-1159 } Email: [ncontiSFSU-at-netscape.net] } --------------- Bettye L. (Smith) Maddux Dept. Biochemistry & Biophysics ALS 2011 Oregon State University Corvallis, OR 97330 Lab: 541.737.1870 Off: 541.737.8018 madduxb-at-onid.orst.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 12:07:30 2004
The University of Chicago has a FEI (formerly Philips) CM120 for sale for $15,000. The instrument is in excellent working condition and includes a Haskris closed circuit water cooling system. For further information contact
Robert Josephs 773 702 1077
186,000 miles a second -- it's not just a good idea -- it's the law
Bob Josephs
http://gingi.uchicago.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 13:06:31 2004
Dear Researchers; Our immunogold workshop in Madison, Wisconsin is being rapidly filled up. If you are interested in attending, contact us asap. The following is the original announcement, Thank you very much.
=======================
Immunogold Workshop Announcement
Dear Researcher:
The Electron Microscopy Facility-Medical School at the University of Wisconsin is hosting a three-day workshop on immunogold techniques from May 24-26, 2004. Dr. Jan Leunissen from Aurion Immunogold Reagents & Accessories, an internationally known expert in the field, will be the instructor for the workshop. The workshop will include lectures, hands-on training, round table discussions, and presentations on applications. Also, participants of the workshop will be able to work on their own samples during the workshop. The workshop main curriculum is detailed below. If you are interested in attending or need more information about the workshop, please contact the workshop technical coordinator Hong Yi by phone (404-727-8692) or email (hyi-at-emory.edu).
MAIN CURRICULUM
The properties of gold particles and their protein conjugates. Theories underlying immunogold labeling protocols. Silver enhancement of gold particles Immunogold labeling on a variety of sample preparations for LM. Immunogold labeling for EM a. Pre-embedding immunogold labeling using ultrasmall gold conjugates and silver enhancement. b. Post-embedding immunogold labeling using conventional colloidal gold conjugates and ultrasmalll gold conjugates. Pre- and post-embedding double immunogold labeling. Background minimization in immunogold labeling Signal amplification in immunogold labeling.
Thanks and we hope to see you in Madison.
Randall Massey Operations Manager UW-Electron Microscope Facility 1300 University Ave. Madison, WI 53706 voice (608)262-2993 Fax (608)262-7306
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 15:38:24 2004
This story was on ABC Evening News last night (Monday April 5) too.
Marc Pypaert wrote:
} ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } A rather strange story read in the BBC online this morning. } A new chemical to become regulated in EM labs in the } US (after uranyl acetate)? Let's hope not! } { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm } } } -- } Marc Pypaert } Department of Cell Biology } Center for Cell and Molecular Imaging } Ludwig Institute for Cancer Research } Yale University School of Medicine } 333 Cedar Street, PO Box 208002 } New Haven, CT 06520-8002 } TEL 203-785 3681 } FAX 203-785 7446 } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 18:19:15 2004
They also could count sodium borhydride and other nasty staff from laboratory shelves. Sergey
At 07:06 AM 4/6/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Today, per request of NRL security, our chemical hygiene officer came by my TEM lab to check that my small amount of osmium tetroxide was present and secure.
} This story was on ABC Evening News last night (Monday April 5) too.
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------------------
} } } A rather strange story read in the BBC online this morning. } A new chemical to become regulated in EM labs in the } US (after uranyl acetate)? Let's hope not! } { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm } } } -- } Marc Pypaert } Department of Cell Biology } Center for Cell and Molecular Imaging } Ludwig Institute for Cancer Research } Yale University School of Medicine } 333 Cedar Street, PO Box 208002 } New Haven, CT 06520-8002 } TEL 203-785 3681 } FAX 203-785 7446 } }
-- -- ********************************************** 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 **********************************************
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Facility Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Mark Pypaert wrote: ======================================================= A rather strange story read in the BBC online this morning. } A new chemical to become regulated in EM labs in the } US (after uranyl acetate)? Let's hope not! } { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm } ======================================================= Based on the sudden media interest in osmium tetroxide, I agree with Mark that burdensome regulations could be placed on another important tool used in electron microscopy.
As an industry, we should be developing standards for security for where osmium tetroxide is being stored in a laboratory setting and suppliers should be establishing standards not only for the storage of OsO4 in their inventory, but procedures for the reporting of suspicious appearing customers. SPI Supplies has had a long standing policy of selling only to institutions (and not to individuals) but this approach "works" only so long as our institutional customers keep materials such as osmium tetroxide in secure locations and in ways that all of the material is accounted for (a "cradle to grave" kind of accounting system). Shortages would have to be explained just as is the case now for laboratories with a US federal tax- free alcohol permit.
I fear that if we don't do this voluntarily and quickly, then regulations will be forced upon us, and without the benefit of input from our industry and this means that the costs of doing research will be increased, perhaps substantially for the shipment and storing of osmium tetroxide.
It would seem to me that developing such standards would be a legitimate role for MSA and other of the world's microscopy societies. But someone has to take some leadership responsibility to start the ball rolling in that direction so that as an industry, we can police ourselves and not have governments dictate to us how it should be done.
Disclaimer: SPI Supplies has been a long time worldwide supplier of osmium tetroxide to electron microscope laboratories.
Chuck
============================================ Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 19:46:23 2004
I note with interest that you keep your nasty staff on laboratory shelves, I would like to introduce a similar policy here in Australia. Cheers,
Mark Blackford Materials and Engineering Science, ANSTO PMB 1, Menai, N.S.W., 2234 Australia
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.
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Wednesday, 7 April 2004 9:35 AM To: Microscopy-at-sparc5.microscopy.com
They also could count sodium borhydride and other nasty staff from laboratory shelves. Sergey
At 07:06 AM 4/6/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
The sudden concern regarding osmium tetroxide probably relates to the following
Poison gas attack foiled in Britain
07.04.2004 By KIM SENGUPTA A planned poison gas attack, with the London underground system as a likely target, has been foiled by the security agencies, it was claimed today.
The plot, by British based supporters of al Qaeda, allegedly involved detonating a combined chemical and explosive "dirty bomb", producing fumes which can choke victims in a confined place.
The conspiracy was uncovered after British and United States intelligence intercepted telephone calls within this country, as well as calls to alleged senior members of the group in Pakistan.
As well as tube trains, passenger terminals at London's Gatwick and Heathrow airports are believed to have been in the list of targets. However, it is believed the plot was exposed before the alleged terror cells had reached the position to carry out an attack.
Security sources said yesterday that the attack involved the use of a chemical called osmium tetroxide to set off a blast.
The substance turns from solid to gas in confined space and is highly corrosive to eyes, skin tissue and lungs, producing a symptom called "dry land drowning." It is believed that a fertiliser-based explosive would have been part of the package.
............
} ------------------------------------------------------------------------------- } } A rather strange story read in the BBC online this morning. } A new chemical to become regulated in EM labs in the } US (after uranyl acetate)? Let's hope not! } { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm } } } -- } Marc Pypaert } Department of Cell Biology } Center for Cell and Molecular Imaging } Ludwig Institute for Cancer Research } Yale University School of Medicine } 333 Cedar Street, PO Box 208002 } New Haven, CT 06520-8002 } TEL 203-785 3681 } FAX 203-785 7446 }
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Ian Hallett HortResearch Mt Albert Research Centre, Private Bag 92 169 Auckland, New Zealand Fax +64 9 815 4201 Telephone +64 9 815 4200 EMail ihallett-at-hortresearch.co.nz
______________________________________________________ The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. ______________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 22:12:37 2004
Having seen several listings of open positions on this list, I thought I would post my CV and let people know that I am looking for positions in microscopy. If anyone is interested, please let me know!
Here is my CV:
D. Scott Snyder 8100 Cambridge #103 Houston, TX 77054 (713)799-9165 dscottsnyder-at-earthlink.net
Education: B. S., Chemistry, University of Idaho (1987) Graduated with a 3.8 GPA
M. S., Biochemistry, Medical College of Wisconsin (1990) Thesis: Interaction of Elongation Factor 3 from the Yeast S. cerevesiae with Azido ATP: Photoaffinity labeled ATP binding sit of EF-3; Proved specificity by competitive enzyme inhibition, completeness of inactivation, stoichiometry, and kinetics of inactivation Coursework: Molecular Genetics, Protein Chemistry, Enzyme Mechanics, Medical Biochemistry, Physical Methods of Biochemistry Rotations: Purification of ubiquitin, enzyme isolation and analysis, affinity labeling of enzyme active sites, western screening for elongation factor 3 clones
Ph. D., Cell Biology with certification in Biological Chemistry, Duke University, 2000 Mentor: Thomas J. McIntosh Dissertation: Structure and Antibiotic Permeability of Bacterial Lipopolysaccharide Bilayers – Determined microbial membrane bilayer structure with XRD, DSC and other analytical techniques then correlated structure with permeability as determined by stopped flow fluorimetry and spectroscopic methods Coursework: Advanced Molecular Biology, Analytical Spectroscopy, Stereochemistry, Cell Biology, Cell Signaling, Bioorganic Chemistry, Genetics Rotations: Lipid A biosynthesis, Lab of Christian Raetz LPS Vaccine formulation, Lab of Thomas McIntosh and MDT, Inc.
Experience Lab Manager, Integrated Microscopy Core, Baylor College of Medicine, Houston, TX Responsible for developing new techniques in microscopy such as FRET, FLIP and spectral imaging applications, training users, maintaining equipment, grant writing to obtain new equipment and general managerial duties in regional microscopy facility. Also involved in research project involving estrogen receptor based signal transduction.
Researcher and Confocal Systems Manager, National Institutes of Health, Rocky Mountain Laboratories, Hamilton MT Responsible for independent research examining the inflammatory response of cultured mammalian cells in response to the TLR2 lipoprotein ligand OspA and the internalization of OspA in such cells. Techniques involved NF kappa B localization assays, as well as TNF and IL-1 ELISAs and extensive live cell confocal microscopy. Also responsible for managing confocal facility. Post Doctoral Fellow, National Institutes of Health, Rocky Mountain Laboratories, Hamilton MT (2000-2001) Project involved determining the mechanism of action of mycolactone, a toxin derived from Mycobacterium ulcerans which is capable of eliminating the inflammatory response as well as inducing cytoskeletal rearrangement and apoptosis in a variety of cell types and is required for virulence in vivo. Techniques involved isolation, derivatization and analysis of mycolactone, mass spectroscopy, cell culture, inhibition of various cell signaling pathways, confocal microscopy, examination of calcium release from the endoplasmic reticulum and apoptosis assays.
M.S. Level Research Associate for Ribi Immunochem Inc., Hamilton, MT (1990-1995) Developed GLP methods for the isolation, fractionation, derivatization and analysis of lipid based inflammatory mediators using HPLC, LC, GC, TLC, NMR, UV/VIS and wet chemical methods Fluorescently labeled and analyzed the cell localization patterns of inflammatory mediators Refined lab bench procedures for industrial setting (scale-up) Developed novel analytical methods for GMP adjuvants and vaccines Assayed effects of inflammatory mediators and derivatives on cell cultures and in animal models using both the ELISA and NO assays Interviewed and trained research technicians Conducted facility tours for investors, lawmakers and the public Received 3 raises and 1 promotion in this position
Other Advanced Training: HPLC Method Development: Lloyd Snyder and John Dolan (LC Resources) HPLC Troubleshooting and Repair: Waters Corporation Interpretation of Mass Spectra: American Chemical Society Advanced Topics in Live Cell Imaging: University of Connecticut Health Sciences Center
Publications SR Hagen, JD Thompson, DS Snyder and KR Myers, Analysis of a Monophosphoryl Lipid A Immunostimulant Preparation from Salmonella R595 by HPLC, Journal of Chromatography, 767 (1997) pages 53-61
DS. Snyder, D Kim and TJ. McIntosh, LPS Bilayer Structure: Effect of Chemotype, Core Mutations, Divalent Cations and Temperature, Biochemistry, v38, number 33 (1999) pages 10758-10767
K Kulkarni, DS Snyder, and TJ McIntosh, Adhesion Between Cerebroside Bilayers, Biochemistry, v38, number 46, (1999) pages 15264-15271
DS Snyder and TJ. McIntosh, The LPS Permeability Barrier: Correlation of Antibiotic Permeabilities with Antibiotic Susceptibilities and Fluorescent Probe Binding Kinetics Biochemistry, v39, number 38 (2000) pages 11777-87
EB Guererro, TJ McIntosh, GR Barclay, DS Snyder, RJ Gibbs, MG Mythen and IR Poxton, Preparation and Preclinical Evaluation of a Novel Liposomal Complete Core Lipopolysaccharide Vaccine, Infection and Immunity, v68 number 11 (2000) pages 6202-8
DS Snyder and PLC Small, Staining of Cellular Mitochondria with LDS-751, JIM, v257 (2001) pages 35-40
DS Snyder and PLC Small, Uptake and cellular actions of mycolactone, a virulence determinant for Mycobacterium ulcerans. Microb Pathog. 2003 Feb;34(2):91-101.
DS Snyder and CF Garon Decreased uptake of bodipy-labelled compounds in the presence of the nuclear stain, DRAQ5.J Microsc. 2003 Sep;211(Pt 3):208-11.
D Welty, DS Snyder. Internalization of OspA in rsCD14 complex and aggregated forms. Mol Microbiol. 2003 Nov;50(3):835-43.
KA Jackson, DS Snyder, MA Goodell, Stem Cell Engraftment: Distinguishing GFP from Autofluorescence. Stem Cells. 2004;22(2):180-7
Patents Vaccine Against Lipopolysaccharide Core, Application # WO 98/51217 by Elliott Bennett Guererro, George Robin Barclay, Ian Raymond Poxton, Thomas James McIntosh and David Scott Snyder (pending)
Method for the Production of 3 O Deacylated 4’ Monophosphoryl Lipid A Kent Myers and Scott Snyder (pending)
Invited Presentations
A Tale of Two Ligands University of North Texas Health Sciences Center at Fort Worth, April 2002
Abstracts DS Snyder, KL Whitaker and KR Myers, Structural and Biological Characterization of Unmodified Re Lipopolysaccharide by Normal Phase Chromatography 1995 FASEB meeting
DS Snyder and TJ McIntosh, Bacterial Antibiotic Susceptibility and Lipopolysaccharide Structure/Permeability, 1999 International Endotoxin Society Meeting
Scholarships, Honors and Awards Richard Asofsky Award for Special Achievement in Equal Employment Opportunity, Hoggsey-Cossey Award, WMA Scholarship, Whitman E Scholarship, 5th Marine Division Scholarship, others available upon request
References Thomas J. McIntosh, Mentor, Phone (919) 684-8950, e-mail mcint001-at-mc.duke.edu Kent Myers, Supervisor at Ribi, Phone (406) 363-6214, e-mail kmyers-at-corixa.com Elliott Bennett Guererro, Director Cardiothoracic Anesthesia, Columbia University and President of MDT, Inc (212) 996-8234, e-mail elliott_guererro-at-smtplink.mssm.edu Pam Small, Supervisor at NIH, Phone (865) 974-4042, e-mail psmall-at-utk.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 23:29:33 2004
The videos of the Tutorials presented at the 2003 MSA meeting in San Antonio are now available. See the current catalog at this URL: http://www.biotech.ufl.edu/sems/newtape00.htm All new titles are available on DVD only. Some VHS tapes of old titles remain. We are no longer producing any new tapes. This is a continuing service of the Education Committee of the Microscopy Society of America. Contact me to order.
Greg Erdos
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 08:32:10 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dmwilliams-at-dow.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 7, 2004 at 08:15:27 ---------------------------------------------------------------------------
Email: dmwilliams-at-dow.com Name: David M. Williams
Question: Does anyone have a good fixative formula for whole body perfusions of rats? I am primarily interested in getting optimal fixation of the periferal nerves for optical and possibly TEM examination. Thank you.
Well yes! It opens up a whole new sphere for bureaucratic interference. But surely, OsO4 is so expensive that each lab knows exactly where every microgram is.
Lesley Weston.
on 06/04/2004 4:34 PM, Sergey Ryazantsev at sryazant-at-ucla.edu wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } They also could count sodium borhydride and other nasty staff from } laboratory shelves. Sergey } } At 07:06 AM 4/6/2004, you wrote: } } } } ---------------------------------------------------------------------------- -} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------------- } } -- } } } } A rather strange story read in the BBC online this morning. } } A new chemical to become regulated in EM labs in the } } US (after uranyl acetate)? Let's hope not! } } { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm } } } } } -- } } Marc Pypaert } } Department of Cell Biology } } Center for Cell and Molecular Imaging } } Ludwig Institute for Cancer Research } } Yale University School of Medicine } } 333 Cedar Street, PO Box 208002 } } New Haven, CT 06520-8002 } } TEL 203-785 3681 } } FAX 203-785 7446 } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } 10833 Le Conte Ave, Room 33-089 } Los Angeles, CA 90095 } } Phone: (310) 825-1144 (office) } (310) 206-1029 (Lab) } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 09:33:21 2004
I was hoping someone could point me in the right direction for finding out about average salaries Electron Microscopist. The average salary for Electron Microscopists just starting out of college with a B.S. and one with several years of experience.
Thank you very much
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 10:09:30 2004
Hi all I agree with these sentiments about developing our own standards, especially as we have the most to gain from this. Maybe we can discuss this at the meeting in Savannah in August for the MSA and in Wolfville, Nova Scotia, Canada in May for the MSC meeting. I would like to be part of this policy making. Elaine
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 11:30:51 2004
Colleagues: Our Bio-Imaging facility has merged with our E-M facility and I now have a Hitachi TEM under my care. Can any one recommend a basic introductory course in electron microscopy. It would be preferable if it was near New York City, or at least in the North East. Thanks
Lloyd Williams
Manager Bio-Imaging Facility Hunter College 695 Park Ave New York, NY 10021
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 11:53:39 2004
We have Leica inverted (DMIRE2) and upright (DMRXA2) microscopes in our Confocal facility that we would like to add live cell imaging capabilities to. At the moment, a heating stage would be adequate. If you are using such a stage (even on a different brand microscope) and are really happy with it, would you please share the manufacturer with me?
Thank you in advance,
Doug
---------------------- Douglas R. Keene Associate Investigator Micro-Imaging Center Shriners Hospital for Children 3101 S.W. Sam Jackson Park Road Portland, Oregon 97239-3009 phone: 503-221-3434 FAX: 503-412-6894
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 12:10:57 2004
I am wanting to localize GFP within cultured fibroblasts at the ultrastructural level (TEM). We are using anti-GFP antibodies provided by Rockland and another provided by Assay Design. We have tried a variety of protocols using prefixation in 0.1% glut and/or rendering the cells permeable with 0.01% saponin. We've also prefixed the cells with 100% methanol (this results in reasonable labeling density but terrible ultrastructure). Our typical protocol utilizes colloidal gold conjugated secondary antibodies.
In any of these protocols I have tried, I am either not satisfied with the preservation of the cells or I am not satisfied with labeling density. Certainly not an unusual quandary for immuno-EM, but surely there is a more satisfactory protocol than the ones I have tried!
My question: Does anyone use a TEM protocol for intracellular labeling that they are happy with?
Many thanks,
Doug
---------------------- Douglas R. Keene Associate Investigator Micro-Imaging Center Shriners Hospital for Children 3101 S.W. Sam Jackson Park Road Portland, Oregon 97239-3009 phone: 503-221-3434 FAX: 503-412-6894
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 12:35:47 2004
I agree with the concerns about 0s04. We also must limit our sales to U.S. agencies, institutions, some industries and other vendors who understand the procedures required when chemicals are involved.
Internally we limit access to all our stains, fixatives, epoxies, etc., even those that are not hazardous. We have close to 50 years experience in handling a wide range of chemicals so if anyone needs any assistance that we might be able to provide please contact us.
Disclaimer: Ladd is a manufacturer and worldwide distributor of EM/Laboratory supplies and chemicals such as Osmium tetroxide.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 12:39:58 2004
It is specific to electron microprobe labs and their managers/technicians, but the following report is available on-line:
Survey of User Fees, Laboratory Support, and Operator Salaries in Electron Microprobe Laboratories
http://research.ou.edu/microprobe/feesurv.pdf
Hope it helps,
Ellery
--------------- Ellery E. Frahm Research Scientist/Manager Electron Microprobe Laboratory Department of Geology & Geophysics University of Minnesota - Twin Cities Lab Website: http://probelab.geo.umn.edu
On 7 Apr 2004, kathy lowe wrote: } I was hoping someone could point me in the right direction for finding out } about average salaries Electron Microscopist. The average salary for } Electron Microscopists just starting out of college with a B.S. and one } with several years of experience. } } Thank you very much
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 13:24:16 2004
By way of background, some of you may recall that there was a meeting of Facility Managers at M&M 2003. The intent to form a formal group to discuss problems such as this one concerning osmium tetroxide as well as many other issues of major interest to facility managers and also of interest to all EM users. The request to form a Focused Interest Group was denied by the MSA Council for reasons that have not yet been fully explained but hopefully will be soon. However, we still hope to meet as a group at M&M2004. I was elected to serve as chair of the group at M&M 2003.
Perhaps this is where the facility manager's group can be of assistance. This would be a reasonable topic for discussion if we meet at M&M2004 providing the MSA council does not want to act prior to that time. I could envision our putting together a draft proposal that would then go to the MSA Council for their discussion/ revision followed by a formal policy statement at the winter council meeting. Does this seem reasonable? Would the Facility Managers like to be involved with this issue?
Please get back to me and I will try to act accordingly.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 4/7/04 11:14 AM, "Elaine Humphrey" {ech-at-interchange.ubc.ca} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Hi all } I agree with these sentiments about developing our own standards, } especially as we have the most to gain from this. Maybe we can } discuss this at the meeting in Savannah in August for the MSA and in } Wolfville, Nova Scotia, Canada in May for the MSC meeting. I would } like to be part of this policy making. } Elaine } } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------------- } } -- } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } Mark Pypaert wrote: } } ======================================================= } } A rather strange story read in the BBC online this morning. } } } A new chemical to become regulated in EM labs in the } } } US (after uranyl acetate)? Let's hope not! } } } { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm } } } ======================================================= } } Based on the sudden media interest in osmium tetroxide, I agree with Mark } } that burdensome regulations could be placed on another important tool used } } in electron microscopy. } } } } As an industry, we should be developing standards for security for where } } osmium tetroxide is being stored in a laboratory setting and suppliers } } should be establishing standards not only for the storage of OsO4 in their } } inventory, but procedures for the reporting of suspicious appearing } } customers. SPI Supplies has had a long standing policy of selling only to } } institutions (and not to individuals) but this approach "works" only so long } } as our institutional customers keep materials such as osmium tetroxide in } } secure locations and in ways that all of the material is accounted for (a } } "cradle to grave" kind of accounting system). Shortages would have to be } } explained just as is the case now for laboratories with a US federal tax- } } free alcohol permit. } } } } I fear that if we don't do this voluntarily and quickly, then regulations } } will be forced upon us, and without the benefit of input from our industry } } and this means that the costs of doing research will be increased, perhaps } } substantially for the shipment and storing of osmium tetroxide. } } } } It would seem to me that developing such standards would be a legitimate } } role for MSA and other of the world's microscopy societies. But someone has } } to take some leadership responsibility to start the ball rolling in that } } direction so that as an industry, we can police ourselves and not have } } governments dictate to us how it should be done. } } } } Disclaimer: SPI Supplies has been a long time worldwide supplier of osmium } } tetroxide to electron microscope laboratories. } } } } Chuck } } } } ============================================ } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } } President 1-800-2424-SPI } } SPI SUPPLIES FAX: 1-610-436-5755 } } PO BOX 656 e-mail:cgarber-at-2spi.com } } West Chester, PA 19381-0656 USA } } Cust.Service: spi2spi-at-2spi.com } } } } Look for us! } } ######################## } } WWW: http://www.2spi.com } } ######################## } } ============================================
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 13:27:28 2004
The New England Society for Microscopy and the Connecticut Microscopy Society announce the annual Spring Symposium, preceded by a workshop on Image Analysis using LISPIX, presented by Dr. David Bright (NIST). The workshop and symposium will be held April 29th. - May 1st. 2004, at the Marine Biological Laboratory, Woods Hole, Massachusetts.
Full details of both events, including registration information, are available on the NESM Web site, accessible at {http://www.msa.microscopy.com/MSALAS/NESM/NESMHome.htm} http://www.msa.microscopy.com/MSALAS/NESM/NESMHome.htm
*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
Colleagues: Our Bio-Imaging facility has merged with our E-M facility and I now have a Hitachi TEM under my care. Can any one recommend a basic introductory course in electron microscopy. It would be preferable if it was near New York City, or at least in the North East. Thanks
Lloyd Williams
Dear Lloyd:
Dr. Hayat used to teach a course at Kean College of New Jersey ( I think it is Kean University, now). It is located in Union, New Jersey; and not too far from NYC.
Also, Montclair State University (in Montclair, New Jersey) used to have an EM course years ago. I don't know if they still offer it.
Finally, Lehigh University in Pennsylvania offers EM mini-courses/workshops (usually held in the summer).
I hope this helps.
Good Luck,
Jackie
------------------------------ Jacqueline Garfield, Electron Microscopist Research & Development Lifecell Corporation One Millennium Way Branchburg, NJ 08876 (908) 947-1182
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 14:45:56 2004
Hi everyone: I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a series of holes within micron (or ideally submicron) accuracy on a strip of metal that has been laser holes drilled into.
I hope to achive this by using the change in z from the top and the bottom of hole using a calibrated stage. Does anyone out there have a device that they had made or purchased at a reasonable price. Are you willing to share your drawings/expertise? Thanks, Michael Coviello UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 15:17:33 2004
} } They also could count sodium borhydride and other nasty staff from } } laboratory shelves.
Dear List, I wonder just how toxic a "dirty bomb" using OsO4 would actually be. The conditions of an explosion often are hypoxic, so OsO4 could be reduced to a non-toxic state. On the other hand, the presence of OsO4 could enhance combustion in the explosion making it more powerful. Maybe those attempting to test such a bomb would then be killed in their own, surprisingly large explosion. If we're very lucky, they might try to make a really dirty bomb using both OsO4 and NaBH4. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 15:24:03 2004
I have had good luck locating various GFP constructs in COS-7 cells using this general technique;
Fix in 4% paraformaldehyde Quench with .1% NaBH4 Permeablize with .1% Triton x-100 Block Molecular Probes' polyclonal anti GFP Aurion GAR ultra small immuno gold Silver enhancement Convention fixation and embedment
Let me know if you want a detailed protocol.
