Having processed some frog skin for TEM, looking at the thick sections I noticed the tissue looks like small balls. Can anyone tell me what this might be? Looking at normal skin such as human tissue this looks completely different. Is there a good book that might describe what these structures are? TIA, Phil 8-{)
We routinly use 4% paraformaldehyde, 2% glutaraldehyde in 0.2M sodium cacodylate buffer at pH 7.2. We probably need a mininium of 500 ml of fixative to perfuse the rat. After perfusion, store whole organs in 3% glut or dice up tissue and store in buffer. Good Luck.
We are in the process of having installed a new JEOL TEM and the service engineer suggested we use an "on-demand" gas regulator to control the dry nitrogen vent gas for the column. This type of regulator allows the pressure in the column to reach 1 atmosphere but no more to prevent over pressuring the column and blowing out vacuum seals, thin windows, etc. I know I have seen such regulators but I cannot find a source. If anyone is using this type of regulator could you please supply a model number and vendor.
Thanks in advance.
Norman Elliott | E-mail: nee-at-lanl.gov Los Alamos National Lab | Fax: 505-665-2104 MST-7 MS E549 | Voice: 505-667-1587 Los Alamos, NM 87545 |
We could not find an inexpensive regulator from gas supply houses, etc. so we use regulators designed for scuba tanks. Still not cheap, but less expensive than the alternaives we found. They have worked well without problem for about 8 years now.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Pathology * * Bowman Gray School of Medicine of Wake Forest University * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
On Tue, 1 Nov 1994, Norman Elliott wrote:
} Dear listreaders, } } We are in the process of having installed a new JEOL TEM and the } service engineer suggested we use an "on-demand" gas regulator to control } the dry nitrogen vent gas for the column. This type of regulator allows } the pressure in the column to reach 1 atmosphere but no more to prevent } over pressuring the column and blowing out vacuum seals, thin windows, etc. } I know I have seen such regulators but I cannot find a source. If anyone } is using this type of regulator could you please supply a model number and } vendor. } } } Thanks in advance. } } } } } Norman Elliott | E-mail: nee-at-lanl.gov } Los Alamos National Lab | Fax: 505-665-2104 } MST-7 MS E549 | Voice: 505-667-1587 } Los Alamos, NM 87545 | } } }
I recently replaced the Wehnelt aperture in my Philips CM12 electron gun. The original aperture had a hole diameter of 0.6 mm and I replaced it with one that has a 0.8 mm dia. A 0.4 mm hole is also available in my column kit.
I know from my experience on an SEM that increasing the hole diameter will increase the beam current output from the gun (which I did to get more counts for x-ray microanalysis).
What else is affected by the hole size - resolution, coherence, or what? How should one decide what diameter to use for this aperture on an SEM or a TEM?
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Reply... RE} EM shielding/ M. Rich Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX: (313)936-3352 jfmjfm-at-umich.edu or John_Mansfield-at-mse.engin.umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
Opinions and experiences here are my own.
We have a field problem in our JEOL 4000EX room. Before all the microscopes were installed the JEOL engineers did a field survey in each room. All was fine. They then installed a 2000FX in one room and got it up and running and then in the adjacent room installed the 4000. Now both reached spec without problem. But, as many of you may know, JEOL had some fearful problems with the guns of the 4000 series. We had really bad filament image flickering and it never seemed to be totally cured (in spite of extensive glow discharging, oops! I mean conditioning). Tracing the problem and trying to blame everything except the gun, lead our engineers (and there were several groups of them!) to test for fields and they notice that there was a substantial field about 5 feet from the scope in the floor. It was below the required level at the objective lens, but that didn't stop them from mentioning it (many times). Turns out that the supply for the 2000 ran under the floor of the 4000 room and the phases were sufficiently unbalanced to produce this field. Now, the 4000 scope had been installed after the 2000 was up and running and so we had taken the resolution tests with that field present and so we didn't think it was a problem. This did not prevent the engineers trying to say that our gun problem was related to that field. The alternative at this point was to admit that the gun was bad and replace it. After a good deal of discussion the gun was replaced. By this time the scope had been down so long that I had asked JEOL Service for a rebate on my service contract since they had not maintained the scope in operating condition for a reasonable portion of the period covered by the contract. The moral of this story is. When you build your scope facility, make sure that you specify where everything goes. And I mean everything, water, power, ground lines, light locations (fluorescent lights can interfere with the scopes, the chokes have quite a field)., everything. If we hadn't had the field in the floor we would not have had to argue about it! It's easier if you don't build in problems for yourself! Don't let the builders or the bean counters change the plans after you have approved them. Our plans were amended and we now have building air instead of our own system and I always have toasting microscopes in the fall and spring when the A/C is in the process of being turned on or off! As I say just my opinion and experiences. I am not trying to get at JEOL, it is just that it took a long time to convince them that it was their gun and not our facility.
The following is an abbreviated version of an offline email dialogue between myself and Paul Nolan of Queen's University regarding his "LaB6 Fix" question. Paul, Nestor and I agreed that this would be a good dialogue to share with the listserver. If anyone has any questions regarding these issues, feel free to contact me.
Best regards, Damon L. Heer. ***************************************************************
I'VE GOT A LaB6 THAT STOPPED EMITING. LOOKS GOOD UNDER THE SEM EXCEPT FOR THE TUNGSTEN BALLS ALL OVER IT! GOING TO TRY TO CLEAN IT IN MY CARBON EVAPORATOR. THINK IT WILL WORK? ANY OTHER IDEAS? (CAN'T AFFORD A NEW ONE)
THANKS, PAUL ----------------------------------------------------- Dear Paul:
You noted that it looks good in the SEM. Do you mean that the crystal still appears to have a reasonable emitting surface remaining? how long did it run before it stopped emitting?
You also noted a build up of tungsten balls on it. Is this build up on the mount or the crystal? I've never seen this kind of contamination on a tip. this is because I mainly deal with our LaB6 cathodes which don't have tungsten in the structure. However, if you are using a Denka cathode, the cement that is used to keep the crystal in the mount may be causing this contamination.
To clean it, we sometimes rub the crystal with a q-tip with a little isopropyl alcohol. If this doesn't remove it, try lightly rubbing it with the wooden stick portion of the q-tip, again with IPA. This basically applies a rather non-abrasive scrub to the crystal. The difficulty here is that both Denka's and Kimball's LaB6 mounts are very fragile and may break under any contact.
Unfortunately, Ed Calomeni is very correct in saying that your best bet is buying a new one.
Good luck! Damon L. Heer ------------------------------------------------------ Damon. There seemed to be no structural damage to the filament ie cracks or defects but the tungsten balls cover the whole filament.
The filament has been in our TEM for about 2 years but has not had too many hours of use . It is a Kimball filament so i dont know if there is any W in the mount or not. I will give it a wipe and look at it in the sem again and see what has happened. In your opinion is this W going to stop the crystal from emitting? (There appears to be no other damage and there is continuity between the mounting posts) ...
Cheers, Paul ------------------------------------- Hi Paul,
In my opinion, if there is no contamination on the cone and flat of the crystal, it should work fine. When an electron microscope is saturated properly, the emission only comes from the flat area.
You noted that the cathode stopped emitting while in use. The reason this usually happens is the emitting flat at the tip of the crystal cone has evaporated to a point. If there still is a flat left on the emitter, I imagine that the flat did become contaminated, but is unnoticeable in the SEM.
I know that a large amount of time is necessary for changing tips in a TEM, so the following recommendation should be weighed against that.
If there remains a reasonable amount of the flat on the crystal, and the W can be cleaned away from the crystal cone and flat, it may be possible to drive off any remaining contamination by operating the cathode at about 0.2 amps above the normal operating filament current for a short period of time. This may recover the cathode.
You mentioned that the flat on the filament was the important part to have clean, well it seems there is no definite flat part on the tip. I was unaware because of my inexeperience with LaB6 that there should be a flat, i assumed it should be pointed (like mine seem to be) Is this filament dead?
cheers paul ----------------------------------- Hi Paul,
Kimball, as well as FEI, uses a "truncated tip" for LaB6 emission. The truncated tip consists of a cone with a flat top that is normal to the optical axis. The "flat" is oriented with the {100} crystal plane, which has been shown to be the best orientation for electron microscope applications. The cone angle and the flat diameter are chosen by the type of application it will be used for. FEI's standard cone angles and flat diameters (and I believe the same goes for Kimball) are as follows:
SEM and Low Res TEM 90 degree full cone angle, 16 micron flat diameter
Hi Res TEM 60 degree full cone angle, 5 micron flat
When the flat is gone, the emission comes from the side of the cone, which isn't the {100} plane and is not normal to the optical axis. The result is poor emission down the column.
If there is no flat left on your tip, I'd say it's dead.
Coincidentally, a colleague and I just wrote an article for our semi-annual newsletter, the FEI Focus, regarding crystal tip issues. If you wish, send me your address and I could send you a copy of this. A reprint also appears in the most recent issue of Microscopy Today.
Cheers, Damon
FEI Company 7451 N.E. Evergreen Parkway Hillsboro, OR 97124-5830
Last year, we installed a JEOL JEM 1210 and were advised to purchase the demand flow regulator for the dry nitrogen. We ended up with a Matheson Model 3421 vacuum regulator which meets JEOL's specs...in fact its one that they recommend. Its pricey at about $400, and is available from any of the industrial gas suppliers, ie AIRCO, etc. However, its maximum inlet pressure is 50 psi, meaning that you can't couple it directly to 3000 psi nitrogen tank. You still must buy a general purpose, dual stage nitrogen regulator, then attach the demand flow regulator to this. The whole setup will end up costing about $600. Incidentally, if you are installing a TEM, then you know that you will have to buy a cylinder of sulfur hexaflouride and a regulator for this. A 100 lb bottle of sulfur hex is about $400, but you can use a nitrogen regulator for its installation if you buy a $15 adaptor. I can supply this information if you need it. Good Luck.
-=buddy=-
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
I have someone interested in the fracture surfaces of choclate. I want to use some type of replication process, as I don't want to put chocolate in my SEM. No, I don't want ESEM, or cryoSEM. I am worried that the solvents of formvar or similar plastics might distort the fracture surface. I need low mag shots, so high resolution replicas are not necessary. thanks in advance, Randy Nessler rnessler-at-emiris.iaf.uiowa.edu
I deleted the original post on this subject, but from what I remember, I think the original poster was looking for a pressure relief valve that tripped at 1 atm, instead of a regulator that would supply 1 atm. We used to have a little pop-off valve that came with a thin window x-ray detector. It was supposed to be installed in the line that bleeds the chamber, so as to prevent a positive pressure from forming in the SEM specimen chamber. A positive pressure ran the risk of rupturing the window. You might call Kevex/Fisons, since this is who supplied the pop-off valve for us. Randy Nessler rnessler-at-emiris.iaf.uiowa.edu
Can anyone point me in the right direction for techniques for making silicon monoxide and aluminum oxide films. A discussion of the resultant surface prop- erties would also be helpful, if such info is written down someplace.
Dear Gib, The wehnelt aperture size and the fields produced in and around the filament and wehnelt determine the locus of points from which electrons emitted by the filament end up being accelerated and sent down the column. The larger the aperture size, 1) the more electrons go down the column, 2) the less cohe- rent the beam, 3) the larger the crossover size, and 4) the more the anode is heated by electrons extracted at angles significantly different from 0 degrees. The best wehnelt aperture size depends on what the instrument is used for. The largest possible size is that for which anode heating becomes a pro- blem. For people like me who do low-dose work and diffraction and whose EDX detector gives high dead time for more intense beams, a smaller aperture is better; for those who are fighting to increase intensities all the time, larger is better. Yours, Bill Tivol
In article nee-at-lanl.gov (Norman Elliott) writes: } } Dear listreaders,
} We are in the process of having installed a new JEOL TEM and the } service engineer suggested we use an "on-demand" gas regulator to control } the dry nitrogen vent gas for the column. This type of regulator allows } the pressure in the column to reach 1 atmosphere but no more to prevent } over pressuring the column and blowing out vacuum seals, thin windows, etc. } I know I have seen such regulators but I cannot find a source.
} Hi Norman. The regulator you need is available at a cost of about $200 from dive shops selling scuba gear. They are just breathing demand valves which allow gas to flow when you suck on the valve but not otherwise. Its important to always have gas available on the pressure side or the microscope vacuum will suck the valve diaphragm to destruction! We have such a system feeding our SEM from the bled off gas in out 175Litre Liquid nitrogen tank, which is the driest gas you can get. If you have a UTW window on your EDS you might install a relief valve on the line so pressure in the microscope won't ever go over atmospheric.
Sulfur hexafluoride is the insulating gas used in high voltage tanks. In the early days, all tanks were filled with dielectric oils, and some of them contained PCB, forcing the manufacturers to find another type oil. As accelerating voltages increased, the manufacturers switched to Freon as a superior insulatant. Of course recently, the EPA has banned the use of most Freons, so now most manufacturers have switched to sulfur hex., which is supposed to be completely inert. The HV tanks don't come pressurized with it, so you have to supply it for the service engineer. As I understand it, SEMs with their lower accelerating voltages still have oil in the tanks.
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
} I recently replaced the Wehnelt aperture in my Philips CM12 electron gun. The } original aperture had a hole diameter of 0.6 mm and I replaced it with one that } has a 0.8 mm dia. A 0.4 mm hole is also available in my column kit. } } I know from my experience on an SEM that increasing the hole diameter will } increase the beam current output from the gun (which I did to get more counts } for x-ray microanalysis). } } What else is affected by the hole size - resolution, coherence, or what? How } should one decide what diameter to use for this aperture on an SEM or a TEM? } } -- } } Gib Ahlstrand, MMS Newsletter Editor } Electron Optical Facility, University of Minnesota, Dept. Plant Pathology } 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 } 612-625-9728 FAX, giba-at-puccini.crl.umn.edu } } We also have a CM12 with LaB6 filament. The general "rule of thumb" is to multiply the Wehnelt distance, ie. distance you screw back the filament into the anode, by 2 1/2 to obtain the Wehnelt aperature size you require. I back off the filament by 200 um. Therefore I require a 500 um. aperature. The lifetime of the filament is just under 1000 hrs.
Hope this gives you some assistance.
Fred Pearson Electron Optics Facility Institute for Materials Research McMaster University Hamilton, Ontario voice (905) 525-9140 x24609 fax (905) 521-2773 eoptics-at-mcmail.cis.mcmaster.ca
} I have someone interested in the fracture surfaces of choclate. I } want to use some type of replication process, as I don't want to put } chocolate in my SEM. No, I don't want ESEM, or cryoSEM. I am worried } that the solvents of formvar or similar plastics might distort the } fracture surface. I need low mag shots, so high resolution replicas are } not necessary. } thanks in advance, } Randy Nessler } rnessler-at-emiris.iaf.uiowa.edu
What will most probably work in your case is one of the dental replicating silicones. These are formulated to make quick, accurate replicas of tooth surfaces and are used in the preparation of ceramic (and other) crownwork. Any dentist should be able to help. We have used these silicones in replicating leaf surfaces, as well as human skin, and the resolution is OK up to at least 1000x mag. There are other (cheaper) types of dental impression materials available, but these water-based formulations give low resolution and are of course not suitable for direct exposure to the SEM vacuum. Have fun.
Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
Message-ID: {MAILQUEUE-101.941102083619.1088-at-anat.uct.ac.za} To: microscopy-at-aaem.amc.anl.gov
In answer to Phil's question regarding frog skin:
What area of the skin are you looking at? If it is only the epidermis, the 'balls' are probably the epidermal cells which have shrunk (a common occurence) during fixation. If it is dermal then you may well be looking at glands situated in the dermal region. If the balls are within the cells then the possibility is that they are the frog equivelant of melanin.
I am unaware of any decent book on frog skin per se. I do have some references which you may find useful if you wish to have them.
O.k., I'll ask for us ignorant folks. Why is a tank of sulfur hexaflouride needed for the installation of a new TEM as was mentioned in responding to the low cost regulator series?
Richard E. Edelmann Electron Microscopy Facility Supervisor Marshall University - School of Medicine Huntington, West Virginia
I am interested to get into contact with those who use an Astromed liquid nitrogen CCD camera, type 3200, on top of any type of microscope to quantify fluorescence images. I would like to exchange experience and test images.
Please mail me directly at : vanderwulp-at-mbl.tno.nl
Hope to hear from you.
PS. Does anybody know of a mailing list used by astronomers because this equipment is frequently used by them.
Thanks for any comment. -- Kees van der Wulp TNO - Nutrition & Food Research Institute INTERNET : vanderwulp-at-mbl.tno.nl Division of Toxicology VOICE : +31 15 843101 PO-Box 5815 FAX : +31 15 843989 2280 HV RIJSWIJK (NL) THE NETHERLANDS
Reply... RE} Sulfur hexaflouride? Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX: (313)936-3352 jfmjfm-at-umich.edu or John_Mansfield-at-mse.engin.umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
On a realted note, how are people managing the switch from Freon to SF6? According to our JEOL Service engineers, the upgrade of our 4000 to SF6 is a freebie as part of our maintenance contract, but they want to stiff us for $20K to upgrade the 2000FX. I have been hoarding a few large cylinders of freon to make sure we can continue to run and service our 2000 for a number of years with freon. What are other folks doing? Jfm.
--------------------------------------
O.k., I'll ask for us ignorant folks. Why is a tank of sulfur hexaflouride needed for the installation of a new TEM as was mentioned in responding to the low cost regulator series?
Richard E. Edelmann Electron Microscopy Facility Supervisor Marshall University - School of Medicine Huntington, West Virginia
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Our JEOL 4000 is affected by magnetic fields due to any TV monitor in the room that is not very new and highly shielded. This includes Mac monitors and the TV monitor for our TV rate camera.
The filament appears to flicker at 3-5 Hz and we are unable to measure a magnetic field at that frequency even near the monitor let alone the microscope! We do measure the 50-60 Hz field at the monitor and the stronger this field the larger the effect on the beam. The effect is greater the closer the monitor is to the scope and the closer to the objective lens.
For best images we turn off the monitors before exposing film. There is an obvious difference!! For some seeing was believing.
Roseann Csencsits Argonne National Laboratory Argonne, Illinois 60439 USA 708-252-4977
this is a reposting- our mail program has not been sending out all correspondence and that which it sends out has had the wrong return address. Sorry for cluttering cyberspace if you've seen this before.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Pathology * * Bowman Gray School of Medicine of Wake Forest University * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
---------- Forwarded message ----------
We could not find an inexpensive regulator from gas supply houses, etc. so we use regulators designed for scuba tanks. Still not cheap, but less expensive than the alternaives we found. They have worked well without problem for about 8 years now.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Pathology * * Bowman Gray School of Medicine of Wake Forest University * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
On Tue, 1 Nov 1994, Norman Elliott wrote:
} Dear listreaders, } } We are in the process of having installed a new JEOL TEM and the } service engineer suggested we use an "on-demand" gas regulator to control } the dry nitrogen vent gas for the column. This type of regulator allows } the pressure in the column to reach 1 atmosphere but no more to prevent } over pressuring the column and blowing out vacuum seals, thin windows, etc. } I know I have seen such regulators but I cannot find a source. If anyone } is using this type of regulator could you please supply a model number and } vendor. } } } Thanks in advance. } } } } } Norman Elliott | E-mail: nee-at-lanl.gov } Los Alamos National Lab | Fax: 505-665-2104 } MST-7 MS E549 | Voice: 505-667-1587 } Los Alamos, NM 87545 | } } }
Our lab is considering building, or purchasing (new or used) an Optical Diffractometer. Does anyone have a list of suppliers, a used instrument, or wishes to share their design? Comments on manufactured models are welcomed. Email directly if you wish to avoid biased opinions.