Cheers,
Tom
-----Original Message----- } From: drk-at-SHCC.org [mailto:drk-at-SHCC.org] Sent: Wednesday, April 07, 2004 12:26 PM To: Microscopy-at-sparc5.microscopy.com
Hello,
I am wanting to localize GFP within cultured fibroblasts at the ultrastructural level (TEM). We are using anti-GFP antibodies provided by Rockland and another provided by Assay Design. We have tried a variety of protocols using prefixation in 0.1% glut and/or rendering the cells permeable with 0.01% saponin. We've also prefixed the cells with 100% methanol (this results in reasonable labeling density but terrible ultrastructure). Our typical protocol utilizes colloidal gold conjugated secondary antibodies.
In any of these protocols I have tried, I am either not satisfied with the preservation of the cells or I am not satisfied with labeling density. Certainly not an unusual quandary for immuno-EM, but surely there is a more satisfactory protocol than the ones I have tried!
My question: Does anyone use a TEM protocol for intracellular labeling that they are happy with?
Many thanks,
Doug
---------------------- Douglas R. Keene Associate Investigator Micro-Imaging Center Shriners Hospital for Children 3101 S.W. Sam Jackson Park Road Portland, Oregon 97239-3009 phone: 503-221-3434 FAX: 503-412-6894
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 16:11:12 2004
Lehigh University in Bethlehem, PA is your closest bet for a formal course in TEM. I took that course and another one years ago in Boston. It's very good, near you and has very good instructors.
Try www.lehigh.edu/microscopy for information and course times. (610) 758-5133
Sincerely,
Paul Beauregard Senior Research Associate Microscopy and Image Analysis
At 12:48 PM 04/07/04 -0400, Lloyd Williams wrote: } } } --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 16:29:54 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kcarson-at-odu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 7, 2004 at 13:47:34 ---------------------------------------------------------------------------
Email: kcarson-at-odu.edu Name: Keith Carson
Organization: Old Dominion University
Title-Subject: [Microscopy] [Filtered] digitizing TEM negatives
Question: I am looking for suggestions on equipment and settings to digitize TEM negatives and produce high quality hard copies from the digital image files.
Dear Debby There is no need to discuss (and provoke therefore) the issue, which does not exist as a scientific matter (are we scientists or politicians?). There are plenty of strict regulations which are already enforced. It's ridiculous, we spent time for such matters. I think, we should keep this place out of political issues. Unsuccessful OsO4 attack in England IS a political matter, not scientific. Sergey
At 11:39 AM 4/7/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Have you tried frozen ultrathin sections (the Tokuyasu method)? This is the ultimate method for this kind of work. Alternatively, you could use lowicryl or similar type of hydrophilic resins. Maybe combined with high-pressure freezing and freeze-substitution for better preservation of morphology.
For the labeling, I have never tried anti-GFP antibodies from Rockland, but I have great results with antibodies from Molecular Probes. Also, we have stopped using gold-conjugated secondary antibodies as these do not always give great labeling efficiency. We prefer to use protein A-gold instead. You might want to try. Best
Marc
On Wednesday, April 7, 2004, at 01:26 PM, Doug Keene wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Hello, } } I am wanting to localize GFP within cultured fibroblasts at } the ultrastructural level (TEM). We are using anti-GFP } antibodies provided by Rockland and another provided by } Assay Design. We have tried a variety of protocols using } prefixation in 0.1% glut and/or rendering the cells } permeable with 0.01% saponin. We've also prefixed the } cells with 100% methanol (this results in reasonable } labeling density but terrible ultrastructure). Our typical } protocol utilizes colloidal gold conjugated secondary } antibodies. } } In any of these protocols I have tried, I am either not } satisfied with the preservation of the cells or I am not } satisfied with labeling density. Certainly not an unusual } quandary for immuno-EM, but surely there is a more } satisfactory protocol than the ones I have tried! } } My question: Does anyone use a TEM protocol for } intracellular labeling that they are happy with? } } Many thanks, } } Doug } } ---------------------- } Douglas R. Keene } Associate Investigator } Micro-Imaging Center } Shriners Hospital for Children } 3101 S.W. Sam Jackson Park Road } Portland, Oregon 97239-3009 } phone: 503-221-3434 } FAX: 503-412-6894 } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 17:11:53 2004
I recommend the Lehigh short course at Lehigh University this June. Check out http:// www.lehigh.edu/microscopy for more information.
Lloyd Williams wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Colleagues: } Our Bio-Imaging facility has merged with our E-M facility and I now have } a Hitachi TEM under my care. Can any one recommend a basic introductory } course in electron microscopy. It would be preferable if it was near New } York City, or at least in the North East. } Thanks } } Lloyd Williams } } Manager Bio-Imaging Facility } Hunter College } 695 Park Ave } New York, NY 10021
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 19:28:39 2004
Sergey; I wish you were right, but I think you can be sure that some knee-jerk bureaucratic bungler will make life miserable for everyone that uses osmium tetroxide in their research. Just remember the weapons of mass deception.....
John Mardinly
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Wednesday, April 07, 2004 2:53 PM To: Microscopy-at-sparc5.microscopy.com
Dear Debby There is no need to discuss (and provoke therefore) the issue, which does not exist as a scientific matter (are we scientists or politicians?). There are plenty of strict regulations which are already
enforced. It's ridiculous, we spent time for such matters. I think, we
should keep this place out of political issues. Unsuccessful OsO4 attack in England IS a political matter, not scientific. Sergey
At 11:39 AM 4/7/2004, you wrote:
} ----------------------------------------------------------------------- ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
c} ------------------------------------------------------------------------------ c} The Microscopy ListServer -- Sponsor: The Microscopy Society of America c} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver c} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html c} -------------------------------------------------------------------------------
c} Hi everyone: c} I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a c} series of holes within micron (or ideally submicron) accuracy on a strip c} of metal that has been laser holes drilled into.
c} I hope to achive this by using the change in z from the top and the c} bottom of hole using a calibrated stage. Does anyone out there have a c} device that they had made or purchased at a reasonable price. Are you c} willing to share your drawings/expertise? c} Thanks, c} Michael Coviello c} UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 02:27:13 2004
does anyone have some informations, or some theory about the charging phenomenon of SEM ?
I know it's because of the secondary electrons yield, producing a dark rectangle on image, when setting a too low voltage (low electron energy) , and a bright one, appearing like a false relief, at a too high voltage (high electron energy). But, is that all?
Where can i find a good bibliography?
thanx.
Sylvain
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 05:53:47 2004
I'm very, very worried about this topic. If this topic is treated as a political matter, it will make the life of researches that work in developmental countries, like me, extremely difficult. We depend on importation to do our research and it is already hard enough to obtain some drugs without the absurd regulations that emerge from time to time due to the politics ignorance and paranoid in what concerns to the scientific matters. It is not rules that will stop terrorists to acquire material that may be used in attacks because these people do not obey rules. They obtain their material in the black market.
As an example, I had to stop working with the effect of aflatoxin in tropical fishes, a serious problem in Brazil, because it became almost impossible to buy the product after some menace of its use by terrorist. Before the application of absurd regulations by the US government, it could take six months to receive the product, now it takes almost two years, after several formularies are filled. Now, I have colleagues that had to agree to received US inspectors in their laboratories asking if they have contact with terrorist groups because they want to buy 10 mg of aflatoxin.
Prof. Dr. Francisco Javier Hernandez Blazquez University of São Paulo Fac. Veterinary Medicine and Animal Sciences Departament of Surgery - Sector of Anatomy Av. Prof. Dr. Orlando Marques de Paiva, 87 05508-000 - São Paulo (SP) - Brasil Tel..55 (11) 3091 1374 Fax 55 (11) 3091 7805 email: fjhblazq-at-usp.br
----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 07, 2004 6:52 PM
Hi, all,
I am looking for an EM and Imaging Lab manager for my lab immediately, at Agriculture and Agri-Food Canada in Kentville, Nova Scotia Canada. Please visit the following website:
http://www.jobs.gc.ca/jobs/p032847e.htm
for more information and instructions for applying.
Kind regards,
Paula.
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 07:14:02 2004
Hi Folks, I tend to agree with Bill Tivol about the use of osmium tetroxide in a 'dirty bomb'. If you consider how reactive the compound is, I would think that the chances of OsO4 actually surviving the delivery explosion in that toxic form would be minimal (a common problem with chemical weapons). It reacts quickly with a variety of things, so the combustion products from an explosion would be likely to change the situation to more of a heavy metal issue than a toxic gas problem. If you look at the way the concept has been presented in the media (especially in the US) there is more emphasis on the fear factor than on the science.
Cheery thoughts for the morning, Robert Simmons
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 07:20:06 2004
Sounds like an AFM [Atomic Force Microscope] may do what you need. The reasonable price part is another matter but you need to define "reasonable."
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: coviello [mailto:coviello-at-mae.uta.edu] Sent: Wednesday, April 07, 2004 6:10 PM To: Microscopy-at-MSA.Microscopy.Com
Hi everyone: I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a series of holes within micron (or ideally submicron) accuracy on a strip of metal that has been laser holes drilled into.
I hope to achive this by using the change in z from the top and the bottom of hole using a calibrated stage. Does anyone out there have a device that they had made or purchased at a reasonable price. Are you willing to share your drawings/expertise? Thanks, Michael Coviello UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 08:05:13 2004
Hi There, I am of need for repair/service on our two PIPS and two DUO-mills from Gatan. Are there any independent service persons that are experts in dealing with these tools? If so, could you please contact me off list to discuss the details. I would like to explore independent route before I ship them to Gatan-Warrendale for repairs.
Thanks,
Jerzy
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 512 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 jerzy.gazda-at-amd.com ******************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 08:15:00 2004
Hi Folks, I tend to agree with Bill Tivol about the use of osmium tetroxide in a 'dirty bomb'. If you consider how reactive the compound is, I would think that the chances of OsO4 actually surviving the delivery explosion in that toxic form would be minimal (a common problem with chemical weapons). It reacts quickly with a variety of things, so the combustion products from an explosion would be likely to change the situation to more of a heavy metal issue than a toxic gas problem. If you look at the way the concept has been presented in the media (especially in the US) there is more emphasis on the fear factor than on the science.
Cheery thoughts for the morning, Robert Simmons
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 08:43:44 2004
Michael, if you can't find a light microscope to suit your needs, most metallographs that I've used have a focus knob with scales graduated down to 1 micron. Coupled with the extremely low depth of field, it can make rather accurate depth measurements by focusing on each plane and measuring the difference. I haven't used it enough to judge the practical accuracy of this technique. It serves us well to measure depth of corrosion pits in metal. Nearly every metallurgical lab has one of these microscopes. You should be able to find one locally.
If this technique doesn't work for you, there's a relatively new instrument developed by my former employer, ERIM International, in Ann Arbor, called a Holomapper. Essentially, it's an expensive instrument that performs two-dimensional surface metrology on parts up to 7" x 7". It does this nearly instantaneously using laser technology with an advertised 0.5 micron uncertainty. This technique would be well-suited to a sample such as yours where you mention all the subject holes are on one sample. One shot collects the information. Analysis would be just a series of mouse clicks using the computer software.
If interested, I can provide contact information. Though ERIM is most interested in selling the equipment, they'd be very willing to direct you to a user to promote wider use of this instrument.
Disclaimer: I have no financial interest in ERIM or its products, I'm just a former employee of the company.
Hi everyone: I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a series of holes within micron (or ideally submicron) accuracy on a strip of metal that has been laser holes drilled into.
I hope to achive this by using the change in z from the top and the bottom of hole using a calibrated stage. Does anyone out there have a device that they had made or purchased at a reasonable price. Are you willing to share your drawings/expertise? Thanks, Michael Coviello UT Arlington
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From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 10:21:04 2004
Boy, is this the question that raises its ugly head. I have on a couple of occassions used zambonies fixative (containing 4% para and picric acid) to fix and immunolabel cells for light and EM. Depending on the antibody you can get very good immunolabeling but the ultrasctructure isn't near as good as if glut was in the mix. I am having trouble remembering if I used saponin or if the picric acid poked sufficient holes to get the labelling done. Can you then fix it hard in EM fix? If I was going into LRWhite to do post embedded labelling the key for improving the ultrastructure for me was to minimize the time spent dehydrating. Due to the light fixation the dehydration extracted a lot of tissue components. I ended up just one minute each, 35%, 50%, 70%, 95% then into 100% for a couple changes and into the infiltration.
However, the next thing I am going to try is light fixation, pelleting, cryoprotection, freezing and ultrathin cryosectioning so the membranes show up. We just got the equipment to do this. If you want to go this direction you are welcome to come up and give it a shot...But not before I get a shot on the new toy. Come to think of it don't you have toys. Have you tried the high pressure freeze and into Lowicryl on these fibroblasts?
Bob Underwood U of Washington Dermatology
On Wed, 7 Apr 2004, Doug Keene wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello, } } I am wanting to localize GFP within cultured fibroblasts at } the ultrastructural level (TEM). We are using anti-GFP } antibodies provided by Rockland and another provided by } Assay Design. We have tried a variety of protocols using } prefixation in 0.1% glut and/or rendering the cells } permeable with 0.01% saponin. We've also prefixed the } cells with 100% methanol (this results in reasonable } labeling density but terrible ultrastructure). Our typical } protocol utilizes colloidal gold conjugated secondary } antibodies. } } In any of these protocols I have tried, I am either not } satisfied with the preservation of the cells or I am not } satisfied with labeling density. Certainly not an unusual } quandary for immuno-EM, but surely there is a more } satisfactory protocol than the ones I have tried! } } My question: Does anyone use a TEM protocol for } intracellular labeling that they are happy with? } } Many thanks, } } Doug } } ---------------------- } Douglas R. Keene } Associate Investigator } Micro-Imaging Center } Shriners Hospital for Children } 3101 S.W. Sam Jackson Park Road } Portland, Oregon 97239-3009 } phone: 503-221-3434 } FAX: 503-412-6894 } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 10:37:24 2004
You might be able to use an AFM (probably a Dimension scope, depending on the size of your strip) in another department at UT-Arlington to measure the holes in your strip. Try engineering, materials science, or the new Institute for Nanoscale Science and Engineering Research and Teaching (INSERT) at UTA to see if there is an AFM on campus.
Bettye Oregon State University
On Apr 8, 2004, at 5:34 AM, Tomic, Peter (Peter) wrote:
} Michael; } } Sounds like an AFM [Atomic Force Microscope] may do what you need. The } reasonable price part is another matter but you need to define } "reasonable." } } Peter Tomic } Agere Systems } Allentown, PA } } -----Original Message----- } } From: coviello [mailto:coviello-at-mae.uta.edu] } Sent: Wednesday, April 07, 2004 6:10 PM } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] LM-measuring depths of holes } } } } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Hi everyone: } I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of } a } series of holes within micron (or ideally submicron) accuracy on a } strip } of metal that has been laser holes drilled into. } } I hope to achive this by using the change in z from the top and the } bottom of hole using a calibrated stage. Does anyone out there have a } device that they had made or purchased at a reasonable price. Are you } willing to share your drawings/expertise? } Thanks, } Michael Coviello } UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 11:07:23 2004
Gee, I would have thought you could have found that much in a can of peanuts off the grocery store shelf!
On 8 Apr 2004 at 8:08, Francisco J. Hernandez Blazquez wrote:
} Now, I have colleagues that } had to agree to received US inspectors in their laboratories asking if } they have contact with terrorist groups because they want to buy 10 mg of } aflatoxin.
All the best,
Andy Buechele, Washington, D.C., U.S.A.
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 12:29:57 2004
Dear Doug, I have done a lot of pre-embedding immuno EM localization of GFP in cultured hippocampal neurons using the microwave and a 5 hour protocol. GFP stands up to glutaradehyde fixation (I use 4% pf and 1% glut in PBS), methanol is too harsh for EM studies.You have to quench the aldehydes with either sodium borohydride or glycine to be able to see your specific fluorescence. I have used anti-GFP from Roche successfully. I permeabilize with 0.025 % to 0.05% saponin -the microwave increases the efficiency of permeabilization. For my secondary, I use Texas Red conjugated fluoronanogold so you can see both the red (anti-GFP) and green fluorescence (GFP) and can check your labeling at the light level before proceeding to EM processing. I silver enhance the nanogold and embed my coverslips in Embed 812 resin. What organelle is the GFP associated with? I will be happy to send you a detailed protocol if you like. Good luck, JoAnn Buchanan
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
Hello, I am wanting to localize GFP within cultured fibroblasts at the ultrastructural level (TEM). We are using anti-GFP antibodies provided by Rockland and another provided by Assay Design. We have tried a variety of protocols using prefixation in 0.1% glut and/or rendering the cells permeable with 0.01% saponin. We've also prefixed the cells with 100% methanol (this results in reasonable labeling density but terrible ultrastructure). Our typical protocol utilizes colloidal gold conjugated secondary antibodies. In any of these protocols I have tried, I am either not satisfied with the preservation of the cells or I am not satisfied with labeling density. Certainly not an unusual quandary for immuno-EM, but surely there is a more satisfactory protocol than the ones I have tried! My question: Does anyone use a TEM protocol for intracellular labeling that they are happy with? Many thanks, Doug
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 12:32:34 2004
You may want to consider doing this through software. We have a product called EFI, other manufacturers have similar products by different names. Essentially, the software acquires images at different focus settings and extracts the focused part. Since the software usually controls the z-motor, each focused piece has a definite height value, and the entire 3-dimensional structure can be extracted. You can then display and measure your object in 3D. The technique does not work for everything, but your application might work. The resolution you can expect is related to the depth of focus of your lens and the number of images you take. With a motorized z-stage, you can do the whole analysis automatically.
If you are interested in getting more information, contact me directly. I'd be more than happy to find out if this works for you, if you can send me a sample.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: coviello [mailto:coviello-at-mae.uta.edu] Sent: Wednesday, April 07, 2004 16:10 To: Microscopy-at-MSA.Microscopy.Com
Hi everyone: I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a series of holes within micron (or ideally submicron) accuracy on a strip of metal that has been laser holes drilled into.
I hope to achive this by using the change in z from the top and the bottom of hole using a calibrated stage. Does anyone out there have a device that they had made or purchased at a reasonable price. Are you willing to share your drawings/expertise? Thanks, Michael Coviello UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 12:44:52 2004
On Apr 8, 2004, at 9:19 AM, Andrew Buechele wrote:
} Gee, I would have thought you could have found that much in a can } of peanuts off the grocery store shelf! } } On 8 Apr 2004 at 8:08, Francisco J. Hernandez Blazquez wrote: } } } Now, I have colleagues that } } had to agree to received US inspectors in their laboratories asking if } } they have contact with terrorist groups because they want to buy 10 } } mg of } } aflatoxin. } } Dear Andrew, 10 mg is a lot of aflatoxin; I'd switch markets. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 14:02:14 2004
Sylvain: I recommend you get yourself the following two books:
Scanning Electron Microscopy: A Student's Handbook by Postek, Howard, Johnson, McMichael Ladd Research (I got my copy from Ladd, but I think other vendors carry it.)
Scanning Electron Microscopy and X-Ray Microanalysis, 3rd edition, by Goldstein, Newbury, Joy, Lyman, et. al. Kluwer academic Press (You can get this one from Kulwer or MSA)
Both of these will answer many questions for you.
sylvain maury wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } hi everyone! } } does anyone have some informations, or some theory about the charging } phenomenon of SEM ? } } I know it's because of the secondary electrons yield, producing a dark } rectangle on image, when setting a too low voltage (low electron energy) } , and a bright one, appearing like a false relief, at a too high voltage } (high electron energy). But, is that all? } } Where can i find a good bibliography? } } thanx. } } Sylvain
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 14:16:08 2004
Does this method work on "soft" surfaces. For example, what does the system do if there are no nice crisp edges to focus on or if there is poor contrast? If at the bottom of Michael's hole or indentation the surface is like a hemisphere, would the system have a tough time hunting for a surface? I've had a similar imaging/measurement problem but AFM resolved the issue within angstroms and without any optical issues like depth of field or sample tilt relative to the objective lens.
Peter Agere Systems
-----Original Message----- } From: Mike Bode [mailto:mb-at-soft-imaging.com] Sent: Thursday, April 08, 2004 1:47 PM To: Microscopy-at-MSA.Microscopy.Com
Michael,
You may want to consider doing this through software. We have a product called EFI, other manufacturers have similar products by different names. Essentially, the software acquires images at different focus settings and extracts the focused part. Since the software usually controls the z-motor, each focused piece has a definite height value, and the entire 3-dimensional structure can be extracted. You can then display and measure your object in 3D. The technique does not work for everything, but your application might work. The resolution you can expect is related to the depth of focus of your lens and the number of images you take. With a motorized z-stage, you can do the whole analysis automatically.
If you are interested in getting more information, contact me directly. I'd be more than happy to find out if this works for you, if you can send me a sample.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: coviello [mailto:coviello-at-mae.uta.edu] Sent: Wednesday, April 07, 2004 16:10 To: Microscopy-at-MSA.Microscopy.Com
Hi everyone: I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a series of holes within micron (or ideally submicron) accuracy on a strip of metal that has been laser holes drilled into.
I hope to achive this by using the change in z from the top and the bottom of hole using a calibrated stage. Does anyone out there have a device that they had made or purchased at a reasonable price. Are you willing to share your drawings/expertise? Thanks, Michael Coviello UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 18:37:12 2004
as I had mentioned in my posting, the method does not work for everything. Transparent samples are of course an issue, and you also have to be able to "look into" the structure, which rules out holes deeper than the working distance of your lens. A hemispherical indentation should not really be a problem, unless it is very smooth and featureless.
I think, soft materials are one example where the optical method has some advantages, as they work without touching the sample, while with an AFM type instrument you "drag" a tip across the surface, and you might have problems with soft materials. The optical technique does not really rely on crisp edges, but rather on some contrast from the material. I don't want to go into a description of the technique here, but if you are interested, you can go to our web site www.soft-imaging.com, then go to Products-"Add-ins"-efi and you'll find an explanation.
An AFM type of instrument can definitely give you results with a much higher resolution (nm), but I understood from Michael's post, that a) he wants to use a microscope, and b) he's looking at micron resolution, not nm. I am also not sure what happens if the holes are tens or hundreds of microns deep, and if you can use an AFM for those aspect ratios.
Otherwise I agree with you, of course. An AFM type instrument is certainly an option that can give you the same or in many cases better results.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Sent: Thursday, April 08, 2004 13:31 To: Mike Bode; Microscopy-at-MSA.Microscopy.Com
Mike;
Does this method work on "soft" surfaces. For example, what does the system do if there are no nice crisp edges to focus on or if there is poor contrast? If at the bottom of Michael's hole or indentation the surface is like a hemisphere, would the system have a tough time hunting for a surface? I've had a similar imaging/measurement problem but AFM resolved the issue within angstroms and without any optical issues like depth of field or sample tilt relative to the objective lens.
Peter Agere Systems
-----Original Message----- } From: Mike Bode [mailto:mb-at-soft-imaging.com] Sent: Thursday, April 08, 2004 1:47 PM To: Microscopy-at-MSA.Microscopy.Com
Michael,
You may want to consider doing this through software. We have a product called EFI, other manufacturers have similar products by different names. Essentially, the software acquires images at different focus settings and extracts the focused part. Since the software usually controls the z-motor, each focused piece has a definite height value, and the entire 3-dimensional structure can be extracted. You can then display and measure your object in 3D. The technique does not work for everything, but your application might work. The resolution you can expect is related to the depth of focus of your lens and the number of images you take. With a motorized z-stage, you can do the whole analysis automatically.
If you are interested in getting more information, contact me directly. I'd be more than happy to find out if this works for you, if you can send me a sample.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: coviello [mailto:coviello-at-mae.uta.edu] Sent: Wednesday, April 07, 2004 16:10 To: Microscopy-at-MSA.Microscopy.Com
Hi everyone: I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a series of holes within micron (or ideally submicron) accuracy on a strip of metal that has been laser holes drilled into.
I hope to achive this by using the change in z from the top and the bottom of hole using a calibrated stage. Does anyone out there have a device that they had made or purchased at a reasonable price. Are you willing to share your drawings/expertise? Thanks, Michael Coviello UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 19:40:52 2004
We've been discussing how it costs us $5,000 U.S. to dispose of a liter of fluid with a gram or two of uranium in it while over in Iraq the U.S. is dumping tons of the stuff.
There's compliance here and compliance there...
-Michael
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 9 00:00:21 2004
I am a private consultant which specializes in training in all phase of electron microscopy. If you would like to have me come to your facility and train you on your equipment, please contact me to discuss details.
Bill McManus President Mt Ogden Scientific Services LLC moss-at-cut.net 435-946-8739
} From: "Lloyd Williams" {Williams-at-genectr.hunter.cuny.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Subject: [Microscopy] Course To Learn Basics of TEM } Date: Wed, 7 Apr 2004 12:48:35 -0400 } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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From MicroscopyL-request-at-ns.microscopy.com Fri Apr 9 10:08:10 2004
I am trying to locate a manual for a Forma Bio-Freezer. Model 8407. It is a chest type -85 freezer. If any one has such could I get a copy?
Greg
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Sat Apr 10 04:47:34 2004
I have just spotted your question and hope I am able to provide a simple explanation?
1. It is not correct to say that an accelerating voltage is too low when viewing an image at a single kV. The only time that this comment may be justified is when, having increased the accelerating voltage, you see the subsurface detail that you desired . In another case when considering EDS the accelerating voltage may be too low to stimulate the specific peak that you may desire. I do not believe there is such a situation that a kV is too low, but of course it may be too low to display the information that you require. In general most operators of non FEG instruments run at too high a kV to truly resolve the specimen surface (i.e. } 5kV)! There are other interesting reasons for contrast changes within images at very low voltages, these relate to secondary electron emission coefficients and are well presented by N.R.Whetton in Methods of Experimental Physics, Vol IV (1962)
2. The dark patch on the image at low accelerating voltages is due to contamination that has deposited on the specimen surface. The contamination is invariably a lower emitter of electrons, thus it shows up as a dark patch or line. You do not see this at a higher kV because the additional voltage causes the beam to penetrate further into the specimen, the sub surface information generated dominating the image hiding the "surface" contamination from your view. Thus to see contamination is an indication that you are seeing the "true surface" of the specimen.
3. Charge on the specimen surface is due to an insufficient earth leakage path in relation to the incident beam current and you are correct in the belief that this may give rise to a bright square on the image.
4. Another visualisation of charge is the bright particle with a black halo around it. Here the charge field that has built up around the particle is preventing the low energy SE escaping, whilst if you look closely the high energy BSE do escape and provide information within the charge (black) cloud.
5. In a charge-discharge situation, often seen on slow scans, the image will dim (through a reduction in SE emitted) as the specimen charges, flashing bright when the system discharges due to the sudden freeing of the SE held under the charge. Thus a progressively darkening area in an image indicates charge, the bright flash across one or more lines indicates discharge!
Good luck with your microscopy.
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
----- Original Message ----- } From: "sylvain maury" {sylvain.maury-at-thalesgroup.com} To: "Microscopy community" {microscopy-at-msa.microscopy.com} Sent: Thursday, April 08, 2004 8:42 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mahtab977-at-yahoo.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, April 9, 2004 at 11:59:09 ---------------------------------------------------------------------------
Question: I'm working on SEMs of bacterial cells attached to various surfaces (eg glass and chitin) and I've been successfully dehydrating my specimens using the basic CPD/CO2 procedure. However I am limited to the number of CPD runs I have time and $$ for. A labmate suggested I try actetone dehydration....is acetone a worthy alternative?
I think one should be grateful rather then sarcastic to anybody that takes seriously any potential threat of terrorism, small and remote as it may be. At least I am, whenever the guard in front of Pizza-Hut, or the movies, or a bus looks into my pockets.
And as Bill Tivol says, 10mg is sometimes a lot of material.
Dr. Uri Admon Israel
----- Original Message ----- } From: "Andrew Buechele" {andrewb-at-vsl.cua.edu} To: {Microscopy-at-sparc5.microscopy.com} ; "Sergey Ryazantsev" {sryazant-at-ucla.edu} Sent: Thursday, April 08, 2004 5:19 PM
Mahtab,
Acetone can work, and so can ethanol, or freeze-drying, or even air, sometimes. The other way is to use hexamethylsilizane (HMDS). This works very well for most bacteria. Procedures and more information can be found in Microscopy Today ( http://www.microscopy-today.com for a table of contents of back issues) and on the Univ. Florida "Tips and Tricks" web site http://www.biotech.ufl.edu/EM/tips/index.html . But briefly: after the final absolute ethanol step, transfer the samples through an EtOH:HMDS series to 100% HMDS. 2:1, 1:2 EtOH:HMDS, 3 X 100% HMDS usually works. Then dry either at room temperature or at 60 deg C. All this IN A FUME HOOD. You do NOT want to breathe HMDS. The rest of the details depend on things like: are the bacteria in suspension or on filters and so forth. Contact me if you have further questions.
Phil
} Email: mahtab977-at-yahoo.com } Name: Mahtab Shahkarami } } Organization: San Jose State University } } Education: Graduate College } } Location: San Jose, CA USA } } Question: I'm working on SEMs of bacterial cells attached to various } surfaces (eg glass and chitin) and I've been successfully } dehydrating my specimens using the basic CPD/CO2 procedure. However } I am limited to the number of CPD runs I have time and $$ for. A } labmate suggested I try actetone dehydration....is acetone a worthy } alternative? } } ---------------------------------------------------------------------------
-- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Sun Apr 11 21:25:44 2004
Francisco J. Hernandez Blazquez wrote: ===================================================== } I'm very, very worried about this topic. If this topic is treated as a } political matter, it will make the life of researches that work in } developmental countries, like me, extremely difficult. We depend on importation } to do our research and it is already hard enough to obtain some drugs without } the absurd regulations that emerge from time to time due to the politics } ignorance and paranoid in what concerns to the scientific matters. It is not } rules that will stop terrorists to acquire material that may be used in attacks } because these people do not obey rules. They obtain their material in the black } market. } } As an example, I had to stop working with the effect of aflatoxin in tropical } fishes, a serious problem in Brazil, because it became almost impossible to buy } the product after some menace of its use by terrorist. Before the application } of absurd regulations by the US government, it could take six months to receive } the product, now it takes almost two years, after several formularies are } filled. Now, I have colleagues that had to agree to received US inspectors in } their laboratories asking if they have contact with terrorist groups because } they want to buy 10 mg of aflatoxin. } =============================================================== I would respectfully like to point out that the matter of export controls over certain potentially dangerous materials is not the result of "absurd regulations" by the US government but the result of consensus agreements through the UN, the IAEA, IATA and other world bodies to which Brazil is also a signor. These regulations may indeed be absurd, but your complaint is with these world bodies, not specifically the US government!
The people who have restricted your access to aflatoxin are the same type of people who have mandated that a $30 bottle of silver paint, something no more "dangerous" than a woman's fingernail polish remover, has to be shipped (to certain countries, including Brazil) by air freight (for several hundred dollars) as "dangerous goods" instead of via FedEx or UPS for a fraction of the amount (as it could be shipped for example, to Japan or to most of the EC countries).
Beryllium TEM grids are restricted for shipment to certain countries because some bureaucrat thought that some rogue nation might purchase a large enough number of beryllium grids to melt down the metal and turn it into some critical part for a nuclear reactor. But these are regulations not of the US government but of one or more of these world bodies. The list of examples is endless.
We as an industry have let this happen because too many of us want to "stay out of politics" but when we think that way, the bureaucrats who have nothing better to do with their time, saddle us with these kinds of onerous situations. I would suggestion we do just the opposite: We have to become politically involved and lobby our legislators to bring some semblance of sanity back to the shipping (and export) regulations. In our own small way , we invite to our company every few years either one of our representatives in Harrisburg or in Washington, DC and explain how government regulations are holding us back from growing and innovating even more. It does have an impact. But this is only a drop in the bucket in terms of what really has to be done in terms of lobbying. Everyone has to be doing it! Worldwide.
In the mean time, so long as researchers using these materials are not going to be willing to keep such materials under some system of strict accountability, with the reporting and explaining of any unaccountable losses, the future would seem to be one where there will indeed be more and more regulation of these materials which will add to everyone's cost of doing research.
I apologize to anyone who might think this an inappropriate topic for discussion on the listserver.
Chuck ============================================ Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Sun Apr 11 22:34:03 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (John_Wahlert-at-baruch.cuny.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Sunday, April 11, 2004 at 19:48:44 ---------------------------------------------------------------------------
Email: John_Wahlert-at-baruch.cuny.edu Name: John H. Wahlert
Organization: Baruch College, CUNY
Education: Graduate College
Location: New York, New York, USA
Question: I would like to repair some old slides of lamprey cross sections. The glue has turned white all around the specimens. Where can I find information about renewing old slides that cannot be replaced?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (akoorts-at-medic.up.ac.za) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, April 12, 2004 at 02:20:53 ---------------------------------------------------------------------------
Question: I am performing immunolocalization and in situ hybridization on ultrathin sections (LR White) of core bone marrow biopsies. I am interested in ferritin subunit expression in the macrophage. It seems as though either the vacuolar contents are extracted or the sections are torn in the vacuolar areas when ultrathin sections are cut. (Fixed: 4% paraformaldehyde, 0.05% glutaraldehyde) Is there perhaps someone that have encountered these problems and are able to make some comments. Any advice would be much appreciated.
University of Connecticut Institute of Materials Science
Staff Position in Transmission Electron Microscopy
The Institute of 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. The Microscopy Laboratory is a user facility which houses the main research microscopes (optical, SEM and TEM) for the IMS. These include a recently commissioned JEOL 2010 UHR FasTEM equipped with EDAX UTW EDS and Gatan EELS/image filter. There is an opening in the Laboratory for a Transmission Electron Microscopy specialist. The appointee will be involved in a range of academic and industrial projects, and will assist in the operation of the Laboratory including performing routine maintenance and training/assisting users of the TEMs.
Candidates should hold a higher degree (MS or PhD) in Materials Science, Physics or a related discipline and must have extensive hands-on experience in a broad range of TEM techniques. Experience in instrument development and/or computer image processing/simulation would be beneficial. This is a fixed-term appointment and is available from July 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 persons 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: Prof. M. Aindow, Institute of Materials Science, University of Connecticut, 97 North Eagleville Road, U-3136, Storrs, CT 06269-3136 USA. Email: m.aindow-at-uconn.edu
Mark Aindow, Associate Professor, Department of Metallurgy and Materials Engineering Institute of Materials Science, 97 North Eagleville Road, Unit 3136 University of Connecticut, Storrs, CT 06269-3136, USA
I expect to have a vacancy, starting immediately (more or less) for a post doc (up to three years) to work on EBSD in the SEM. The ideal candidate will have prior experience of three kinds: experimental electron microscopy, dynamical diffraction theory and programming. Although the project is in SEM, experimental experience in TEM would be equally suitable. The aim of the work is to advance the technique of EBSD. Prior work has advanced the ability of EBSD to measure strain. Recent work has shed light on the formation of EBSD patterns through energy filtering. The next steps are a) to further the use of EBSD to measure strain and to apply it to electromigration, b) to develop better spatial resolution in EBSD through energy filtering and c) to write software to simulate EBSD patterns.
Candidates interested in this post please contact me. If you have sent me email enquiring about a job and have got a reply saying I have no vacancies (which was true at the time), please feel free to send your details again - I have deleted the information you sent previously.
Alwyn Eades -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 12 08:01:58 2004
} I am limited to the number of CPD runs I have time and $$ for. A } labmate suggested I try actetone dehydration....is acetone a worthy } alternative? } } --------------------------------------------------------------------------- NO. You may have good success using Hexamethyldisilazane (HMDS). Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 12 08:11:56 2004
The woman who shares my lab space is the caretaker of the histology slide collection for the medical students here. She has about 250 sets of slides in her care. At the end of each course, when the boxes are turned in, she goes through them all and selects out those whose mounting medium has dried out, clouding the specimen as you described. She has been very successful in refurbishing these slides simply by soaking them (sometimes for days) in a staining jar filled with xylene. At some point, the old coverglass drops off. If the staining has faded, she restains them, then remounts them with fresh media and a new coverglass. Most of these slides were originally mounted with either Canada Balsam or Permount, and she stills prefers Permount to remount them. Try it with one or two slides. Good luck, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 12 12:54:36 2004
I have a client that is having difficulty keeping their fly heads in their thin sections. The chiton tends to seperate from the surrounding plastic. The head is taken off the fly, probosis removed and then processed more or less using a typical proceedure into epoxy.
any ideas.
Robert J. Kayton, Ph.D. C.R.O.E.T., L606 Oregon Health & Science Univ. kayton-at-ohsu.edu 503-494-2504-Office 503-703-3938-Cell www.ohsu.edu/croet/facilities/emicroscopy
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 12 13:31:05 2004
Chuck; Excellent note, but there is an even more insidious thing coming in the way of export controls: I had to have an investigation to determine if hiring a summer intern (in the US) who happened to be from China (a Ph.D. student at Berkeley) required an export license. Being student from China is a category that covers a lot of people, and when the legality of hiring somebody can be covered by export control law, these bureaucratic tentacles have extended too far.
John Mardinly Intel
-----Original Message----- } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] Sent: Sunday, April 11, 2004 8:44 PM To: MICROSCOPY BB
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Francisco J. Hernandez Blazquez wrote: ===================================================== } I'm very, very worried about this topic. If this topic is treated as a } political matter, it will make the life of researches that work in } developmental countries, like me, extremely difficult. We depend on importation } to do our research and it is already hard enough to obtain some drugs without } the absurd regulations that emerge from time to time due to the politics } ignorance and paranoid in what concerns to the scientific matters. It is not } rules that will stop terrorists to acquire material that may be used in attacks } because these people do not obey rules. They obtain their material in the black } market. } } As an example, I had to stop working with the effect of aflatoxin in tropical } fishes, a serious problem in Brazil, because it became almost impossible to buy } the product after some menace of its use by terrorist. Before the application } of absurd regulations by the US government, it could take six months to receive } the product, now it takes almost two years, after several formularies are } filled. Now, I have colleagues that had to agree to received US inspectors in } their laboratories asking if they have contact with terrorist groups because } they want to buy 10 mg of aflatoxin. } =============================================================== I would respectfully like to point out that the matter of export controls over certain potentially dangerous materials is not the result of "absurd regulations" by the US government but the result of consensus agreements through the UN, the IAEA, IATA and other world bodies to which Brazil is also a signor. These regulations may indeed be absurd, but your complaint is with these world bodies, not specifically the US government!
The people who have restricted your access to aflatoxin are the same type of people who have mandated that a $30 bottle of silver paint, something no more "dangerous" than a woman's fingernail polish remover, has to be shipped (to certain countries, including Brazil) by air freight (for several hundred dollars) as "dangerous goods" instead of via FedEx or UPS for a fraction of the amount (as it could be shipped for example, to Japan or to most of the EC countries).
Beryllium TEM grids are restricted for shipment to certain countries because some bureaucrat thought that some rogue nation might purchase a large enough number of beryllium grids to melt down the metal and turn it into some critical part for a nuclear reactor. But these are regulations not of the US government but of one or more of these world bodies. The list of examples is endless.
We as an industry have let this happen because too many of us want to "stay out of politics" but when we think that way, the bureaucrats who have nothing better to do with their time, saddle us with these kinds of onerous situations. I would suggestion we do just the opposite: We have to become politically involved and lobby our legislators to bring some semblance of sanity back to the shipping (and export) regulations. In our own small way , we invite to our company every few years either one of our representatives in Harrisburg or in Washington, DC and explain how government regulations are holding us back from growing and innovating even more. It does have an impact. But this is only a drop in the bucket in terms of what really has to be done in terms of lobbying. Everyone has to be doing it! Worldwide.
In the mean time, so long as researchers using these materials are not going to be willing to keep such materials under some system of strict accountability, with the reporting and explaining of any unaccountable losses, the future would seem to be one where there will indeed be more and more regulation of these materials which will add to everyone's cost of doing research.
I apologize to anyone who might think this an inappropriate topic for discussion on the listserver.
Chuck ============================================ Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 12 19:06:58 2004
We use Cytovision(v2.75) software by Applied Imaging to detect the ScFISH probes on metaphase chromosomes, while capturing the probe signal it has an option in the software to increase the exposure time of the camera . Usually, we have to increase the exposure time of the camera to be able to see the ScFISH probes.
Currently, we are working on developing an automated microscopy system, for that I need to figure out, how to increase the exposure time of the camera using the following system configuration.
Operating system : Windows NT ICPCI frame grabber card (Coreco Imaging Inc.,) COHU camera ( 4912-5010) ITEX-ICPCI (v3.1.0.0) software
Does anybody have an idea on how to control the exposure time of the camera using ITEX-ICPCI (v3.1.0.0) software. I guess it should be on the on lines of triggered acquisition, but I dont have much idea on how to do it. Please reply me with your valuable suggestions.
Thanks in advance for your help.
Regards, Pavan Yanala pkymw5-at-umkc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 12 23:11:25 2004
Add 120 metric tons of depleted uranium spread over Kosovo in former Yugoslavia by American forces. Sergey
At 06:56 PM 4/8/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Charles USA has veto in UN, if they don't like something, they veto it, so US share complete responsibility for every exportation international law. From another hand, I don't see much willingness from US to cooperate with UN in many cases (like Iraq), so there is definitely double standards: US smartly use UN to "cover up" unpopular decisions (like discrimination in export) if it's in favor of US, otherwise they simply ignore UN resolutions, international laws and perform on their own. Your posting is too politically correct to be interesting. I am sorry. This communication reminds to me the worse time at Soviet Union, when you have to be very politically correct... what is happening with freedom of speech in this country and does this country is trying to reproduce mistakes of USSR? In the beginning of this discussion, I was trying to express the opinion, that our forum should be out of politics, nobody supports me, so I decided to express my opinion as a person who could compare and who has a freedom to see things free from political correctness ) I am not a US citizen). If this forum is going to be an arena for political correctness, I'll leave this place immediately, I had enough in USSR. Sergey
At 08:44 PM 4/11/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
we have an inverted microscope Zeiss Axiovert 200 and use the POCMini System from Zeiss. In general we use it with glass slides with a diameter of 30mm to observe cells. The device reaches the adjusted temperature very quickly is very stable during measurements.
Regards, Mario
Mario Brameshuber Single Dye Tracing Institute for Biophysics Johannes Kepler University of Linz Altenbergerstra?e 69, 4040 Linz Austria phone: +43 732 2468 9288
-----Ursprungliche Nachricht----- Von: drk-at-SHCC.org [mailto:drk-at-SHCC.org] Gesendet: Mittwoch, 07. April 2004 19:09 An: Microscopy-at-sparc5.microscopy.com Betreff: Heated stage for LM
---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
We have Leica inverted (DMIRE2) and upright (DMRXA2) microscopes in our Confocal facility that we would like to add live cell imaging capabilities to. At the moment, a heating stage would be adequate. If you are using such a stage (even on a different brand microscope) and are really happy with it, would you please share the manufacturer with me?
Thank you in advance,
Doug
---------------------- Douglas R. Keene Associate Investigator Micro-Imaging Center Shriners Hospital for Children 3101 S.W. Sam Jackson Park Road Portland, Oregon 97239-3009 phone: 503-221-3434 FAX: 503-412-6894
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 13 07:45:18 2004
While this started out as a discussion on the problems getting Osmium due to various exportation rules (which I had no problem with discussing), it is now starting to wander off topic.
Regardless of how one feels about politics in the world, that subject does not belong on this forum. There are a number of open forums on the Internet that you can us for these purposes. This is not one of them.
It is time to end this thread , and let us return to the subject at hand namely: Microscopy & Microanalysis.
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 13 08:27:23 2004
Quoting Robert Kayton {kayton-at-ohsu.edu} : } I have a client that is having difficulty keeping their fly heads in } their thin sections. The chiton tends to seperate from the surrounding } plastic. The head is taken off the fly, probosis removed and then } processed more or less using a typical proceedure into epoxy. } } Robert J. Kayton, Ph.D. } C.R.O.E.T., L606 } Oregon Health & Science Univ. } kayton-at-ohsu.edu } 503-494-2504-Office } 503-703-3938-Cell } www.ohsu.edu/croet/facilities/emicroscopy
Robert, I have had this same problem with pupae and adults, and have tried many things but nothing worked well. The best results were obtained by doing all the trimming with a glass knife, taking only thin sections off the block at a time, less pressure at the epon/cuticle interface. The cuticle at the bottom of the block should sectioned off so that the knife hits tissue.
In that the head is "lumpy", the first sections will still tend to pop out of the plastic. If there is a need to look at these, it is helpful to collect the thick sections on water, either in a boat or a drop of water at the edge of the glass knife and pick up the material with a loupe like EMS sells. I keep the water at the top of the knife by drawing a line of finger nail polish (old and cheap are fine) across the knife about 5 mm from the top edge. The polish sticks much better than wax. Time must be allowed to let the polish dry, so I do several at a time usually after I break the knives so they will be ready when I need them.
As the material gets large enough to pick up with a hair, I just transfer that to the slide and forget the excess plastic.
I am looking forward to others suggestions also.
Pat Connelly Dept of Biology Univ. of Pennsylvania Philadelphia, PA psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 13 08:27:24 2004
Quoting Robert Kayton {kayton-at-ohsu.edu} : } I have a client that is having difficulty keeping their fly heads in } their thin sections. The chiton tends to seperate from the surrounding } plastic. The head is taken off the fly, probosis removed and then } processed more or less using a typical proceedure into epoxy. } } Robert J. Kayton, Ph.D. } C.R.O.E.T., L606 } Oregon Health & Science Univ. } kayton-at-ohsu.edu } 503-494-2504-Office } 503-703-3938-Cell } www.ohsu.edu/croet/facilities/emicroscopy
Robert, I have had this same problem with pupae and adults, and have tried many things but nothing worked well. The best results were obtained by doing all the trimming with a glass knife, taking only thin sections off the block at a time, less pressure at the epon/cuticle interface. The cuticle at the bottom of the block should sectioned off so that the knife hits tissue.
In that the head is "lumpy", the first sections will still tend to pop out of the plastic. If there is a need to look at these, it is helpful to collect the thick sections on water, either in a boat or a drop of water at the edge of the glass knife and pick up the material with a loupe like EMS sells. I keep the water at the top of the knife by drawing a line of finger nail polish (old and cheap are fine) across the knife about 5 mm from the top edge. The polish sticks much better than wax. Time must be allowed to let the polish dry, so I do several at a time usually after I break the knives so they will be ready when I need them.
As the material gets large enough to pick up with a hair, I just transfer that to the slide and forget the excess plastic.
I am looking forward to others suggestions also.
Pat Connelly Dept of Biology Univ. of Pennsylvania Philadelphia, PA psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 13 08:33:22 2004
I have followed with some interest the line of discussion re: osmium tetroxide. I say some, because the discussion went from one subscriber sharing the news that Osmium Tetroxide was going to become strictly regulated to a great heated political debate.
I believe in freedom of speech, but I do not think the ListServer is a forum for such discussion. Let's leave the politics out of this server and keep to the facts. The issue of Osmium Tetroxide has been more than discussed and unless someone has some real facts re: the regulation of it's usage, please communicate among yourselves.
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 13 08:48:58 2004
I had a similar problem with fruit fly legs. Two possible solutions (which I could not try in the study I was involved with) are 1. treating with a chitinase or 2. taking tissue immediately after molting so the chitin is still relatively soft. Also check out papers in Nature 184:1584, 1959 (Beckel) and Trans. Am. Micros. Soc. 74:197-201, 1955 (DeGiusti and Ezman).
Geoff
Robert Kayton wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 13 08:53:31 2004
Announcing a 5 day intensive AFM short course entitled "AFM and Other Scanned Probe Microscopies". It is being taught at N.C. State University in Raleigh, NC by Prof. Phil Russell, Dr. Joe Griffith and others from June 21 -25, 2004.
This one-week short course on atomic force microscopy has evolved from the numerous Scanned Probe Microscopy courses developed and taught by Prof. Russell over the past 2 decades. It is designed for technicians, scientists, engineers and researchers. The course includes lectures and laboratories with hands-on time using a variety of scanning probe microscope (SPM) systems.
For more information got to www.ncsu.edu/aif/afmcourse
I think quite a few people supported your initial attempt to keep politics out of this forum. There are other newsgroups for that sort of thing.
Lesley Weston.
on 12/04/2004 9:51 PM, Sergey Ryazantsev at sryazant-at-ucla.edu wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Charles } USA has veto in UN, if they don't like something, they veto it, so US share } complete responsibility for every exportation international law. From } another hand, I don't see much willingness from US to cooperate with UN in } many cases (like Iraq), so there is definitely double standards: US smartly } use UN to "cover up" unpopular decisions (like discrimination in export) if } it's in favor of US, otherwise they simply ignore UN resolutions, } international laws and perform on their own. Your posting is too } politically correct to be interesting. I am sorry. This communication } reminds to me the worse time at Soviet Union, when you have to be very } politically correct... what is happening with freedom of speech in this } country and does this country is trying to reproduce mistakes of USSR? In } the beginning of this discussion, I was trying to express the opinion, that } our forum should be out of politics, nobody supports me, so I decided to } express my opinion as a person who could compare and who has a freedom to } see things free from political correctness ) I am not a US citizen). If } this forum is going to be an arena for political correctness, I'll leave } this place immediately, I had enough in USSR. Sergey } } At 08:44 PM 4/11/2004, you wrote: } } } } ---------------------------------------------------------------------------- -} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------------- } } -- } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } Francisco J. Hernandez Blazquez wrote: } } ===================================================== } } } I'm very, very worried about this topic. If this topic is treated as a } } } political matter, it will make the life of researches that work in } } } developmental countries, like me, extremely difficult. We depend on } } importation } } } to do our research and it is already hard enough to obtain some drugs } } without } } } the absurd regulations that emerge from time to time due to the politics } } } ignorance and paranoid in what concerns to the scientific matters. It is } } not } } } rules that will stop terrorists to acquire material that may be used in } } attacks } } } because these people do not obey rules. They obtain their material in the } } black } } } market. } } } } } } As an example, I had to stop working with the effect of aflatoxin in } } tropical } } } fishes, a serious problem in Brazil, because it became almost impossible } } to buy } } } the product after some menace of its use by terrorist. Before the } } application } } } of absurd regulations by the US government, it could take six months to } } receive } } } the product, now it takes almost two years, after several formularies are } } } filled. Now, I have colleagues that had to agree to received US inspectors } } in } } } their laboratories asking if they have contact with terrorist groups } } because } } } they want to buy 10 mg of aflatoxin. } } } } } =============================================================== } } I would respectfully like to point out that the matter of export controls } } over certain potentially dangerous materials is not the result of "absurd } } regulations" by the US government but the result of consensus agreements } } through the UN, the IAEA, IATA and other world bodies to which Brazil is } } also a signor. These regulations may indeed be absurd, but your complaint } } is with these world bodies, not specifically the US government! } } } } The people who have restricted your access to aflatoxin are the same type of } } people who have mandated that a $30 bottle of silver paint, something no } } more "dangerous" than a woman's fingernail polish remover, has to be shipped } } (to certain countries, including Brazil) by air freight (for several } } hundred dollars) as "dangerous goods" instead of via FedEx or UPS for a } } fraction of the amount (as it could be shipped for example, to Japan or to } } most of the EC countries). } } } } Beryllium TEM grids are restricted for shipment to certain countries because } } some bureaucrat thought that some rogue nation might purchase a large enough } } number of beryllium grids to melt down the metal and turn it into some } } critical part for a nuclear reactor. But these are regulations not of the } } US government but of one or more of these world bodies. The list of } } examples is endless. } } } } We as an industry have let this happen because too many of us want to "stay } } out of politics" but when we think that way, the bureaucrats who have } } nothing better to do with their time, saddle us with these kinds of onerous } } situations. I would suggestion we do just the opposite: We have to become } } politically involved and lobby our legislators to bring some semblance of } } sanity back to the shipping (and export) regulations. In our own small way } } , we invite to our company every few years either one of our representatives } } in Harrisburg or in Washington, DC and explain how government regulations } } are holding us back from growing and innovating even more. It does have an } } impact. But this is only a drop in the bucket in terms of what really has } } to be done in terms of lobbying. Everyone has to be doing it! Worldwide. } } } } In the mean time, so long as researchers using these materials are not going } } to be willing to keep such materials under some system of strict } } accountability, with the reporting and explaining of any unaccountable } } losses, the future would seem to be one where there will indeed be more and } } more regulation of these materials which will add to everyone's cost of } } doing research. } } } } I apologize to anyone who might think this an inappropriate topic for } } discussion on the listserver. } } } } Chuck } } ============================================ } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } } President 1-800-2424-SPI } } SPI SUPPLIES FAX: 1-610-436-5755 } } PO BOX 656 e-mail:cgarber-at-2spi.com } } West Chester, PA 19381-0656 USA } } Cust.Service: spi2spi-at-2spi.com } } } } Look for us! } } ######################## } } WWW: http://www.2spi.com } } ######################## } } ============================================ } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } 10833 Le Conte Ave, Room 33-089 } Los Angeles, CA 90095 } } Phone: (310) 825-1144 (office) } (310) 206-1029 (Lab) } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } }
-- Lesley Weston.