Thanks Fred Pearson fax (905) 521-2773 McMaster University Hamilton Ontario, Canada eoptics-at-mcmail.cis.mcmaster.ca
I need advice, recommendations, information about video microscopy systems; I want to buy a system to use in my undergraduate classes (gen. zool., invert. zool.) to observe small, live animals. I especially need specific brands, models that others have used successfully in similar situations. The students themselves will be using the system and making and editing videos--so the equipment needs to be fairly sturdy yet enable good quality images. At this time I have price quotes from a number of sources (Carolina Biological, Fisher, Nikon, Olympus) but of course each salesperson recommends his/her system! Thanks for any advice. P.M. Biesiot Dept. Biol. Sci. Univ. Southern Mississippi Hattiesburg, MS 39406-5018 pbiesiot-at-whale.st.usm.edu
} } O.k., I'll ask for us ignorant folks. Why is a tank of sulfur } hexaflouride needed for the installation of a new TEM as was } mentioned in responding to the low cost regulator series? } SF6 is used in only some new TEMs. Hitachi uses SF6 in the guns of all TEMs that go to 200KV and over. And as of yet only in the HV tanks of the FE-TEMs. Its used as an insulator, instead of oil or Freon. Cal-Hitachi service
For years we used a basic optical bench with a low power laser and lenses we scrounged from microscopes and enlargers and cameras. The best idea is to buy two 2 meter benches and use an optically flat mirror (astronomy supply house stock) to bend the pattern back so you can view the pattern as you move the negative around. The best viewing/recording device is an old 4x5 camera of some sort. You don't even need one with a working shutter or light tight bellows as you will be using the diffractometer in the dark. You can then easily use polaroid 45 materials for hard copy of the patterns. BUT you will have to be cautious about viewing the pattern and always e.g. have the zero order beam blocked with a metal patch. I have the scars to show how careful you need to be.
email me for further information. Mel Dickson } Thanks } Fred Pearson} fax (905) 521-2773} McMaster University} Hamilton Ontario, Canada} eoptics-at-mcmail.cis.mcmaster.ca
The "small balls" structures might be mucous glands. See fig. 5 in Hauser F et al 1990. Expression of spasmolysin (FIM-A.1): an integumentary mucin from Xenopus laevis. Exp Cell Res 189: 157-162 - it shows a phase constrast micrograph of Xenopus skin with mucous glands that might be interpreted as "small balls".
On Tue, 1 Nov 1994, rutledge phil wrote:
} Having processed some frog skin for TEM, looking at the thick sections I } noticed the tissue looks like small balls. Can anyone tell me what this } might be? Looking at normal skin such as human tissue this looks completely } different. Is there a good book that might describe what these structures } are? } TIA, } Phil } 8-{) }
Hello, I have been approached by a first grade teacher at Pond Cove Elementary School who wants to instill an early love for microscopy in the children. Are there any free or low-cost dissecting microscopes out there?? Thanks, Ada Ada Olins Univ. Tennessee-Oak Ridge GSBMS Biology Division, ORNL P.O. Box 2009 Oak Ridge, TN 37831-8077
Michel et al., I have used methylamine tungstate quite successfully for seing some very fine filaments on the surface of tooth decay bacteria. I follow Wyatt et al., 1988. Oral Microbiol Immunol 3:162-168. If this journal is not widely available outside dental schools just send a psotal address or fax # and I can get a copy to you.
Being a scientist doesn't insure good spelling, any more than being in a company insures courtesy.
A. K. Christensen
---------------------------------
On Thu, 3 Nov 1994, John Mardinly, 5-2346, Page 322-6490, SC2-24 wrote:
} Folks; } What I want to know is why there are so many of you out there claiming } to be scientists yet can't get the spelling of fluoride any better than } "flouride"? } } John Mardinly } Intel MATTEC }
A few weeks ago a person posted a note about having a SuperCard stack about SEM's and offered the FTP site for it........I replyed to him and asked for the site name but never got any reply.....could somebody help me out? It would be great for teaching and training.....
Thanks
--
E.Todd Voiles *** Facility Manager Central Facility for Electron Microscopy University of Nebraska tvoiles-at-unlinfo.unl.edu
Our Hitachi has a "branch" in the vent line with a pressure relief valve which then allows us to use a standard regulator - would that be a suitable solution to the problem?
We have used a Polaron optical diffractometer for several years. It came as a complete package: bench, lenses, laser, holder for negatives, etc. The addresses were:
Polaron Equipment Ltd. 53-63 Greenhill Crescent Watford Business Park Watford Hertfordshire WD1 8QS
Polaron Instruments, Inc. 2293 Amber Drive Line Lexington Industrial Park Hatfield, PA 19440 (215)822-2665
Russell E. Cook Electron Microscopy Center for Materials Research Materials Science Division Argonne National Laboratory Argonne, Illinois 60439 USA
Good Friday Morning, Ada Olins (Univ. of TN) asks for (inexpensive) microscope sources suitable for young students. In this broad area, I would like to introduce the "DiscoveryScope" to readers. It is "an inexpensive, exceptionally durable viewing system for exploring small things. It's 1-inch focal length lens produces beautiful wide field, high resolution views of the small invertebratres, tiny plants, protist, and inorganic subjects. Special chamber and holders are provided for viewing both terrestrail and aquatic organisms. Not just for beginners, DiscoveryScope makes a useful "spotting scope" for any microscopist who collects samples from the field, particularly if macro photography is intended." Discovery Scope Inc PO Box 607 Green Valley, AZ 85622 Tel/Fax: (800)398-5404 I have no financial interest in this product (unfortunately) - and but submit that it is a nifty device! Don Grimes, Microscopy Today
Microscopy & Microanalysis Public Domain Software Library site in a few weeks. Brendon Griffin will be visiting ANL in November and will be bringing a copy along. The FTP site is:
WWW.AMC.ANL.GOV
It currently hosts the EMMPDL, MASLIB, Test Images, and PDL Software all for Microscopy&Microanalysis.
Your Friendly Neighborhood SysOp (who can sometimes spell)
John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu Time: 14:24
Date:11/4/94 URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html NC EMAL
I currently have a working copy that Brendon sent me and I will make it available when he gives me the OK. Warning, it is an 11 megabyte file and requires at least 5 megabytes to run. It runs only on a Mac.
Does anybody have a suggestion for a shutter for a Nikon Diaphot light source? We're looking for the following requirements: - easy to mount on upper (halogen) or lower (mercury) lamp input - computer controlled via NIH-Image (or IP-Lab Spectrum) - cheap!
The two main applications are: - time lapse w/ fluorescence - collecting random fields w/ fluorescence
At this point we don't need selectable filters, just a shutter, although we wouldn't mind getting various filter positions as part of the deal.
anybody know an FTP site with an MSDOS program for displaying TIFF files? Archie and veronica turn up lots of pictures but no programs.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
Jay Jerome asked about TIF viewers for PC machines...
There are currently NO TIFF viewers in the ANL Library for PC's. but the ANL FTP site has GIF viewers for PC machines (both DoS and WinDoze) the are in the following path:
ANLAEM Software Library ANL Shareware Library Freeware Shareware PC Viewers
The site address is: WWW.AMC.ANL.GOV The programs are called: VGIF.exe, VUGIF.exe, WINGIF.exe, CSHOW.exe
The PC commerical program PhotoStyler certainly opens TIFF files, and so do a lot of other commerical programs (PhotoShop etc...).
As a note of warning. Some PC TIFF reading programs will not read TIFF files that were created on a MacIntosh. Most Mac programs that I've used, on the other hand, are intelligent enough to recognize the differences and translate appropriately. Also not all TIFF readers can handle the full TIFF specification. Many are coded assuming that 8 bit gray scale and/or 24bit (8x3) color is the end product. If you have 12,16 ... bit scientific data many (but not all) will not display properly. Make sure you take this into account if your looking at something other than photographic quality images.
There are also the usual suite of Translator programs which will convert between the different image formats. Those programs are usually cheaper than the full blown image processing/editing programs. By purchasing one of these translator programs you could then use the Shareware Viewers....
We have been using a 286 PC-based Kontron IBAS-II with IBAS 2.0 software for image analysis for several years with only middling success.
We am now assisting some researchers with semi-quantitative analysis of the silver grains from their in situ hybridization studies. They are interested in purchasing a Macintosh system to run the NIHIMAGE program you have developed. Their interest primarily stems from the following paper: Lucas, L.R., Mize, R.R. and Harlan, R.E. Semiquantitative analysis of in-situ hybridization results using IMAGE software: a rapid method for counting reduced silver grains over mRNA-positive cells. J. Neuroscience Methods 52:101-109 (1994). They have asked us to help them determine what computer and video hardware they need to set up their system.
Is anyone willing to suggest the required hardware for an adequate Mac system to run NIHIMAGE? We want neither a Volkswagen nor a Cadillac, just a good level system with a Macintosh they can also use for other purposes. Do you recommend a Quadra or a PowerPC? Which model? We assume that 8 MB RAM is minimum and 16 MB is better. Do you have any particular video camera and frame grabber your recommend? (We currently have a DAGE MTI CCD72 video camera). Our computer network manager has a nubus VideoSpigot frame grabber available if it would be of use. We assume it is best to run the program using two monitors. Any recommendations for the video monitor?
We've noticed from reading some of the information on the microscopy usenet newsgroup that a PowerPC version of NIHIMAGE is being developed. Is the PowerPC version tested enough for a "novice" user? We have about 5 years of image analysis experience on the IBAS system so we know much of the terminology and the steps involved in image analysis. We just son't have a clue about the Mac...
Any assistance you can provide would be greatly appreciated.
Aloha, Tina Weatherby Carvalho Marilyn F. Dunlap Biological EM Facility Pacific Biomedical Research Center University of Hawaii at Manoa 1993 East-West Road Honolulu, HI 96822 (808)956-6151 or (808)956-6251 or FAX (808)956-4768 e-mail: dunlap-at-ahi.pbrc.hawaii.edu tina-at-ahi.pbrc.hawaii.edu
} } { {Michael Rich, rich-at-egr.msu.edu } { {Michigan State University asks about cheap magnetic shielding.. Michael, there is no such thing because of the physics of magnetic fields. Radio frequency interference CAN be excluded by making a Faraday cage - a complete box - using copper mesh or even fine chicken wire mesh. BUT to excludelow frequency magnetic fields you must use solid sheets of some ferromagnetic material. Any gaps between sheets must be covered over by strips of similar material. To entirely shield a room the six faces of the room must have the shield applied (yes that includes the floor and ceiling). The material must either be a high permeability material like the alloy hipernom or it may be ordinary steel sheet but much thicker. To shield a complete room costs $40 - 50,000. Much of the cost lies in the fact that the construction should be done just right to obtain best shielding. Try looking up the yellow pages for a shielding supplier. I have an old address for one: Amuneal Manufacturing Corp, 4737 Darrah St. Philadelphia Pa 19124.
Recently I have been using PVP (polyvinylpyrrolidone) in various techniques but to date I have had no luck. Firstly I have tried PVP with LR Gold resin (as recommended by the LR technical blurb), I have also tried adding it my fixative to increase osmotic pressure (as recommended in the paper I have been following to localise the enzyme Thiamine Pyrophosphatase) and finally I have tried it when infusing tissue with cryoprotectant for cryoultramicrotomy (as recommended by Tokuyasu)
In these papers they talk of "dissolving" the PVP in the various solutions but my PVP will not dissolve!!! I am left with a fine white suspension which quickly settles out when stirring is stopped.
Question: Should the PVP dissolve (and perhaps leave a clear solution) or is the fine suspension I have normal? (It blocks up canulas when perfusing, it makes it difficult to see the tissue when processing).
Note: The Merck index states that PVP is soluble in alcohols and water - have I got the wrong one?
Many thanks in anticipation.
Allan Mitchell
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
} anybody know an FTP site with an MSDOS program for displaying TIFF } files? Archie and veronica turn up lots of pictures but no programs.
Jay, Try Graphic Workshop (gws70c.zip), which is located for example at the ftp.uni-mannheim.de (134.155.50.51): /disk2/systems/msdos/graphics
Regards,
Petr +-------------------------------------------+-------------------------+ | Dr. Petr Schauer | tel.: (+42 5) 41321246 | | Head of Electron Microscopy Laboratory | fax : (+42 5) 41211168 | | ACADEMY OF SCIENCES OF THE CZECH REPUBLIC | E-mail: petr-at-isibrno.cz | | INSTITUTE OF SCIENTIFIC INSTRUMENTS | petr-at-mrkev.vabo.cz | | Kralovopolska 147, CZ-612 64 Brno | | | Czech Republic | | +-------------------------------------------+-------------------------+
We used tons of sulfur hexafluoride when I supervised a lab testing image intensifiers. We would flood bare tubes with it, so breathed a lot of it. It is non-toxic, and can only make you pass out if it excludes the oxygen. Sulfur hex actually quenches sparks as it decomposes in the arc. As for fluorine remaining in the tank this is highly unlikely, since it would react with anything it came into contact with.
As for spelling, I have come to the rule of thumb for fluorine, fluorescence etc. that if it spells flour it is wrong. I still have fond feelings for a professor who gently pointed this out to me.
The balls seen might be 'membrane blisters' as written up in scanning vol 1 p 166-173 1978 E. Shelton & Mowczko the basic idea is that OsO4 sometimes creates '? blebs 'like little balloons on the surface of cells, these were up to 6 microns long but mainly 1-2 microns or sometimes smaller *(they say .2 to .6) in a variety of cells and conclude that glutaraldehyde a lone is best foir fixation. Since many people have not found this, it may only be certain tissues that do this in most people's routine handling of fixation. Alan Pooley Marine science rutgers nj
Concerning spelling errors etc (flouride vs fluoride etc) may I suggest to all the gentlemen and lady scientists that we quietly suggest corrections of errors ti individuals and avoid blaring them over the net. ( I expect an individual correction for my mistyping of 'to' in the previous l Alan Pooley marine sci sem lab rutgers nj
Hey folks: A reply to Allen Mitchell -- I think you're problem might lie in the molecular weight of the PVP you're using, or the temperature you're trying to dissolve it at, or both. We routinly use PVP with a m.w. of 40,000 (Sigma cat. PVP-40) in fixatives for in situ perfusion of rat livers -- 2% w/v takes a while (several minutes) to go into sol'n, but it does go in. I also make a Tokuyasu-style infiltration medium for cryosectioning -- that uses PVP m.w. 10,000 (Sigma PVP-10) but it is dissolved into NaCO3-phospate (100g of PVP into 30ml of liquid!) and must be done by alternating between a 60C oven and a sonicator full of hot water. Feel free to E-mail me (gmartin-at-welchlink.welch.jhu.edu) for particulars of preparing this stuff (it's a real witch's brew). I can't speak as to the particulars of PVP added to LRGold -- I've always used LRGold straight-up -- but I would advise against putting you're specimens into a sol'n cloudy with undissolved PVP.
Take Care, Greg Martin Dept Cell Biology and Anatomy Johns Hopkins School of Medicine
On Mon, 7 Nov 1994, Richard Easingwood wrote:
} A question from Allan Mitchell } } Recently I have been using PVP (polyvinylpyrrolidone) in various techniques } but to date I have had no luck. } Firstly I have tried PVP with LR Gold resin (as recommended by the LR } technical blurb), I have also tried adding it my fixative to increase } osmotic pressure (as recommended in the paper I have been following to } localise the enzyme Thiamine Pyrophosphatase) and finally I have tried it } when infusing tissue with cryoprotectant for cryoultramicrotomy (as } recommended by Tokuyasu) } } In these papers they talk of "dissolving" the PVP in the various solutions } but my PVP will not dissolve!!! I am left with a fine white suspension } which quickly settles out when stirring is stopped. } } Question: Should the PVP dissolve (and perhaps leave a clear solution) or } is the fine suspension I have normal? (It blocks up canulas when perfusing, } it makes it difficult to see the tissue when processing). } } Note: The Merck index states that PVP is soluble in alcohols and water - } have I got the wrong one? } } Many thanks in anticipation. } } Allan Mitchell } } Richard Easingwood } Department of Anatomy and Structural Biology, } P.O. Box 913 } University of Otago, } Dunedin, New Zealand. } Fax:64-3-479 7254 } Telephone:64-3-479 7301 } } }
} Reply to: RE} PVP solubility } } Richard, } Could you send me the reference for the use of PVP, i.e. the Tokuyasu ref? } } Thanks---Mike } } Mike_Schwartz-at-qm.yale.edu } Fax 203-785-5263 } Voice 203-785-4324 } Michael Schwartz, Ph.D. } Associate Professor } Section of Neurobiology } Yale University School of Medicine } 333 Cedar St. } New Haven, CT 06510
Mike, the reference is Tokuyasu, K.T.(1989) Use of poly(vinylpyrrolidone) and poly(vinyl alcohol) for cryoultramicrotomy.Histochemical Journal 21, 163-171. I will fax a copy to you if you like.
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
One last (?) word on spelling errors: "It's a damn small mind that can think of only one way to spell a word." - Andrew Jackson (seventh President of the U.S.)
} A question from Allan Mitchell } } Recently I have been using PVP (polyvinylpyrrolidone) in various techniques } but to date I have had no luck. } Firstly I have tried PVP with LR Gold resin (as recommended by the LR } technical blurb), I have also tried adding it my fixative to increase } osmotic pressure (as recommended in the paper I have been following to } localise the enzyme Thiamine Pyrophosphatase) and finally I have tried it } when infusing tissue with cryoprotectant for cryoultramicrotomy (as } recommended by Tokuyasu) } } In these papers they talk of "dissolving" the PVP in the various solutions } but my PVP will not dissolve!!! I am left with a fine white suspension } which quickly settles out when stirring is stopped. } } Question: Should the PVP dissolve (and perhaps leave a clear solution) or } is the fine suspension I have normal? (It blocks up canulas when perfusing, } it makes it difficult to see the tissue when processing). } Note: The Merck index states that PVP is soluble in alcohols and water - } have I got the wrong one? } Many thanks in anticipation. } } Allan Mitchell } } Richard Easingwood } Department of Anatomy and Structural Biology, } P.O. Box 913 } University of Otago, } Dunedin, New Zealand. } Fax:64-3-479 7254 } Telephone:64-3-479 7301
Dear Allan, We make up sucrose/PVP as a cryo-protectant using the following technique. It finallly goes into solution after being stirred overnight. Dissolve 30g PVP in 80ml water at approx 60C, add sucrose (79g) and 20 ml 0.5M PBS. Stir in coldroom overnight. Store at 4C. The solution is a clear yellow. Much like a good Hunter Chardonnay. Hope this helps. Regards, Gerald Little.
Dr Gerald J. Little | Ph (61 49) 215618 The Neuroscience Group | Discipline of Anatomy | Fax (61 49) 216903 Faculty of Medicine and | Health Sciences | The University of Newcastle | Email ANGJL-at-medicine.newcastle.edu.au Australia, 2308 |
MR-Received: by mta ISHTAR.MUAS; Relayed; Tue, 08 Nov 1994 05:49:55 -0400 (EDT) MR-Received: by mta ISHTAR; Relayed; Tue, 08 Nov 1994 05:49:55 -0400 (EDT) Disclose-recipients: prohibited
We have been using several different fluorescein based dyes to measure pH gradient within epidermal cells of roots from maize seedlings in situ. Treatment of the tissue with snarf-am under the appropriate conditions leads to a rather uniform labeling of both the vacuole and cytoplasm. Therefore, the snarf in these cells should be in pH environments ranging from pH 5 in the vacuole to almost pH 8 in to cytoplasm. We would like to be able to measure this pH gradient. The pH dependence graph of snarf in the Molecular Probes book makes one think it should be able to detect pH changes throughout this range. We have a calibration curve using a ratio of two excitation wavelengths of 420 and 475 nm at a constant emission wavelength of 580 + 30 nm. However, this ratio varies only between pH 7 and 9. We have experimented with another calibration curve using a ratio intensities with excitation and emission wavelengths of 420 nm and 580 nm compared to that obtained at exciation and emission wavelength of 480 and 620 nm. With standard solutions, this ratio shows pH-dependent changes from pH 5 to 9 but the majority of the change is between pH 7 and 8. However, the calibration does not look as clean when one tries to do the calibration by clamping cells at various pH by various means. Any suggestions would be appreciated. Thank You for Your Reply in advance, Dave Brauer plant Physioogist dbrauer-at-arserrc.gov
If your simply interested in viewing Tiff images (and many other formats) on a PC I would suggest Norton Desktop for Windows (by Symantec). The Norton Veiwer does offer limited image manipulation such as zooming. If your like me and have basically been forced from DOS to the Windows environment for $90 Norton Desktop greatly improves windows and has alot more to offer than just viewing and shuffling files (though as a replacment for Windows Filemanager alone I would recomend it). I don't know if the lastest version of Norton Commander has added Tiff viewing compatablity - but I wouldn't doubt it - and it runs under DOS without windows.