3512 W. 31st Avenue Vancouver, B.C. V6S 1X9
(604) 263 3122
lesley-at-vancouverbc.net
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 14 09:01:06 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bwareham-at-utah.gov) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, April 12, 2004 at 16:25:49 ---------------------------------------------------------------------------
Email: bwareham-at-utah.gov Name: Beverly Wareham
Organization: Utah Veterinary Diagnostic Lab
Title-Subject: [Microscopy] [Filtered] TEM film Development
Question: I am looking for a facility that would be willing to develop Kodak 4489 film. Please include pricing and turn around time. Thank you, Beverly Wareham
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ottagonosole-at-tiscali.it) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 14, 2004 at 08:54:22 ---------------------------------------------------------------------------
Email: ottagonosole-at-tiscali.it Name: giovanni de caro, md
Organization: associazione bimbononno onlus - italia
Question: We are a no profit organization devoted to youngsters education; we have founded a small volunteer operated natural sciences museum in southern Italy . Please see our website for details on our activities at : http://web.tiscali.it/bimbononno Recently we obtained a dismissed Leitz- AMR 1000 scanning electron microscope, which was in working condition when turned off in november, 2003. we have the manulas and electronic schemes. We are seeking for an expert in the installation and operation of this instrument in order to help us in performing reinstallation and recalibration at our lab.
Thank you
Giovanni De Caro, MD Director Museo laboratorio di Scienze Naturali "S.Eugenio de Mazenod"- Ripalimosani (Campobasso) Italia
Many people have studied drosophila tissues by TEM despite the embedding difficulties. I have done many thin sections of the eye and the epoxy usually separates from the eye surface boundary. I just collected the section parts that contain the tissue onto a grid with or without a film support. If you need to examine areas near the surface boundary a film support is usually necessary.
Larry Ackerman Keck Advanced Microscopy Lab Dept. of Biochemistry & Biophysics University of California San Francisco 600-16th Street, Room S101, Box 2140 San Francisco, CA 94158 (for postal mail use 94143)
415-476-4441 FAX 415-514-4142
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 14 14:34:22 2004
Well, I got caught unexpectedly with a lab tour and was asked about cutting thin sections. I later found out the thickness I "guesstimated" would cut a nitrogen atom in half. So how thin can a section of organic material be cut with a ultra-microtome? Can metals be thinned even thinner/
Thanks for the help and I promise I will not ask about osmium!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 14 18:13:35 2004
I'm having a problem with my CCD camera on the TEM. When working with an unprocessed (not gain or dark count corrected), dark (no beam on the CCD), 1024x1024 image - I get a non-linear horizontal profile of the image. In other words, the intensity stays constant from pixels 0 - 600, then the profile increases linearly from pixel 600 to 1024 (slope ~ 0.3 pixel value/pixel number) from the left to the right of the image. I can rotate the CCD on the optic axis of the TEM, and this problem persists in the same location (increasing from left to right) regardless of the rotation of the camera. This problem is seen in all images, but is more prevalent in images with } 10 sec exposure time.
Has anyone ever seen this problem before? If so, how did you correct for it?
Instruments being used: Camera is a Proscan HSC 2, 1024x1024 on axis cooled high speed slow scan CCD camera on a LEO 912 EFTEM (120 kV with LaB6).
Any help would be SUPER!!
Thanks for your help,
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 02:46:11 2004
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 07:26:01 2004
I am interested in knowing what scanner(s) and negative holders people are using to scan 8 X 10 cm TEM negatives. We are having some problems with our current setup.
Thank you, Maureen Petersen Ohio Agricultural and Research Center/OSU Wooster, Ohio --
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 08:26:47 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (karthins-at-labindia.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, April 15, 2004 at 03:42:37 ---------------------------------------------------------------------------
We're planning to buy a TEM for our Histology lab. Does anybody know web sources for different models, comparing spesifications, prices and reviews. We're especially interested in Hitachi 7600, Jeol JEM-1011 and LEO Libra 128. Does anybody have experience with these microscopes? If you inform us, we'd be greatly appreciated. Thanks in advance
Dr. Necat Yžlmaz
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 08:41:00 2004
I am looking for PC-based image analysis software that can find, count and measure particles. The particles may be either spheres or vesicles, so the software must be able to find/measure/count both types. I would also like the software to report actual diameters as opposed to equivalent sphere diameter, though doing both is acceptable. Finally, I need it to be able to do statistics on the analyses that it performs.
If you can provide the names of some software packages that meet these criteria, as well as the company that makes it and contact information, it would be a great help to me. Thanks!
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 08:49:53 2004
if you rotate the camera and the problem persists in the same direction, it is most likely not the camera that is defective. When you rotate the camera, do you also rotate the scintillator? If so, it cannot be the scintillator either, and it must be the setup of your microscope. Is your microscope well aligned?
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: William Stratton [mailto:wgstratton-at-wisc.edu] Sent: Wednesday, April 14, 2004 17:08 To: Microscopy
Hello all,
I'm having a problem with my CCD camera on the TEM. When working with an unprocessed (not gain or dark count corrected), dark (no beam on the CCD), 1024x1024 image - I get a non-linear horizontal profile of the image. In other words, the intensity stays constant from pixels 0 - 600, then the profile increases linearly from pixel 600 to 1024 (slope ~ 0.3 pixel value/pixel number) from the left to the right of the image. I can rotate the CCD on the optic axis of the TEM, and this problem persists in the same location (increasing from left to right) regardless of the rotation of the camera. This problem is seen in all images, but is more prevalent in images with } 10 sec exposure time.
Has anyone ever seen this problem before? If so, how did you correct for it?
Instruments being used: Camera is a Proscan HSC 2, 1024x1024 on axis cooled high speed slow scan CCD camera on a LEO 912 EFTEM (120 kV with LaB6).
Any help would be SUPER!!
Thanks for your help,
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 09:24:00 2004
Ultra-thin sections are generally 40-120nm thick, with 60-80nm the dominate range for most Biologicals.
There are also "super-thin" sections 4 - 20nm.
But atomic spacings are much smaller than either of these, ( personally not being a true materials person) atomic sizes seem to range from ~ 0.028 - 0.27nm. And I believe that sucrose is 0.8nm across.
} } Well, I got caught unexpectedly with a lab tour and was asked about cutting thin } sections. I later found out the thickness I "guesstimated" would cut a nitrogen } atom in half. So how thin can a section of organic material be cut with a } ultra-microtome? Can metals be thinned even thinner/ } } Thanks for the help and I promise I will not ask about osmium! } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 } } } 330-668-2235 Ext. 238 } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 09:37:47 2004
I have a user who uses our sputter coater to coat porous membranes with gold or silver. His question is how deep the gold/silver can travel into his membrane pores. The pores are 60 micron deep and the pore diameter is 200 nm. Is it directly proportional to the number of times he coats or is it totally random ?
Thanks in advance.
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://www.cs.nmsu.edu/~biology/Faculty%20&%20Staff/Ghoshroy/Ghoshroy.htm http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 09:38:21 2004
At 08:38 AM 04/15/2004 -0400, maureen petersen wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Maureen -
We are having very good luck using a Microtek ScanMaker 8700 and SilverFast AI software. Of course the TEM film is an odd size. Therefore we use the "one-size-fits-all" glass holder. It's a little tedious to place the negatives properly, but I've gotten good at it. We FTP the images to those who need them and they are really pleased with the results. Bear in mind however that we are a diagnostic medical service and generally prints do not have to be made. But our results are good enough to make diagnoses directly off the monitor.
Joiner
Joiner Cartwright, Jr., Ph.D. Director, Electron Microscope Lab Department of Pathology Baylor College of Medicine
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 11:36:54 2004
We have an Epson Perfection 3200 Photo and we just put the negatives right on the screen, because there isn't a holder that will take them. Works fine.
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Joiner Cartwright, Jr., Ph.D. [mailto:joiner-at-bcm.tmc.edu] Sent: Thursday, April 15, 2004 10:56 AM To: maureen petersen; Microscopy-at-MSA.Microscopy.Com
At 08:38 AM 04/15/2004 -0400, maureen petersen wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Maureen -
We are having very good luck using a Microtek ScanMaker 8700 and SilverFast AI software. Of course the TEM film is an odd size. Therefore we use the "one-size-fits-all" glass holder. It's a little tedious to place the negatives properly, but I've gotten good at it. We FTP the images to those who need them and they are really pleased with the results. Bear in mind however that we are a diagnostic medical service and generally prints do not have to be made. But our results are good enough to make diagnoses directly off the monitor.
Joiner
Joiner Cartwright, Jr., Ph.D. Director, Electron Microscope Lab Department of Pathology Baylor College of Medicine
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 11:45:21 2004
Just about every image analysis package should be able to do what you are asking for. Start with something free, like NIH-Image (the Java version will run on your PC). If you need more, but still a lot less expense than a package like Image Pro Plus, try using Photoshop with Fovea Pro or The Image Analysis Toolkit (ReindeerGraphics.com), which can definitely measure those features and report actual diameter (as well as inscribed circle size, circumscribed circle size, etc., depending on how round they really are). The built-in statistics are modest (mean, variance, skew, kurtosis, regression, etc.) but you can always dump the data into a spreadsheet of stats program.
======= In a message dated 4/15/04 11:15:10 AM, Brian.Kirkmeyer-at-iff.com writes:
} I am looking for PC-based image analysis software that can find, count } and } measure particles. The particles may be either spheres or vesicles, so } } the software must be able to find/measure/count both types. I would also } } like the software to report actual diameters as opposed to equivalent } sphere diameter, though doing both is acceptable. Finally, I need it to } } be able to do statistics on the analyses that it performs. } } If you can provide the names of some software packages that meet these } } criteria, as well as the company that makes it and contact information, } it } would be a great help to me. Thanks!
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 12:35:19 2004
Image Pro Plus by Media Cybernetics provides a full range of utilities for capturing, communicating, processing, measuring, analyzing, archiving, reporting, and printing.
Nicola Ranieri, Microscopy/Image Analysis US FDA Forensic Chemistry Center 6751 Steger Drive Cincinnati, Ohio 45237-3097 (513) 679-2700 X253 (513) 679-2761 FAX nranieri-at-ora.fda.gov
-----Original Message----- } From: Brian.Kirkmeyer-at-iff.com [mailto:Brian.Kirkmeyer-at-iff.com] Sent: Thursday, April 15, 2004 10:01 AM To: Microscopy-at-MSA.Microscopy.Com
I am looking for PC-based image analysis software that can find, count and measure particles. The particles may be either spheres or vesicles, so the software must be able to find/measure/count both types. I would also like the software to report actual diameters as opposed to equivalent sphere diameter, though doing both is acceptable. Finally, I need it to be able to do statistics on the analyses that it performs.
If you can provide the names of some software packages that meet these criteria, as well as the company that makes it and contact information, it would be a great help to me. Thanks!
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 12:43:11 2004
You mention that a dedicated STEM or TEM/STEM (200-300kV) can achieve ~0.8A resolution.
I think instead of "a" that should be "two". As far as I know, only Steve Pennycook's and Phil Batson's dedicated STEMs have reached this figure. For TEMs the number is similar. One would be the high-voltage FEG of Tonamura, and we took the One-Angstrom Microscope at Berkeley to 0.78A in 2001.
Regards, Mike
principe wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } A SEM-based STEM at 30kV can achieve subnanometer resolution (~0.8nm). This } is typically a solid state detector that integrates with an ultra-high } resolution SEM. } } A dedicated STEM or TEM/STEM (200-300kV) can achieve ~0.8A resolution. } } Regards, } Ed } } ******************************************* } Edward Principe, Ph.D. } LEO Electron Microscopy } Applications Development Scientist } principe-at-leo-usa.com } 415-420-4299 (cell) } 650-595-5516 (fax) } ******************************************* } } ----- Original Message ----- } } From: "Patton, David" {David.Patton-at-uwe.ac.uk} } To: "michael shaffer" {michael-at-shaffer.net} } Cc: {Microscopy-at-MSA.Microscopy.com} } Sent: Tuesday, March 30, 2004 7:15 AM } Subject: [Microscopy] STEM } } } } } } } -------------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------------- } ----- } } } } Does STEM (scanning transmission electron microscopy) give } } better resolution than the best FE SEMs? } } } } Dave } } } } On Tue, 30 Mar 2004 10:13:01 -0330 michael shaffer } } {michael-at-shaffer.net} wrote: } } } } } } } } } } } } -------------------------------------------------------------------------- } ---- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -------------------------------------------------------------------------- } ----- } } } } } } Breck Bowles writes ... } } } } } } } Sue Tyler wrote:- } } } } } } } } } Could you tell me if there is such a thing as a LM system } } } } } that will project high quality images at high magnification } } } } } that will also transmit to the web? I welcome all comments. } } } } } } } It gets technically easier with a live slide if you view on a colour } } } } TV monitor the same feed that is going to the Net. At the simplest, } } } } buy a webcam, yank the lens out, and your microscope eyepiece, } } } } assuming you can find an old ... } } } } } } It seems to me you could employ a Nikon Coolpix setup. That is, the } CP99x } } } series and I believe the CP5000 provide for NTSC (and PAL) TV output. } This } } } video signal could then go into a video input as provided by many video } } } input cards. Forgive me if I don't have the time to research current } } } possibilities and manufacturers' model numbers, but the possibility has } } } existed for some time. } } } } } } hth & cheerios ... shAf :o) } } } Avalon Peninsula, Newfoundland } } } www.micro-investigations.com } } } } } } } } } } } } } } } } } } This incoming email to UWE has been independently scanned for viruses } and any virus detected has been removed using McAfee anti-virus software } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } } } } } } } This email has been independently scanned for viruses and any virus } detected has been removed using McAfee anti-virus software } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 13:32:30 2004
Thank you for your reply to the Ir coater question. Your email reply brought up an issue I was trying to resolve about metal sputter coatings. You seem to have a lot of nice paperwork handy and background on the subject. So I have a question that I have never seen answered.
I used a small desk top sputter coater in my lab to coat samples for SEM and it even had uses in TEM. It was not a high resolution coater but it worked for me in TEM. It had a Au-Pd target and one could sputter a coating onto a polymer substrate, for example. After thin sectioning the polymer material, one could determine size of the sputtered metallic particles. In the TEM, they looked more like bright nanoparticle spheres.
Everyone talks about fine grained Pt, Ir, Cr, etc.
So did you or anyone else on the list ever see published or determine what particle size 'fine grained' was for Cr, Ir, W, Ta, Pt, Os, or any other sputter coated metals or alloys that manufacturers sell with their sputter coaters?
As you can imagine, it is not that difficult to determine these sizes in a TEM with it's higher magnifications and levels of resolution. All you basically need is a microtome, the proper target, and a TEM.
Any help with this question about 'fine grain sizes' would be of interest to me.
Sincerely,
Paul Beauregard Senior Research Associate 144 Chapel View Drive Greensburg, PA 15601 (724) 834-2247
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 13:57:05 2004
I second that emotion - we use the Epson Perfection 3200 Photo Scanner, too. It works great.
Beth
} We have an Epson Perfection 3200 Photo and we just put the negatives } right on } the screen, because there isn't a holder that will take them. Works } fine. } } Peggy Sherwood } Lab Associate, Photopathology } Wellman Laboratories of Photomedicine (W224) } Massachusetts General Hospital } 55 Fruit Street } Boston, MA 02114 } 617-724-4839 (voice mail) } 617-726-6983 (lab) } 617-726-3192 (fax) } msherwood-at-partners.org }
} I am interested in knowing what scanner(s) and negative holders people } are using to scan 8 X 10 cm TEM negatives. We are having some problems } with our current setup. } } Thank you, } Maureen Petersen } Ohio Agricultural and Research Center/OSU } Wooster, Ohio
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
As stated before, as good as all Image Analysis software packages will be capable of doing this. Next to ImageJ (NIH-image) which runs in Java written macros and also runs on Mac, and Image Pro Plus, I'd like to add one more: Zeiss KS300/400 software, soon being replaced by the new Axiovision 4, whcih will enable users to write programs (macros) with visual basic. Gives you lots of open doors towards the future. How about comparing Image Pro Plus and Zeiss-software, no idea, but both have a price, ImageJ (or Image NIH) is free downloadable...but does not offer backup as commercial software (allthough, there's also a good mailinglist to exchange ideas/questions, just as for the Axiovision/KS-software!). Best regards,
Sven Terclavers
-----Original Message----- } From: Brian.Kirkmeyer-at-iff.com [mailto:Brian.Kirkmeyer-at-iff.com] Sent: Thursday, April 15, 2004 16:01 PM To: Microscopy-at-MSA.Microscopy.Com
I am looking for PC-based image analysis software that can find, count and measure particles. The particles may be either spheres or vesicles, so the software must be able to find/measure/count both types. I would also like the software to report actual diameters as opposed to equivalent sphere diameter, though doing both is acceptable. Finally, I need it to be able to do statistics on the analyses that it performs.
If you can provide the names of some software packages that meet these criteria, as well as the company that makes it and contact information, it would be a great help to me. Thanks!
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 16:14:30 2004
a very good point! I might have misunderstood the "persisting in the same direction" part. Let's make this very clear:
If you have a pattern on the computer screen due to some defect or artifact, and you rotate the camera, and afterwards the pattern is the same in size and direction ON THE COMPUTER SCREEN, it is due to the camera system, including optics (or fiber-optics) and phosphor screen. If you rotate the camera and the pattern rotates also, the problem lies with a part that was not rotated. This could be the microscope, but also the optics or scintillator, depending on whether these parts rotate with the camera or not.
Sorry for any confusion my earlier posting might have created.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] Sent: Thursday, April 15, 2004 15:15 To: Mike Bode
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 16:31:11 2004
I have carried out a number of experiments with my students during SEM courses to try to determine the best route to use when trying to sputter coat "into" holes.
Conventional surface coating techniques do not work. We found that the best way to coat inside holes was to use the best vacuum that the sputter coater could reach and "force" it to coat using whatever gas remained. A better mean free path means "straight line" deposition and my reason for the success!
My deduction was that under conventional sputter coating procedures we rely upon the mean free path being poor to improve the scatter and hence the ability to coat rough surfaces fairly evenly. When trying to deposit a coat inside a hole the poor mean free path prevents the deposit travelling into the hole.
I have no proof of my theory other than when students are asked to try and coat a 2mm deep 1mm diameter hole the only way to ensure metal penetrates the hole that they have found is using the technique set out above; comments?
Happy sputtering
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com ----- Original Message ----- } From: {sghoshro-at-NMSU.Edu} To: "MSA Listserve" {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, April 15, 2004 3:56 PM
I work with semiconductor devices that have rather opposite dimensions. They are typically 1u-2u deep and 0.5u in diameter. The holes are vias between metal interconnect layers. The vias are like wine glasses or flat bottomed V-shape or U-shape.
What I have observed with Au is that it winds up being a honeycomb of Au filaments rather than an amorphous layer. The honeycomb cells are rather consistent in dimensions from top, down holes and at the bottom of the holes. I think it is safe to say that all sputter coatings lie on the surface--they do not penetrate the specimen.
If Au is coated again, there is a cross hatch of honeycombs. Sometimes the alignment is significant and other times, not so. I use low mT values (80-85mT) and low current (20 mA). With better vacuum, the coating is less ballistic since the Ar ions are not as aggressive, as I figure the situation to be. Perhaps a plasma person could state this more elegantly than I. But it seems that any slight change in vacuum materially affects the characteristics of the Au coating. And in particular, I would characterize it more like a web than as a coating.
What I do know is that if I switch to Au/Pd (60/40) or Pt, the coating is amorphous and cannot be easily seen. Au coating can be seen at about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. However, Au/Pd and Pt look totally different than Au alone. Au/Pd and Pt is more granular rather than honeycomb. And it will look this way at 350KX and greater.
Some metallurgical specimens I have imaged are very gross. E.g., .5mm diameter, 10 mm deep. Au/Pd coating in a Denton Desk II works fine. Tilted at 5 degrees, I can look down the entire extent of the hole. Max mag in this situation is { 5KX. Settings: 85mT, 20mA, 40 seconds. This seems to be about 80A of coating (give or take). In amorphous or aligned specimens, EDS will pick up the coating. Either delete from peak ID or increase KV to have it obfuscated by volumetric interaction.
For no known obvious reason, I would not use Ag. Well, it may oxidize. For the above reasons, I would not use Au. A high vacuum, low current sputter of Au/Pd ought to work for this application. Please let us know how it works out in the end.
gary g.
At 07:56 AM 4/15/2004, you wrote:
} Hi fellow listers, } } I have a user who uses our sputter coater to coat porous membranes with } gold or silver. His question is how deep the gold/silver can travel into } his membrane pores. The pores are 60 micron deep and the pore diameter is } 200 nm. Is it directly proportional to the number of times he coats or is } it totally random ? } } Thanks in advance. } } Soumitra } } ************************************************************* } Soumitra Ghoshroy } College Associate Professor, Biology } Director, Electron Microscopy Lab } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } Tel: 505-646-3268 (office), 646-3283 (lab) } Fax: 505-646-3282 } e-mail:sghoshro-at-nmsu.edu } http://www.cs.nmsu.edu/~biology/Faculty%20&%20Staff/Ghoshroy/Ghoshroy.htm } http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 20:22:18 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (martha-at-uta.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, April 15, 2004 at 15:21:35 ---------------------------------------------------------------------------
Email: martha-at-uta.edu Name: Martha Gracey
Organization: University of Texas at Arlington
Education: Graduate College
Location: Arlington, Texas USA
Question: I need help fixing a liquid sample of liposomes for SEM. The cells have no additive in the control and a protein added to the experimental line. After looking at these, I will then have to add a gold antibody for SEM again. Can you help direct me to a reference. I tried air dting and exposing to Osmium vapors and got lousy results. Thanks Martha
} I am looking for PC-based image analysis software that can find, count and } measure particles. The particles may be either spheres or vesicles, so } the software must be able to find/measure/count both types. I would also } like the software to report actual diameters as opposed to equivalent } sphere diameter, though doing both is acceptable. Finally, I need it to } be able to do statistics on the analyses that it performs.
We have analySIS from Soft Imaging System. It can do all you ask and then some; it's really powerful. Today I was trying to show someone how to do some analyses with the free ImageJ, which is really great for the price, plus they could do it at their own computer. However, they were unwilling to write or manage macros, and we were having trouble getting it to identify, count and measure the cells we wanted, so I went back to analySIS. Problem solved.
Aloha, Tina
**************************************************************************** * 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 MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 21:05:44 2004
The best sections I ever was able to made weres 10 nm thick proven by independent methods. It was a sections from ribosomal crystals parallel to the crystallographic planes (it was another story how to do it). I guess 10 nm is a limit for biological samples embedded into the plastic by conventional technique. I would imagine, someone could do thinner sections of uniform material like metal or even plastic (frozen?). I would think, the limit here would be their stability for manipulation - how to mount them on EM grid and made flat... You definitely could not cut atom in half, because "cutting" is actually splitting between atoms (forces between atoms are weaker than inside). Thickness of sections depends more from the knife than ultra-microtome (if instrument is in perfect shape of coarse). Diatome knifes are certified down to 20 nm I believe. Instrument itself more responsible for reproducibility - how many sections of similar thickness you may produce in the row. You may produce very good thin section even on the very old hardly used instrument; you may produce 20 sections of the same quality in the row only on the very good, stable machine... No osmium tetroxide for while, but still, if somebody have problems with that stuff, let me know.... Have a great day. Sergey
At 07:41 AM 4/15/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
It's a mailinglist I started myself, as comparing to the ImageJ mailinglist. I announced the existence of it here, but only once, and indeed, maybe I should've done it more! Although, Zeiss knows of it, even some people from Zeiss are members and we are having a discussion to move the mailinglist to a Zeiss-server, in combination of an online download-page, where you can download all (small) macro's Zeiss, I and maybe others, have written ourselves. If you need advice or a macro, just write a message to: KS300-at-topica.com after subscription. To subscribe: KS300-subscribe-at-topica.com
That's all! Hope to see you soon as a 'member' :-) And don't worry about tons of emails, this list is actually not that often used, but I must say, every question has been answered and solved...
Best regards,
Sven
-----Original Message----- } From: Philip Oshel [mailto:peoshel-at-wisc.edu] Sent: Friday, April 16, 2004 16:01 PM To: Sven.Terclavers-at-med.kuleuven.ac.be
LEHIGH MICROSCOPY SCHOOL
Lehigh University, Bethlehem, PA U.S.A. June 6-18, 2004
* Scanning Electron Microscopy and X-ray Microanalysis
* Introduction to SEM and EDS for the New SEM Operator
* Analytical Electron Microscopy Analysis for TEM Specimens
* Atomic Force Microscopy and Other Scanned Probe Microscopes
* Characterization of Nanostructures
* Focused Ion Beam Instrumentation and Applications
* Problem Solving with SEM and X-ray Microanalysis
* Quantitative X-ray Microanalysis of Bulk Specimens and Particles
* Particle Characterization, Preparation, Microscopy, and Analysis
See course descriptions at: {http://www.lehigh.edu/microscopy} www.lehigh.edu/microscopy
For more information contact Sharon Coe at 610.758.5133 {mailto:Sharon.coe-at-Lehigh.edu} Sharon.coe-at-Lehigh.edu
******************************** Sharon L. Coe Events Coordinator Lehigh Microscopy School 5 East Packer Avenue Bethlehem, PA 18015 610-758-5133 (phone) 610-758-4244 (fax) {mailto:slc6-at-lehigh.ed} slc6-at-lehigh.ed {http://www.lehigh.edu/microscopy} www.lehigh.edu/microscopy
********************************
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 16 11:06:38 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jjwolo-at-uic.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, April 16, 2004 at 10:49:29 ---------------------------------------------------------------------------
Question: I am trying to find a copy of the instruction manual for an Olympus Inverted Microscope Models IM, IM-2 (early 80's models). Anything would be helpful. The Olympus company has not responded to my e-mails.
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Martha The problem with liposomes is that when you dehydrate in alcohols, they dissolve so normal preparation techniques do not work.
The best approach you can take with SEM is to use cryo-SEM. We have a cryo stage on a our field emission SEM and have used it for all sorts of lipids and waxes that could not be looked at conventionally. The sample has to be "cracked open" and the surface water sublimated off.
As for the immunogold treatment, we have been having some success with using immunononogold and then gold enhancement. Then using the backscatter detector to pick up the gold deeper in the cells. Using a mix of the secondary electron detector and the backscatter detector, you can see the gold even if it is inside cells because the backscatter detector reaches deeper than the secondary electron detector.
Both these techniques will be taught at the third International Cryo EM course here in June. (see the website http://www.emlab.ubc.ca )
You might want to consider plunge freezing and cryo TEM. We will also have a Vitrobot and a Leica plunge freezer at the cryo course. We have a TEM with a cryo stage. You can bring your own sample of liposomes and try out all the techniques including hgih pressure freezing. If you check out the poster you will see a picture taken at last year's course of some hela cells which were infected with chlamidia, high pressure frozen, cracked open and looked at with cryo SEM. Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 16 15:36:12 2004
We recently started using Epson Perfection 3200 Photo flatbed scanner and just like Beth we are quite happy with its performance. We use the negative holder that came with the scanner for scanning TEM negatives.
Soumitra
No financial interest in Epson, just a happy user.
} } } ----------------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------------- -- } } I am interested in knowing what scanner(s) and negative holders } people are using to scan 8 X 10 cm TEM negatives. We are having some } problems with our current setup. } } Thank you, } Maureen Petersen } Ohio Agricultural and Research Center/OSU } Wooster, Ohio } -- } }
Soumitra Ghoshroy Ph.D College Associate Professor, Biology Director, Electron Microscopy Lab New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office) 505-646-3283 (lab) Fax: 505-646-3282
From MicroscopyL-request-at-ns.microscopy.com Sat Apr 17 09:04:30 2004
I would like to find out whether Image-Pro Plus or another software package allows you to obtain the real sphere diameter from a microtomed section. Since the microtome does not necessarily cut the sphere at the largest section, a correction needs to be applied. I looked into stereology, but it would be nice to have this included in the software together with the count/measure/statistics of the particles. If this is not included in the software, is it accurate enough to use the following equation: d_actual=(4 x d_measured)/PI ?
Thank you!
Steven Swier Institute of Materials Science University of Connecticut.
_________________________________________________________________ Vraag van de week: Welk soort project zou jij financieel ondersteunen? http://www.msn.be/microsoft/potential/default.asp
From MicroscopyL-request-at-ns.microscopy.com Sun Apr 18 10:37:09 2004
In a message dated 4/18/04 10:49:26 AM, stevenswier-at-hotmail.com writes:
} I would like to find out whether Image-Pro Plus or another software package } } allows you to obtain the real sphere diameter from a microtomed section. } } Since the microtome does not necessarily cut the sphere at the largest } } section, a correction needs to be applied. I looked into stereology, but } it } would be nice to have this included in the software together with the } count/measure/statistics of the particles. If this is not included in the } } software, is it accurate enough to use the following equation: d_actual=(4 } x } d_measured)/PI ? }
Fovea Pro (www.reindeergraphics.com) does include the stereological measurements and calculations for spheres seen in section, and you can even run the Photoshop-compatible plugins within Image Pro Plus. Be aware, however, that this method (inherently) depends critically on several assumptions: 1) that the particles are in fact spheres - surprisingly small deviations have major influences on the results 2) that the sections are thin,and you do not have "overprojection" problems 3) that small polar caps are visible (or that you can specify the extent to which they are not) 4) that you have lots of data, because the propagation of errors tends to (greatly) magnify the statistical fluctuations due to counting.
The sphere unfolding technique was very popular back in the 60's, but modern wisdom places instead an emphasis on newer stereological techniques that emphasize the design of specimen preparation techniques that avoid the sources of bias and produce superior results. Read "Practical Stereology, 2nd Edition" by Russ and Dehoff, or "Unbiased Stereology" by Howard and Reed, for more details (or look up papers by Cruz-Orive in past issues of the Journal of Microscopy).
And lastly, no, your equation is wrong and won't work anyway.
From MicroscopyL-request-at-ns.microscopy.com Sun Apr 18 19:51:03 2004
Gary Gaugler wrote: ==================================================================== .............What I do know is that if I switch to Au/Pd (60/40) or Pt, the coating is amorphous and cannot be easily seen. Au coating can be seen at about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. However, Au/Pd and Pt look totally different than Au alone. Au/Pd and Pt is more granular rather than honeycomb. And it will look this way at 350KX and greater. ==================================================================== Could you tell us the measurement used to conclude it is an amorphous coating? So far as I know, one does see a grain, even with Pt or Au/Pd irrrespective of the brand of sputter coater used.
Chuck
============================================ Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Sun Apr 18 20:53:08 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (johnpell-at-temple.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, April 18, 2004 at 19:54:26 ---------------------------------------------------------------------------
Email: johnpell-at-temple.edu Name: John Pell
Organization: Temple University
Title-Subject: [Microscopy] [Filtered] artifacts and blunders/Royal Rife
Question: I am an undergraduate student of biology at Temple University working on a pet research project on Royal Rife's microscopes for a microscopy class. I noticed a post on the Microscopy Listserve under the subject "artifacts and blunders" that contained an oblique reference to Rife. I would like to have more information characterizing Rife's blunders and the types of optical artifacts he saw through his scopes. Most of the websites with information about Rife are maintained by individuals or organizations concerned with mythologizing Rife's miraculous discovery of a cure for cancer using "vibrational energy." A critical discussion of the optical principles Rife may have used or misused does not seem to be a priority for them. If someone would point me towards such a discussion, or some of the information that could be used to construct such a discussion, I would be much obliged.
Using an FEI Sirion SFEG, the coating is amorphous. The "measurement" is looking at the image via the SEM. Plain Au is honeycombed. Would you like a sample of this? See what you can see at 350KX. What would you use to do this yourself? Let's compare images. If it would be productive, of course.
How do we define amorphous versus non-amorphous at this mag? This is further complicated by the manner in which the coating is deposited. There are way too many variables. Perhaps I simplify the topic.
So, what do you want to see? I can produce pix at 350KX of Au and Au/Pd coatings. If these are a big surprise to you, I can provide specimen images. Alternatively, show me what you have at this same mag. Produce Au and Au/Pd and Pt images. That pretty well covers the normal sputter coating environment.
I am always open to new views on this subject.
gary g.
At 07:12 PM 4/18/2004, you wrote:
} -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Gary Gaugler wrote: } ==================================================================== } .............What I do know is that if I switch to Au/Pd (60/40) or Pt, the } coating is amorphous and cannot be easily seen. Au coating can be seen at } about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. } However, Au/Pd and Pt look totally different than Au alone. Au/Pd and Pt is } more granular rather than honeycomb. And it will look this way at 350KX and } greater. } ==================================================================== } Could you tell us the measurement used to conclude it is an amorphous } coating? So far as I know, one does see a grain, even with Pt or Au/Pd } irrrespective of the brand of sputter coater used. } } Chuck } } ============================================ } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } President 1-800-2424-SPI } SPI SUPPLIES FAX: 1-610-436-5755 } PO BOX 656 e-mail:cgarber-at-2spi.com } West Chester, PA 19381-0656 USA } Cust.Service: spi2spi-at-2spi.com } } Look for us! } ######################## } WWW: http://www.2spi.com } ######################## } ============================================
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 07:12:25 2004
If you are truely producing amorphous Au this would be very significant. I must admit to being somewhat skeptical.
An image of a near feature less "surface" is not sufficient evidence to call something amorphous.
The best way to answer this question would be to prepare a TEM cross-section. Then using either HREM or Electron Diffraction from the coating you could verify if the coating is crystalline or not.
Nestor
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From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 08:33:51 2004
Contamination control of airborne dust, although not of concern for massive samples or non-trace analysis, is of concern often enough at our laboratory: Particularly with unknowns and when we are micromanipulating a one-of-a-kind particle that can be confused with contaminants; when using filtration procedures for trace quantities of particulate samples that involve the filtering of large volumes of air; when conducting ultra-trace component analysis within a microgram of a powder; or when developing certain standard reference materials. Unwanted dust can add many days to sample preparation or analysis, or can jeopardize a sample or an analysis, making it an expensive problem if it occurs. For these reasons we sometimes use cleanrooms and contamination control protocols with specially trained staff, sometimes learning what is needed the hard way.
How prevalent is the need for contamination control at other facilities, knowing there will be differences in environments and analytical goals? It would be interesting if others would care to share experiences about the significance of ambient dust in specimens, and what approaches to control seem adequate for their particular case.
I've been tasked with pulling together sensible specifications for a new computer to capture digital images from our pol scope. Many options appear to be straight forward: More RAM, - 512 MB seems to be enough. More HD storage, - As Daisy use to say, you can never be too rich, too thin or have enough storage. I think 120GB is enough. CD burner at 48X seems enough
but which monitor? Should I look for a fast response time, say 16ms, to help me focus? Are flat screens better for viewing than the more traditional monitors? Do I need more then 17 inch diagonal screen? Is the integrated Intel 3D Extreme Graphics video card a good choice for photomicrography?
Any suggestions for a good driver to capture my images? I intend to use photoshop to do any post imaging processing that needed, but I would like to be able to do a little tweaking to maximize my images when I capture them.
I have also inhered a Microimaging Video system and a RGB Automatic Camera, but I have no instructions or user manual. If anyone knows a web site or has a copy they would like to share I would be grateful!
Any advice, opinions or alternative suggestions are welcome!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 08:49:35 2004
Thank you for all of your tips. I got some great responses with some really good ideas to solve this problem.
To clear up some questions: All images that I see this problem with are acquired when the beam is off (no intensity on the CCD), the images are also raw (not dark count or gain corrected). The problem is not with an artifact present in all CCD images, but rather the horizontal line profile of the image (a measure of the average pixel value across the image horizontally) increases from left to right.
What the solution may be: From responses I've received it may be that the scintallator may have corroded from prolonged exposure to condensation/contamination within the TEM column. We're going to try and clean it off to rid ourselves of the problem.
Thanks again!
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 10:41:38 2004
Indeed, amorphous likely means different things to different folks and in different situations. From a metallurgical or atomic standpoint, it would mean that it lacks a definitive or distinct crystalline structure--i.e., no lattice. On the other hand, amorphous could mean that it is shapeless, of no definite form or organization or methodical arrangement.
OK, does this mean no form or organization at the eye ball level? At the magnifying glass level? LM, SEM, TEM level? I think that I "see" the problem of definition and ability to define it.
The general consensus seems to be one of grains. A honey comb shaped coating of Au would have the metal portions containing or consisting of grains of Au. Thus, the honey comb has organization and methodical arrangement--not amorphous. And the Au metal would have grains--not amorphous. Therefore, sputtered Au is not amorphous.
Now to discuss Au/Pd. This material sputters totally differently than Au alone. The coating is like a layer of grains of sand. These dots, if you will, may touch one another in places and not in others. Unlike reflowed metal, the dots are distinct and quite 3-D. Is this an amorphous layer? If each dot was one grain, and each grain was not touching each other grain, is the coating amorphous or not? Does continuity play into the definition?
At damascene vias, the ILD is definitely amorphous. These areas etch totally differently than areas away from the vias. It's interesting to see that areas away from vias exhibit some degree of amorphousness. Why the via areas are more strongly amorphous, I do not yet know. This is something I am working on at the moment.
If I can find some Au and Au/Pd pix that I can post, I will do so. I have the pix somewhere and I have specimens (all over the place). If there is interest, I will work on getting them.
gary g.
At 05:34 AM 4/19/2004, you wrote: } Gary } } If you are truely producing amorphous Au this would } be very significant. I must admit to being somewhat } skeptical. } } An image of a near feature less "surface" is not sufficient } evidence to call something amorphous. } } The best way to answer this question would be to prepare } a TEM cross-section. Then using either HREM or Electron Diffraction from } the coating you could verify if the coating is crystalline } or not. } } } Nestor } } } } } ------------------------------------------------------------------------- } ----- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------- } ------ } } } } Using an FEI Sirion SFEG, the coating is amorphous. } } The "measurement" is looking at the image via the SEM. } } Plain Au is honeycombed. Would you like a sample of } } this? See what you can see at 350KX. What would you } } use to do this yourself? Let's compare images. } } If it would be productive, of course. } } } } How do we define amorphous versus non-amorphous } } at this mag? This is further complicated by the } } manner in which the coating is deposited. There } } are way too many variables. Perhaps I simplify } } the topic. } } } } So, what do you want to see? I can produce pix } } at 350KX of Au and Au/Pd coatings. If these are } } a big surprise to you, I can provide specimen images. } } Alternatively, show me what you have at this same } } mag. Produce Au and Au/Pd and Pt images. That pretty well } } covers the normal sputter coating environment. } } } } I am always open to new views on this subject. } } } } gary g. } } } } } } At 07:12 PM 4/18/2004, you wrote: } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } Gary Gaugler wrote: } } } ==================================================================== } } } .............What I do know is that if I switch to Au/Pd (60/40) or Pt, the } } } coating is amorphous and cannot be easily seen. Au coating can be seen at } } } about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. } } } However, Au/Pd and Pt look totally different than Au alone. Au/Pd and } Pt is } } } more granular rather than honeycomb. And it will look this way at } 350KX and } } } greater. } } } ==================================================================== } } } Could you tell us the measurement used to conclude it is an amorphous } } } coating? So far as I know, one does see a grain, even with Pt or Au/Pd } } } irrrespective of the brand of sputter coater used. } } } } } } Chuck } } } } } } ============================================ } } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } } } President 1-800-2424-SPI } } } SPI SUPPLIES FAX: 1-610-436-5755 } } } PO BOX 656 e-mail:cgarber-at-2spi.com } } } West Chester, PA 19381-0656 USA } } } Cust.Service: spi2spi-at-2spi.com } } } } } } Look for us! } } } ######################## } } } WWW: http://www.2spi.com } } } ######################## } } } ============================================
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 10:45:03 2004
I had never heard of Rife until some guy got on this list last year raving about why all of us so-called scientists had ignored Rife's contributions to science/microscopy/medicine.When I asked him what exactly Rife's contributions were, I got a few websites (long since deleted) but no real evidence or references in the literature. I did read one website which I suspect a Google search could bring up. Once I waded through 5-6 pages of unsubstantiated claims, I did find some discussion of his microscope. While the complexity of the instrument was somewhat impressive, the images it delivered were not. I think the "optical principles" involved were largely products of Rife's imagination. My general impression of the reaction of working scientists to people like Rife is one of dismissal. Since the evidence offered is usually imaginary or artifactual, those who work in the field in question don't bother to refute the "evidence" (most of us have better things to do). Unfortunately, this leads the gulible to accuse the scientific community of a conspiracy since it can't "refute" the work of the soon to be demi-god. The web has made the propagation of theories about Rife much easier. Good luck with your project.
Geoff
by way of MicroscopyListserver wrote:
} Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (johnpell-at-temple.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, April 18, 2004 at 19:54:26 } --------------------------------------------------------------------------- } } Email: johnpell-at-temple.edu } Name: John Pell } } Organization: Temple University } } Title-Subject: [Microscopy] [Filtered] artifacts and blunders/Royal Rife } } Question: I am an undergraduate student of biology at Temple University working on a pet research project on Royal Rife's microscopes for a microscopy class. I noticed a post on the Microscopy Listserve under the subject "artifacts and blunders" that contained an oblique reference to Rife. I would like to have more information characterizing Rife's blunders and the types of optical artifacts he saw through his scopes. Most of the websites with information about Rife are maintained by individuals or organizations concerned with mythologizing Rife's miraculous discovery of a cure for cancer using "vibrational energy." A critical discussion of the optical principles Rife may have used or misused does not seem to be a priority for them. If someone would point me towards such a discussion, or some of the information that could be used to construct such a discussion, I would be much obliged. } } Thanks, } } John Pell } } } --------------------------------------------------------------------------- } } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 10:55:22 2004
We recently produces a very thin coating of Ni and TiO2 that was sputtered simultaneously using a Edwards sputtering system. According to other publications these should be Polycrystalline. In the Tecnai 12 the diffraction the pattern was amorphous. We were pleased since the polycrystalline samples we proved can be introduced by beam damage (Remove C1 aperature) and that introduced a polycrystalline sample. Beautiful artefact. Still were not convinced that the film was amorphous. The sample was examined at fei Netherlands on the Technai G2 equipped with a FEG gun, a beauty. It proven to be nano crystals. We are still waiting for the final data set from fei. I would not be surprised if the Gold film follows the same logic.
-----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-microscopy.com] Sent: Monday, April 19, 2004 2:35 PM To: microscopy-at-ns.microscopy.com
Gary
If you are truely producing amorphous Au this would be very significant. I must admit to being somewhat skeptical.
An image of a near feature less "surface" is not sufficient evidence to call something amorphous.
The best way to answer this question would be to prepare a TEM cross-section. Then using either HREM or Electron Diffraction from the coating you could verify if the coating is crystalline or not.
Nestor
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 11:38:15 2004
TEM Analyst Position --------------------- A position is open for a Transmission Electron Microscopy Analyst in the Process Characterization Group at International SEMATECH in Austin, Texas.
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Our toolset includes two 300 keV TEM's (one FEG, one LaB6), with SUTW-EDX, GIF, multiple HAADF-STEM and CCD cameras. Our sample preparation toolset includes several PIPS and Duomill tools as well as 2 FEI FIB systems.
This is a temporary position (1 year renewable contract) with benefits. Preference made to qualified persons willing to work off-shift.
Interested parties should reply offline to Brendan.Foran-at-Sematech.Org -------------------
Sincerely, Brendan ----------------------------------------------- Brendan Foran, Ph.D. Senior Member Technical Staff Transmission Electron Microscopy Group Leader Process Characterization Lab- ATDF International SEMATECH phone (512) 356-3936 -----------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 11:41:30 2004
I would definitely add more RAM---at least a gigabyte, I would think. Some of the images I work with are over 140MB and a few images running in Photoshop can gobble memory like there's no tomorrow. And if you're doing video, it's even worse. I would also get the highest speed processor I could afford. The 2.8 gigahertz processors are fairly reasonable, and since I built my computer I think the 3.0+ GHZ processors are starting to come down. In terms of the monitor, I'm told that CRT's still have a thin edge over flat panels for image quality, plus a huge advantage in price, but the flats are getting better all the time. BIG monitors are definitely nicer, though, and that's where CRTs can save large amounts of money over flat panels. Finally, I would consider a DVD or combo burner to increase storage capacity for your images. Big files fill up a CD pretty quickly.
Hope this helps. Good luck.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Frank.Karl-at-degussa.com [mailto:Frank.Karl-at-degussa.com] Sent: Monday, April 19, 2004 8:35 AM To: microscopy-at-msa.microscopy.com
I've been tasked with pulling together sensible specifications for a new computer to capture digital images from our pol scope. Many options appear to be straight forward: More RAM, - 512 MB seems to be enough. More HD storage, - As Daisy use to say, you can never be too rich, too thin or have enough storage. I think 120GB is enough. CD burner at 48X seems enough
but which monitor? Should I look for a fast response time, say 16ms, to help me focus? Are flat screens better for viewing than the more traditional monitors? Do I need more then 17 inch diagonal screen? Is the integrated Intel 3D Extreme Graphics video card a good choice for photomicrography?
Any suggestions for a good driver to capture my images? I intend to use photoshop to do any post imaging processing that needed, but I would like to be able to do a little tweaking to maximize my images when I capture them.
I have also inhered a Microimaging Video system and a RGB Automatic Camera, but I have no instructions or user manual. If anyone knows a web site or has a copy they would like to share I would be grateful!
Any advice, opinions or alternative suggestions are welcome!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 13:52:35 2004
Get the fastest P4 you can afford. 2.66GHz or 3GHz. Get at least 1GB DDR RAM. The motherboard will likely have three DIMM slots, so put in three 512MB PC2700 DDR DIMMs--1.5GB. Its FSB should handle these.
Put in two hard drives. Dual 120G is good. Set up D: as Photoshop's scratch disk. This way PS will work very fast since it does not have to use C: for work and scratch. Plus, I tend to keep data on D: and programs on C:. This way, backup is done mostly of D:.
For optical, I think that optimum is Plextor Premium along with Panasonic DVR-106D DVD-R/RW-CD-R/RW. If you don't get the DVD burner, get a DVD-ROM drive at least. The DVD burner is excellent for big backups. Also get Roxio's newest Version 7 burning software. It is a major improvement over Version 6. The 106D will write 4X DVDs. And it also becomes a backup burner for CDs.
I'd recommend not using on-board video. If it fails, you are dead. Get an Nvidea/ASUS GeForce FX5200 8X AGP 128MB. Fast and offloads CPU a lot. Integrated usually means on-board rather than a separate board. If separate, it is probably OK.
Also, go with WindowsXP Pro over Win2K Pro. XP is a new OS rather than an update of 2K. Very stable and reliable. 2K is good too but XP is later and has more features.
Flat screen CRTs are my favorite. I use a Sony Multiscan E500 21". There are newer models. For images, it is great. For my camera capture system, I use Samsung 191t TFT/LCD. PS7 and image processing is done on the CRT system. If you will have only one system, I recommend CRT. The 21" units are HEAVY! Refresh rate is 80Hz. No flicker.
gary g.
At 06:34 AM 4/19/2004, you wrote:
} I've been tasked with pulling together sensible specifications for a new } computer to capture digital images from our pol scope. Many options appear } to be straight forward: } More RAM, - 512 MB seems to be enough. } More HD storage, - As Daisy use to say, you can never be too rich, } too thin or have enough storage. I think 120GB is enough. } CD burner at 48X seems enough } } but which monitor? } Should I look for a fast response time, say 16ms, to help me focus? } Are flat screens better for viewing than the more traditional monitors? } Do I need more then 17 inch diagonal screen? } Is the integrated Intel 3D Extreme Graphics video card a good choice for } photomicrography? } } Any suggestions for a good driver to capture my images? I intend to use } photoshop to do any post imaging processing that needed, but I would like } to be able to do a little tweaking to maximize my images when I capture } them. } } I have also inhered a Microimaging Video system and a RGB Automatic Camera, } but I have no instructions or user manual. If anyone knows a web site or } has a copy they would like to share I would be grateful! } } Any advice, opinions or alternative suggestions are welcome! } } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 } } } 330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 14:06:09 2004
you need to be VERY careful if you try to do something with the Phosphor. It can be damaged beyond repair very easily.
You mention that you get a signal from the camera with the beam off. What is the exposure time? Usually the phosphor is covered with a thin metallic layer to provide conductivity and block photons. One of the reasons for your "shadow" might be an uneven thickness of this layer. What do you see if you darken the TEM room completely and increase exposure time until you see something?
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: William Stratton [mailto:wgstratton-at-wisc.edu] Sent: Monday, April 19, 2004 08:11 To: Microscopy
Hello all,
Thank you for all of your tips. I got some great responses with some really good ideas to solve this problem.
To clear up some questions: All images that I see this problem with are acquired when the beam is off (no intensity on the CCD), the images are also raw (not dark count or gain corrected). The problem is not with an artifact present in all CCD images, but rather the horizontal line profile of the image (a measure of the average pixel value across the image horizontally) increases from left to right.
What the solution may be: From responses I've received it may be that the scintallator may have corroded from prolonged exposure to condensation/contamination within the TEM column. We're going to try and clean it off to rid ourselves of the problem.
Thanks again!
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 15:25:24 2004
One of our users has come to us with a sticky problem.
Her siRNA transfections look great in live cells, but after fixation and immunostaining there are red dots everywhere, all over the coverslip (polylysine coated), all over the cells, and in the cells.
Anybody else encounter this problem?
Thanks. ____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 20:39:25 2004
The question about whether coatings deposited using a sputter coater are 'amorphous' or 'crystalline' seems to be in dispute here. On an atomic level, if you can make an amorphous metal then we should be able to image it in our high resolution TEM (JEOL 4000EX) and not find the orderly arrangement of atoms in the layers. For gold deposited on amorphous (definitely) silicon, we can see easily at 800kX (5MX final mag) that the gold 'islands' of about 5nm diameter are composed of crystalline gold with each atom being able to be identified and counted in the orderly structure. We do the same thing at interfaces in semiconductor devices to note the changes of atomic structure where two metal system combine. It should also be noted that if the layer is thin enough, the diffraction pattern is hard to find (very weak signal) but with a long enough dwell time you will definitely see that the metal layer has atomic level crystalline structure.
The argument that seems to be being presented here is semantic, not scientific. Whether or not a layer appears as a 'honeycomb' has nothing to do with the fact that the structure of the 'honeycomb' is crystalline or amorphous. For the purposes of SEM the ideal is to have an 'amorphous' layer from the standpoint that at the magnifications involved there is no contribution of 'ordered' structures from the coating. This has absolutely nothing to do with whether or not the atomic level structures are crystalline or amorphous. We routinely coat samples with whatever material is best suited for the images we want to obtain. That might be gold, Au/Pd, Cr, Ag, Ti or whatever else seems to work. For Auger work we even use deposited Li to eliminate charging without contributing significantly to the spectral data. For SEM you are striving to achieve a coating that allows imaging of your actual sample without adding 'artifacts' that will confuse the interpretation of your data. If that means that you deposit a gold/palladium or chromium layer with such fine structure that your SEM image sees only the information from your sample but doesn't present an artifact then, by all means, call it 'amorphous'. A more distinctive description might, however, be 'uniform' at the magnification used. By this definition the actual atomic level crystallinity of the layer is not called into question. If you really want to know about the atomic scale ordering then you need to move on to HRTEM since SEM is not sufficient to see the atomic scale features. You could also use atomic force microscopy to 'see' the atomic structure of your surface coating.