If you need additional image manipulation or another "viewer" WordPerfect 6.0 ($130 - educational discount) handles Tiff format with no problem, and allows limited image manipulation (zooming, cut, pasting, contrast,brightness) and printing. I would assume that most other current word processors should have similar features.
But for "real" image processing you need to spend much more money for the dedicated image processing programs. You might want to check out the article "High-end Image Editing Software" in Computer Shopper, October 1994, 14: 548-558, and the article "Graphical Voodoo" in the same issue p.610-622 which offers an excellent review of the various imaging formats pros and cons.
Richard E. Edelmann Electron Microscopy Facility Supervisor Marshall University - School of Medicine Huntington, West Virginia
Position available A postdoctoral position is available within the Structural Biology Imaging Center of the University of Pittsburgh Medical School. The focus of the position will be to study the interactions/role of spectrin like proteins in muscle, brain and other tissue systems. Specifically dystrophin and alpha actinin. The principal approach will be to use a variety of high resolution light and electron microscopic techniques (confocal, Immuno-EM, etc) to define where, when, and in association with which proteins these specific molecules and isoforms of these molecules are found. The applicant should have experience/interest in modern microscopic methods and be willing to learn other techniques as needed.
If you are, or know of someone who may be interested please contact the director of the lab listed below Simon C. Watkins Director SBIC 840 Scaife Hall University of Pittsburgh Pittsburgh PA 412-648-3051 email Swatkins-at-Pitt.edu
Its alright, Bill. In the spirit of academic freedom & creativity I'm perfectly willing to accept your spelling of "Copland". Or to put it to you as a quote from our oatmeal poet-lorryate[sic], Bob Dylan, "Don't think twice, its alright......Babe".
In message {9411072352.AA03570-at-MOLE.mmm.com} writes: } One last (?) word on spelling errors: } "It's a damn small mind that can think of only one way to spell a word." } - Andrew Jackson (seventh President of the U.S.)
Gib
P.S. - to follow soon. Must go aline the CM12 for 120kV.
I agree with Scott, Adobe Photoshop is the best way to view/manipulate images, but for the Mac, there is a shareware program called GIFconverter 2.3.2 which will open any picture file and convert to any known format you can think of.
"You can't help respecting anybody who can spell TUESDAY, even if thy don't spell it right; but spelling isn't everything. There are days when spelling Tuesday simply doesn't count." A.A. Milne. from The House at Pooh Corner.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
John F. Mansfield North Campus Electron Microbeam Analysis Lab. 2455 Hayward Ann Arbor MI 48109-2143 Tel: (313)936-3352 Email: jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html Time: 12:59 Date: 11/8/94
Graphic Converter is better & supports more formats.
--------------------------------------
Ron
------------------ RFC822 Header Follows ------------------ Received: by mse.engin.umich.edu with SMTP;8 Nov 1994 12:20:25 -0400 Received: by judgmentday.rs.itd.umich.edu (8.6.9/2.2) with X.500 id LAA22095; Tue, 8 Nov 1994 11:18:29 -0500 Received: from AAEM.AMC.ANL.GOV by judgmentday.rs.itd.umich.edu (8.6.9/2.2) with SMTP id LAA22076; Tue, 8 Nov 1994 11:18:23 -0500
I am looking for feed back from anyone out there listening.
I am wondering how UNIX, Windows NT and OS/2 compare overall but especialy how the latter two compare with UNIX. I am looking into upgrading operating systems. In general the hardware platform will be a 586/90MHz PCI/EISA Bus, 16 Meg RAM, 1 GB diskspace, eithernet, etc.. I would like to go with UNIX (Novell or SCO), but I am concerned about possible problems with running the vast array of software written for the windows world in the X-windows environment and if the windows software would run better under Windows NT (Or OS/2).
Does anyone out there have any experiences to share? General preferences? Considerations or comments?
If you E-mail directly I'll try and coallate the info and either post it as one message or make it available to anyone who'd like it.
Thanks in advance.
Richard E. Edelmann Electron Microscopy Facility Supervisor Marshall University - School of Medicine Huntington, West Virginia
Message-Id: {9411082037.AA25076-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Re: format converters, was Orig-Author: {"John Mansfield" {John_Mansfield-at-mse.engin.umich.edu} }:ddn:wpafb ----------------------------------------------------------- John F. Mansfield North Campus Electron Microbeam Analysis Lab. 2455 Hayward Ann Arbor MI 48109-2143 Tel: (313)936-3352 Email: jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html Time: 12:59 Date: 11/8/94
Graphic Converter is better & supports more formats.
Dear Peter, I actually run a screen recoating service from the E.M. Unit here at very reasonable rates. If you are interested in any further information contact me direct.
Cheers,
Tony Romeo
Tony Romeo Internet: tony-at-emu.su.oz.au Electron Microscope Unit Madsen Building F09 Telephone: +61 2 351 2351 The University of Sydney Facsimile: +61 2 552 1967 Sydney, NSW 2006 Australia
Message-Id: {1994Nov08.163926.3183459-at-mail.magic.ca} To: microscopy-at-aaem.amc.anl.gov
Hey Ron, I think you have the programmes mixed up, GraphicConverter v2.0 by Thorsten Lemke converts most any graphic file. In addition you can do a little image processing.
I'm trying to embed plant material is some type of matrix that will allow me to make fairly thick sections, say 25 - 50 um.
I've used paraffin and the tissue starts to fracture when section thickness gets greater than 20 - 25 um. Frankly besides embedding the tissue in plastic and then grinding it down I'm not sure what else to do.
Bob, I don't know about Graphic Converter that John was talking about, but most images that are passed around these days (that I know of) are either in TIFF, GIF, or JPEG format. This program will convert between those and a a handful of others. I think i pulled it off of an Apple shareware site. I think the U. of Texas has one. If you can't find it, I could try sending it to you compacted. But check out the one Jahn Mansfield was talking about first.
We use Adobe Photoshop (both PC and Mac) and have found it very powerful and useful. It is however expensive (at least in Australia). JASC produce a Windows product called Paint Shop Pro which is not as expensive (around $70 USD) but can read and write many (around 40) image formats including most of the more common TIFF formats. It also has contrast/brightness controls, gamma correction and some filters.
I think, though I'm not sure, that there is a shareware version available.
According to the manual the address and 'phone number is:
JASC Inc. 10901 Red Circle Drive, Suite 340 Minnetonka, MN 55343 (612) 930-9171
I have no interest in the company, I just use the software!!
Other programs which may be of use are Image Alchemy (PC) or NIH Image (Mac).
Good Luck!!
##################################################################### ********************** * Between the idea * 0------* And the reality * } ---|--- { * Between the motion * | * And the act * / \ * Falls the Shadow * _/ \_ * T.S. Eliot * ********************** Colin Veitch Tel + 61 (0)52 27 5611 CSIRO Division of Wool Technology Tel + 61 (0)52 27 5891 (dir.) P.O. Box 21 Fax + 61 (0)52 27 5657 BELMONT Vic 3216 Australia
There have recently been several items posted concerning magnetic shielding of EM rooms. Several people have pointed out that this is an expensive and generally impractical thing to attempt, and they are right.
Something that is often not appreciated is that you don't actually "shield" magnetic fields in the sense of "blocking" them. Instead you "shunt" the fields around the area you are trying to protect. The commonly used shielding materials have extremely high initial permeability which means that magnetic fields will travel through them in preference to other routes. In other words, you don't build a "dam" in the way of the field, but instead you provide a "sponge" to channel it around the area to be shielded.
This sounds fairly simple until you realize that the high permeability of the material means that you can also easily channel magnetic fields INTO the area you are trying to shield if the enclosure is not a properly designed magnetic circuit. Just sticking a sheet of "mu-metal" in the way of the field is likely to make things worse. Holes or open joints can also create "lensing" effects.
Even in the best of cases, all of the magnetic flux won't flow through the path provided by the "shield" -- some amount of field always fringes out into the surrounding area. So at best, all you can do is attentuate the fields (not eliminate them entirely). The more efficient the circuit you provide for the fields to travel in, the greater the attenuation. However, as enclosures increase in size, more of the field lines will find it advantageous to take a "short cut" across the enclosed free-space region. For a simple cylindrical field, the attenuation varies as the permeability times thickness of the shield, divided by its diameter. In practical terms, this says it's pretty easy to shield a small region, but to get comparable shielding of a room, for example, the shield has to be quite thick (or consist of multiple isolated layers). Obviously, it is best to shield the smallest possible volume, which is why you want the shielding designed into your EM, rather than applied as an afterthought.
By way of reference, the initial permeability of a typical 80/20 (Ni/Fe) shielding material will be in excess of 20,000, as compared to several hundred for "soft" iron (free space is 1.0). Oriented silicon steel, as used in transformers, has IP of about 1000 and is often a good shielding material.
The above remarks apply principally to the case of lower frequency fields (such as the omnipresent 60 Hz). For high frequency fields one gets substantial eddy-current shielding just by using a good electrically conductive enclosure, such as aluminum.
There is real art to designing a good shield, and for all but the simplest cases, one is well advised to seek the assistance of someone who does it routinely. I'm not such a person, but have had good luck in the past working with Advance Magnetics (219) 223-3158, whose catalog also contains a lot of helpful application information (I have no connection to this firm). There are also a number of other good companies in this business.
By the way, I have been looking for years for an inexpensive PC-based program which would be useful for designing magnetic shields -- ideally something in the freeware/shareware realm. If someone out there knows of such a thing (or knows of someone who might), I'd appreciate hearing about it.
Another good shareware alternative for viewing TIFF, or just about anything else, is Paintshop Pro. It claims to handle three or so types of compression for TIFF files, up to 32 bit colour depth etc. I don't know whether it will cope with _all_ TIFF files but it works for everything I have tried it with.
Good Luck
Doug
+------------------------------------+ | Dr Douglas Arrell | | Mechanical Performance and Joining | | Institute for Advanced Materials | | 1755 ZG Petten | | Netherlands | | {ARRELL-at-JRC.NL} | | Tel. (+31) 2246 5287 | | Fax (+31) 2246 1917 | +------------------------------------+
I have been questioned as to the validity of my inquiry of PC Operating systems on this BBS, let me explain.
I am setting up a local network system to intergrate my EM Facility. This platform will be the backbone server for the exchange of data (Images from LM, LSM, TEM, SEM, EDS, etc), such that users can manipulate their images at a variety of sites. It is being designed to be networked directly to microscopes which use DOS, Windows, and X-Windows. This net will also allow for sellection between a variety of hardcopy output devices. It is also the intention that this system serve as an internet node allowing for the exchange of images across the net.
The platfrom will also be used for tasks such as E-mail, word processing, statistical analysis.
The microscope/networking end is covered. However I was interested in collecting further info for the operating system to function under. Since the users of this BBS are very familar with computer systems and complex software (i.e. high end imaging software) I thought this was a good place for this question.
I am sorry if you disagree.
Yes, this type of a system could be purchased from a vendor out there for however many $10k's (and for those of you who can afford it great) but the I know many labs out there are like mine in which we need to do everthing we can to save cost, and still get the best we can afford.
Richard E. Edelmann Electron Microscopy Facility Supervisor Marshall University - School of Medicine Huntington, West Virginia
A number of people have commented recently on how difficult it is to shield stray fields, suggesting that one needs to shield a whole room at the cost of } $10K. I would like to point out that simple, and cheap shielding with a little Mumetal has, EXPERIMENTALY, worked in at least two cases I know of: a) The old Cambridge HREM where a small shield was introduced in the column. (Ask Dave Smith now at ASU for further details.) b) Our own HREM here at Northwestern.
True, such a simple shield may not completely cure the problem, but for a few hundred dollars it is well worth the attempt! A very simple test that my students devised was to wave a small magnet near the microscope to test what regions were sensitive.
I should also comment that most people build small (cheap) shields around LEED systems, and most surface science systems operating with low energy electrons use shields with holes in them (for flanges). The VG STEMs also have a shield above the objective which has holes in it.
} Dear All, } } Anyone out their re-coat their phosphor screens? } } How does one go about it? Are the supplies available in South Africa? } } The screen I am proposing to coat has an alluminium base, about 12 cm } diameter, for a Hitachi TEM. } } Many thanks for any advice. } } Peter Richards. } } _______________________________________________________________ } } } -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- } -at- -at- } -at- Peter D. G. Richards -at- } -at- Dept Anatomy and Cell Biology -at- } -at- UCT Medical School -at- } -at- Observatory -at- } -at- 7925 -at- } -at- RSA -at- } -at- Tel: 021-406 6285. -at- } -at- Internet: retep-at-anat.uct.ac.za -at- } -at- -at- } -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- } Peter: Recoating screens is a fairly simple thing to do. I used to recoat mine all the time using a technique used by a now defunct company called AEI. I used to use a phosphor put out by JEOL. Always got beautiful screens. Their phosphor gave me a screen with good contrast levels. If you can give me a fax number, I will fax a copy of the technique to you. I still have some of the phosphor and could send some to you if there isn't any problem with the mail system getting it to you. Let me know. Phil
This is to add on to the message posted by Colin Veitch: } } JASC produces a Windows product called Paint Shop Pro which is not as expensive (around $70 USD) but can read and write many (around 40) image formats including most of the more common TIFF formats. { {
This is a very nice product. It is shareware and can be downloaded from Compuserve, AOL, etc. The registration fee is $69. It lists at least 20 discrete formats (GIF,TIF,JPG, etc) and seems to handle them all smoothly, including converting between.
JASC also sells a companion Windows product called "Image Commander" which allows you to setup "catalogs" of images and view them as a page of thumbnail pictures or select a particular one for viewing. It has provisions for organizing the catalogs, annotating the images, etc., and would look like a very useful thing for anyone handling lots of digitized micrographs. This is also shareware, and the registration is $39.
I have no interest in the company, but have had good experience with both products.
Message-Id: {1994Nov09.134725.3200862-at-mail.magic.ca} To: microscopy-at-anlemc.msd.anl.gov
S
Hey Ron, I think you have the programmes mixed up, GraphicConverter v2.0 by Thorsten Lemke converts most any graphic file. In addition you can do a little image processing.
I am gathering and storing away much of the information on this bulletin board regarding networking of microscopes and image storage systems. I, for one, appreciate the inquiries of people such as Richard Edelmann regarding computers and operating systems for microscopy imaging. Also, I have noticed in the last week that my spelling has improved vastely.
Nancy Smith Director, EM Lab CSU,Hayward Hayward, CA 94542
Although you may end up using it inelegantly, e.g. via telnet and ftp, probably the way to go is: o Unix machine with tons of disk space as a server when images are stored temporarily o stand alone PCs (or Macs, etc.) have their own removable media and users MUST purchase their own disks for storage.
We have found that attempting to network PCs (some DOS, some windows, some OS2), UNIX, OS7, and Macs is nearly impossible. The only platform that appears to be compatible with all the other platforms is UNIX.
Subject: Time:5:05 AM OFFICE MEMO Re:Spelling Fluorine Date:11/10/94
Like the professor who pointed out that if the first five letters spelled 'flour' it was wrong, I had a biochemistry professor who told us to split the word into two syllables, and if it came out 'flo-urine', it was wrong.
Our department has available, for the cost of moving, a Hitachi HS-8 TEM. It is a one owner instrument and has been maintained on manufacturer's service contract since installation. It is currently fully operational. We will be moving this microscope very soon.
Please contact me directly for more information.
John chandler-at-lamar.ColoState.EDU Fort Collins CO
I recently spend 4 weeks in the Netherlands learning microscopy for yeast samples-only morphology and immunogold techniques. Now I am trying to set up our lab in America to do these techniques. I have three major problems-limited experience, no one at my institute can help me, and having learned in Europe I have no knowledge of good American suppliers for EM stuff.
Here is what I need: Right now we have a sorvall MT2-B "Porter-Blum" ultra-microtome. We are looking for a table for it (vibration resistant), as well as for a new ultramicrotome.
I also would like to know a good general supplier (other than EM sciences) for things like grids, grid holders, capsule holders, etc....
I also would like to know where people get good quality formaldehyde, glutaraldehyde, potassium permanganate, and ethanol ( in other words the basic chemicals needed).
Thanks for your help. Kim Wilson-Oregon Graduate Institute
In article {MAILQUEUE-101.941108141914.288-at-anat.uct.ac.za} , RETEP-at-anat.uct.ac.za writes: } Dear All, } Anyone out their re-coat their phosphor screens? } How does one go about it? Are the supplies available in South Africa? } The screen I am proposing to coat has an alluminium base, about 12 cm } diameter, for a Hitachi TEM. } } Many thanks for any advice. } } Peter Richards.
Dear Peter, We routinely recoat our screens, and, if the phosphor is available, the rest of the supplies will be easy. The steps are: 1) Make } 500 ml 0.5% gel- atin solution, 2) Filter with type GS 0.22 mu millipore -at- ~30 C, 3) Suspend 10.6 g (for ~45 g/cm**2 and ~170 cm**2 screen size, your mileage may vary) phosphor in 500 ml gelatin soln, 4) Warm and degas the suspension while, 5) Clean screen blank, 6) Glow discharge blank for 15 min, 7) Place blank in funnel so that it is horizontal and so that there is an area between the edge of the blank and the wall of the funnel (our screen blank is not completely circular, but has two straight sides cut along chords, you may have to build a frame to achieve this geometry), 8) Attach a hose to the bottom of the funnel, and clamp it off, 9) Fill the funnel with 50% ethylene glycol to a level just above the bottom face of the blank and make sure there are no air bubbles trap- ped beneath the surface, 10) Pour the phosphor suspension down a glass rod and move the rod around to avoid pileup on any one area, 11) Cast a film of collo- dion diluted 1:100 in n-butyl acetate on the surface of the liquid by deposi- ting drops with a Pasteur pipette until the entire surface is covered, 12) When the n-butyl acetate has evaporated, sweep the film aside with a wedge of filter paper; repeat until the liquid surface is free of debris, 13) open the clamp on the hose and allow the liquid to drain slowly--~2 drops/sec--until the level is below the bottom surface of the blank, 14) Place a clean sheet of paper over the top of the funnel and draw air through the hose and past the screen; leave overnight, and 14) Remove the screen from the funnel, check for evenness of coating and lack of foreign objects (eyebrows etc.), then bake at 60 C for 24 hours. Good luck, it takes a little practise, but the blanks can, of course, be reused. Yours, Bill Tivol
Fluorine being on the scene, I take my chance once more to ask again a question stayed without any response since two months :
"How to avoid the almost systematic decomposition of inorganic fluoride compounds under the electron beam during a TEM experiment (JEOL 2010) ? Any suggestion appreciated (inversely as the cost)."
The question may be formulated in another way : Has somebody ever worked with some success on inorganic fluorides by TEM ?
An element of response is in "The electron microscopic observation of phase transition in KFeF4, RbFeF4, and RbVF4," by R. Deblieck, J. Van Landuyt & S. Amelinckx, J. Solid State Chem. 59, 379-387 (1985). The problem with inorganic fluorides is electrical conduction : "After washing in benzene, in order to remove tape residues, the thinned plates are mounted on 3-mm copper grids by means of silver paste to ensure good thermal and electrical contact. In spite of these precautions electrical conduction remains a problem and substantial charging phenomena still occur under the electron bombardment for observation. Moreover, all materials are subject to damage by electron irradiation..... The investigated compounds prove to be sensitive to decomposition under electron irradiation, probably through temperature-driven evaporation of fluorine."
Other experiences on inorganic fluorides by TEM ? Suggestions ?
Microscopists do not like fluorine, this may be why there are some spelling problems.
Armel Le Bail - Fluoride Lab - armel-at-ONE.univ-lemans.fr
Message-Id: {9411101344.AA06245-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: TEM and inorganic fluorides Orig-Author: {lebail-at-cnrs-imn.fr}:ddn:wpafb ----------------------------------------------------------- Hi,
Fluorine being on the scene, I take my chance once more to ask again a question stayed without any response since two months :
"How to avoid the almost systematic decomposition of inorganic fluoride compounds under the electron beam during a TEM experiment (JEOL 2010) ? Any suggestion appreciated (inversely as the cost)."
The question may be formulated in another way : Has somebody ever worked with some success on inorganic fluorides by TEM ?