Drew Hirt Materials Research Laboratories, Inc. Tel (800) 424-1776 Fax (330) 750-0776 drew-at-hirt.com http://www.mrllab.com
on 4/19/04 11:57 AM, Gary Gaugler at gary-at-gaugler.com wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Nestor: } } Indeed, amorphous likely means different things } to different folks and in different situations. } From a metallurgical or atomic standpoint, it } would mean that it lacks a definitive or distinct } crystalline structure--i.e., no lattice. On the } other hand, amorphous could mean that it is shapeless, } of no definite form or organization or methodical } arrangement. } } OK, does this mean no form or organization at the } eye ball level? At the magnifying glass level? } LM, SEM, TEM level? I think that I "see" the } problem of definition and ability to define it. } } The general consensus seems to be one of grains. } A honey comb shaped coating of Au would have the } metal portions containing or consisting of grains } of Au. Thus, the honey comb has organization and } methodical arrangement--not amorphous. And the } Au metal would have grains--not amorphous. Therefore, } sputtered Au is not amorphous. } } Now to discuss Au/Pd. This material sputters } totally differently than Au alone. The coating } is like a layer of grains of sand. These dots, } if you will, may touch one another in places and } not in others. Unlike reflowed metal, the dots } are distinct and quite 3-D. Is this an amorphous } layer? If each dot was one grain, and each grain } was not touching each other grain, is the coating } amorphous or not? Does continuity play into the } definition? } } At damascene vias, the ILD is definitely amorphous. } These areas etch totally differently than areas } away from the vias. It's interesting to see that } areas away from vias exhibit some degree of } amorphousness. Why the via areas are more strongly } amorphous, I do not yet know. This is something } I am working on at the moment. } } If I can find some Au and Au/Pd pix that I can } post, I will do so. I have the pix somewhere and } I have specimens (all over the place). If there } is interest, I will work on getting them. } } gary g. } } } } } } At 05:34 AM 4/19/2004, you wrote: } } Gary } } } } If you are truely producing amorphous Au this would } } be very significant. I must admit to being somewhat } } skeptical. } } } } An image of a near feature less "surface" is not sufficient } } evidence to call something amorphous. } } } } The best way to answer this question would be to prepare } } a TEM cross-section. Then using either HREM or Electron Diffraction from } } the coating you could verify if the coating is crystalline } } or not. } } } } } } Nestor } } } } } } } } } ------------------------------------------------------------------------- } } ----- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------- } } ------ } } } } } } Using an FEI Sirion SFEG, the coating is amorphous. } } } The "measurement" is looking at the image via the SEM. } } } Plain Au is honeycombed. Would you like a sample of } } } this? See what you can see at 350KX. What would you } } } use to do this yourself? Let's compare images. } } } If it would be productive, of course. } } } } } } How do we define amorphous versus non-amorphous } } } at this mag? This is further complicated by the } } } manner in which the coating is deposited. There } } } are way too many variables. Perhaps I simplify } } } the topic. } } } } } } So, what do you want to see? I can produce pix } } } at 350KX of Au and Au/Pd coatings. If these are } } } a big surprise to you, I can provide specimen images. } } } Alternatively, show me what you have at this same } } } mag. Produce Au and Au/Pd and Pt images. That pretty well } } } covers the normal sputter coating environment. } } } } } } I am always open to new views on this subject. } } } } } } gary g. } } } } } } } } } At 07:12 PM 4/18/2004, you wrote: } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } } } Gary Gaugler wrote: } } } } ==================================================================== } } } } .............What I do know is that if I switch to Au/Pd (60/40) or Pt, the } } } } coating is amorphous and cannot be easily seen. Au coating can be seen at } } } } about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. } } } } However, Au/Pd and Pt look totally different than Au alone. Au/Pd and } } Pt is } } } } more granular rather than honeycomb. And it will look this way at } } 350KX and } } } } greater. } } } } ==================================================================== } } } } Could you tell us the measurement used to conclude it is an amorphous } } } } coating? So far as I know, one does see a grain, even with Pt or Au/Pd } } } } irrrespective of the brand of sputter coater used. } } } } } } } } Chuck } } } } } } } } ============================================ } } } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } } } } President 1-800-2424-SPI } } } } SPI SUPPLIES FAX: 1-610-436-5755 } } } } PO BOX 656 e-mail:cgarber-at-2spi.com } } } } West Chester, PA 19381-0656 USA } } } } Cust.Service: spi2spi-at-2spi.com } } } } } } } } Look for us! } } } } ######################## } } } } WWW: http://www.2spi.com } } } } ######################## } } } } ============================================ } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 21:14:33 2004
Whoa, I did not intend for this to get way off base, out of hand, perverted, convoluted, complicated, ...fill in the adjectives.
Post: This is the point of issue. There is a fine structure of the coating. At low mag, it cannot be seen. At high mag, it can be seen. Is it an "artifact?" No. It is real. But then, we need to define an artifact in the context of coating, et. al. As Nestor and Charles said, it is an issue of grains. This is bounded. We can deal with this.
If the SEM sees information of the coating, then what? It is not an artifact. Furthermore, what does "uniform" mean? It seems that your posting says that in one case the coating is amorphous but in another it is not...based on mag and artifacts. But "uniform" is counter-amorphous... sigh.
I think we are headed towards making a mountain out of a mole hill. Perhaps we should let this dog lay. If not, Nestor will keep us on track. Notwithstanding, I will do some EBSD studies later this year and report back to the list.
gary g.
At 07:01 PM 4/19/2004, you wrote: } To All: } } [snip] If that means that you deposit a } gold/palladium or chromium layer with such fine structure that your SEM } image sees only the information from your sample but doesn't present an } artifact then, by all means, call it 'amorphous'. A more distinctive } description might, however, be 'uniform' at the magnification used. By this } definition the actual atomic level crystallinity of the layer is not called } into question. If you really want to know about the atomic scale ordering } then you need to move on to HRTEM since SEM is not sufficient to see the } atomic scale features. You could also use atomic force microscopy to 'see' } the atomic structure of your surface coating. } } Drew Hirt } Materials Research Laboratories, Inc. } Tel (800) 424-1776 } Fax (330) 750-0776 } drew-at-hirt.com } http://www.mrllab.com } } } on 4/19/04 11:57 AM, Gary Gaugler at gary-at-gaugler.com wrote: } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } --} - } } } } Nestor: } } } } Indeed, amorphous likely means different things } } to different folks and in different situations. } } From a metallurgical or atomic standpoint, it } } would mean that it lacks a definitive or distinct } } crystalline structure--i.e., no lattice. On the } } other hand, amorphous could mean that it is shapeless, } } of no definite form or organization or methodical } } arrangement. } } } } OK, does this mean no form or organization at the } } eye ball level? At the magnifying glass level? } } LM, SEM, TEM level? I think that I "see" the } } problem of definition and ability to define it. } } } } The general consensus seems to be one of grains. } } A honey comb shaped coating of Au would have the } } metal portions containing or consisting of grains } } of Au. Thus, the honey comb has organization and } } methodical arrangement--not amorphous. And the } } Au metal would have grains--not amorphous. Therefore, } } sputtered Au is not amorphous. } } } } Now to discuss Au/Pd. This material sputters } } totally differently than Au alone. The coating } } is like a layer of grains of sand. These dots, } } if you will, may touch one another in places and } } not in others. Unlike reflowed metal, the dots } } are distinct and quite 3-D. Is this an amorphous } } layer? If each dot was one grain, and each grain } } was not touching each other grain, is the coating } } amorphous or not? Does continuity play into the } } definition? } } } } At damascene vias, the ILD is definitely amorphous. } } These areas etch totally differently than areas } } away from the vias. It's interesting to see that } } areas away from vias exhibit some degree of } } amorphousness. Why the via areas are more strongly } } amorphous, I do not yet know. This is something } } I am working on at the moment. } } } } If I can find some Au and Au/Pd pix that I can } } post, I will do so. I have the pix somewhere and } } I have specimens (all over the place). If there } } is interest, I will work on getting them. } } } } gary g. } } } } } } } } } } } } At 05:34 AM 4/19/2004, you wrote: } } } Gary } } } } } } If you are truely producing amorphous Au this would } } } be very significant. I must admit to being somewhat } } } skeptical. } } } } } } An image of a near feature less "surface" is not sufficient } } } evidence to call something amorphous. } } } } } } The best way to answer this question would be to prepare } } } a TEM cross-section. Then using either HREM or Electron Diffraction from } } } the coating you could verify if the coating is crystalline } } } or not. } } } } } } } } } Nestor } } } } } } } } } } } } } ------------------------------------------------------------------------- } } } ----- } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } ------------------------------------------------------------------------- } } } ------ } } } } } } } } Using an FEI Sirion SFEG, the coating is amorphous. } } } } The "measurement" is looking at the image via the SEM. } } } } Plain Au is honeycombed. Would you like a sample of } } } } this? See what you can see at 350KX. What would you } } } } use to do this yourself? Let's compare images. } } } } If it would be productive, of course. } } } } } } } } How do we define amorphous versus non-amorphous } } } } at this mag? This is further complicated by the } } } } manner in which the coating is deposited. There } } } } are way too many variables. Perhaps I simplify } } } } the topic. } } } } } } } } So, what do you want to see? I can produce pix } } } } at 350KX of Au and Au/Pd coatings. If these are } } } } a big surprise to you, I can provide specimen images. } } } } Alternatively, show me what you have at this same } } } } mag. Produce Au and Au/Pd and Pt images. That pretty well } } } } covers the normal sputter coating environment. } } } } } } } } I am always open to new views on this subject. } } } } } } } } gary g. } } } } } } } } } } } } At 07:12 PM 4/18/2004, you wrote: } } } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } } } } } Gary Gaugler wrote: } } } } } ==================================================================== } } } } } .............What I do know is that if I switch to Au/Pd (60/40) or } Pt, the } } } } } coating is amorphous and cannot be easily seen. Au coating can be } seen at } } } } } about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. } } } } } However, Au/Pd and Pt look totally different than Au alone. Au/Pd and } } } Pt is } } } } } more granular rather than honeycomb. And it will look this way at } } } 350KX and } } } } } greater. } } } } } ==================================================================== } } } } } Could you tell us the measurement used to conclude it is an amorphous } } } } } coating? So far as I know, one does see a grain, even with Pt or Au/Pd } } } } } irrrespective of the brand of sputter coater used. } } } } } } } } } } Chuck } } } } } } } } } } ============================================ } } } } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } } } } } President 1-800-2424-SPI } } } } } SPI SUPPLIES FAX: 1-610-436-5755 } } } } } PO BOX 656 e-mail:cgarber-at-2spi.com } } } } } West Chester, PA 19381-0656 USA } } } } } Cust.Service: spi2spi-at-2spi.com } } } } } } } } } } Look for us! } } } } } ######################## } } } } } WWW: http://www.2spi.com } } } } } ######################## } } } } } ============================================ } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 21:28:32 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mmckillip-at-smurfitlcom) from on Monday, April 19, 2004 at 11:39:07 ---------------------------------------------------------------------------
Email: mmckillip-at-smurfitlcom Name: M McKillip
Organization: Smurfit Stone Container Corp
Title-Subject: [Microscopy] [Filtered] MListserver:Embedding polymer films
Question: Is there a method to enhance polymer film bonding to a Poly/Bed 812 embedding medium. Frequently the thin films release from the embedding medium during microtoming.
My intent was to try to put this into some kind of reasonable perspective. An 'artifact' in SEM is anything induced by the sample prep that is not representative of the actual sample you are studying. The whole point of development in coating systems has historically been to reduce the artifacts that are induced. The argument about whether to use gold (the age-old standard) or gold-palladium addresses the fact that gold alone produces grains (structures) that are visible at relatively low magnifications (let's say about 20kX). With Au-Pd you can get to higher magnifications before this becomes a problem. Are they really there in your image? Yes. Are they representative of your sample? No. That's why they are called 'artifacts'. If you are seeing the coating then, yes, there is a fine structure of the coating, whether it is 'grains' or 'honeycombs' or anything else. The goal of the coating is that there is no fine structure that can be misinterpreted as a characteristic of the actual sample. Again, none of this discussion has anything to do with 'crystallinity' and the use of 'amorphous' is apparently ambiguous in this case. I suggested 'uniform' but, again, all of your discussions so far have been aimed at suggesting that the 'smooth', 'uniform' and 'essentially invisible' coating on your sample is 'amorphous'. This is an attempt to get to the basic issue and leave out the semantics and confusion that seem to have crept in.
Drew Hirt Materials Research Laboratories, Inc. Tel (800) 424-1776 Fax (330) 750-0776 drew-at-hirt.com http://www.mrllab.com
on 4/19/04 10:35 PM, Gary Gaugler at gary-at-gaugler.com wrote:
} Whoa, I did not intend for this to get way } off base, out of hand, perverted, convoluted, } complicated, ...fill in the adjectives. } } Post: This is the point of issue. There is a fine structure } of the coating. At low mag, it cannot be seen. At } high mag, it can be seen. Is it an "artifact?" No. } It is real. But then, we need to define an artifact } in the context of coating, et. al. As Nestor and } Charles said, it is an issue of grains. This is } bounded. We can deal with this. } } If the SEM sees information of the coating, then what? } It is not an artifact. Furthermore, what does "uniform" } mean? It seems that your posting says that in } one case the coating is amorphous but in another } it is not...based on mag and artifacts. But "uniform" } is counter-amorphous... sigh. } } I think we are headed towards making a mountain out } of a mole hill. Perhaps we should let this dog lay. } If not, Nestor will keep us on track. Notwithstanding, } I will do some EBSD studies later this year and report } back to the list. } } gary g. } } } At 07:01 PM 4/19/2004, you wrote: } } To All: } } } } [unsnip]
The question about whether coatings deposited using a sputter coater are 'amorphous' or 'crystalline' seems to be in dispute here. On an atomic level, if you can make an amorphous metal then we should be able to image it in our high resolution TEM (JEOL 4000EX) and not find the orderly arrangement of atoms in the layers. For gold deposited on amorphous (definitely) silicon, we can see easily at 800kX (5MX final mag) that the gold 'islands' of about 5nm diameter are composed of crystalline gold with each atom being able to be identified and counted in the orderly structure. We do the same thing at interfaces in semiconductor devices to note the changes of atomic structure where two metal system combine. It should also be noted that if the layer is thin enough, the diffraction pattern is hard to find (very weak signal) but with a long enough dwell time you will definitely see that the metal layer has atomic level crystalline structure.
The argument that seems to be being presented here is semantic, not scientific. Whether or not a layer appears as a 'honeycomb' has nothing to do with the fact that the structure of the 'honeycomb' is crystalline or amorphous. For the purposes of SEM the ideal is to have an 'amorphous' layer from the standpoint that at the magnifications involved there is no contribution of 'ordered' structures from the coating. This has absolutely nothing to do with whether or not the atomic level structures are crystalline or amorphous. We routinely coat samples with whatever material is best suited for the images we want to obtain. That might be gold, Au/Pd, Cr, Ag, Ti or whatever else seems to work. For Auger work we even use deposited Li to eliminate charging without contributing significantly to the spectral data. For SEM you are striving to achieve a coating that allows imaging of your actual sample without adding 'artifacts' that will confuse the interpretation of your data. If that means that you deposit a } } gold/palladium or chromium layer with such fine structure that your SEM } } image sees only the information from your sample but doesn't present an } } artifact then, by all means, call it 'amorphous'. A more distinctive } } description might, however, be 'uniform' at the magnification used. By this } } definition the actual atomic level crystallinity of the layer is not called } } into question. If you really want to know about the atomic scale ordering } } then you need to move on to HRTEM since SEM is not sufficient to see the } } atomic scale features. You could also use atomic force microscopy to 'see' } } the atomic structure of your surface coating. } } } } Drew Hirt } } Materials Research Laboratories, Inc. } } Tel (800) 424-1776 } } Fax (330) 750-0776 } } drew-at-hirt.com } } http://www.mrllab.com } } } } } } on 4/19/04 11:57 AM, Gary Gaugler at gary-at-gaugler.com wrote: } } } } } } } } } } } } } ---------------------------------------------------------------------------- -} } - } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } ---------------------------------------------------------------------------- } } --} - } } } } } } Nestor: } } } } } } Indeed, amorphous likely means different things } } } to different folks and in different situations. } } } From a metallurgical or atomic standpoint, it } } } would mean that it lacks a definitive or distinct } } } crystalline structure--i.e., no lattice. On the } } } other hand, amorphous could mean that it is shapeless, } } } of no definite form or organization or methodical } } } arrangement. } } } } } } OK, does this mean no form or organization at the } } } eye ball level? At the magnifying glass level? } } } LM, SEM, TEM level? I think that I "see" the } } } problem of definition and ability to define it. } } } } } } The general consensus seems to be one of grains. } } } A honey comb shaped coating of Au would have the } } } metal portions containing or consisting of grains } } } of Au. Thus, the honey comb has organization and } } } methodical arrangement--not amorphous. And the } } } Au metal would have grains--not amorphous. Therefore, } } } sputtered Au is not amorphous. } } } } } } Now to discuss Au/Pd. This material sputters } } } totally differently than Au alone. The coating } } } is like a layer of grains of sand. These dots, } } } if you will, may touch one another in places and } } } not in others. Unlike reflowed metal, the dots } } } are distinct and quite 3-D. Is this an amorphous } } } layer? If each dot was one grain, and each grain } } } was not touching each other grain, is the coating } } } amorphous or not? Does continuity play into the } } } definition? } } } } } } At damascene vias, the ILD is definitely amorphous. } } } These areas etch totally differently than areas } } } away from the vias. It's interesting to see that } } } areas away from vias exhibit some degree of } } } amorphousness. Why the via areas are more strongly } } } amorphous, I do not yet know. This is something } } } I am working on at the moment. } } } } } } If I can find some Au and Au/Pd pix that I can } } } post, I will do so. I have the pix somewhere and } } } I have specimens (all over the place). If there } } } is interest, I will work on getting them. } } } } } } gary g. } } } } } } } } } } } } } } } } } } At 05:34 AM 4/19/2004, you wrote: } } } } Gary } } } } } } } } If you are truely producing amorphous Au this would } } } } be very significant. I must admit to being somewhat } } } } skeptical. } } } } } } } } An image of a near feature less "surface" is not sufficient } } } } evidence to call something amorphous. } } } } } } } } The best way to answer this question would be to prepare } } } } a TEM cross-section. Then using either HREM or Electron Diffraction from } } } } the coating you could verify if the coating is crystalline } } } } or not. } } } } } } } } } } } } Nestor } } } } } } } } } } } } } } } } } ------------------------------------------------------------------------- } } } } ----- } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } } To Subscribe/Unsubscribe -- } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } ------------------------------------------------------------------------- } } } } ------ } } } } } } } } } } Using an FEI Sirion SFEG, the coating is amorphous. } } } } } The "measurement" is looking at the image via the SEM. } } } } } Plain Au is honeycombed. Would you like a sample of } } } } } this? See what you can see at 350KX. What would you } } } } } use to do this yourself? Let's compare images. } } } } } If it would be productive, of course. } } } } } } } } } } How do we define amorphous versus non-amorphous } } } } } at this mag? This is further complicated by the } } } } } manner in which the coating is deposited. There } } } } } are way too many variables. Perhaps I simplify } } } } } the topic. } } } } } } } } } } So, what do you want to see? I can produce pix } } } } } at 350KX of Au and Au/Pd coatings. If these are } } } } } a big surprise to you, I can provide specimen images. } } } } } Alternatively, show me what you have at this same } } } } } mag. Produce Au and Au/Pd and Pt images. That pretty well } } } } } covers the normal sputter coating environment. } } } } } } } } } } I am always open to new views on this subject. } } } } } } } } } } gary g. } } } } } } } } } } } } } } } At 07:12 PM 4/18/2004, you wrote: } } } } } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } } } } } } } Gary Gaugler wrote: } } } } } } ==================================================================== } } } } } } .............What I do know is that if I switch to Au/Pd (60/40) or } } Pt, the } } } } } } coating is amorphous and cannot be easily seen. Au coating can be } } seen at } } } } } } about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. } } } } } } However, Au/Pd and Pt look totally different than Au alone. Au/Pd and } } } } Pt is } } } } } } more granular rather than honeycomb. And it will look this way at } } } } 350KX and } } } } } } greater. } } } } } } ==================================================================== } } } } } } Could you tell us the measurement used to conclude it is an amorphous } } } } } } coating? So far as I know, one does see a grain, even with Pt or Au/Pd } } } } } } irrrespective of the brand of sputter coater used. } } } } } } } } } } } } Chuck } } } } } } } } } } } } ============================================ } } } } } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } } } } } } President 1-800-2424-SPI } } } } } } SPI SUPPLIES FAX: 1-610-436-5755 } } } } } } PO BOX 656 e-mail:cgarber-at-2spi.com } } } } } } West Chester, PA 19381-0656 USA } } } } } } Cust.Service: spi2spi-at-2spi.com } } } } } } } } } } } } Look for us! } } } } } } ######################## } } } } } } WWW: http://www.2spi.com } } } } } } ######################## } } } } } } ============================================ } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 22:37:59 2004
I'm in violent agreement with you. What is of concern to me is that as I/we move to extreme sub-micron dimensions such as 0.18u and below, how does the coating affect what we see and can present?
The problem is that at 0.5u and larger, the coatings are not all that significant. However, for extreme sub-micron feature sizes, I'm worried about the coating. At what point does the coating just exist and then become an artifact and then cloud the specimen? Heisenberg uncertainty plays into this...perhaps.
At 350KX and above, Au is not invisible...IMO. Au/Pd is visible but looks different. Pt is pretty much invisible. Os seems like a very ideal coating. At 90nm, what is a good coating???
As semiconductor feature sizes continue to reduce, how does this affect our procedures for analyzing them? How do we analyze the specimens without having to explain coatings? If we have to explain them, it puts our findings into question...or argumentation enters into the picture. This is not good.
gary g.