An element of response is in "The electron microscopic observation of phase transition in KFeF4, RbFeF4, and RbVF4," by R. Deblieck, J. Van Landuyt & S. Amelinckx, J. Solid State Chem. 59, 379-387 (1985). The problem with inorganic fluorides is electrical conduction : "After washing in benzene, in order to remove tape residues, the thinned plates are mounted on 3-mm copper grids by means of silver paste to ensure good thermal and electrical contact. In spite of these precautions electrical conduction remains a problem and substantial charging phenomena still occur under the electron bombardment for observation. Moreover, all materials are subject to damage by electron irradiation..... The investigated compounds prove to be sensitive to decomposition under electron irradiation, probably through temperature-driven evaporation of fluorine."
Other experiences on inorganic fluorides by TEM ? Suggestions ?
Microscopists do not like fluorine, this may be why there are some spelling problems.
Armel Le Bail - Fluoride Lab - armel-at-ONE.univ-lemans.
If necessary, you can add additional NuBus cards to your Mac by adding a NuBus expansion chassis. I have one made by Second Wave which gives me an additional three slots (as well as a power supply and connections for an internal hard disk drive) on my Mac IIci. There are also six slot chasses available.
Second Wave can be contacted at: 9430 Research Blvd., Echelon II, Suite 260 Austin, TX 78759-6541 512-343-9661 512-343-9663 (FAX) Applelink: D0864
I have no connection to Second Wave, I'm just a satisfied customer.
Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2005 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA ----------------------------------------------------- (313) 936-1550 (voice) **** Next time, **** (313) 763-4690 (FAX) *** take the *** chender-at-umich.edu (e-mail) **** train! **** ----------------------------------------------------- On Thu, 10 Nov 1994, Steve Bill 5355/5128/5642 wrote:
} } We are currently using a HP Laserjet 4M to output TEM images from our Gatan } Slow-scan camera (mac based). What I'm looking for is an enhancement that will } improve the output of images from this printer. I've heard of some grey-scale } sofware upgrades and some add-in cards that are supposed to boost the resolution } or improve the grey-scale rendition of the HP laser printers but I don't have any } specific information or pointers to any of them. } } The Mac by the way has no NUBUS slots available (all three are taken) so anything } we consider must either be software-based or hardware added to the printer itself. } } Thanks, } } Steve Bill }
Dear Armel et al., My experience with fluorides is limited to organics, but the problem of radiation damage is still serious with these fluorides as well. The best procedure IMHO is to use as low a dose as possible--especially for focusing, etc., where the electrons are causing damage, but not being recorded. We use our fairly crude, but sensitive, video system to scan grids for areas of interest, focusing, etc., and LoDose or SO163 for image recording. I have found that if the lens column is aligned well, the wobbler can be used to focus quickly within one click of the fine objective control (=.99 micron). A subsequent through-focus series should yield good images. This works at mags up to 200,000, and for low dose images, I have had my best luck with somewhat lower mags than this. I use a 30 micrometer C2 aperture with the wehnelt bias on the lowest setting and the C1 lens maximally excited. This produces a beam current in the picoamp range, where charging, heating, etc. are minimized (it helps to use 1.2 MV electrons if you have them). Remember, alignment of the lens column is critical--check and correct it the same day you take the pictures. Good luck. Yours, Bill Tivol
In response to: "How to avoid the almost systematic decomposition of inorganic fluoride compounds under the electron beam during a TEM experiment...?"
You might try cryo EM, that is lowering the temperature of the specimen towards that of liquid nitrogen or liquid helium. This preserves visible structure for a time in biological specimens, presumably due to a decreased vapor pressure of the products of radiolysis.
We are struggling with imaging droplets of liquid fluorinated hydrocarbons, by freezing the drops on a substrate and viewing them with the low-temperature scanning electron microscope at -180 C. At low temperature, large (1mm diameter) drops are resistant to the 10kV beam, small (1 m) droplets sublime. Interestingly, even after a ca. 100A gold coat the fluorocarbon drops appear brighter than adjacent aqueous structures, perhaps due to charging or the differing secondard electron coefficients of fluorocarbon and water.
Jacob Bastacky, MD 1-116 Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
Steve Bill 5355/5128/5642 {sbill-at-asdg.enet.dec.com}
Steve,
Information about LaserPrinter Enhancements:
PC based controllers are available from the following companies;
LaserMaster Corporation - WinJet Product Line (Windows Only) Talltree Systems- JLaser? Greylaser? (? if still selling) DPTech - ? (not sure if they are still selling units) XLI Corp - ?(not sure of Name) Should be selling controllers only No complete products including engine.
Mac Based products; I am currently not aware of any in a controller or software only basis.
Complete Printer subsystem for High-Res output -- PC & MAC & Unix*
LaserMaster ImagePrinter 1800 DPI (#2 Version) Appletalk, Eithertalk, TCP/IP, Novel Netware, Parallel (HS), Serial communications on unit. 24 meg Ram 66 Mhz Multi-Tasking Pipeline Processor 240 Meg Hard Drive internal Pipeline Assoc. Postscript Level 2 interpreter & PCL 4 Languages 8 1/2 X 11" to 12.5 X 19" paper size on line.
*unix is only for TCP/IP standards on limited System OS
This is only 1 of our printers avaiable. I do work for LM in the Scientific and Medical industries and can help you if you would like. Contact Greg Begin LM Imaging at 1-800-950-6363 Ext: 3207 or Mail address of gregb-at-sales.LMT.com I will be out at the American Heart Assoc. meeting next week and then the Radiological Society of North American durring the last week of Nov. Greg Begin LM Imaging Div. LaserMaster Corporation /\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\
} Date sent: Thu, 10 Nov 94 09:59:20 EST } From: Steve Bill 5355/5128/5642 {sbill-at-asdg.enet.dec.com} } To: microscopy-at-aaem.amc.anl.gov } Copies to: sbill-at-asdg.enet.dec.com } Subject: Laser printer enhancements?
} } We are currently using a HP Laserjet 4M to output TEM images from our Gatan } Slow-scan camera (mac based). What I'm looking for is an enhancement that will } improve the output of images from this printer. I've heard of some grey-scale } sofware upgrades and some add-in cards that are supposed to boost the resolution } or improve the grey-scale rendition of the HP laser printers but I don't have any } specific information or pointers to any of them. } } The Mac by the way has no NUBUS slots available (all three are taken) so anything } we consider must either be software-based or hardware added to the printer itself. } } Thanks, } } Steve Bill }
After so many requests for the procedure, it is a little cheaper to put it out on the microscopy board. So, here goes for those who want it. I get my phosphor from JEOL, cat.#423-011(green) or 423-010(yellow). These are old cat. numbers, they might have changed. You can call the parts dept. at (508) 536-2342 and ask for Maria Ramos.
1. Withdraw the old screen from the microscope, taking care to avoid depositing any loosened phosphor in the viewing chamber. 2. Remove the old coating from the screen plate by washing in acetone. The cleaned plate must be free from all particles of matter and the surface must be free from blemishes and scratches. 3. Prepare a sufficient volume of 4% (w/v) suspension of phosphor powder in acetone containing about 1% of collodion. To make collodion use: 4grams parlodion, 25ml 100% ETOH, 75ml ether., Any grade of collodion that is suitable for specimen preparation can be used. The total volume must be sufficient to fill the selected dish with liquid to a depth of about 1cm above the surface of the screen plate in position on the bottom of the dish. 4. Agitate the suspension vigorously then pour it rapidly into the dish. 5. Wait about 5 seconds to allow large particles to settle and violent swirling to cease. 6. Slide the screen smoothly into the liquid, preferably without scraping the bottom of the dish. 7. Cover the dish with a piece of plexiglass with a small (1/4"-1/2") hole drilled in it and leave it to settle (I usually let it sit overnight). When the remaing liquid is clear, draw off the liquid by inserting a suction tube through the hole in the lid. It is extremely important not to disturb the screen or the liquid above the screen in any way as this is being done. Draw off the liquid steadly and slowly then remove the suction tube. 8. Leave the screen to dry without any further disturbance of any kind. Do not lift the lid to inspect the screen until the powder is quite dry because a slight change in drying conditions can produce a visible mark on the damp surface. 9. When the screen is dry, remove the screen and wipe off any excess phosphor from the back and sides of the screen plate. 10. Put screen back into the microscope.
Note: A newly-coated screen will de-gas for a short time when it is first placed in the microscope. Pumping times may therefore be longer than normal at first.
A very small particle size is desirable for high resolution screens and it may be found advantageous to agitate the phosphor suspension ultrasonically before putting it in the dish. Screen resolution is also affected by the thickness of the phosphor layer and by light scattering within it.
It may take a few tries to get the hang of it, but when I had to recoat my screens this technique worked rather well. Fortunately, I no longer have to recoat my screens.
According to a OSHA publication (1978), liquid containing Osmium tetroxide should be absorbed in vermiculite, dry sand or similar material and then disposed off in a sealed container in a secured sanitary landfill.
Greetings, There was an interesting paper in the Recent Microscopy Today (october) in which different ways of neutralizing liquid osmium waste/spills are tested. They find corn oil-soaked kitty litter to be the best treatment of all, with pure corn oil not far behind.
Return-Path: ert-at-leka.hut.fi Received: from leka.hut.fi (leka.hut.fi [130.233.224.22]) by santra.hut.fi (8.6.8.1/8.6.7) with ESMTP id QAA07915 for {listserver-at-anlemc.msd.gov} ; Thu, 10 Nov 1994 16:19:57 +0200 Received: (from ert-at-localhost) by leka.hut.fi (8.6.8.1/8.6.7) id QAA110591 for listserver-at-anlemc.msd.gov; Thu, 10 Nov 1994 16:19:44 +0200
dear Nestor,
Microscopists, I will like to participe in the network of the microscopists.
My email: eero.ristolainen-at-hut.fi
Address: Helsinki University of Technology Center for Chemical Analysis FIN-02150 Espoo, Finland
Michael, Have you considered: 1) freezing sectioning on a sliding (=sledge) microtome or 2) a vibratome (I've had my best luck with 10% gelatin fixed 16-20 hr in 10% formaldehyde) Phil Oshel poshel-at-luc.edu
With budgetary restictions on the sciences these days, many of us are no longer able to attend the National Microscopy Society of America (MSA) meeting. We are limited in the ability to present our work, share our ideas and research, or meet with our colleagues due purely to financial limitations.
Shams Ghoniem of the Iowa Microscopy Society has suggested that MSA consider awarding grants to professional and scientific staff similar to the program currently in place for students. This recommendation, put forward by Peter Ingram, LAS/MSA president, was met with a lukewarm response by the MSA council. I believe that MSA would consider such a proposal if enough support by the MSA membership is provided. MSA would benefit by increased attendance at meetings, more incentive to join MSA, as well as in supporting education and professional development of their members.
I am requesting therfore that letters be sent fo Peter Ingram in support of this proposal. We need letters from across the country including letters from faculty who are willing to match an MSA award. These letters need to be sent by the first of January to be influential at the next MSA committee meeting.
His address is: Peter Ingram LAS Director Research Triangle Institute Box 12194 3040 Cornwallis Road Research Triangle Park, NC 27709
E-mail: ingram-at-rcc.rti.org
I thank you for your attentive ear and for your letters!
I understand that there are sources for particularly sharp AFM tips. The normal AFM tip has a radius on the order of 10's of nanometers and isn't very useful for "deep" contours. Does onyone out there know of a source for sharper tips.
Daniel L. Callahan Department of Mechanical Engg. and Materials Science Rice University dlc-at-owlnet.rice.edu
Group - Preface: I find myself "semi-retired" (meaning that I have more free time than most of you), yet interested in microscopy (our newsletter "Microscopy Today") and rather alert when it comes to data bases. Result: I am considering building the equipment listing data base that has been discussed. My THOUGHT: To build a data base with two sections: Section 1: A simple listing of members - names, companies, addresses, tel. & eMail address, etc. To include all interested, not necessarily only those identified with equipment. Hard copy of this section could, of course, be easily provided. Section 2: A listing of equipment categories. I.E., I would expect SEM, ESEM, etc. to be categories. Each category to be a "field" in which we c ould list individual products - which can be searched on. Then, perhaps, to provide the data base on disks for those interested. Perhaps there is a way that one can access the data base by eMail? Before I consider getting serious, I have two questions: 1) Would the above solve the need as discussed? Or is there other and/or dif ferent detail that should be included? Applications? 2) I happen to use "Q&A" as my data base software. In Q&A, items can be separated in a keyword field with semicolons and then searched on. I can export to any other data base software. I do not know if other data base software will allow this keyword search? Rather than swamping the listserver with comments from the whole membership, may I suggest that only those who have previously commented on this subject publish their response to the full readership. Others might just send their opinions to me. Assuming that the concept is OK, or close, the next step is to ask your help in defining categories. I will ask for that assistance should the project be a "go". Regards, Don Grimes, Microscopy Today
My computer address at Cambridge has been changed. It is noe pe13-at-cus.cam.ac.uk Will you please make the necessary alteration at your end so I may continue to get all the good stuff on the MSA file server.
unsubscribe eliesh.oneil-at-gtri.gatech.edu Elizabeth G. (Eliesh) O'Neil Research Scientist I Georgia Tech Research Institute Electro-Optics, Environment, and Materials Laboratory Materials Analysis Center 925 Dalney Street, Baker 271 Atlanta, Georgia 30332-0827 Phone: (404) 853-0590 FAX: (404) 894-5073 Email: eliesh.oneil-at-gtri.gatech.edu
Please do not forget the User Facilities and equipment lists and DATABASE that Sandy Silvers has compile and distributes via her local society. It is called the MSA TECH FORUM FACILITY DIRECTORY. The last update that I received from Sandy was dated 7/94. This is a working DATABASE with the following information...
Facilities ] Phone Lists State Listings Equipment Listings.
The contact phone number listing is : 706-546-3471
We have called Vincent and Associates at (800) 828-6972 regarding their products. We are waiting for specifications and pricing via snail mail. Has anybody experience with operating one of their shutters (e.g. D122 or iD123) from a serial port, specifically from within NIH-Image? They make it sound very simple, but we thought we'd find out whether anybody has experience.
The Ludl system, at first glance, seems far more expensive. Is it significantly better? And the same question as above regarding the Vincent shutter.
We're waiting to hear from Scientific Imaging Syatems too.
Group - In response to my recent "offer" to develop an appropriate data base, I note the comment from Nestor where he recommends the data base as developed by Sandy Silvers. I accept that as a "non-recommendation" to work with me. I should point out that the "meat" of Sandy's data base is two listings: 1) An "equipment listing" including type/model, year installed, whether under a service contract, and user charge rates. 2) An "ancillary equipment listing" including quantity, manufacturer, type, model #s, etc. There is no way (I BELIEVE) to do a search on specific instruments or applications - and come up with a list of others with a specific interest. I should also point out that Sandy's system has sections for: 1) Type of Facility (biology, material science, core univ., etc. 2) List of personnel and users. 3) Other services provided 4) Number of users per year. Kindly understand that I have NO INTEREST in competing against Nestor and his wishes - we (including I) owe him too much. And, considering the amount of work involved, I have no interest in accomplishing a proper, searchable, data base without the full support of the "management" of the listserver. As a result, I withdraw my offer - with thanks to you all that have expressed interest and support. Don Grimes, Microscopy Today
I know of one case where sheilding was successful. The Field emmission sem at cell biology, Yale Medical school had all the usual problems until a layer of (I thought it was brass sheet but am not sure now) metal was applied on all 6 surfaces of the room at ???? cost. I don't have his email but Phillip Male (pronounced Mal) at cell biology yale university is the one to contact, the shielding DID work! alan pooley marine sci rutgers univ nj
Does anyone have names and phone numbers for suppliers of standards for my EDX analysis. I need to know the compositions to see how well my setup is doing it's work.
Cheers from Sunny Vancouver. Dan Henne Simon Fraser University henne-at-sfu.ca
Sorry If I gave the wrong impression. I have no problems with either Don or Sandy running a DATABASE on equipment. I just wanted to make sure that Sandy's work over the last few years was remembered.
Don, your more than welcome to continue to use this forum for suggestions and/or comments. I apologize if I gave the wrong impression....
As I Mentioned on the net earlier, we have an old JEOL model JSM-2 SEM with a Nuclear Diodes/ EDAX model 508 EDS system, built in 1968. It has a manual vacuum valving system and the power supply is vacuum tube based. It was refurbished in the mid 80's and the EDS was repaired. Although in good operating condition a year ago when last used, we have no further use for it. We have permission to donate it to a non-profit charitable institution or University. Anyone wanting to acquire it would have to come here to Minneapolis, pack it up and ship it at their own expense. JEOL will not be able to supply parts for it, so you're on your own there. Only qualified organizations may apply; no individuals or for-profit businesses. Any requests can be sent to me at the e-mail address below. If you're seriously interested, please contact me ASAP. We must move it by the end of the year. By the way, thanks for all the advice given to me earlier.
Mike Boucher Boucher-at-tcrca.usbm.gov ***************************************************************** Michael L. Boucher Sr. Boucher-at-TCRCA.USBM.GOV Geology-Mineralogy/Chemistry Labs Ph 612-725-4614 Twin Cities Research Center Fax 612-725-4527 U.S. Bureau of Mines Center 725-4500 Department of Interior 5629 Minnehaha Avenue South Minneapolis, MN 55417-3099 U.S.A. *****************************************************************
I need some expert advice on the preparation of positive-image transparencies. I typically use 35 mm slides in my own presentations, but one of my collaborators insists on overhead transparencies. I have seen composite overheads which appear to have positive images on film attached to them. So my questions are:
Are these made by exposing light-sensitive film on an enlarger?
What type of film is used? (Manufacturer, Stock No., Sizes available)
What are the common exposure times and f-stops on the enlarger?
What are the developer/fixer chemical ratios and developing/fixing times?
A prompt response would be appreciated since this project was thrown into my lap on short notice with an even shorter deadline. BTW, I tried a laser xerox of a print (HRTEM image) onto a transparency and the results were unacceptable.
Cheers,
David A. Howell MSM Dept. Michigan State University E. Lansing, MI howelld-at-egr.msu.edu
Color laser copiers working in B/W mode give very good quality reproductions, and are far more convenient than printing methods. Most Kinko's have a copier -- you may have to work with them to make sure that they give good quality results, and many Universities also have an appropriate copier.
Very often when I am cryo-sectioning , the sections curl so badly that they are useless. This was the reason for purchasing the static-line ionizer. This really did not help the curling problem, although it served other purposes. My sections are mostly polymer-elastomer blends, primarily nylon based. If you have any suggestions I would be eternally greatful. Also if anyone has suggestions on how to pick up the samples off of the cryo knife that would also help. Right now we are using a very fine sewing needle, but something even thinner yet ridgid would be better
Thank you
Andrea Monisera. --------- End of Original Message ---------
I have seen the eyelash hairs of Dalmatians (dogs) used for this. they are forked at the end and work very well for picking up cryosections
You can take your slide to a digitizing scanning device and then feed that image to print it or to have it printed on an overhead transparency. This is done in some photo departments.
Nina Allen.
} } I need some expert advice on the preparation of positive-image } transparencies. I typically use 35 mm slides in my own } presentations, but one of my collaborators insists on } overhead transparencies. I have seen composite overheads } which appear to have positive images on film attached to them. } So my questions are: } } Are these made by exposing light-sensitive film on an } enlarger? } } What type of film is used? (Manufacturer, Stock No., Sizes } available) } } What are the common exposure times and f-stops on the enlarger? } } What are the developer/fixer chemical ratios and developing/fixing } times? } } A prompt response would be appreciated since this project was } thrown into my lap on short notice with an even shorter } deadline. BTW, I tried a laser xerox of a print (HRTEM image) onto a } transparency and the results were unacceptable. } } Cheers, } } David A. Howell } MSM Dept. } Michigan State University } E. Lansing, MI } howelld-at-egr.msu.edu }
} I am in search of a US supplier for polyester wax. This product is } produced by a company in the UK. } } Please respond directly to my e-mail address } } TIA } } Skip } melsen-at-MICROBIO.emory.edu } I expect there might be enough interest for a general reply to the list, if anyone knows, since there is an article in the new Biotechnoques [17(5):846-848] by L.L Richardson and M. Dym about polyester wax embedding for LM level immuno. They cut at 5-10 micorns at 4 degrees in a cryostat and ethanol dehydrate and air dry to get the sections to adhere to slides. The supplier they list is BDH, Poole, UK. I've never cut the stuff, but their pictures look decent. I have heard of cutting polyester wax on a sliding microtome with a funnel of dry ice above the block so the cool vapor keeps the sections from melting. If anyone has experience with cutting this wax or info about U.S. suppliers, I'd be interested in hearing about it.