At 08:44 PM 4/19/2004, you wrote: } Gary: } } My intent was to try to put this into some kind of reasonable perspective. } An 'artifact' in SEM is anything induced by the sample prep that is not } representative of the actual sample you are studying. The whole point of } development in coating systems has historically been to reduce the artifacts } that are induced. The argument about whether to use gold (the age-old } standard) or gold-palladium addresses the fact that gold alone produces } grains (structures) that are visible at relatively low magnifications (let's } say about 20kX). With Au-Pd you can get to higher magnifications before } this becomes a problem. Are they really there in your image? Yes. Are } they representative of your sample? No. That's why they are called } 'artifacts'. If you are seeing the coating then, yes, there is a fine } structure of the coating, whether it is 'grains' or 'honeycombs' or anything } else. The goal of the coating is that there is no fine structure that can } be misinterpreted as a characteristic of the actual sample. Again, none of } this discussion has anything to do with 'crystallinity' and the use of } 'amorphous' is apparently ambiguous in this case. I suggested 'uniform' } but, again, all of your discussions so far have been aimed at suggesting } that the 'smooth', 'uniform' and 'essentially invisible' coating on your } sample is 'amorphous'. This is an attempt to get to the basic issue and } leave out the semantics and confusion that seem to have crept in. } } Drew Hirt } Materials Research Laboratories, Inc. } Tel (800) 424-1776 } Fax (330) 750-0776 } drew-at-hirt.com } http://www.mrllab.com } } } } on 4/19/04 10:35 PM, Gary Gaugler at gary-at-gaugler.com wrote: } } } Whoa, I did not intend for this to get way } } off base, out of hand, perverted, convoluted, } } complicated, ...fill in the adjectives. } } } } Post: This is the point of issue. There is a fine structure } } of the coating. At low mag, it cannot be seen. At } } high mag, it can be seen. Is it an "artifact?" No. } } It is real. But then, we need to define an artifact } } in the context of coating, et. al. As Nestor and } } Charles said, it is an issue of grains. This is } } bounded. We can deal with this. } } } } If the SEM sees information of the coating, then what? } } It is not an artifact. Furthermore, what does "uniform" } } mean? It seems that your posting says that in } } one case the coating is amorphous but in another } } it is not...based on mag and artifacts. But "uniform" } } is counter-amorphous... sigh. } } } } I think we are headed towards making a mountain out } } of a mole hill. Perhaps we should let this dog lay. } } If not, Nestor will keep us on track. Notwithstanding, } } I will do some EBSD studies later this year and report } } back to the list. } } } } gary g. } } } } } } At 07:01 PM 4/19/2004, you wrote: } } } To All: } } } } } } [unsnip] } } The question about whether coatings deposited using a sputter coater are } 'amorphous' or 'crystalline' seems to be in dispute here. On an atomic } level, if you can make an amorphous metal then we should be able to image it } in our high resolution TEM (JEOL 4000EX) and not find the orderly } arrangement of atoms in the layers. For gold deposited on amorphous } (definitely) silicon, we can see easily at 800kX (5MX final mag) that the } gold 'islands' of about 5nm diameter are composed of crystalline gold with } each atom being able to be identified and counted in the orderly structure. } We do the same thing at interfaces in semiconductor devices to note the } changes of atomic structure where two metal system combine. It should also } be noted that if the layer is thin enough, the diffraction pattern is hard } to find (very weak signal) but with a long enough dwell time you will } definitely see that the metal layer has atomic level crystalline structure. } } The argument that seems to be being presented here is semantic, not } scientific. Whether or not a layer appears as a 'honeycomb' has nothing to } do with the fact that the structure of the 'honeycomb' is crystalline or } amorphous. For the purposes of SEM the ideal is to have an 'amorphous' } layer from the standpoint that at the magnifications involved there is no } contribution of 'ordered' structures from the coating. This has absolutely } nothing to do with whether or not the atomic level structures are } crystalline or amorphous. We routinely coat samples with whatever material } is best suited for the images we want to obtain. That might be gold, Au/Pd, } Cr, Ag, Ti or whatever else seems to work. For Auger work we even use } deposited Li to eliminate charging without contributing significantly to the } spectral data. For SEM you are striving to achieve a coating that allows } imaging of your actual sample without adding 'artifacts' that will confuse } the interpretation of your data. If that means that you deposit a } } } gold/palladium or chromium layer with such fine structure that your SEM } } } image sees only the information from your sample but doesn't present an } } } artifact then, by all means, call it 'amorphous'. A more distinctive } } } description might, however, be 'uniform' at the magnification } used. By this } } } definition the actual atomic level crystallinity of the layer is not } called } } } into question. If you really want to know about the atomic scale ordering } } } then you need to move on to HRTEM since SEM is not sufficient to see the } } } atomic scale features. You could also use atomic force microscopy to } 'see' } } } the atomic structure of your surface coating. } } } } } } Drew Hirt } } } Materials Research Laboratories, Inc. } } } Tel (800) 424-1776 } } } Fax (330) 750-0776 } } } drew-at-hirt.com } } } http://www.mrllab.com } } } } } } } } } on 4/19/04 11:57 AM, Gary Gaugler at gary-at-gaugler.com wrote: } } } } } } } } } } } } } } } } } } } ---------------------------------------------------------------------------- } -} } - } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } ---------------------------------------------------------------------------- } } } --} - } } } } } } } } Nestor: } } } } } } } } Indeed, amorphous likely means different things } } } } to different folks and in different situations. } } } } From a metallurgical or atomic standpoint, it } } } } would mean that it lacks a definitive or distinct } } } } crystalline structure--i.e., no lattice. On the } } } } other hand, amorphous could mean that it is shapeless, } } } } of no definite form or organization or methodical } } } } arrangement. } } } } } } } } OK, does this mean no form or organization at the } } } } eye ball level? At the magnifying glass level? } } } } LM, SEM, TEM level? I think that I "see" the } } } } problem of definition and ability to define it. } } } } } } } } The general consensus seems to be one of grains. } } } } A honey comb shaped coating of Au would have the } } } } metal portions containing or consisting of grains } } } } of Au. Thus, the honey comb has organization and } } } } methodical arrangement--not amorphous. And the } } } } Au metal would have grains--not amorphous. Therefore, } } } } sputtered Au is not amorphous. } } } } } } } } Now to discuss Au/Pd. This material sputters } } } } totally differently than Au alone. The coating } } } } is like a layer of grains of sand. These dots, } } } } if you will, may touch one another in places and } } } } not in others. Unlike reflowed metal, the dots } } } } are distinct and quite 3-D. Is this an amorphous } } } } layer? If each dot was one grain, and each grain } } } } was not touching each other grain, is the coating } } } } amorphous or not? Does continuity play into the } } } } definition? } } } } } } } } At damascene vias, the ILD is definitely amorphous. } } } } These areas etch totally differently than areas } } } } away from the vias. It's interesting to see that } } } } areas away from vias exhibit some degree of } } } } amorphousness. Why the via areas are more strongly } } } } amorphous, I do not yet know. This is something } } } } I am working on at the moment. } } } } } } } } If I can find some Au and Au/Pd pix that I can } } } } post, I will do so. I have the pix somewhere and } } } } I have specimens (all over the place). If there } } } } is interest, I will work on getting them. } } } } } } } } gary g. } } } } } } } } } } } } } } } } } } } } } } } } At 05:34 AM 4/19/2004, you wrote: } } } } } Gary } } } } } } } } } } If you are truely producing amorphous Au this would } } } } } be very significant. I must admit to being somewhat } } } } } skeptical. } } } } } } } } } } An image of a near feature less "surface" is not sufficient } } } } } evidence to call something amorphous. } } } } } } } } } } The best way to answer this question would be to prepare } } } } } a TEM cross-section. Then using either HREM or Electron Diffraction } from } } } } } the coating you could verify if the coating is crystalline } } } } } or not. } } } } } } } } } } } } } } } Nestor } } } } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------------- } } } } } ----- } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } } } } To Subscribe/Unsubscribe -- } } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } ------------------------------------------------------------------------- } } } } } ------ } } } } } } } } } } } } Using an FEI Sirion SFEG, the coating is amorphous. } } } } } } The "measurement" is looking at the image via the SEM. } } } } } } Plain Au is honeycombed. Would you like a sample of } } } } } } this? See what you can see at 350KX. What would you } } } } } } use to do this yourself? Let's compare images. } } } } } } If it would be productive, of course. } } } } } } } } } } } } How do we define amorphous versus non-amorphous } } } } } } at this mag? This is further complicated by the } } } } } } manner in which the coating is deposited. There } } } } } } are way too many variables. Perhaps I simplify } } } } } } the topic. } } } } } } } } } } } } So, what do you want to see? I can produce pix } } } } } } at 350KX of Au and Au/Pd coatings. If these are } } } } } } a big surprise to you, I can provide specimen images. } } } } } } Alternatively, show me what you have at this same } } } } } } mag. Produce Au and Au/Pd and Pt images. That pretty well } } } } } } covers the normal sputter coating environment. } } } } } } } } } } } } I am always open to new views on this subject. } } } } } } } } } } } } gary g. } } } } } } } } } } } } } } } } } } At 07:12 PM 4/18/2004, you wrote: } } } } } } } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } } } } } } } } } Gary Gaugler wrote: } } } } } } } ==================================================================== } } } } } } } .............What I do know is that if I switch to Au/Pd (60/40) or } } } Pt, the } } } } } } } coating is amorphous and cannot be easily seen. Au coating can be } } } seen at } } } } } } } about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. } } } } } } } However, Au/Pd and Pt look totally different than Au alone. Au/Pd and } } } } } Pt is } } } } } } } more granular rather than honeycomb. And it will look this way at } } } } } 350KX and } } } } } } } greater. } } } } } } } ==================================================================== } } } } } } } Could you tell us the measurement used to conclude it is an amorphous } } } } } } } coating? So far as I know, one does see a grain, even with Pt or } Au/Pd } } } } } } } irrrespective of the brand of sputter coater used. } } } } } } } } } } } } } } Chuck } } } } } } } } } } } } } } ============================================ } } } } } } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } } } } } } } President 1-800-2424-SPI } } } } } } } SPI SUPPLIES FAX: 1-610-436-5755 } } } } } } } PO BOX 656 e-mail:cgarber-at-2spi.com } } } } } } } West Chester, PA 19381-0656 USA } } } } } } } Cust.Service: spi2spi-at-2spi.com } } } } } } } } } } } } } } Look for us! } } } } } } } ######################## } } } } } } } WWW: http://www.2spi.com } } } } } } } ######################## } } } } } } } ============================================ } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 03:17:16 2004
Hi Frank, Your new computer should have a minimum of one gigabyte RAM. Imaging processing applications are memory hungry. A digital video capture card maintains the integrity of your data. If you convert digital to analog and back to digital loss of image quality may occur. ATI Radion makes a good combined video-capture card, some people prefer a separate capture card. This will also have the fast connections should you upgrade your camera at some time in the future. Two hard drives allow you to keep your programs and applications separate from large image and other data files. This speeds up your system and decreases maintenance (like defragmenting). It makes backup easier and your files are on a separate device if your 'C' drive fails. DVD burners have come down in price (under $200.) and can store over 4gB . Sony just released their 4th generation 8X DVD burners. They developed the DVD format and have a reputation for reliability. (Be aware that there are two DVD formats, "+" and "-". Most current brands and models can use both types of media.)
Bob Carter 2000 Bayshore Road Lopez Island, WA 98261
-----Original Message----- } From: Frank.Karl-at-degussa.com [mailto:Frank.Karl-at-degussa.com] Sent: Monday, April 19, 2004 6:35 AM To: microscopy-at-msa.microscopy.com
I've been tasked with pulling together sensible specifications for a new computer to capture digital images from our pol scope. Many options appear to be straight forward: More RAM, - 512 MB seems to be enough. More HD storage, - As Daisy use to say, you can never be too rich, too thin or have enough storage. I think 120GB is enough. CD burner at 48X seems enough
but which monitor? Should I look for a fast response time, say 16ms, to help me focus? Are flat screens better for viewing than the more traditional monitors? Do I need more then 17 inch diagonal screen? Is the integrated Intel 3D Extreme Graphics video card a good choice for photomicrography?
Any suggestions for a good driver to capture my images? I intend to use photoshop to do any post imaging processing that needed, but I would like to be able to do a little tweaking to maximize my images when I capture them.
I have also inhered a Microimaging Video system and a RGB Automatic Camera, but I have no instructions or user manual. If anyone knows a web site or has a copy they would like to share I would be grateful!
Any advice, opinions or alternative suggestions are welcome!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 08:31:08 2004
Does anyone have a fixation procedure to recommend for E. coli? These cells are being used as a control for some more hazardous strains and must be chemically fixed in a containment hood prior to being brought to our facility.
The E. coil cells were fixed using 2.5% glut + 2% PAF in 0.1M Cacodylate buffer, pH 7.4, (with added 1mM CaCl2, 2mM MgCl2.6H20, 0.25% NaCl) followed by osmium post-fix, ETOH dehydration and embedding in 812 generic resin or 812 generic:spurr mixture. This has produced cells with obvious collapse of the plasma membrane at the poles of the cells. The pulling away of the plasma membrane from the cell wall gives the initial appearance of large vacuoles at either end of the cell. Other bacteria strains, such as listeria, have been well preserved using this protocol.
However, E. coli cells that were fixed in the same primary fix-buffer combination and then plunge frozen do not show the same artifact. In this case the cells were freeze-substituted in acetone-osmium and then embedded in Spurr resin. I was surprised at the difference since both samples were initially fixed identically. This seems to indicate that the difference might not be related to osmolarity of the primary fix as I initially suspected.
I might note that these cells were grown in liquid culture. The above protocol works fine with cells grown on agar.
Older references suggest using acrolein for primary fixation. I prefer to not use this chemical due to it¹s hazardous nature. Other older references utilize vernal-acetate buffer which is also no longer desirable due to it¹s classification as a restricted substance requiring special permissions to purchase.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 09:06:40 2004
I don't know if you made this clearer at an earlier stage, but a major factor in handling and storage will be the resolution and quality of images that you capture from your microscope. There will be great difference between a 1 mega pixel image captured/stored in JPEG format and 5-6 mega pixel image captured/stored in TIF or RAW format - a factor of 50x or more.
If you are after bigger formats then DVD writers must be a better prospect than CD writers especially as they should be able to write to CDs as well. But if you're archiving to these disks you would need to consider that none are considered as archival quality. I think most experts suggest that for 5-10 year storage you should maintain at least one duplicate CD or DVD disks ie if one goes wrong after use then check the unused backup.
I would also point out that there are really 5 DVD formats (DVD+R, DVD-R, DVD+RW, DVD-RW and DVD RAM). Although it is the slowest of the 5 formats, DVD RAM does guarantee 100,000x writes compared with 1,000x for the others so it should be the nearest to archival. I believe this is because of a more meticulous error checking system. At present I think Iomega and LG are the only manufacturers of writers for all of the formats. But the big problem is that because DVD RAM disks are more expensive and slower, they are less popular in the West (a bit more popular in Japan I think). So if you used a system like that you may want RAM for internal use and say DVD+R for customers or other users.
I hope I have got all my facts correct and haven't just muddied the waters.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences University of Sunderland Tyne & Wear UK
e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Bob Carter {bob-at-rockisland.com}
Perhaps "the answer" is to use a microscope that allows high-resolution imaging of your structures without any coating, through proper choice of low voltage and/or Variable Pressure imaging conditions ( in VP mode, nitrogen/air ionized by the secondaries generated by the primary beam acts as both the neutralizing gas and as an "internal scintillator"). Through both improved low-voltage imaging resolution and VP mode, high-resolution secondary electron imaging on uncoated "charge challenged" samples is achievable on the latest generation secondary electron microscopes.
Regards, Ed
*********************** Edward L. Principe, Ph.D. Applications Development Scientist Zeiss SMT, Inc. (formerly LEO Electron Microscopy) ***************************** ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "Materials Research Laboratories, Inc." {mrllab-at-raex.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Monday, April 19, 2004 8:59 PM
Hello Microscopists !
First I looked on the MSA website for electron microscopy service labs in the Cleveland, Ohio metropolitan area - and even with Google. All to no avail. So now I'm beating the drums on the Microscopy Listserver.
The specimens will be some broken wheel bolts - about the size you'd have holding your car's wheels on. Mebbe 9/16 inch diameter and an inch and a half long. Both fractography and metallographically prepared cross sections. Mebbe microhardness, too. Ability to EDX or WDS carbon would be a plus. This would be a common-ground examination as part of a legal case. We're trying to piggyback the lab work onto the on-site exam; hence the need for a lab in the Cleveland area. Don't be bashful. I'll submit results to the firm that's organizing the party through my own counsel/client.
Best regards, George Langford, Sc.D. Amenex Associates, Inc. http://wwww.amenex.com/
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 12:07:39 2004
pick your favourite fixation method. the bacterium will be inactivated by formaldehyde, formaldehyde:glutaraldehyde, or glutaraldehyde. this has always worked for us, anyway
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 12:08:10 2004
Hi Debby, All the bacteria that I have done like a lower pH. (was microscopist for Microbiology Dept at Univ. of Illinois and looked at all the bugs that came in the Dept for various types of research) That is why veronal acetate buffer was used for so long because it buffers well at pHs of 6.8 or so. It however, as you pointed out, is now restricted. We get a different appearance of the walls when we do a strictly Os fix and one involving both G and Os especially obvious in the Gram Negative bacteria. The traditional "Kellenberger" method puts Os directly in the liquid growth media and pellets the bugs down then resuspends them in fresh Os. It appears you cannot do that however because of the necessary pre-treatment for safety purposes. We get fairly good preservation however using either Os only or both G and Os. We always do an en bloc UA stain after the Osmium because of the free nucleic acids which stain nicely. I assume you are putting the bugs in agar? Perhaps you might want to send me an image. Have you peeked at the bugs in SEM to see what their surface looks like with a simple G, Os, dehydration, and CPD or freeze drying run? That would also give you an idea of how crenulated the surface might be expected to be. Don't know if any of the above helped, but just thought I would toss in my two cents.
Good Luck, Judy
Debby Sherman wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Does anyone have a fixation procedure to recommend for E. coli? These cells } are being used as a control for some more hazardous strains and must be } chemically fixed in a containment hood prior to being brought to our } facility. } } The E. coil cells were fixed using 2.5% glut + 2% PAF in 0.1M Cacodylate } buffer, pH 7.4, (with added 1mM CaCl2, 2mM MgCl2.6H20, 0.25% NaCl) } followed by osmium post-fix, ETOH dehydration and embedding in 812 generic } resin or 812 generic:spurr mixture. This has produced cells with obvious } collapse of the plasma membrane at the poles of the cells. The pulling away } of the plasma membrane from the cell wall gives the initial appearance of } large vacuoles at either end of the cell. Other bacteria strains, such as } listeria, have been well preserved using this protocol. } } However, E. coli cells that were fixed in the same primary fix-buffer } combination and then plunge frozen do not show the same artifact. In this } case the cells were freeze-substituted in acetone-osmium and then embedded } in Spurr resin. I was surprised at the difference since both samples were } initially fixed identically. This seems to indicate that the difference } might not be related to osmolarity of the primary fix as I initially } suspected. } } I might note that these cells were grown in liquid culture. The above } protocol works fine with cells grown on agar. } } Older references suggest using acrolein for primary fixation. I prefer to } not use this chemical due to it1s hazardous nature. Other older references } utilize vernal-acetate buffer which is also no longer desirable due to it1s } classification as a restricted substance requiring special permissions to } purchase. } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 13:13:57 2004
George , Try Smithers in Akron (330-762-7441 ARDL also in Akron at 330-794-660 Noveon between Akron and Cleveland at 216-447-5000.
Good luck
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
"George Langford, To: Microscopy-at-MSA.Microscopy.com Sc.D." cc: "George Langford, Sc.D." {amenex-at-amenex.com} {amenex-at-amene Subject: [Microscopy] Commercial SEM labs in the x.com} Cleveland, OH area ?
04/20/2004 11:48 AM Please respond to "George Langford, Sc.D."
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Microscopists !
First I looked on the MSA website for electron microscopy service labs in the Cleveland, Ohio metropolitan area - and even with Google. All to no avail. So now I'm beating the drums on the Microscopy Listserver.
The specimens will be some broken wheel bolts - about the size you'd have holding your car's wheels on. Mebbe 9/16 inch diameter and an inch and a half long. Both fractography and metallographically prepared cross sections. Mebbe microhardness, too. Ability to EDX or WDS carbon would be a plus. This would be a common-ground examination as part of a legal case. We're trying to piggyback the lab work onto the on-site exam; hence the need for a lab in the Cleveland area. Don't be bashful. I'll submit results to the firm that's organizing the party through my own counsel/client.
Best regards, George Langford, Sc.D. Amenex Associates, Inc. http://wwww.amenex.com/
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 13:33:38 2004
Although we have always kept OsO4 wrapped up to protect it from light because someone somewhere said that it was light sensitive, now I'm wondering if that is indeed true or not. I personally cannot really remember if it is, or if this is just a superstition. How do you people store your OsO4, and is it really light sensitive or not?
Garry
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From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 14:04:23 2004
Last week I posted an inquiry regarding an instruction manual for an older model Olympus microscope, and noted that Olympus America had not responded to my e-mails from their web site. Thanks to those who responded, and I must add that Olympus America responded immediately to my posting on this microscopy site. In fact, Olympus provided three e-mail responses, phone calls, a manual, some needed technical information and additional support links. They were terrific. Good job.
John J. Wolosewick, Ph.D. Department of Anatomy and Cell Biology M/C 512 University of Illinois at Chicago 808 S. Wood Street Chicago, Illinois 60612
jjwolo-at-uic.edu 312-996-6022
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 16:35:04 2004
Materials Research Laboratories, Inc. 290 North Bridge Street Struthers, OH 44471 800 424-1776 http://www.mrllab.com/
} } George , } Try Smithers in Akron (330-762-7441 } ARDL also in Akron at 330-794-660 } Noveon between Akron and Cleveland at 216-447-5000. } } Good luck } } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 } } } 330-668-2235 Ext. 238 } } } } "George } Langford, To: Microscopy-at-MSA.Microscopy.com } Sc.D." cc: "George Langford, Sc.D." {amenex-at-amenex.com} } {amenex-at-amene Subject: [Microscopy] Commercial SEM labs in the } x.com} Cleveland, OH area ? } } 04/20/2004 } 11:48 AM } Please } respond to } "George } Langford, } Sc.D." } } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } } Hello Microscopists ! } } First I looked on the MSA website for electron microscopy service labs } in the Cleveland, Ohio metropolitan area - and even with Google. All } to no avail. So now I'm beating the drums on the Microscopy Listserver. } } The specimens will be some broken wheel bolts - about the size } you'd have holding your car's wheels on. Mebbe 9/16 inch diameter } and an inch and a half long. Both fractography and metallographically } prepared cross sections. Mebbe microhardness, too. Ability to EDX } or WDS carbon would be a plus. This would be a common-ground } examination as part of a legal case. We're trying to piggyback the } lab work onto the on-site exam; hence the need for a lab in the } Cleveland area. Don't be bashful. I'll submit results to the firm } that's organizing the party through my own counsel/client. } } Best regards, } George Langford, Sc.D. } Amenex Associates, Inc. } http://wwww.amenex.com/ } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 16:58:31 2004
Quoting Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} : } Although we have always kept OsO4 wrapped up to protect it from light } because someone somewhere said that it was light sensitive, now I'm } wondering if that is indeed true or not. I personally cannot really } remember if it is, or if this is just a superstition. How do you people } store your OsO4, and is it really light sensitive or not? } } Garry +++++++++++++ Garry, I store the OsO4 in the refrigerator and I KNOW that the light is off because there is no light bulb in there!
Pat Connelly Dept. of Biology Univ. of Pennsylvania
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 16:58:27 2004
Quoting Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} : } Although we have always kept OsO4 wrapped up to protect it from light } because someone somewhere said that it was light sensitive, now I'm } wondering if that is indeed true or not. I personally cannot really } remember if it is, or if this is just a superstition. How do you people } store your OsO4, and is it really light sensitive or not? } } Garry +++++++++++++ Garry, I store the OsO4 in the refrigerator and I KNOW that the light is off because there is no light bulb in there!
Pat Connelly Dept. of Biology Univ. of Pennsylvania
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 20:47:01 2004
Debby I did not have problems with E.coli fixation using 1.5% GA in PBS 1-2h on ice. Your fixation looks over killing to me (for how long, overnight I guess?). I don't see any reason to use formaldehyde for EM if antigenity is not a critical issue. Usually people use formaldehyde for immuno-EM in combination with low GA concentration (so formaldehyde partially substitute GA) . Over fixation by itself may cause the shrinkage and other artefacts. Sergey.
At 06:52 AM 4/20/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pamarone-at-bdsinternational.com) from on Tuesday, April 20, 2004 at 06:39:53 ---------------------------------------------------------------------------
Question: Does anyone have any information or references regarding staining/labelling for SEM? I would like to follow a liposome-based structure through the digestive tract, through its metabolism and would like to have a technique for labelling such a structure. Thank you
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (curtlane2003-at-yahoo.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, April 20, 2004 at 22:41:46 ---------------------------------------------------------------------------
Email: curtlane2003-at-yahoo.com Name: curt lane
Organization: lane assoc.
Title-Subject: [Microscopy] [Filtered] MListserver:sem service labs
Question: I am trying to get a list of sem service labs currently operating in northern california. thank you
We do outside SEM work and we are located in Euclid. You can visit our website at www.atclabs.com for more information. We have 2 SEM digitally equipped with PGT systems which helps us to produce more presentable photos EDS and WDS analysis. We also have a full metallography, chemistry, mechanical, and X-ray labs. For more information please contact Joe Radisek at 216-692-5456 e-mail: Radisek-at-argo-tech.com or myself.
Regards,
Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
"George
Langford, To: Microscopy-at-MSA.Microscopy.com Sc.D." cc: "George Langford, Sc.D." {amenex-at-amenex.com} {amenex-at-amene Subject: [Microscopy] Commercial SEM labs in the x.com} Cleveland, OH area ?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (le_thiec-at-nancy.inra.fr) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 21, 2004 at 08:28:31 ---------------------------------------------------------------------------
Email: le_thiec-at-nancy.inra.fr Name: Le Thiec
Organization: INRA
Title-Subject: [Microscopy] [Filtered] MListserver: WDS and cryo
Question: Dear all
I would like to analyse the element composition of biological samples also I am looking for a scanning electron microscope (field emission will be better) with cryo system and WDS system. it will be possible for me to go everywhere in the world. If you know a laboratory which has a SEM with WDS and cryo, please contact me. Thank you in advances. Didier
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (fremingt-at-fhcrc.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 21, 2004 at 08:43:33 ---------------------------------------------------------------------------
Question: Does anyone have sucessful results fixing and processing samples of conidia of aspergillus for sem? We have tried many methods, but instead of being round, they have places where the walls seem to have indented. We have tried 2% gluteraldehyde fixation, with and without post-fixing in osmium. We have dehydrated in both alcohols and acetone and used HMDS and nothing changes the result.
Any help would be appreciated.
F. Remington fremingt-at-fhcrc.org Fred Hutchinson Cancer Research Center
Due to the dissolution of a lab, 13 steel histology knives are available for free to any educational and/or non-profit organization/lab/personnel. They are not available to any commercial outlet.
Description - 12 specimens, 12 cm blades, most likely stainless steel, date from approximately 1940-1960, Each in box. Look decent, most likely will require some sharpening, though they are sharp to the finger.
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Daniel Geiger -- ***************************************************************************************
Daniel L. Geiger, Ph.D. Research Associate Santa Barbara Museum of Natural History Adjunct Assistant Professsor, University of Southern California, Los Angeles
Mailing Address: Molecular Systematics Lab, Natural History Museum of Los Angeles County 900 Exposition Blvd., Los Angeles, CA 90007, USA phone: (213) 763 3431 fax: (213) 748 4432 NEW e-mail geiger-at-vetigastropoda.com NEW www http://www.vetigastropoda.com
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 11:30:57 2004
Try vapor fixing in osmium vapor. Put you material in a small container (I use a glass Petri dish sealed with double parafilm) with a few drops of 4% osmium tetroxide. Leave it for about 72 hours at RT in a hood, then air dry. Aspergillus conidia tend to dimple a bit naturally, but this will stabilize them as well as,any method, and better than most. Robert
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 12:07:47 2004
After many years of living with osmicated 'fridges, I tried an experiment by keeping the osmium tetroxide at the back of the chemical hood at room temperature.
We only keep about 50ml at a time and the chemical hood is always on, so there didn't seem to be any vapour hazard (unlike the smelly 'fridge).
After 5 years of doing this, I can say that there seems to be no harm in keeping 2% osmium tetroxide at room temperature and in full light every day. Now keep the osmium tetroxide in full light and at room temperature and we have the added bonus of a clean fridge.
Hope this is useful.
PS. Thanks Nestor.
Regards,
Paul Webster.
Paul Webster, Ph.D. House Ear Institute 2100 West Third Street Los Angeles CA 90057 phone (213) 273 8026 fax (213) 413 6739 email: pwebster-at-hei.org
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 12:35:05 2004
This is a quote from Advanced Inorganic Chemistry, 2nd Ed. by Cotton and Wilkinson on page 1005 under tetroxides. This is a well known book. "Above ~180°, RuO4 can explode, giving RuO2 and O2, and it is decomposed slowly by light; OsO4 is more stable in both respects." The later statement suggests or implies that OsO4 can also be decomposed by light because it does NOT say, "; OsO4 is very stable and does not decompose or explode."
So I always wrapped OsO4 in Al foil as a minimum and kept all of it in a glass container tray is case of a broken ampoule. OsO4 ampoules are not something you want out in the open on a bench anyway. For potential RuO4 spills, use a crystallizing dish lined and padded on the bottom with paper towels. Unsaturated corn oil is recommended for OsO4 but it's a mess to clean up. Yuk!
The article in the Sept-Oct 2002 issue of Microscopy Today on page 20 would indicate that the reaction rates are indeed quite different. I sent you an email reprint of the discussion of some of the chemistry of Os and Ru and their reaction indicators in this article.
Paul Beauregard Senior Research Associate Microscopy and Image Analysis
At 01:54 PM 04/20/04 -0500, Garry Burgess wrote: } } } --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 15:04:29 2004
I would like to remind researchers that osmium made up in distilled water is very stable at room temperature. However, if some of you make up a 1% in buffer and store as such then definitely refrigerate your solution because reduces at a faster rate. We have noticed this in our laboratory and now only make up 2% aqueous and dilute with an equal amount of buffer when we need it. We store it in the hood in a glass stoppered 100 ml wide mouth bottle which is enclosed in a small bell jar.
Karen Bentley
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
-----Original Message----- } From: Webster, Paul [mailto:PWebster-at-hei.org] Sent: Wednesday, April 21, 2004 1:29 PM To: MSA listserver submission (E-mail) Cc: Nestor-at-Zaluzec.Com; Zaluzec-at-aaem.amc.anl.gov
Hi,
After many years of living with osmicated 'fridges, I tried an experiment by keeping the osmium tetroxide at the back of the chemical hood at room temperature.
We only keep about 50ml at a time and the chemical hood is always on, so there didn't seem to be any vapour hazard (unlike the smelly 'fridge).
After 5 years of doing this, I can say that there seems to be no harm in keeping 2% osmium tetroxide at room temperature and in full light every day. Now keep the osmium tetroxide in full light and at room temperature and we have the added bonus of a clean fridge.
Hope this is useful.
PS. Thanks Nestor.
Regards,
Paul Webster.
Paul Webster, Ph.D. House Ear Institute 2100 West Third Street Los Angeles CA 90057 phone (213) 273 8026 fax (213) 413 6739 email: pwebster-at-hei.org
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 18:12:25 2004
William; If you see this problem with the beam off, it is certainly NOT your phosphor causing the problem. This noise is typical of a CCD readout when the CCD is a frame read-out, not an interline readout and when thermal noise is excessive due to a failure of the cooling system.
John Mardinly Intel
-----Original Message----- } From: William Stratton [mailto:wgstratton-at-wisc.edu] Sent: Monday, April 19, 2004 7:11 AM To: Microscopy
Hello all,
Thank you for all of your tips. I got some great responses with some really good ideas to solve this problem.
To clear up some questions: All images that I see this problem with are acquired when the beam is off (no intensity on the CCD), the images are also raw (not dark count or gain corrected). The problem is not with an artifact present in all CCD images, but rather the horizontal line profile of the image (a measure of the average pixel value across the image horizontally) increases from left to right.
What the solution may be: From responses I've received it may be that the scintallator may have corroded from prolonged exposure to condensation/contamination within the TEM column. We're going to try and clean it off to rid ourselves of the problem.
Thanks again!
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 19:36:20 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jlflow2-at-uky.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Wednesday, April 21, 2004 at 16:07:08 ---------------------------------------------------------------------------
Email: jlflow2-at-uky.edu Name: Jen Flowers
Organization: University of Kentucky, Dept. of Plant Pathology
Education: Graduate College
Location: Lexington, KY, USA
Question: Hi, I am looking for a contact that is familiar with viewing conifer tissue using confocal microscopy. I plan on looking at a fungal pathogen growing through pine tissue for part of my dissertation research. Any help would be greatly appreciated
Hello, Anyone have a Med 020 sputter coater/evaporator I could get some training on? We bought a used unit and the manual doesn't have a very good section on how to procedureally use it. Thanks, any advice appreciated.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 23:21:36 2004
Just a quick email to say thank you to all who answered my earlier posting and who gave of their time and knowledge. After much fiddling with my obselete PC I finally got some software working and managed to export my images.
Regards George
George Theodossiou Physicist / Electron Microscopist
AMCOR Research and Technology Ph: +61 3 9490 6135 Fax: +61 3 9499 4295 Mobile: 0409 568 840 email: George.Theodossiou-at-amcor.com.au
************************************************************************ CAUTION - This message may contain privileged and confidential information intended only for the use of the addressee named above. If you are not the intended recipient of this message you are hereby notified that any use, dissemination, distribution or reproduction of this message is prohibited. If you have received this message in error please notify AMCOR immediately. Any views expressed in this message are those of the individual sender and may not necessarily reflect the views of AMCOR. ************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 00:00:32 2004
Hi Jen We had a project which looked at a fungus infecting pine.
The fungus was transfected with GFP. The pine autofluoresced in the red and the green. We ballanced up the autofluorescence and then with the software on the Bio-Rad Radiance subtracted the red from the green just leaving the fungus GFP in the green which we could then overlay with the red autofluorescence. This was published as "The use of the green fluorescent protein as a biomarker for sapstain fungi" by S.Lee, S.H. Kim and C. Breuil For. Path. 32 (2002) 153-161
The Bio-Rad Radiance worked very well on this project but if you have access to a spectral imaging system, you can also eliminate the autofluorescence using software.
Elaine
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From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 12:57:20 2004
I am looking to acquire (beg, borrow, purchase) a Philips (now FEI) multiple sample holder PW6598 for a CM12-TEM. It allows for three TEM grids to inserted at the same time.
Any suggestions would be appreciated.
thanks and regards,
Jim
********************************************************* Dr. Jim Quinn james.quinn-at-sunysb.edu Materials Science 631-632-6663 FAX:8052 SUNY at Stony Brook http://www.matscieng.sunysb.edu/ Stony Brook, New York 11794 - 2275 *********************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 14:18:28 2004
I have never fixed human tissue overnight in Osmium Tetroxide, or even left it in lower concentration alcohols, for fear of ultrastructural details being leached away during the night, but lately I'm left wondering if perhaps one could get away with fixing animal [human] tissue in 1% Osmium Tetroxide overnight in the fridge. Prior to this, we've always fixed in Osmium for half an hour to an hour.
Does anyone have any experience with overnight fixation of animal tissue in 1% Osmium Tetroxide in water overnight? It would be a great time saver to me if I knew for certain that this wouldn't adversely affect the ultrastructure of the tissue.
Garry
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 14:38:54 2004
Try checking the booklet Identification of Common Aspergillus Species by Dr. Maren A. Klich , mycologist from our facility, published by Centraalbureau voor Schimmelcultures, Utrecht (2002). Different species were imaged in a XL 30 ESEM using either 3% glutaraldehyde in 0.05m cacodylate buffer, ethanol dehydration, and CPD or 2% Osmium vapor fixation and either air-drying or freeze-drying. Most images of conidia and ascospores were adequate, but we had less success with preserving structure on the conidial heads. Now that we have the Peltier cold stage accessory, I intend to image different Aspergillus species uncoated in wet mode under various vacuum conditions. As "they" say "one size does not fit all"!
Bruce F. Ingber, Biologist Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124 bingber-at-srrc.ars.usda.gov 504-286-4270 phone 504-286-4419 fax
} } } "Robert Simmons" {rsimmons-at-gsu.edu} 04/21/04 11:52AM } } } ------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Try vapor fixing in osmium vapor. Put you material in a small container (I use a glass Petri dish sealed with double parafilm) with a few drops of 4% osmium tetroxide. Leave it for about 72 hours at RT in a hood, then air dry. Aspergillus conidia tend to dimple a bit naturally, but this will stabilize them as well as,any method, and better than most. Robert
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (fremingt-at-fhcrc.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 21, 2004 at 08:43:33 ---------------------------------------------------------------------------
Question: Does anyone have sucessful results fixing and processing samples of conidia of aspergillus for sem? We have tried many methods, but instead of being round, they have places where the walls seem to have indented. We have tried 2% gluteraldehyde fixation, with and without post-fixing in osmium. We have dehydrated in both alcohols and acetone and used HMDS and nothing changes the result.
Any help would be appreciated.
F. Remington fremingt-at-fhcrc.org Fred Hutchinson Cancer Research Center
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 14:48:30 2004
I'm trying the Nikon software ACT-1 that manages the Nikon DXM1200 digital camera. The software has room for only 8 custom scales. I want to set predefined scales for all stops in my stereo zoom, and all objectives in my compound microscope, which makes much more than 8.
Have you figured out a workaround for this? I will appreciate any help!
Cheers, Martin
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 15:59:05 2004
Hi Garry, We routinely leave our tissues in 75% alcohol overnight in the refrigerator. It doesn't shrink or swell the tissue. Perhaps if one is interested in lipids it might be an issue but we have compared it with tissues that are done straight through, and don't see a difference in animal or plant tissue. When we leave it in osmium overnight however we get maceration of the tissue. We actually use that fact to clean the cytoplasmic stuff out of cells for cryofracturing for SEM. Now that we do microwaving however we can do the entire process in one short morning. Good Luck, Judy
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95201 209-954-5284 209-954-5600 jmurphy-at-deltacollege.edu
Garry Burgess wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I have never fixed human tissue overnight in Osmium Tetroxide, or even left } it in lower concentration alcohols, for fear of ultrastructural details } being leached away during the night, but lately I'm left wondering if } perhaps one could get away with fixing animal [human] tissue in 1% Osmium } Tetroxide overnight in the fridge. Prior to this, we've always fixed in } Osmium for half an hour to an hour. } } Does anyone have any experience with overnight fixation of animal tissue in } 1% Osmium Tetroxide in water overnight? It would be a great time saver to } me if I knew for certain that this wouldn't adversely affect the } ultrastructure of the tissue. } } Garry } } This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 17:58:58 2004
Our lab always fixed in 2.5% Glutaraldehyde overnite. We have fixed fat specimens in OsO4. What type(s) of specimens does your lab work with? Sara Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} wrote:
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I have never fixed human tissue overnight in Osmium Tetroxide, or even left it in lower concentration alcohols, for fear of ultrastructural details being leached away during the night, but lately I'm left wondering if perhaps one could get away with fixing animal [human] tissue in 1% Osmium Tetroxide overnight in the fridge. Prior to this, we've always fixed in Osmium for half an hour to an hour.
Does anyone have any experience with overnight fixation of animal tissue in 1% Osmium Tetroxide in water overnight? It would be a great time saver to me if I knew for certain that this wouldn't adversely affect the ultrastructure of the tissue.
Garry
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
__________________________________ Do you Yahoo!? Yahoo! Photos: High-quality 4x6 digital prints for 25¢ http://photos.yahoo.com/ph/print_splash
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 23 02:16:05 2004
Hi all, I am looking for single crystal Quartz and Sapphire substrate, having a very high optical purity. I need those for single molecule / nanoparticle fluorescence microscopy experiments at low temperature. Does any-body know of a company (or companies) that makes them in various dimensions and thickness and with a proven optical purity. (I tried to get some info from Marketech International Inc., but they've ignored my email messages thus far) I would appreciate any help. Thanks, Eli
-- *************************************** Eli Rothenberg Institute of Chemistry, the Farkas Center for Light Induced Processes and the Center for Nanoscience and Nanotechnology The Hebrew University of Jerusalem Jerusalem 91904, Israel TEL.:972-2-6586814 FAX: 972-2-5618033 http://chem.ch.huji.ac.il/~nano/ ****************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 23 03:34:36 2004
Eli, have you tried Goodfellow at www.goodfellow.com? They might be able to help you.
Ed Komarnicki MacDermid Inc.
eli rothenberg {elir-at-chem.ch.huji.ac.il} 04/23/04 03:37 AM
To microscopy {Microscopy-at-MSA.Microscopy.Com} cc
Subject Quartz and Sapphire substrates.
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi all, I am looking for single crystal Quartz and Sapphire substrate, having a very high optical purity. I need those for single molecule / nanoparticle fluorescence microscopy experiments at low temperature. Does any-body know of a company (or companies) that makes them in various dimensions and thickness and with a proven optical purity. (I tried to get some info from Marketech International Inc., but they've ignored my email messages thus far) I would appreciate any help. Thanks, Eli
-- *************************************** Eli Rothenberg Institute of Chemistry, the Farkas Center for Light Induced Processes and the Center for Nanoscience and Nanotechnology The Hebrew University of Jerusalem Jerusalem 91904, Israel TEL.:972-2-6586814 FAX: 972-2-5618033 http://chem.ch.huji.ac.il/~nano/ ****************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 23 08:57:44 2004
(a plain-text reply without plots for the benefit of the microscopy list-server)
} From your plots it seems that you need a FEG to reach resolutions beyond about 3 A, But I can’t see any great advantage up to about 5 A resolution. Is that correct? (and what is all the excitement about then?)
Philip
-----Original Message----- } From: Lucken, Uwe [mailto:uwl-at-nl.feico.com] Sent: 23 April 2004 15:03 To: Philip Koeck; 3dem-at-ucsd.edu; microscopy-at-sparc5.microscopy.com Cc: Max.Sidorov-at-Amd.Com
It might be safer to leave the tissue in buffer overnight after the initial aldehyde fixation, rather than in OsO4. As you say, the OsO4 step would only add 30-60 minutes in the morning.
Lesley Weston.
on 22/04/2004 12:39 PM, Garry Burgess at GBurgess-at-exchange.hsc.mb.ca wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } } I have never fixed human tissue overnight in Osmium Tetroxide, or even left } it in lower concentration alcohols, for fear of ultrastructural details } being leached away during the night, but lately I'm left wondering if } perhaps one could get away with fixing animal [human] tissue in 1% Osmium } Tetroxide overnight in the fridge. Prior to this, we've always fixed in } Osmium for half an hour to an hour. } } Does anyone have any experience with overnight fixation of animal tissue in } 1% Osmium Tetroxide in water overnight? It would be a great time saver to } me if I knew for certain that this wouldn't adversely affect the } ultrastructure of the tissue. } } Garry } } This e-mail and/or any documents in this transmission is intended for the } address(s) only and may contain legally privileged or confidential } information. Any unauthorized use, disclosure, distribution, copying or } dissemination is strictly prohibited. If you receive this transmission in } error, please notify the sender immediately and return the original. }
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 23 10:24:44 2004
"Microanalysis - A New Tool in Combating Terrorism" by Dennis Ward of the Federal Bureau of Investigation will be the keynote presentation at the Spring meeting of the Central States Microscopy and Microanalysis Society on Friday, May 7 at the University of Missouri, Columbia. Other presentations include "Scanning Probe Microscopy Under Controlled Environments" by Dr. Shije Wu of Molecular Imaging Corporation, "Explosives and Explosives Residue Identification" by William Randle of the Missouri State Highway Patrol Crime Laboratory, and additional talks on microscopy applications in the biological and materials sciences. There will also be a poster display area and vendors' exhibits.
This event is hosted by the UMC Electron Microscopy Core Facility and will begin at 8 a.m. on May 7th, 2004 in the Adams Conference Center of the College of Veterinary Medicine. Morning and afternoon snack breaks are included and registration is FREE! A catered lunch will be available for $10.00 per person (reservations required), and we ask that those wanting lunch please let us know ASAP (!!!) so we can get a firm number to give to the caterer.
For more information, directions, and registration forms please contact me at the address below.
Hope to see you there!
Randy Tindall, President Central States Microscopy and Microanalysis Society Web: http://treefrog.cvm.uiuc.edu/~lam/csmms/
Electron Microscopy Core 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/
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 23 10:54:49 2004
I would also add that for x-ray or EELS microanalysis, which some biologists - albeit a few(!) carry out and actually call "structural biology, FEGs can be important since one obtains a far higher electron flux per unit area than with LaB6 thus improving precision of measuements. In some cases of detection and mapping of rather low subcellular contents in small regions (in membranes for example) it can be crucial.
Peter
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-- Peter Ingram Sr. Physicist Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
----- Original Message ----- } From: pamarone-at-bdsinternational.com (by way of MicroscopyListserver)
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (david.mahoney-at-baesystems.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, April 23, 2004 at 11:28:27 ---------------------------------------------------------------------------
Email: david.mahoney-at-baesystems.com Name: Dave Mahoney
Organization: BAE SYSTEMS
Title-Subject: [Microscopy] [Filtered] New SEM
Question: Iím not sure members can comment on equipment but Iím currently in the market for a new SEM I have narrowed he choices down to JEOL model JSM-6460LV or a Hitachi S-3600N. If anyone has any comments, good or bad, on reliability, performance, image quality, and service issues all comments would be greatly appreciated. Thank you BAE SYSTEMS Dave Mahoney 250 Knotter Drive Cheshire, CT 06489
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (skoval-at-uwo.ca) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, April 23, 2004 at 15:54:21 ---------------------------------------------------------------------------
Email: skoval-at-uwo.ca Name: Susan Koval
Organization: University of Western Ontario
Title-Subject: [Microscopy] [Filtered] MListserver: Philips EM300 parts
Question: I am looking for replacement lens for an old, but still needed, Philips EM300. Specifically, I need the intermediate and projector lens coils and housings. If anyone has an EM300 they are using for replacement parts, and can provide the above items, please contact me.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rinaldop-at-uol.com.br) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, April 23, 2004 at 17:34:05 ---------------------------------------------------------------------------
Email: rinaldop-at-uol.com.br Name: Rinaldo Pires dos Santos
Organization: UFRGS - Porto Alegre - RS - Brazil
Title-Subject: [Microscopy] [Filtered] Problems when dehydrate plant tissues
Question: I am working with the ultrastructure of the gametophyte and sporophyte of Frullania brasiliensis (a folious hepatophyte). After the aldehyde and osmium fixation, when I dehydrate with alcohol or acetone (10, 30, 50, 70, 90, 100; 15-30 minutes each), the samples shrink. Some sugestion? Thank you. Rinaldo P. Santos
In addition to state of the art "scopes" from API, AutoQuant, Bio-Rad, III, Nikon, Perkinelmer, and Zeiss students at the 2004 UBC Live-cell course can expect to use several really new 3D microscope systems:
1. Olympus will bring their new Fluoview 1000 and also a new disk scanner (www.olympusamerica.com/seg_section/seg_product.asp?p=5&sc=1&product=962)
2. Visitech are planning to bring their new Vt"eye" AOD video-rate confocal system (www.vt-eye.info/) as well as a Yokogawa-based system.
3. Jenalab will bring their DERMALINSPECT 2-photon in-situ optical biopsy system (www.jenlab.de/english/products/Derma_Inspect/derma_inspect.html).
and
4. LaVisionBiotech will be bring their TrimScope, high-speed multifocal, multiphoton microscope. (www.lavisionbiotec.de/start_product.html)
Now, some more good news:
Several of the students who were originally accepted to attend the UBC Live-Cell Course have now found that they will not be able to attend after all.
This means that we now have room for about 3 more students.
If you are interested, please go to: (www.3dcourse.ubc.ca) and look around. The application form can be found at (www.3dcourse.ubc.ca/application.htm)
Cheers,
Jim Pawley -- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU 3D Microscopy of Living Cells Course, June 12-24, 2004, UBC, Vancouver Canada Info: www.3dcourse.ubc.ca/ Applications due by March 15, 2004
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 26 06:47:51 2004
could anyone please suggest and explain reasonable values for the minimum number of points to be grouped as a grain as well as for the minimum misorientation between two points in EBSD/OIM.
Regards.
Andreas Meyer.
&-] ############################### &-]
Moritz Andreas Meyer Dipl.-Ing (FH) MSc. Sr. Materials Analyst SEM Materials Analysis Department
We have contacted Nikon, and have an answer for the scales question: Sorry, only eight.
I solved the problem writing a simple Visual Basic script for IMatch to place scales on digital images. The script process a batch of images, reading the scale codes between [] form the file name (it could take that information from somewhere else).
Regards, Martin
At 02:28 AM 4/23/2004, you wrote: } Hi Martin } } I don't have the answer to your problem but I also use the DXM1200 and ACT } software. If anyone comes up with an answer could you let me know? } } Thanks in advance } } Gareth } Med vänliga hälsningar/With best regards } } Gareth } } ***Come to Stockholm in 2004 to the 26th IFBLS congress*** Take a look at } http://www.vardforbundet.se/ifbls2004 } } http://www.ki.se/biomedlab } e-mail Gareth.Morgan-at-labmed.ki.se } } Tel +46 8 5858 1038 } Fax +46 8 5858 7730 } } Gareth Morgan MPhil MSc FIBMS, } Department of Laboratory Medicine (Labmed), } Karolinska Institute, } Karolinska University Hospital at Huddinge, F46 } SE 141 86 Stockholm } Sweden } } OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet } för klinisk patologi och cytologi. } } NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. } Clinical Histo- and Cytopathology Laboratory. }
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 26 09:59:15 2004
I think I have to be a bit more specific. The question is whether a FEG is worth the extra investment for the sort of work we are planning.
On the one hand we image individual proteins with a magnification of 20K- 50K and defocus between 1000 nm and 2000 nm. There we want to get resolutions between 5 and 10 Angstrom. Judging from Uwe’s plot that shouldn’t be a problem with a LaB6.
On the other hand we image 2D crystals at about 60K magnification aiming at a resolution of about 2 to 3 Angstrom. Here we work much closer to focus (500 nm), which should improve the envelope of the LaB6 quite a bit. (It would be nice, Uwe, if you could send me a plot for 500 nm defocus for comparison. I can’t reproduce your plots with the parameters you give.)
For these two types of application, is there any point in investing in a FEG? } From what I’ve seen so far there doesn’t seem to be.
Yours sincerely,
Philip
-----Original Message----- } From: max.sidorov-at-spansion.com [mailto:max.sidorov-at-spansion.com] Sent: 23 April 2004 18:42 To: uwl-at-nl.feico.com; Philip.Koeck-at-biosci.ki.se; 3dem-at-ucsd.edu; microscopy-at-sparc5.microscopy.com
Hi,
I would like to build an eyepiece with a c-mount thread. I already have the eyepiece but without lens. But when I look through it I only can see part of what I can see through a 10x eyepiece. I have no vignetting. The image looks nice. But it looks like magnified.
Can anybody tell me which kind of lens I have to insert into my eyepiece in order to see aproximately what I can see with a 10x WF eyepiece.
Anneliese Schmaus
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 26 11:25:00 2004
I wonder if you have recommendations for limits in the length of image file names. For example, it happened to me that during ftp transfers some long file names were truncated to 64 characters + extension.
I always tried to limit the most important information in the first 6 or 8 characters, but this cannot always be done.
May be you know of a practical name size that will work safely in most cases using windows - mac - ftp - linux - internet - etc.
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 26 13:22:52 2004
We had the same problem in our lab, during transfer from pc to a Mac-network (everyone works with Macs, except Microscopy...). I had to reduce the filenames to 27 characters, but by using codes, it's always possible to trace back some photos. Here's the text I sent around in the lab for users of the mciroscopes, hope it helps you out! Best regards,
Sven Terclavers ____________________________________________________________________________ ____________ Scoobidoo is the Microscopy-server which is used to archive images. Here you can find out how to save and archive your image-files.
Every microscope user gets a personal folder on the server to store his/her images. Some folders are already created, if you don't have one yet, please contact me. Within this folder, you have to create subfolders according to the tissue you're taking photos of followed by the year, e.g. folder 'Sven Terclavers', subfolders 'Heart_2002', 'Limb_2002', 'Heart_2003' etc.
In these folders you can now store your images (preferably in High Quality JPEG-format, at least 300 dpi) of the corresponding tissue. To be able to quickly find the data about a photo (microscope, magnification, dye, …) we can use 27 characters (without the format-extension) to name the photos. So from now one, please always use the next configuration to store image-files:
RRRRRRmmmmTTssssDDD_Mxxxaaa.zzz
RRRRRR = description of the research (MMI, AB , PlGF, …) mmmm = mouse number TT = mouse type (WT, HO, HE) ssss = slide and/or coupe-number DDD = dye (you will find a list of abbreviations on the last page) M = microscope (Z [Zeiss], L [Leica], C [Confocal]) xxx = magnification aaa = number of the photo (in case you take more than one photo of a tissue) zzz = file-format (preferably JPEG)
· e.g. MMI1306HO0416TBM_Z40x002.jpg o MMI = Mouse myocard infarct o 1306 = mouse number o HO = homozygot o 0416 = slide 04, coupe 16 o TBM = thrombomodulin staining o Z40x = Zeiss, 40x magnification o 002 = second image
The first part of the name makes it possible to find the slide in case a new photo has to be taken. With the second part, followed after the underscore, it is possible to find what microscope and magnification was used to be able to add a scale bar afterwards. In some situations, when working on the Leica-microscope, you might need to add more characters to the magnification (because of the extra 2 magnification-lenses). Then you can use the extra characters from the research-description RRRRRR, mouse-number or photo-number. ____________________________________________________________________________ ___________
-----Original Message----- } From: Martin Ramirez [mailto:ramirez-at-amnh.org] Sent: Monday, April 26, 2004 18:27 PM To: microscopy-at-msa.microscopy.com
Hi all,
I wonder if you have recommendations for limits in the length of image file names. For example, it happened to me that during ftp transfers some long file names were truncated to 64 characters + extension.
I always tried to limit the most important information in the first 6 or 8 characters, but this cannot always be done.
May be you know of a practical name size that will work safely in most cases using windows - mac - ftp - linux - internet - etc.
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 26 15:02:14 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mocherla-at-eng.fsu.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, April 26, 2004 at 12:53:13 ---------------------------------------------------------------------------
Email: mocherla-at-eng.fsu.edu Name: supriya
Organization: Florida State University
Education: Graduate College
Location: Tallahassee, Florida,
Question: I would like to observe liposomes under TEM. I found a paper which talks about using cryo-TEM ( Ref: Almgren et al, 2000. The procedure described there is a bit difficult. Can anybody help me in getting this real straight by using TEM or SEM.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cjreinbold-at-bcsew.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, April 26, 2004 at 14:42:39 ---------------------------------------------------------------------------
Email: cjreinbold-at-bcsew.edu Name: Corbett
Organization: The Blood Center
Education: Graduate College
Location: Milwaukee, WI, USA
Question: Dear Microscopist,
The problem I'm having is the presence of a dark ring in the center of field. The instrument we're using is a Zeiss Axioskop upright with Plan-Neofluar 100X/1.30 oil ojective, with a mercury bulb having approx. 87hrs for FITC fluorescence. The sample is a cultured cell line using Falcon 8 well slides and using antiquench for mounting. I have performed the easy steps of cleaning and aligning with no clearing of the problem. Any suggestions with this problem are welcomed. CR-