------------------------------------------------------------------ |Glenn Holm *mime mail ok* Internet:karuzis-at-wccf.mit.edu | |M.I.T Dept. of Brain + Cog. Sci. This VAX doesn't do NeXTmail | |Cambridge, MA 02139 "Real Neuroscientists don't do gels!" | ------------------------------------------------------------------
Reply... RE} MSA staff support Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX: (313)936-3352 jfmjfm-at-umich.edu or John_Mansfield-at-mse.engin.umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
This is a reply to the original message posted to the Microscopy List Server that is included below. These opinions are mine own and do not necessarily relect those of my employer.
As a University employee who has to watch every dollar, I frequently find it is expensive to go to conferences. However, I do not think I should get handouts from MSA. I think that MSA money is better spent encouraging students (grade school, high school and both undergraduate and graduate university students) to get interested in various aspects of microscopy. The outreach programs in schools, the MSA scholoarships and bursaries for attending our conferences are much more important and have an impact on a greater number of people than sending a few senior people to the meetings. There are too many people wanting to milk money out of MSA for their own benefit, rather than the good of the community as a whole. As I say, just my "humble" opinion. John Mansfield.
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Dear fellow microscopists,
With budgetary restictions on the sciences these days, many of us are no longer able to attend the National Microscopy Society of America (MSA) meeting. We are limited in the ability to present our work, share our ideas and research, or meet with our colleagues due purely to financial limitations.
Shams Ghoniem of the Iowa Microscopy Society has suggested that MSA consider awarding grants to professional and scientific staff similar to the program currently in place for students. This recommendation, put forward by Peter Ingram, LAS/MSA president, was met with a lukewarm response by the MSA council. I believe that MSA would consider such a proposal if enough support by the MSA membership is provided. MSA would benefit by increased attendance at meetings, more incentive to join MSA, as well as in supporting education and professional development of their members.
I am requesting therfore that letters be sent fo Peter Ingram in support of this proposal. We need letters from across the country including letters from faculty who are willing to match an MSA award. These letters need to be sent by the first of January to be influential at the next MSA committee meeting.
His address is: Peter Ingram LAS Director Research Triangle Institute Box 12194 3040 Cornwallis Road Research Triangle Park, NC 27709
E-mail: ingram-at-rcc.rti.org
I thank you for your attentive ear and for your letters!
Kathy Walters secretary, Iowa Microscopy Society
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Reply... RE} } Linking pc to Mac Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX: (313)936-3352 jfmjfm-at-umich.edu or John_Mansfield-at-mse.engin.umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
I think the easiest way of linking a PC to a Mac over Ethernet would be to put ethernet cards in both machines (I think you have a Quadra with built in E-net and so you are all set there. Get a thin net connector for the Quadra and configure the board for the PC for thin net. If you have an EISA bus PC I would suggest getting a NE3200 (I think it is) 32 bit card that should give you the fastest throughput. However if you are on a really tight budget then you can get a 16bit card from 3Com for less than $200. You can just connect the two computers with one strip of thin net cable and terminate both ends. Then I would use public domain or shareware TCP/IP software for each machine. The Mac OS System 7.5 comes with MacTCP and you can use Fetch, NCSA Telnet or XferIt (or the commercial program Versaterm) to run ftp on the Mac and NCSA telnet to run ftp on the PC (there may be other better implementations of the TCP/IP stack for the PC that are shareware, I am not sure.) Fetch is really nice, you could put the PC into server mode (run ftpd) and use the nice interface of Fetch to put or get your files. If you are using only those two machines you can use pretty much any IP address you like for the two machines. If you put them on the Internet though, you will need to get IP addresses allocated by your local network administrator. Hope this helps. John Mansfield --------------------------------------
} cytogen-at-tigger.jvnc.net wrote: } } } Hello- } } I have a pc-based system to read in digital medical image data. I transfer } } these as TIFF or Interfiles to a Quadra 950 for processing and display and } } eventually for submission to the FDA. At the moment I use MacLinkPC to } } transfer the files over a serial line. At the rate of 57,600 baud, 100Mb } } of image data takes all night to transfer. What is the EASIEST way to take } } advantage of the Quadra's built-in Ethernet capability to speed up this } } transfer? I would prefer a solution which did not include taking a lengthy } } network administrator's course. Any suggestions will be appreciated. } } } } John G Wolodzko } } (jwolodzko-at-cytogen.com) } } Cheapest and fastest might be to use a hard-disk cartridge drive on both } the pc and Mac, then use sneaker-net! } } Roger Pyle
I've used the Syquest cartridge/sneaker-net route and it may be the fastest. A less expensive solution may be the ethernet and "PhoneNet PC" software (sold as part of Timbuktu from Farallon). I have an Asante EN2000+(thin-net) ethernet card on a PC running PhoneNet and a "friendly-talk" thin-net adapter on the Quadra. I create a "server" share folder using system 7 Appleshare on the Quadra and use PhoneNet to make the PC a client on the Quadra's share folder.(You can't make the PC a server using PhoneNet). With the proper priveliges set up on the Quadra you copy your files into it's share folder. For easiest running I would call Farallon and ask them what their default ethernet card is and buy that with the Timbuktu software. Installation and setup is then straightforward without "taking a lengthy network administrator's course". Lots of Luck, Fran Tanzella Sr. Chemist, SRI International
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I haven't used the negative reversal procedure in a few years to make sheet film into transparencies. It wasn't cost effective to keep it up and never gave me the quality that I liked.
The method I now use is to use a dye-sublimination printer to write directly to "transparency" film. This produces photographic quality output. Of course this means you need to find someone with a dye-sub printer. Typical cost/print is about $2-$3 for an 8x10 transparency. The printers themselves run about $10K.
If you have a good laser-printer (at least 300 dpi) and a graphics/image processing program that does reasonable dithering (Adobe Photoshop for example) then you can print to "laser printer transparency" film. This give marginally acceptable prints but I wouldn't want to use these unless there was no other alternative. The better laser printers support a "photo-grade" mode (the name varies depending upon the manufacturer) which greatly improves the gray-scale dithering pattern, but it still won't compete with dye-subs (in my opinion).
The very high end printers (~2700+dpi) like the Linotype Imagesetters give results which compare with dye subs. These are typically found in graphic arts departments. If you look with an eye piece you can still see the dither pattern but it's pretty small.
There are also the ink-jet printers which depending upon the graphic do okay jobs. I tend not to use these for continuous tone images but prefer them on solid fill graphics. Mainly due to their lower cost/print ~$1 for an 8x10.
But all of this assumes that you have the images/slides already in some computer and ready for printing. If not then you'll have to go across to your graphic arts department and have them shoot an 8x10 negative and do the reversal for you. Last time I checked the price for this (here at ANL) was about $12/transparency.
This likely doesn't solve your problems but at least you can ask around and see if the printers are handy. I would be surprized if there were none on your campus.
Sorry Subscribers, I'm still having a system problem. You will likely continue to get doubling of some messages that donot clear the queues before a crash happens. I'm looking into the problem.
We are looking for a good home for a fully operational Philips 300G TEM. New HT tank stable operating at 20,40,60,80, and 100 kV. Both biological stage and goneometer stage. Several specimen holders (single-tilt, double-tilt, tilt-rotate). Planning on surplusing soon (Univ. of Wash., Seattle, WA).
Interested parties contact by email: merock-at-u.washington.edu
} I am in search of a US supplier for polyester wax. This product is } produced by a company in the UK. } } Please respond directly to my e-mail address } melsen-at-MICROBIO.emory.edu
Try Polysciences, Warrington, PA or Aldrich Chemical Company, Milwaukee, WI. I haven't used polyester in years, but both stocked it as of 1988. Aldrich sold 5 gal and up quantities and a wide range of mol wts; I had gotten two 1-gal samples from them that lasted me years. (See also my paper - 1987, JEM Technique 6:35-41). --Jon
Jon L. Norenburg {norenbur-at-onyx.si.edu} Invertebrate Zoology, National Museum of Natural History Smithsonian Institution, Washington, DC 20560 Voice 301-238-3508, Fax 301-238-3361
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Has anyone had experience rebuilding an SEM electron gun?
Our Hitachi S-570 SEM has had emission current fluctuations which I traced to the gun cable-gun connection inside the gun. Trouble is, the connections are surrounded by a white, opaque (silicone?) potting compound. When I press down on the potting compound, my emission current troubles disappear, thus I know that the connection is shaky inside.
I'm guessing that I can carefully cut away the old potting compound, expose and repair the cable to gun connections, and finally repot the gun. Not knowing exactly what I am getting into, I thought I would ask for some net wisdom before the first slice.
BTW, the instrument is 9 years old and a new gun and cable assembly runs about $5K from Hitachi. There's my incentive!
Thanks,
Carl
Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2005 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA ----------------------------------------------------- (313) 936-1550 (voice) **** Next time, **** (313) 763-4690 (FAX) *** take the *** chender-at-umich.edu (e-mail) **** train! **** ----} (While you still can!) -----------------------------------------------------
} Does anyone have names and phone numbers for suppliers of standards } for my EDX analysis. I need to know the compositions to see how well } my setup is doing it's work. }
Another source is: Cannon Microprobe 1041 NE 100th St. Seattle, WA 98125 (206) 522-9233 Attn: Bart Cannon
They can make a SEM mount with your choice of standards from a list of hundreds of minerals/materials. The prices are also reasonable.
Disclaimer: I have not purchased the standards, nor do I have any association with Cannon Microprobe. Just passing along some info I recently came across.
Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2005 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA ----------------------------------------------------- (313) 936-1550 (voice) **** Next time, **** (313) 763-4690 (FAX) *** take the *** chender-at-umich.edu (e-mail) **** train! **** -----------------------------------------------------
Thank you for putting forward your comments, so that I may effectively clairify my position.
I strongly feel that University Professional and Scientific staff (P&S) are often overlooked for their contributions to microscopy. Being a part of the University community, I can only speak from that perspective. I do not know how easy or difficult it is for staff from industry or goverment receive support from their employers. I do know that University P & S staff as a rule have very little control over use of grant money for travel, and also have much smaller salaries with which to fund themselves to go to meetings. I do not suggest that MSA pick up the tab for all expenses, but funding from a $10,000 source could go a long way towards improving someone's chances of obtaining matching funds from their employers or other university sources. As an example, there could be twenty-five $400 awards. I may be accused of thinking small, but what I originally had in mind was $2000 total for the program (perhaps five $400 awards) with a simple waiving of registration fees for another 5 people. Why couldn't we try this as apiolet program for one year? If no one applies it hasn't cost MSA a cent.
As Microscopists we cannot afford to be stagnant. There has to be continuing education so that we are able to provide meaningful assistance to our employers, and the many opportunities available at the annual MSA meeting are the best source for this nessecary growth in ourselves.
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When cutting cryo-sections it is necessary to have an anti-roll plate on the knife. The best probe to use to pick up frozen sections, is an eye lash stuck to an applicator stick. An anti static gun may help to reduce the charge build up in the block-section-knife region. If all else fails read the appropriate chapter in "Low Temperature Microscopy and Analysis" by Patrick Echlin, Published by Plenum in 1992.
Good luck with one of the most frustrating activities in cryomicroscopy
Patrick Echlin Director, Multi-Imaging Laboratory School opf Biological Sciences CambridgeOn Mon, 14 Nov -1 kayton-at-ohsu.edu wrote:
} ------------ Original Message ------------- } } } } Very often when I am cryo-sectioning , the sections curl so badly } that they are useless. This was the reason for purchasing the static-line } ionizer. This really did not help the curling problem, although it served } other purposes. My sections are mostly polymer-elastomer blends, primarily } nylon based. If you have any suggestions I would be eternally greatful. Also } if anyone has suggestions on how to pick up the samples off of the cryo } knife that would also help. Right now we are using a very fine sewing } needle, but something even thinner yet ridgid would be better } } } Thank you } } Andrea Monisera. } --------- End of Original Message --------- } } I have seen the eyelash hairs of Dalmatians (dogs) used for this. they are } forked at the end and work very well for picking up cryosections } } } Bob Kayton } OHSU } Portland , Oregon } } E-mail address kayton-at-ohsu.edu }
Nestor writes: * * The method I now use is to use a dye-sublimination printer * to write directly to "transparency" film. This produces photographic * quality output. Of course this means you need to find someone * with a dye-sub printer. Typical cost/print is about $2-$3 for * an 8x10 transparency. The printers themselves run about $10K. * There is a dye-sub printer (PrimeraPro) available from: Fargo Electronics 7901 Flying Cloud Dr. Eden Prairie, MN 55344 (800) 258-2974 which lists for $1895.00 with 8X10 prints costing about $3-$4 each. There is also a 4X5 format available at a little over $1 per print, you just have to switch the ribbon and feed tray. I have tested this printer for wax thermal transfer, dye-sub prints and dye-sub transparencies. Although it may not be the same quality you will find form the $10K printer, it appears to me to give you } 95% of the performance for less than 20% of the price.
Keith A. Kelley Miles Research Center kelley-at-mrc.com
There is a software package written by David Bright called MacLispX which might be what your thinking about. I've never heard of Isaac. MacLispX is available downloadable from the EMMPDL FTP site (WWW.AMC.ANL.GOV) check out the ANL Shareware/Mac Shareware/Imaging/MacLispX area
For information about the ISSAC system, contact Michael Postek at NIST, Gaithersburg. 301-975-2299
John
} } } There is, I think, a software package from NIST called ISAAC. } It is supposed to handle images with more pixels and more bits } than other programs for acquiring SEM images. } 1 Does anyone know where to get more information on this? } 2 Does anyone have experience using it? } } If you want a giggle, read the piece in New Scientist (November 5 1994, } page 21), which is where I heard about it. } } Alwyn Eades } Alwyn Eades } Center for Microanalysis of Materials } University of Illinois
ABSTRACTS FOR THE JOURNAL OF MICROSCOPY, VOLUME 176 PART 2, NOVEMBER 1994. **************************************************************************
Journal of Microscopy, Vol. 176, Pt. 2, November 1994, pp. 85-98.
High performance X-ray detection in a new analytical electron microscope
C E Lyman, J I Goldstein, D B Williams, D W Ackland, S von Harrach, A W Nicholls & P J Statham, Materials Science & Engineering, Lehigh University, Whitaker Laboratory, 5E Packer Avenue, Bethlehem PA 18015-3195, USA
SUMMARY
X-ray detection by energy-dispersive spectrometry in the analytical electron microscope (AEM) is often limited by low collected X-ray intensity (P), modest peak-to-background (P/B) ratios, and limitations on total counting time (t) due to specimen drift and contamination. A new AEM has been designed with maximization of P, P/B and t as the primary considerations. Maximization of P has been accomplished by employing a field- emission electron gun. X-ray detectors with high collection angles, high-speed beam blanking to allow only one photon into the detector at a time, and simultaneous collection from two detectors. P/B has been maximized by reducing extraneous background signals generated at the specimen holder, the polepieces and the detector collimator. The maximum practical t has been increased by reducing specimen contamination and employing electronic drift correction. Performance improvements have been measured using the NIST standard Cr thin film. The 0.3 steradian solid angle of X-ray collection is the highest value available. The beam blanking scheme for X-ray detection provides 3-4 times greater throughput of X-rays at high count rates into a recorded spectrum than normal systems employing pulse-pileup rejection circuits. Simultaneous X-ray collection from two detectors allows the highest X-ray intensity yet recorded to be collected from the NIST Cr thin film. the measured P/B of 6300 is the highest level recorded for an AEM. In addition to collected X-ray intensity (cps/nA) and P/B measured on the standard Cr film, the product of these can be used as a figure- of-merit to evaluate instruments. Estimated minimum mass fraction (MMF) for Cr measured on the standard NIST Cr thin film is also proposed as a figure-of-merit for comparing X-ray detection in AEMs. Determinations here of the MMF of Cr detectable show at least a threefold improvement over previous instruments.
Journal of Microscopy, Vol. 176, Pt. 2, November 1994, pp. 99- 109.
The preparation of cryosections from plant tissue: an alternative method appropriate for secondary ion mass spectrometry studies of nutrient tracers and trace metals
Dennis B Lazof, Jack G Goldstein, Thomas W Rufty, Cynthia Suggs & Richard W Linton, USDA-ARS, Crops Research Laboratory, PO Box 1168, Oxford NC 27565, USA
SUMMARY
A method involving cryostat sectioning (10 micrometre thickness) and freeze-drying is presented for the preparation of plant tissue for microanalytical studies. The method is well suited for semi-quantitative imaging by secondary ion mass spectrometry (SIMS) and offers significant advantages over bulk freeze-dried or freeze-substitution preparations. Segments of corn or soybean root (5mm) are quench-frozen, embedded externally, sectioned in a cryostat (10 micrometre), pressed onto ultrapure Si and slowly freeze-dried. Images of these sections with secondary electron microscopy and SIMS indicated good morphological preservation. It was possible to section tissues of a wide developmental range, as well as roots varying sixfold in diameter. SIMS images are presented which demonstrate the ability to detect and localize nutrient tracers, such as Rb+, following brief exposures (10 min) to the intact plant. Likewise, a toxic metal (Al) was localized in root tissue after brief exposure ( {1 day) of the intact plant root to micromolar external concentrations. Elemental redistribution during processing was minimal, as demonstrated most explicitly by the lack of movement of loosely bound Ca from the outer cell walls into the adjacent embedding material. Preservation of compositional differences between cellular content and cell wall was supported by a semi-quantitative treatment of SIMS images.
Journal of Microscopy, Vol. 176, Pt. 2, November 1994, pp. 110- 120.
Vitrification of aqueous suspensions from a controlled environment: an improved plunging device
B J Battersby, J C W Sharp, R I Webb & G T Barnes, Department of Chemistry, The University of Queensland, Queensland 4072, Australia
SUMMARY
In the process of vitrifying aqueous suspensions for cryo- transmission electron microscopy, water is solidified without crystallization. Vitrification can be achieved by rapidly plunging an aqueous thin film into a liquid cryogen. The preparation of aqueous thin films prior to vitrification must be performed in an environmental cabinet at controlled temperature and humidity in order to prevent evaporation and temperature- induced phase changes in the thin film. The device described here incorporates several important features which make the apparatus simpler and more convenient to use than similar devices described in the literature. One of these features includes the use of a totally enclosed environmental cabinet in which the grid, sample, micropipette and absorbent paper are equilibrated before thin- film preparation. Other features include a cryogen dewar on a swing arm for easy refilling, a guillotine shutter which is used to trigger the plunger electrically and a semi-automatic system which facilitates rapid transfer of the vitrified specimen from liquid propane to liquid nitrogen for storage and reduces handling of the specimen. To demonstrate the utility of the device, results showing the influence of temperature on the morphology of phospholipid vesicles are presented. A commercial cryotransfer apparatus (which is used for the transportation of the vitrified specimen to the electron microscope cold stage) has been modified to reduce the possibility of reversion of the vitreous phase to the crystalline ice phases.
Journal of Microscopy, Vol. 176, Pt. 2, November 1994, pp. 121- 131.
An atomic force microscope for cytological and histological investigations
Tullio Mariani, Antonio Musio, Carlo Frediani, Isabella Sbrana & Cesare Ascoli, Istituto di Biofisica - CNR, Via Lorenzo 26, 56127 Pisa, Italy
SUMMARY
An atomic force microscope (AFM) specifically designed for cytological and histological studies and able to operate on the same scale of the highest optical magnification is described. The AFM is a non-invasive instrument; it operates on samples which do not require any kind of treatment and it can produce information that supplements and completes the information given by traditional microscopical methods. The apparatus has been used to image fixed human chromosomes and to investigate the action of trypsin during the staining for banding. First results showed that banding patterns very similar to G-banding pre-exist to staining and to trypsin treatment in human metaphase chromosomes, and that the trypsin treatment induces a structural collapse in the chromatin. The instrument was also used on thin sections of plant tissue and gave promising results. Experience confirmed that AFM is a suitable tool for this kind of investigation, and proved the importance of developing AFMs specifically designed for routine use in cytology and histology, conceived for non- specialized users.
Journal of Microscopy, Vol. 176, Pt. 2, November 1994, pp. 132- 142.
Unstained and in vivo fluorescently stained bacterial nucleoids and plasmolysis observed by a new specimen preparation method for high power light microscopy of metabolically active cells
Eduard Kellenberger & Cornelia Kellenberger van der Kamp, Institut de Genetique et Biologie, Microbienne (IBGM), Universite de Lausanne, Rue Cesar Roux 19, CH-1005 Lausanne, Switzerland
SUMMARY
Microscope slides were coated with a layer of gelatin, the thickness of the gelatin increasing linearly along the long axis. The bacterial suspension is applied to the dried gelatin and covered by a coverslip. The medium is absorbed by the gelatin and thus the cells applied against the coverslip. By this method, cultures of concentrations below 100 000 000 cells/ml provide statistically relevant numbers for observation without prior concentration steps. It is easier to apply than the existing methods for the observation of bacterial nucleoids by phase contrast imaging. Because the cells are maintained in growing conditions the method is useful for the vital fluorescence DAPI- staining of various bacterial species and for observations of plasmolysis and its reversal at different physiological conditions and extracellular osmolalities. The previously generally assumed view that the plasmolytic changes of the cell morphology are immediate upon the hyperosmotic shock and are rapidly repaired when the cell is able to metabolize actively was confirmed; this is in contrast to some recent claims.
Journal of Microscopy, Vol. 176, Pt. 2, November 1994, pp. 143- 151.
Fibre optic scrambling in light microscopy: a computer simulation and analysis
Frederick B Reitz & Len Pagliaro, Center for Bioengineering WD-12, University of Washington, Seattle, Washington 98195, USA
SUMMARY
Optical fibres bent in two mutually perpendicular planes have proven useful for randomizing illumination in light microscopes. These optical scramblers can increase the resolution and/or contrast obtained with several modes of light microscopy. Here computer simulations are used to investigate several factors affecting light randomization in curved optical fibres in order to further the theoretical basis for scrambler design. Light passing through 90 degree bends of optical fibre of varying radii of curvature was modelled by ray tracing in two dimensions, and scrambling mechanisms were observed. The effects of varying the position and angle of entry of light on the phase and direction of propagation of emergent light were determined. It was found that (a) thorough scrambling does not necessarily require high numerical aperture entry of light into the fibre, (b) considerable order persists after a single 90 degree bend of an idealized fibre and (c) a higher degree of scrambling (at the cost of transmission efficiency) is achieved in more tightly curved fibres. The pathlength variations introduced by scrambling proved smaller than typical laser coherence lengths, requiring temporal scrambling (vibrating the fibre).
Journal of Microscopy, Vol. 176, Pt. 2, November 1994, pp. 152- 157.
Edge detection, three-dimensional cell boundary reconstruction and volume and surface area estimation from differential interference contrast images
C M Kenyon, M Yanai & P T Macklem, Meakins Christie Laboratories, McGill University, Montreal, Canada H2X 2P2
SUMMARY
Three-dimensional (3-D) cell morphology is important for the understanding of cell function and can be quantified in terms of volume and surface area. Differential interference contrast (DIC, or Nomarski) imaging can enable cell edges to be clearly visualized in unstained tissue due to the slight difference in refractive index between aqueous media and cytoplasm. DIC is affected in only one direction - the direction of the optical shear. A 1-D edge detector was used in that direction with a scale length equal to that of an in-focus edge to highlight cell boundaries. By comparison with the signal from the edge detector on an out-of-focus slice, the in-focus slices could be segmented and, after noise suppression, cell outlines obtained. A voxel paradigm was used to calculate cell volume and differential geometry was used for surface area estimation. We applied this approach to obtain 3-D information by optical sectioning of motile Amoeba proteus.
Journal of Microscopy, Vol. 176, Pt. 2, November 1994, pp. 158- 166.
Nuclear diffuseness as a measure of texture: definition and application to the computer-assisted diagnosis of parathyroid adenoma and carcinoma
A J Einstein, J Barba, P D Unger & J Gil, The Lillian & Henry M Stratton -, Hans Popper Department of Pathology, Box 1194, One Gustave L Levy Place, New York NY 10029, USA
SUMMARY
A measure of texture, the nuclear diffuseness, was formulated for use in biological classification, and specifically to characterize quantitatively chromatin texture. Nuclear diffuseness corresponds to the amount of local intensity variation in the digitized image of a nuclear profile. As a setting in which to test the efficacy of nuclear diffuseness as a diagnostic tool, the identification of parathyroid adenoma and carcinoma was considered. Digitized images of sections of parathyroid chief cell nuclei were obtained from 16 biopsies, and the nuclear diffuseness, as well as other morphometric descriptors, were computed. With just the average nuclear diffuseness and average nuclear profile area, jackknife (leave- one-out) classification using an artificial neural network was able to diagnose correctly and unambiguously the condition (normal, parathyroid adenoma, or parathyroid carcinoma) in 15 of 16 cases. In one case, the neural network assigned a higher weight to the correct diagnosis, but was unable to distinguish between normal and adenoma conclusively.
Journal of Microscopy, Vol. 176, Pt. 2, November 1994, pp. 167- 177.
Estimation of the transport properties of polymer composites by geodesic propagation
R Bremond, D Jeulin, P Gateau, J Jarrin & G Serpe, Centre de Geostatistique, Ecole Nationale Superiere des Mines, 35 Rue Saint-Honore, F 77305 Fontainebleau, France
SUMMARY
The purpose of this study is to estimate the transport coefficients (diffusion, permeability) of polymer composites from two-dimensional images of these media (polyethylene/polyamide alloys) obtained by the scanning electron microscope. The main characteristics of the media investigated are a wide morphological variability (nodular and lamellar media) and transport properties that are very different according to the phase. The computation tools used must accordingly have a wide range of validity. The idea of the proposed algorithm is to calculate the distance from an edge of the image, with a modified distance according to the phase crossed. The results obtained are compared with those obtained by finite difference integration of Fick's laws, and with permeability measurements of hydrocarbons in the materials.
Tim Prusnick asked in a post to sci.techniques.microscopy about info on EM. Dear Tim, For technical aspects on how EM's work and as a referrence, I reccom- mend Principles of Electron Optics by P.W. Hawkes and E. Kasper from Academic Press (St Edmundsbury Press Ltd. in the UK). Yours, Bill Tivol
Tony Romeo Internet: tony-at-emu.su.oz.au Electron Microscope Unit Madsen Building F09 Telephone: +61 2 351 2351 The University of Sydney Facsimile: +61 2 552 1967 Sydney, NSW 2006 Australia
Tony Romeo Internet: tony-at-emu.su.oz.au Electron Microscope Unit Madsen Building F09 Telephone: +61 2 351 2351 The University of Sydney Facsimile: +61 2 552 1967 Sydney, NSW 2006 Australia
I would to know if there are any training courses specifically for the preparation of biological materials for examination in a SEM and TEM.
Sorry, I cannot be specific about the type of samples, I just need to know about general techniques (embedding, fixing, staining etc).
Are there any held by the RMS for example?
As I am in materials characterisation centre, the staff (including me!) have little experience of the preparation techniques. In particular hands on training would be a must.
Keith Moulding
Materials Characterisation And Preparation Centre, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Our interests are to descriminate the interdiffusion of methacrylate = resins into mineralized substrates using TEM. At present, we are unable = to distinguish resin interdiffusion depth versus epoxy embedding media = following decalicification. Ideas are to find specific stains for the = resins, or, maybe label the methacrylate with a reactive colloidal gold. = Any suggestion on how to descriminate these phases would be appreciated = very much. Help needed,,,thank you
A member of our organization has been asked to make a presentation related to EM in education. Knowing that this is a topic near to the hearts of many on this list, we thought we might ask for some input.
(1) As part of the presentation, we intend to list some of the principal areas where EM has made important contributions. Specifically, we're trying to identify areas where a discovery of large societal significance owes principally to EM. We have a few ideas, but are looking for others. Suggestions ?
(2) We're looking for the names of people who might have had experience in using EM in educational programs (as opposed to research) at the secondary and/or college levels, or have a strong interest in promoting same.
People with suggestions can contact me directly or via this list-server.
Our interests are to descriminate the interdiffusion of methacrylate resins into mineralized substrates using TEM. At present, we are unable = to distinguish resin interdiffusion depth versus epoxy embedding media = following decalicification. Ideas are to find specific stains for the = resins, or, maybe label the methacrylate with a reactive colloidal gold. = Any suggestion on how to descriminate these phases would be appreciated = very much. Help needed,,,thank you
In article YBerta-at-ms-mail.chemse.gatech.edu writes: } } } We need to replace the gold target on our ISI sputter coater; SPI is out of } the targets right now and Electron Microscopy Sciences doesn't carry targets } for ISI. }
Any competent local gold worker can make you a target much cheaper than the specialist suppliers. Many of Sydneys jewellers have manufacturing capacities and can easily make me a target any specified diameter and thickness. Better still, they will accept an old perforated target and add gold to restore a solid sheet. Gold palladium however is too hard and has too high a melting point for local manufacturers to rework.
We just had a 100 lb tensile stage delivered from Ernest Fullam to fit on our Leica/Cambridge S360. Is there any one out there who has one fitted on an S360? We need some feedback on practical fitting. The main problem is the plate that adapts the drive motor to the left side port has no flat ground on its edge to clear the door hinge. Our workshop can easily grind the flat, but I'd like a clue where the (eccentrically located) motor should be located. Looks as if 12 O'clock to 3 O'clock might suit. Any other feedback on operation will be welcome as the manual is a minimalist document.
An oldi, but very nice article is written by Naziri et al. 1972: In situ superplasticity experiments in the 1 million volt electron microscope. J. Microscopy, 97, 229 - 238
Hopes this helps.
Cheers, Timon
} Hello! } } I wonder if anyone can help me: } } I just got the title of a lecture I need to give in } connection to my Ph.D. 'defence' in 14 days..: } } "In Situ Experiments in the Transmission Electron Microscope" } } I have found some references in the ICEM-94 proceedings } and some late numbers of Ultramicroscopy.... } } -Does anyone have some other references? } } -Are you working with this, I am very interested in getting } some ideas!!! } } -What are the main problems? -What can be studied? } } etc... } } You may respond directly to me or through the news-group! } } I am waiting.. } } Randi. } } email: randih-at-imf.unit.no
------------------------------------------------ Timon Fliervoet, Timonf-at-earth.ruu.nl, Faculty of Earth Sciences, Department of Geology, Utrecht University, P.O. Box 80.021 3508 TA Utrecht, the Netherlands. tel: ++31 30 - 535054, fax: ++31 30 - 537725
Randi, A recent issue of the Materials Research Society bulletin (MRS Bulletin,June 1994,Volume XIX, No.6) is devoted almost entirely to materials science research now being done using in situ electron microscopy.
________________________________________________________________ Tom Fellers Florida State University Center for Materials Research and Technology Tallahassee, FL 32306-4000 (904) 644-6559 fellers-at-phy.fsu.edu
Does anyone know of a video board for the Mac which will allow acquisition of the next video field after an external trigger? The boards I've seen (Scion LG-3 and Neotech) will allow display of a single field, but even or odd has to be specified. What I need is a board which will acquire the next field whether it is the even or odd field. I understand the early image grabber boards did work this way. My problem is that I need to acquire an image which is illuminated by a single srobe flash (on the order of 10 nsec duration). Both Illumination and acquisition are triggered from an external source which is not controlled. I've tried synchronizing the strobe to flash twice (once at each video field), but this results in to much motion artifact. If I grab and display an entire video frame, I get an excessive amount of flicker. If I grab an entire frame but display only one field, about half of the time I display non-illuminated images. If I wait for the next even or odd field, I can have up to a 30 msec delay, which is unacceptable.
Any information about video boards or other solutions to this problem will be appreciated. At this point I'm open to almost any suggestion. Thanks.
Dave DeFily defily-at-tamu.edu Medical Physiology, Texas A&M
There is no room on the microscopy forum for crude jokes of this type. It is, in my opinion, very unprofessional to send jokes of this type across a public forum of this calibre. We should try to keep all information and questions related to microscopy and not to jokes that will offend a lot of individuals. Let's try to act professional from now on! Phil 8-{(
I didnot find the joke posted by Calvert either funny or appropriate. He is now history and has been removed from the listserver. I do not want to start a moderated system because I don't have the time, however I will not hesitate to stomp on things like that.
I also agree that those jokes ARE NOT appropriate for this server. They had absolutely nothing to do with microscopy to say the least.
Peling
-------------------------------------------------------------- Peling Melville peling-at-amnh.org Interdepartmental Laboratories American Museum of Natural History
Several years ago, Leitz was purchased by Cambridge, which in turn was purchased by someone else, etc. The company that includes what was Leitz is now called Leica. An address from 1992 is: Leica Inc., 111 Deer Lake Road, Deerfield, Illinois 60015, Tel: 708-405-0123, Fax: 708-405-0147. The world headquarters for microscopes is (again, from some time ago): Leica Mikroskopie und Systeme GmbH, Ernst Leitz-Strasse, P.O. Box 2040, W 6330 Wetzlar 1, Tel: 64 61-29-0, Fax: 64 41-29-33 99. You can probably find a more convenient source in the Washington D.C. area.
--------------------------------
On Mon, 21 Nov 1994, Mark O. Walderhaug wrote:
} Gentle List Readers, } I have recently been given a Leitz Laborlux 11 Microscope. } It presently only has one objective lens and I'm interested } populating the turret with more objectives. I'm having a } hard time finding any information about Leitz. Am I looking } in the wrong place, or has it been swallowed up by another } manufacturer? } Would some kind person point me in the right direction to } find out about getting objectives for this microscope? } Thank you for you patience and kindness. } } Mark O. Walderhaug voice: 202 205-4682 fax: 202 401-7740 } Microbial Ecology Branch HFS-517 Food and Drug Administration } 200 C St. S.W., Washington, DC 20204 USA BITNET: MOW-at-BFD } Internet: mow-at-vm.cfsan.fda.gov or mow-at-bfd.ssw.dhhs.gov }
On Mon, 21 Nov 1994, Mark O. Walderhaug wrote: } Gentle List Readers, } I have recently been given a Leitz Laborlux 11 Microscope. } It presently only has one objective lens and I'm interested } populating the turret with more objectives. I'm having a } hard time finding any information about Leitz. Am I looking } in the wrong place, or has it been swallowed up by another } manufacturer? } Would some kind person point me in the right direction to } find out about getting objectives for this microscope? } Thank you for you patience and kindness.
Gentle List Poster,
Altho' I am not entirely too kind, I will answer your question... Yes, Leitz has been swallowed up "buy" another. Cambridge Instruments and Wild Leitz "merged" in April of 1990. Their new name, at least in the US, is Leica. You may have luck at, according to the letter they sent me:
Leica Inc. 111 Deer Lake Road Deerfield, IL 60015 [may have changed to 60090??? sorry, don't know...] (708) 405-0123 fax: (708) 405-0147 [or: (708) 405-0030???]
The different info within the [..] are from my American Laboratory (February 1994) Buyers' Guide. If anyone desires to subscribe (free to qualified individuals) to American Laboratory, they can be reached at:
Question: is this Laborlux 11 you received a newer or older model of the Laborlux S [1989 model]? Just curious...
You are quite welcome, and hope this helps you. -Rob ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, | Rob Tayloe | MSM Spelunkers Club /\v/\ | | Metallographic Lab. | Missouri Speleological Survey /\v/\ | | Rolla Research Center | Bat Conservation International /\v/\ | | U.S. Bureau of Mines | Missouri Cave & Karst Conservancy | | tayloe-at-rorc.usbm.gov | National Speleological Society #32993 /\v/\ | | (314) 364-3169 x247 | American Cave Conservation Association | ''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''
Two years ago I was fortunate enough to attend the International Conference on Current Trends in Immunomicroscopy at the George Washington University Medical Center, where Dr.Gwen Childs gave a great talk on In Situ Hybridization. At that time her email address was "GCHILDS-at-UTMBEACH". I'm sure she'd be of help. References at that time were:
Childs et al, Molecular Endocrinology 1:926-932 (1987). Hoeffler et al, Histochem J 18:597-604 (1986). Childs et al, Endocrinology 130:335-344 (1992). " " " " 131: (July,1992)
You might need to read this book: "Dynamic Experiments In The Electron Microscope",E.P.Butler,K.F.Hale; Practical Methods in Electron Microscopy Vol 9 (ed. A.M.Glauert); North-Holland P.C.;1981. In general, in situ experiments could reveal oriented phase transition (topotaxy....) and structural disorder (domain, superstructure, modulation..).
Mr-Received: by mta PETVAX.MUAS; Relayed; Tue, 22 Nov 1994 07:15:55 -0400 Mr-Received: by mta PETVAX; Relayed; Tue, 22 Nov 1994 07:15:56 -0400 Mr-Received: by mta SRVR01; Relayed; Tue, 22 Nov 1994 07:16:57 -0400 Disclose-Recipients: prohibited
} Perhaps Mr. Calvert should have spent more time and energy composing his } "real message". } } Perhaps Mr. Calvert simply typed the wrong address for this posting.
Another option might be that he left his machine unattended and it wasn't him at all. Applications such as Eudora do not require a password to _send_ messages (well, it doesn't on our system, anyway) and, theoretically, _anyone_ could send messages purporting to be me (or anyone else).
Thankfully, we don't seem to have anyone that puerile here :-)
If it was indeed an attempt at humour, then it is to be condemned, but the whole thing seems so _unlikely_ that I, at least, am not jumping to hasty conclusions.
Anyway, may as well add another tuppence worth while I'm here...
Concerning in-situ experiments: One suggestion might be to contact Gatan (say) for a brochure of available TEM stages. This may provide a range of heating, cooling, straining, whatever stages with suggested and actual experiments. There are innumerable heating/cooling/straining studies out there.
Slightly more off the beaten track these days - considerable radiation damage research has been performed in-situ: One of the most elegant (and difficult!) being by TJ Black, ML Jenkins, MA Kirk (and possibly IM Robertson, CA English ?) in the early 1980's in which disordered zones created by cascade collapse during ion bombardment were imaged in weak(ish)-beam dark-field using a superlattice reflection of initially-ordered Cu3Au in-situ in the Argonne Tandem/HVEM. This was done using a liquid He stage.
Ahh.. found these refs.. TI: COLLAPSE OF DEFECT CASCADES TO DISLOCATION LOOPS. AU: Kirk_MA Robertson_IM Jenkins_ML English_CA Black_TJ Vetrano_JS JN: Journal of Nuclear Materials 1987 Vol.149 No.1 pp.21-28
TI: DISPLACEMENT CASCADE COLLAPSE IN Cu//3Au AT LOW TEMPERATURES. AU: Black_TJ Jenkins_ML Kirk_MA JN: Institute of Physics Conference Series 1984 No.68 pp.343-346
There will be earlier ones, but BIDS doesn't go back before 1983 and I don't have the relevent file to hand. See the above for refs. I seem to remember there was a Proc. Roy. Soc. article on this (?), and it wouldn't surprise me if there was something in Phil. Mag. A as well...
Hope this helps - you certainly have a large field to cover/select from!
-------------------------------------------------------------------------- Dr. S.L. King Dept. Electronics and Electrical Engineering, University College London Torrington Place, London WC1E 7JE England
We are attempting to remove the aluminum disk from beneath the magnetic layers on a hard disk. Given the problems with polishing aluminum mechanically, the decision was made to try chemical methods first. I realize that there are plenty of sources on chemical thinning and electropolishing, but what I'm looking for is good advice on what will work readily to eliminate 100 microns to 1 mm of alloyed aluminum and leave the NiP and higher layers alone. We have tried heated gallium with no great success (perhaps too low of Al solubility for our mass). Any assistance would be appreciated.
Daniel L. Callahan Department of Mechanical Engg. and Materials Science Rice University dlc-at-owlnet.rice.edu
Help! We just bought a Sony UPD-7000 dye-sub printer for our computer facility, and don't have the printer driver for the Mac. Does anyone have it and can share, if it's not illegal, or have any suggestions as to where to get it? I've tried all the Sony reps I can find, to no avail. We have the printer, so I think we would have the rights to the driver as well. I'm interested in either a Chooser document or a Photoshop export module. thanks
Judging from the answers posted, it is not clear what type of in situ experiments are in question. I personally posted a book title on immuno-in situ directly to the user asking question. Please be specific when theme is ambigous. Never assume that only users of your own background are reading the message! Thanks.
************************************************************ *Cesar D. Fermin, Ph.D \|*|/ Fax (504) 587-7389 * *Tulane Medical School /|*|\ Answ. Mach.(504) 584-261 * *Pathology/SL79 \|*|/ Secretary (504) 584-2436 * *New Orleans, La 70 112 /|*|\ Lab (504) 5841 * *Fermin-at-TMC.Tulane.edu -} Director of Morphological Services* ************************************************************
I am looking for some advice on methodology to use to study the internal structure of a paperboard laminate which will undergo edge wick, permeation experiments. In this experiment, four fresh cuts are made on a square sheet of a paper laminate which is then placed into an aqueous bath. Theoretically, the fluid should be equally absorbed by the laminate assuming its fiber matrix is randomly oriented -- this does not seem to be the case with one of our samples. We must identify why this one sample performs differntly.
We will use microtomy to prepare cross-sections of this laminate and our FEG SEM to evaluate its internal structural orientation. From my experience working with paper samples, I know the visual artifacts may be subtle without the help of a staining system. Does anyone have any recommendation of a (heavy metal/or other type-cellulosic) stain which could be used to track the degree of permeation either optically or with the the backscattered electron detector on our SEM?
Any other thoughts about evualting the morphology of our laminate would be greatly appreciated.
Happy Holidays,
Dr. Lisa Detter-Hoskin Sr. Research Scientist Materials Analysis Center Georgia Tech Research Institute Atlanta, GA 30332-0827
Message-Id: {m0rA1ZK-0007KIC-at-stjohns.ohsu.edu} Message-Version: 2 } To: Microscopy-at-AAEM.AMC.ANL.GOV
Has anyone else out there had a problem with little bubbles showing up on their 0.5 to 1 micron sections from Spurrs blocks? These are very small, and only appear after I coverslip my slide. The most frustrating characteristic is that the bubbles are not consistantly present. Suggestions are appreciated.
Dear Daniel, Since aluminum is complexed by OH-, a mild base may do the job. I'm not sure what is alloyed with the aluminum on a hard disk, but maybe it could be complexed as well--if it is magnesium, a phosphate will probably do the job. Good luck. Yours, Bill Tivol
We also have been having major problems with holes appearing in our ultrathin sections of Spurr resin embedded specimens (nematodes). We're not sure we've solved this yet but the problem may be due to the resin dissolving material from some plastic containers or if the blocks are polymersied uncovered some of the components volatalise off, resulting in the holes. Has anyone any other suggestions?
____________________ David Wharton Department of Zoology P.O. Box 56 Dunedin Tel (064) (03) 479 7963 Fax (064) (03) 479 7584
I am looking for some advice on methodology to use to study the internal structure of a paperboard laminate which will undergo edge wick, permeation experiments. In this experiment, four fresh cuts are made on a square sheet of a paper laminate which is then placed into an aqueous bath. Theoretically, the fluid should be equally absorbed by the laminate assuming its fiber matrix is randomly oriented -- this does not seem to be the case with one of our samples. We must identify why this one sample performs differntly.
We will use microtomy to prepare cross-sections of this laminate and our FEG SEM to evaluate its internal structural orientation. From my experience working with paper samples, I know the visual artifacts may be subtle without the help of a staining system. Does anyone have any recommendation of a (heavy metal/or other type-cellulosic) stain which could be used to track the degree of permeation either optically or with the the backscattered electron detector on our SEM?
Any other thoughts about evaluating the morphology of our laminate would be greatly appreciated.
Happy Holidays,
Dr. Lisa Detter-Hoskin Sr. Research Scientist Materials Analysis Center Georgia Tech Research Institute Atlanta, GA 30332-0827
} Judging from the answers posted, it is not clear what type of in situ } experiments are in question. I personally posted a book title on immuno-in } situ directly to the user asking question. Please be specific when theme is } ambigous. Never assume that only users of your own background are reading the } message! Thanks.
Agreed, when I first saw the words in-situ I thought of hot stage experiments or on-line inspection. Experiments typical in the materials testing arena or even materials processing. Please provide more detail regarding in-situ experiments, it is a very broad term. . .
P. Joyce
Peter J. Joyce Graduate Research Assistant - Materials Science & Engineering University of Texas at Austin (512) 471-5723
From the other post in this thread, I thought the problem was coverslip media bubbles. I must not have read it carefully enough. My best luck for infiltrating specimens has been to leave the specimens in a 50%ETOH/50% Spurrs mixture overnite. The next day, I change to 100% resin, about four times in eight hours. It doesn't seem to matter how they are polymerized, in Beem capsules that are open or closed. Randy Nessler rnessler-at-emiris.iaf.uiowa.edu
On Wed, 23 Nov 1994, David Wharton wrote:
} We also have been having major problems with holes appearing in our } ultrathin sections of Spurr resin embedded specimens (nematodes). We're not } sure we've solved this yet but the problem may be due to the resin } dissolving material from some plastic containers or if the blocks are } polymersied uncovered some of the components volatalise off, resulting in } the holes. Has anyone any other suggestions? } } } ____________________ } David Wharton } Department of Zoology } P.O. Box 56 } Dunedin } Tel (064) (03) 479 7963 } Fax (064) (03) 479 7584 }
Reply... RE} Coating screens Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX: (313)936-3352 jfmjfm-at-umich.edu or John_Mansfield-at-mse.engin.umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
This response is a little late, but we send our screens to the same people that JEOL do (or at least did). I believe the company is: Grant Scientific Corporation Electron Microscopy Products Division 1385 Rock Island Road Gilbert, South Carolina 29054 803-892-2841 Contact: Dunkelberger, Dana We have had good luck with them and get our 2000, 4000, and EM420 screens recoated there. Hope this helps.
} On Tue Nov 22 12:12:01 PST 1994, Robert Kayton,MAC,CROET wrote: } } } Has anyone else out there had a problem with little bubbles showing up on } } their 0.5 to 1 micron sections from Spurrs blocks? These are very small, = and } } only appear after I coverslip my slide. The most frustrating characterist= ic } } is that the bubbles are not consistantly present. =7FSuggestions are } } appreciated. } }
If there are bubbles in the resin from mixing (likely) it is also possible to remove them by centrifuging. This avoids the problems of placing uncured resin in a vacuum system (messy and the hardener generally is more volatile and you may pump sufficient hardener off to prevent full curing).
Norman Elliott | E-mail: nee-at-lanl.gov Los Alamos National Lab | Fax: 505-665-2104 MST-7 MS E549 | Voice: 505-667-1587 Los Alamos, NM 87545 |
-Thank you all for answers conserning in-situ experiments!
I have my background in materials science, and I guess that was what my committee thought about when giving me the title 'in situ experiments in the transmission electron microscope', but it has been interesting to see that 'in situ' can mean different things (hybridisation) as I also saw in my litterature seach.. I need to mention something about this..
It was exactly the point to find out what people thought about when seeing the title above.. (I am free to say whatever I want under the given title, and need to pick out something..)
Due to heavy construction work adjacent to our EM facility, that is expected last for the next three years, I am looking into the possibility of installing vibration isolation platforms for 2 or 3 of our instruments. I am aware of TMC (Technical Manufacturing Corporation, Peabody, MA) that manufactures such platforms. Are there other manufacturers of vibration isolation platforms for EMs? Also, I would be grateful to hear comments from persons who have installed the plaforms in their facilities. Thanks!
My friend and I are trying to do simple indexing of some TEM electron diffraction patterns we obtained from an aluminum alloy. On asking around, we were refered to this group for info on NIHimage software. We are graduate students of physical metallurgy. So, if any good soul out here knows how we can get NIHimage or any other user-friendly and affordable software to help us complete our work, we shall appreciate the assistance.
} We also have been having major problems with holes appearing in our } ultrathin sections of Spurr resin embedded specimens (nematodes). We're not } sure we've solved this yet but the problem may be due to the resin } dissolving material from some plastic containers or if the blocks are } polymersied uncovered some of the components volatalise off, resulting in } the holes. Has anyone any other suggestions? } } } ____________________ } David Wharton } Department of Zoology } P.O. Box 56 } Dunedin } Tel (064) (03) 479 7963 } Fax (064) (03) 479 7584 } Dave: have you tried vacuum infiltration with Spurrs? On difficult to infiltrate specimens, I leave the specimen in 3:1(Spurrs:Propylene Oxide) overnight under vacuum then I leave it in pure resin under vacuum until I get ready to embed it that afternoon. I then cure it in a vacuum oven for 8 hours(I use receipe A in the Spurrs handout for the resin). Hope this helps! Phil
PS: I leave the caps off of my specimen bottles when I leave them in 3:1 overnight.
I am sorry to hear that some people have had trouble with delays in filling their orders for my book 'Vacuum Methods in Electron Microscopy'. I have checked with the publisher, Portland Press, and have been assured that orders placed with the following should be handled promptly: 1. In the U.S.A. and Canada: Portland Press, Ashgate Publishing Co., Old Post Road, Brookfield, VT 05036-9704 Ph: 802-276-3162 Fx: 802-276-3837 2. In the UK and Europe: Portland Press, Commerce Way, Colchester, CO2-8HP, UK Ph: 0206-796351; Fx: 0206-799331 3. In Japan: OBK Marketing Services, T's Building 3F, 1-38-11 Matsubara, Setagaya-ku, Tokyo 156 Ph: 03-5300-1658 Fx: 03-5300-1615
I will be very much interested in having comments on the book from any of you that buy it, to see how well you think I have succeeded in presenting the information electron microscopists need about their vacuum systems. Wilbur C. Bigelow
Back in the 1960s Professor L. O. Brockway and several of his graduate students carried out a series of rather elegant experiments in which they recorded the nucleation and growth of oxide particles on single-crystal thin films of several metals in a heated gas reaction stage inside a JEOL JEM-6A TEM using a cine camera. Here are references to some of their papers: J. Electrochem. Soc. Vol 121, No. 11, Nov. 1974, p. 1534 (1974); ibid, vol. 119, p.899 (1972); "Proc. Symp. Funds. of Gas-Surface Interact", p. 147, Academic Press,(1967); J. Appl. Phys. Vol 37, p. 2703 (1966); ibid, Vol 34, p.921 (1963); "Single Crystal Films" M. H. Francombe & H. Sato, Eds., Macmillan (1964). Good luck with your talk - hope you pass your exam successfully.
Regarding your problem of tiny bubbles in Spurrs.....
You dont say if the bubbles are all over, or only in the blank resin or the tissue....
- the mixing process can introduce bubbles: After mixing the resin, centrifuge it - try 5-10 min at high speed on a table top centifuge using 15 or 50 ml conical tubes, depending on the volume of resin you are preparing
- If the problem is only within the tissue, try infiltrating under vacuum
Studies by transmission electron microscopy certainly made a unique, and extremely important, contribution to elucidating the microstructural changes that occur during the heat treatment of steels. This was a matter of fundamental importance in the field of physical metallurgy, because it served as a foundation for understanding metallurgical heat treating processes in general, and lead to the improvement of these processes and the production of improved metal and alloy products in many instances. Much of this work was carried out by Subcommittee XI, of Committee E4, of the ASTM in the early 1950s, and is summarized by a report by Bill Grube which was published in "Revue Universelle des Mines", Series 9, Vol. XII, No. 10 (1956) and in the "Proc. of the ASTM" Vol. 56 (1956). I have a copy of the first of these articles that I can loan you, if you are interested in seeing it. Wilbur C. Bigelow (bigelow-at-umich.edu)
In response to the apology offered to the listserver by Dave. Apology accepted, at least by me.
Bummer it had to happen to you, but these sort of fopas are a part of the system. And we all need to get used to the limitations of this new form of communication. We all need help SPELLING, and we all say things which may be offensive (at least to some group in our society), and now that our communication is being recorded and disseminated via the information superhighway we better brace ourselves for some high speed cyber-collisions. After all how many of us know enough about this INTERNET system to be confident that Dave was the person who sent the jokes? there are a lot of hidden doors in the NET.
Let's all be a little more understanding of our human counterparts, someday we may ask for the same.
Last night there was a news story on hackers changing a doctors medical records, several PAP smear tests were reversed via the INTERNET, some kind of practical joke (not so funny to those patients "misdiagnosed").
} Due to heavy construction work adjacent to our EM facility, that is } expected last for the next three years, I am looking into the possibility of } installing vibration isolation platforms for 2 or 3 of our instruments. I am } aware of TMC (Technical Manufacturing Corporation, Peabody, MA) that } manufactures such platforms. Are there other manufacturers of vibration } isolation platforms for EMs? Also, I would be grateful to hear comments from } persons who have installed the plaforms in their facilities. Thanks! } M.V.: At one time I had 2 AEI 801 E.M.s and the department decided to redo the department from the ground floor(where I was) up. I put rubber pads with a hardness of 80 durometers under the scopes and had no problem with vibration. They had to jackhammer a pit in a room next to the microscope and I still could use the scope while they worked. Of course, I did this only if it was a diagnostic path. case. For routine research using the EM I used the scope whenever they weren't digging. The noise would drive you batty. So, I found other uses for my time. This may help or you may have a condition where you have to have these platforms. I try the least expensive (the pads were 4"x4" and cost me at the time less than $10.00) way to do things like this first. Good luck! Phil 8-{)
Hi, sorry about the blank post: fat fingers. Here's the real one.
Greetings! =09I am teaching a TEM course this Fall and am having considerable=20 difficulty with two areas:1) the students are finding it very hard to=20 stigmate the objective lens properly, and 2) we are having very poor=20 success with a negative staining protocol using bacteria and T-even=20 bacteriophage. I=D5m hoping that those of you who have taught others to=20 properly align a TEM might send me input on their stigmation=20 technique/instruction. We are using a JEOL JEM 100S, stigmating at 50K=20 (the mini-lens assembly is defective, preventing stigmation at higher=20 mags). The resulting negatives clearly show that the astigmatism has not= =20 been corrected. I have worked individually with the three students and,=20 while I seem to be able to stigmate the lens correctly, they remain very=20 frustrated by their inability to do so. I now wonder if there are other=20 (better) teaching techniques that I might try. In brief, I stigmate the=20 lens using a holey grid with the beam spread to obtain the most coherent=20 illumination, working at the interface between focus and just=20 over-focus. I find the black fringe inside the hole easier to=20 visualize. Once I find the point of focus, I bring the image to just=20 over-focus and determine which of the two (X,Y) stigmators seems to have=20 the greater effect. Then, one at a time, I adjust the stigmators to=20 yield an even fringe within the hole, as I shift back and forth from=20 focus to just over-focus. I encourage the students to be patient and=20 take their time, and to work with the room and panel lights off, so that=20 the image can be viewed distinctly. Any and all comments will be=20 appreciated. =09I won=D5t go into detail regarding the negative staining protocol,=20 as I=D5m not sure that it=D5s necessary or relevant to many of the readers.= =20 However, our problem seems to be that we are not obtaining the negative=20 image that one expects (with either 2% PTA, pH 7, or 2% UA in Bacitracin,= =20 filtered). We have tried freshly prepared solutions, new bacterial=20 cultures, variations in application of the stains (drop vs. flotation)=20 and still seem to have little success. Anyone out there using these or=20 similar techniques on bacteria or virus? Again, I=D5d appreciate comments= =20 from all who have insight or experience with these techniques. I have=20 read the appropriate texts and articles and can=D5t seem to pin down the=20 likely source of our problems. =09Finally, I have been given an Ilford Ilfoprint Mark II Super 12=20 wet print processor and now would like to find a few parts for it. =20 Specifically, chemicals (activator and stabilizer) and the two bottles=20 that function as solution reservoirs. Can=D5t afford to buy the dry to dry= =20 version. Anybody able to point me to a source? Kodak Polycontrast paper= =20 is used for the majority of the prints from our darkroom. =09Thanks for your time and assistance! =09=09Dwight
Dwight Beebe IRBV, Dept. de sciences biologiques=09=09beebed-at-ere.umontreal.ca Universite de Montreal=09=09=09=09Voice:514-872-4563 4101, rue Sherbrooke est=09=09=09FAX:514-872-9406 Montreal, PQ H1X 2B2 Canada
} grammatical, and word-omission errors. Not to beat a man while he is } down, but there is no reason not to take advantage of one of the greatest } benefits of writing in a text editor: the chance to view one's work, } read it over, and modify it if necessary for clarity. This is especially } true in a public forum.
Don't forget that while some people might be writing brilliant prose on Windows or Mac machines beautifully ethernetted to the World, others (like me) are struggling with ancient Unix editors (vi, pico) hoping to get a message finished and sent off before the software decides to insert spurious characters or the modem throws a wobbly, kicking us off line, or someone else in the office picks up the 'phone, inserting a series of odd characters which might, or might not, mean something to the computer, including, perhaps, even posting something. I hope no-one judges the contents of the message purely on its presentation - very much a symptom of the eighties and nineties (at least in this country, it seems).
this message is for an annoucement to microscopists in materials sciences, concerning a set of programs, "SHRLI-SIMPLY" ((c) T. EPICIER and M.A. O'KEEFE), running on PCs, and which is available on FTP.UNIV-LYON1.FR, in PUB/DOS/HRTEM (usual login - anonymous - and binary transfer). As can be expected from its title, this package is running SHRLI ((c) M.A. O'Keefe, 1980), which allows HRTEM simulations to be conducted (multislice method, 128x128 FFT). But is allows more than this : building cells /supercells (crystalline or 'non-crystalline' nature), drawing/modifiying structures, calculating 'geometrical' reciprocal lattice sections and/or dynamical diffraction patterns, helping the user to index S.A.D. patterns, plotting Pendellosung curves, calculating CTF, numerical diffractograms (including astigmatism defect), comparing calculated and digitized images,.. Although the programs are running under DOS, a graphical user-interface makes any operation (calculation, screen display, files selections, print,...) very easy. This version of S-SIMPLY is freeware, and has limitations ; however, all options are correctly (hopefully) working. Further information is available in the README.TXT and INSTALL.BAT files, accompanying the SIMPLY1.ZIP and SIMPLY2.ZIP files. Obviously, feel free to contact me for any comments and/or criticisms regarding these programs....
P.S. recent publications illustrating (in a very limited way) the use of S-SIMPLY are : - "Superlubricity of molybdenum disulphide", J.M. MARTIN et al, Phys. Rev. B, 48, 14, 10 583 - "Quantitative HREM analysis of potassium incorporation into cordierite", T. EPICIER et al, Proceed. ICEM 13 (Editions de Physique, Paris), 1994, p. 391 - "Benefits of HREM for the study of metal-ceramic interfaces", T.E. and C. ESNOUF, J. Phys. III France, 4, (1994), 1811. ______________________________________________
Thierry EPICIER GEMPPM-502, INSA de Lyon, 69621 VILLEURBANNE, France tel : (33) 72 43 84 94 (83 85) FAX : (33) 72 43 85 28 Email : EPICIER-at-CISMSUN.UNIV-LYON1.FR
this message is for an annoucement to microscopists in materials sciences, concerning a set of programs, "SHRLI-SIMPLY" ((c) T. EPICIER and M.A. O'KEEFE), running on PCs, and which is available on FTP.UNIV-LYON1.FR, in PUB/DOS/HRTEM (usual login - anonymous - and binary transfer). As can be expected from its title, this package is running SHRLI ((c) M.A. O'Keefe, 1980), which allows HRTEM simulations to be conducted (multislice method, 128x128 FFT). But is allows more than this : building cells /supercells (crystalline or 'non-crystalline' nature), drawing/modifiying structures, calculating 'geometrical' reciprocal lattice sections and/or dynamical diffraction patterns, helping the user to index S.A.D. patterns, plotting Pendellosung curves, calculating CTF, numerical diffractograms (including astigmatism defect), comparing calculated and digitized images,.. Although the programs are running under DOS, a graphical user-interface makes any operation (calculation, screen display, files selections, print,...) very easy. This version of S-SIMPLY is freeware, and has limitations ; however, all options are correctly (hopefully) working. Further information is available in the README.TXT and INSTALL.BAT files, accompanying the SIMPLY1.ZIP and SIMPLY2.ZIP files. Obviously, feel free to contact me for any comments and/or criticisms regarding these programs....
P.S. recent publications illustrating (in a very limited way) the use of S-SIMPLY are : - "Superlubricity of molybdenum disulphide", J.M. MARTIN et al, Phys. Rev. B, 48, 14, 10 583 - "Quantitative HREM analysis of potassium incorporation into cordierite", T. EPICIER et al, Proceed. ICEM 13 (Editions de Physique, Paris), 1994, p. 391 - "Benefits of HREM for the study of metal-ceramic interfaces", T.E. and C. ESNOUF, J. Phys. III France, 4, (1994), 1811. ______________________________________________
Thierry EPICIER GEMPPM-502, INSA de Lyon, 69621 VILLEURBANNE, France tel : (33) 72 43 84 94 (83 85) FAX : (33) 72 43 85 28 Email : EPICIER-at-CISMSUN.UNIV-LYON1.FR
this message is for an annoucement to microscopists in materials sciences, concerning a set of programs, "SHRLI-SIMPLY" ((c) T. EPICIER and M.A. O'KEEFE), running on PCs, and which is available on FTP.UNIV-LYON1.FR, in PUB/DOS/HRTEM (usual login - anonymous - and binary transfer). As can be expected from its title, this package is running SHRLI ((c) M.A. O'Keefe, 1980), which allows HRTEM simulations to be conducted (multislice method, 128x128 FFT). But is allows more than this : building cells /supercells (crystalline or 'non-crystalline' nature), drawing/modifiying structures, calculating 'geometrical' reciprocal lattice sections and/or dynamical diffraction patterns, helping the user to index S.A.D. patterns, plotting Pendellosung curves, calculating CTF, numerical diffractograms (including astigmatism defect), comparing calculated and digitized images,.. Although the programs are running under DOS, a graphical user-interface makes any operation (calculation, screen display, files selections, print,...) very easy. This version of S-SIMPLY is freeware, and has limitations ; however, all options are correctly (hopefully) working. Further information is available in the README.TXT and INSTALL.BAT files, accompanying the SIMPLY1.ZIP and SIMPLY2.ZIP files. Obviously, feel free to contact me for any comments and/or criticisms regarding these programs....
P.S. recent publications illustrating (in a very limited way) the use of S-SIMPLY are : - "Superlubricity of molybdenum disulphide", J.M. MARTIN et al, Phys. Rev. B, 48, 14, 10 583 - "Quantitative HREM analysis of potassium incorporation into cordierite", T. EPICIER et al, Proceed. ICEM 13 (Editions de Physique, Paris), 1994, p. 391 - "Benefits of HREM for the study of metal-ceramic interfaces", T.E. and C. ESNOUF, J. Phys. III France, 4, (1994), 1811. ______________________________________________
Thierry EPICIER GEMPPM-502, INSA de Lyon, 69621 VILLEURBANNE, France tel : (33) 72 43 84 94 (83 85) FAX : (33) 72 43 85 28 Email : EPICIER-at-CISMSUN.UNIV-LYON1.FR
} In response to Dave Calvert's apology, I would to add my two cents about a } common problem in electronic correspondence: lack of editing...
Also, many mail programs have cancellation commands which, if used soon enough (e.g. before the letter gets out of the spool), will allow you to kill the thing before it gets out the door. This won't save you from "next morning regrets," but it can help you out of that feeling you get when you hear the door slam and you realize you've just left your keys in the car...
A comment was recently posted regarding the appropriatness of the listserver as an avenue for a "fast literature review". I just wish to put in my two cents worth on the subject. I think first of all, a few facts about myself are appropriate. I'm 31 years old, and recently received my Doctoate in Germany (6 months ago, to be exact). I am also a real fan of doing proper literature reviews, and when somebody approaches me with a problem, and for one reason or another I know they haven't done a thorough literature review, I get quite upset. This said, I see absolutely nothing wrong with using the listserver as a means of obtaining information for such an exercise. After all, I think the main point behind these comprehensive type examinations is to teach the student how to make best use of his or her time to solve a specific problem on short notice. Therefore, I think the use of the listserver is a very clever idea. It also has obviously brought whole new insights into in-situ experiments. As a materials scientist, I would never have come to the idea that such experiments can also play a significant role in biological research, as has now been pointed out to me. Furthermore, I believe that most scientists, as a result of the changing times, are finding themselves involved in many different projects (upon completion of their university theses). I think very few of us, if any, have the time to do complete literature reviews on all of them, even though all of us would dearly love to such. A number of years ago, this situation might have been different (I admit, I can't say for certain), when scientists at laboratories perhaps had only one or two major projects to work on.
I think it would be very interesting to hear what other scientists with more experience think about this. I would also like to kindly ask Nestor to step in if he believes this is inappropriate for the listserver.
I'm afraid my recent message about "SHRLI-SIMPLY" was sent 3 or 4 times.... My apologize for that, it was not a marketing strategy !!! ______________________________________________
Thierry EPICIER GEMPPM-502, INSA de Lyon, 69621 VILLEURBANNE, France tel : (33) 72 43 84 94 (83 85) FAX : (33) 72 43 85 28 Email : EPICIER-at-CISMSUN.UNIV-LYON1.FR
Message-ID: {MAILQUEUE-101.941126114645.320-at-eye1.eye.ufl.edu} To: microscopy-at-aaem.amc.anl.gov
Dear Phil: We process retinas for histology, TEM, and SEM: frog, chicken, mouse, rat, rabbit, pig, cow and human. We alter our processing techniques slightly for the species and size, and for what we want to end up with (immunolabelling, EDS, straight morphology, etc.). You are welcome to contact me at: eclausnz-at-eye1.eye.ufl.edu Evelyn Clausnitzer U of Florida Ophthalmolgy
Message-ID: {MAILQUEUE-101.941126115206.352-at-eye1.eye.ufl.edu} To: microscopy-at-aaem.amc.anl.gov
Does anyone have a good procedure for the fixation, processing and mounting of whole mammalian retinas? TIA Phil
Dear Phil: We process retinas of several species and for a variety of purposes. Please contact me at eclausnz-at-eye1.eye.ufl.edu for details. Evelyn Clausnitzer U of Florida Ophthalmolgy
I'm looking for anyone that has parts for a Philips 500 series SEM system. I'm in the process of modifying the secondary electron detector system and am looking for as much as possible of a second SEI detector system (grid, scintillator, pmt...) to save the cost of building a complete new detector. If you've got any of these items and no longer need them can you please touch base with me off-line.
} Dwight Beebe } IRBV, Dept. de sciences biologiques=09=09beebed-at-ere.umontreal.ca } Universite de Montreal=09=09=09=09Voice:514-872-4563 } 4101, rue Sherbrooke est=09=09=09FAX:514-872-9406 } Montreal, PQ H1X 2B2 Canada
Hi Dwight. We have a lot of experience in teaching students both microscopy and preparation. First can I suggest you think what is most important for the class you are taking. Virus morphology, preparation or microscope operation? For our virology class, we make a very heavy concentration of T4 phage with a sucrose density gradient centifugation so they deal with a suspension known to have ample virions. (And we check it before hand). We use 1 p.c PTA with bacitracin. I use parlodion filmed grids coated with carbon, then exposed to ion discharge within a few hours of use to guarantee wettability. The students apply the phage suspension to the filmed grids and allow adsorption for say 5 minutes. Then they wash away the sucrose by touching the grid on three drops of distilled water ( 1% sodium acetate if you want to add some ions). Then they add a drop of PTA. For best negative staining they should blot off the stain almost at once as with prolonged contact you get some positive staining as well.
We let each student put in one of their grids and look around. We try to let them experience some of the excitement of seeing something they don't understand first, then understand it by say turning up the magnification or finding a virion. They get prompted with questions like "what do you see? describe it to us? what do you think it is? etc. They take a picture (or two) each. I don't bother getting them to do astigmatism as its only a 4 hour class (all the microscopy you can learn in 4 hours!).
I only teach astigmatism correction when the students are intended to become independent operators. This occurs in one on one coaching in microscope use. I TELL students about astigmatism in introductory lectures but I dont expect them to absorb much except the name and that its an ever attendant problem while focussing. In fact I introduce the stigmators as being additional focus controls for two cylindrical lenses to correct defects in the main lens. I use the model of human astigmatism being corrected with additional lenses.
I gave away using holes for correction about 25 years ago. Even moving from one hole to the next on a grid can change the environment enough to introduce a little astigmatism and I assert that changing grids is not acceptable. Putting in a separate grid was all right when you moved the aperture by thumping the column with a rubber hammer, but not now.
I teach astigmatism correction by observing the structure of the phase granularity you can see near focus. For REALLY GROSS astigmatism (most often introduced by someone giving the knobs a good old wind) I use a bit of the edge of the grid and correct on the fringes there. A TV system on the column is great for demonstrating these effects.
For phase granularity correction; wind the objective focus to and fro and watch the background structure. If it goes from streaks one way to streaks the other, there is astigmatism. Find the objective focus that gives the "best" focus. Use ONE astigmatism control like a focus control; wind it to and fro and find the "best" focus. Repeat with the other astigmatism control. Repeat the whole cycle, starting with the objective until the background structure no longer makes streaks, but makes circles that vary in size as the focus is changed. With experience you can make nearly perfect correction and even recognise a required amount of defocus. At real focus the granularity becomes vanishingly small (phase contrast at a minimum) and everything looks a little fuzzy.
Please send feed back on how useful you find these ideas.
all the best in casting microscopic pearls of wisdom
Practice makes perfect, that is the only recommendation I have for astigmating the microscope. In my many years, I have always prefered using a 1% ammonium molybdate solution for negative staining. It is not a dense as PTA or UA but it seems to be a better "overall" stain. What are your samples in? If in a sucrose gradient (for virus) the sucrose interfers with the negative staining, also are your bacteria in a growth media? After adhereing the samples to a carbon-coated grid, try a brief rinse in dH2O, than stain for 10-15 sec. If lack of sample on grids is the problem, try using a glow discharged grid. No help for you with the print processor parts.
Forwarded message: } From dast-at-msc.cornell.edu Wed Nov 23 18:56:00 1994 } Reply-To: dast-at-msc.cornell.edu } From: dast-at-msc.cornell.edu } Message-Id: {199411232355.AA11766-at-jordi.msc.cornell.edu} } X-Originated-From: jordi.msc.cornell.edu } X-Msc-Version: IDA Client - Sparc } Subject: Re: InGap on GaAs (fwd) } To: hunt-at-msc.cornell.edu } Date: Wed, 23 Nov 1994 18:55:49 +0000 (GMT) } Cc: dieguez-at-iris1.fae.ub.es } In-Reply-To: {199411231425.AA25253-at-borg.msc.cornell.edu} from "hunt-at-msc.cornell.edu" at Nov 23, 94 09:25:08 am } X-Mailer: ELM [version 2.4 PL21] } Mime-Version: 1.0 } Content-Type: text/plain; charset=US-ASCII } Content-Transfer-Encoding: 7bit } Content-Length: 2130 } } Angel, } } See comments below - in CAPS } } } } } Forwarded message: } } From vaytek-at-INS.INFONET.NET Sat Nov 19 05:53:29 1994 } X-Originated-From: lynx.msc.cornell.edu } X-Msc-Version: IDA Server } Date: Fri, 18 Nov 1994 16:47:23 CST } From: vaytek-at-INS.INFONET.NET } To: microscopy-at-aaem.amc.anl.gov } Message-Id: {00987A8C.867A916E.336-at-INS.INFONET.NET} } Subject: InGap on GaAs } } From: MX%"dieguez-at-iris1.fae.ub.es" 16-NOV-1994 18:20:14.00 } To: VAYTEK } CC: } Subj: } } Return-Path: {dieguez-at-iris1.fae.ub.es} } Received: from AAEM.AMC.ANL.GOV by INS.INFONET.NET (MX V4.1 AXP) with SMTP; } Wed, 16 Nov 1994 18:20:11 CST } Date: Wed, 16 Nov 94 07:48:46 +0100 } From: dieguez-at-iris1.fae.ub.es (Angel Dieguez Barrientos) } Message-ID: {9411160648.AA09675-at-iris1.fae.ub.es} } Subsject: CTEM - Need information about lines parallel in a layer } Apparently-To: microscopy-at-aaem.amc.anl.gov } } } My question is about a group of parallel lines that I have } observed in a sample of InGaP on GaAs, where the substrate is } misoriented 10 degrees. } } SOUNDS LIKE (001). USUALLY MISORIENTED TOWARDS 110 BUT NEEDS } TO BE SPECIFIED. } } The lines observed appear more clearly } when I tilt the sample about 30 degrees and I see in the 111 } direction (I am talking about a cross-section). Due that the } thickness of the layer is 2 microns, I think that the lines are } not due to moiree fringes provided for the superposition of the } substrate and the layer. On the other hand, because of the layer } is clearly observable, I think that the lines not correspond } to fresnel fringes due to thickness variations. } If it can help you, this system is ordered by not much (CuPt } structure). The lines are parallel to the substrate in a 011 cross- } -section and tilted about 12 degrees in a 0-11 cross-section. } } } COULD IT BE MISFIT DISLOCATIONS ? WHAT IS THE COMPOSITION OF THE } INGAP ? IS IT A LATTICE MATCHED COMPOSITION ? IF YES, HAS THE } COMPOSITION BEEN CHECKED WITH SOME OTHER METHOD (RBS, KNOWLEDEGABLE } X-RAY ? ) } } } } } } I need your help!!!!. } Please help me!!!!!!. } } Sinceresly yours A. Dieguez. } } } ieter, } I thought you might be able to suggest something. } This is from the microscopy listserver. } John Hunt }
I'd be curious to know how many of you that are experiencing problems with= Spurr's have had these difficulties (or any others) since the additive was= changed in the NSA of American manufacture. You might remember that I= requested help with problems using Quetol 651 last year--it appears now= that the NSA modification is the culprit: I changed to FLUKA NSA and have= just produced blocks identical to what I had prior to June of 1993. I= can/will provide anyone interested in details a complete workup and= explanation as to how I finally solved the mystery. Grace Kennedy UCSD
Dear Dwight, I, too, have trouble with stigmating our HVEM. Some of this comes from lower contrast at higher voltage, and some is just lack of skill. I have found it necessary to shoot sets of photos, first for the coarse adjustment (which usually does not change--unless someone has done something major) and then for the angle and magnitude of the fine. A significant help has been using our crude-but-sensitive video system in conjunction with a small condenser aperture and a large (admits 0.3 nm) objective aperture. Using the small beam gives high coherence--which you also found useful--but the image is too faint to cor- rect the astigmatism from the phosphor screen. Using the large obj. aper. im- proves the contrast without disturbing the fringe (we only get ~0.5 nm resolu- tion, so a 50 mu aper. is big enough for us; your mileage may vary.). Under these conditions, I am able to adjust the stigmator *reasonably* well. Obviously, these procedures might not be appropriate for your instrument and/or purpose. Good luck; I will be interested in any good ideas which come out of this discussion. Yours, Bill Tivol
EMS sent out a letter earlier this year explaining that the formula for the= additive in NSA which keeps Spurr's blocks light colored has been changed. = Althought Stacie K. from EMS swears that the change is quite recent, and my= Quetol problems started in June of 1993, I nevertheless purchased NSA from= FLUKA--My Quetol problem appears to be completely solved. I just ran 4= test blocks with two different accelerators with identical results--perfect= blocks. In the course of the last year and a half, I have tested/run down= virtually everything you can think of to solve this problem, with no= progress until now. I was just wondering if the Spurr's bubble problem= could be related to this change in additive; after all, Spurr's contains a= component remarkable similar in structure to Quetol. Any ideas???? Grace
In addition to Grace Kenndy's comments about NSA, I was wondering how many people are having trouble with section contamination in the last year or so. I've personally been having trouble and no of a few others which have been getting uniform peppering on their sections. The tourble doesn't seem to be the urnayl acetate, nor lead citrate (a variety of recepies and batches yield the same results). But resins using MNA instead of NSA do not seem to have this problem. Could the new NSA formulation be the source of the problem?
Richard E. Edelmann Electron Microscopy Facility Supervisor Marshall University - School of Medicine Huntington, West Virginia
Teaching them how to stigmate. How ambitious. You seem to be doing the right thing and have the key words,"time" and "patience". Aren't those two of the most important concepts in teaching EM, from embedding and sectioning through photography?
Teaching EM can be frustrating, as it is sometimes difficult to get students out of the mindset that all they need do for a course is read, listen, and write. Now they are in a situation where they have to plan ahead, apply concepts to real situations, and realize that the outcome is dependent on hundreds of techniques, chemicals, physical situations, and equipment conditions, any one of which can be screwed up with or without the students' knowledge.
It is rewarding when students return (as they occasionally do) and acknoledge that later success in graduate school, medical school, and jobs was aided by having taken EM. Some specifically use EM in research and work, while others no longer use it as a technique, but still have improved their situations in less tangible ways because of it.
In recent weeks I've had occasion to find that two of the labs I work with have been using formaldehyde stock solutions that were 3-4 years old. I would like to put together a memo to all the other labs I work with alerting them to the need to use fixatives of a more "recent" vintage. I have looked in several histology texts to see if I can find a guideline for when formaldehyde (either 37% stock, or 10% buffered) become too old to use, but they haven't been much help. Does someone have a reference they can refer me to or could you pass on what your guidelines are?
My experience is primarily in TEM and I know that concentrated glutaraldehyde tends to polymerize over time and we never let our 3% glut. exceed 4 months in age. I've learned that formaldehyde gets more acidic over time, but when is it too far gone?
Thanks for you assistance.
Douglas W. Cromey, M.S. Cell Biology and Anatomy Arizona Health Sciences Center 1501 N. Campbell Ave. Tucson, AZ 85724 (602)626-2824 dcromey-at-ccit.arizona.edu
The same discussion has been going on in our lab with regards to 4% paraformaldehyde. We have noticed a definite change in as little as one week after fix preparation, and marked changes after 2 weeks. Our fixatives are generally kept no longer than 4 weeks. The changes we notice are that with transcardiac perfusion, the body gets very rigid within 5 min. when using fix less than 1 week old. It takes longer for body stifeening after 1 week and athe body remains semi-limp after two weeks, but the brain histology still looks fine after overnight immersion. So it might be that there is some diminuition of fixative strength, but not so much that early postmortem changes are not arrested, and it takes longer for tho9rough fixation to occur. Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
Formaldehyde that is prepared from paraformaldehyde powder is normally made fresh just before use since it very poor shelf life. However, it is a relatively pure form of formaldehyde and is used usually with glutaraldehyde as a fixative for EM studies. Commercially available 37% formaldehyde solution on the other hand has impurities including methanol and therefore is not good for EM studies. But it can be readily used for studies at the light microscopy level. It is quite stable compared to the formaldehyde generated from paraformaldehyde powder. I had the opportunity a few years ago to compare results obtained from cells fixed with a new and a 2 year-old stock of the 37% formaldehyde (for the fluorescent-localization of F-actin with Rhodamine/Phalloidin). I could not detect any difference.
Message-Id: {m0rCdbG-0007KOC-at-stjohns.ohsu.edu} Message-Version: 2 } To: Microscopy-at-AAEM.AMC.ANL.GOV
RESEARCH ASSISTANT
Faculty Research Assistant Electron Microscopy-1.0 FTE. Requires BS in biological science (MS preferred),, training in biological lab instrumentation and protocol. Experience in EM tissue preparation is required. Responsible for histotechniques. Send Letter, curriculum vitae and names, addresses, and phone numbers of three references to :
Benita J. Pinz, Executive Assistant College of Veterinary Medicine OSU Corvallis, OR 97331-4801
Whenever possible we only make enough fixative for that day. That goes for glut., paraformaldehyde or glut/para mixes. If we need to store it before use we keep it frozen. We have not done any longevity test but it seems to be good at least one month. We usually use it up before any longer time. We do the same with 8% glut vials taht we have opened and not used completely. We seal the top with parafilm and then place the vial in a screw top jar with a good seal.
} Whenever possible we only make enough fixative for that day. That goes for } glut., paraformaldehyde or glut/para mixes. If we need to store it before use } we keep it frozen. Greg, You keep the glutaraldehyde frozen? I had understood that that was inappropriate, but I would be happy to be convinced otherwise. I've not researched this but I also have never run into a lab where this is done. I do TEM on a very infrequent basis, but often need to do fixation on very short notice. So, any improvement in shelf-life of reagents would be a real boon to me. Another win for the "net"! :-) --Jon
Jon L. Norenburg {norenbur-at-onyx.si.edu} Invertebrate Zoology, National Museum of Natural History Smithsonian Institution, Washington, DC 20560 Voice 301-238-3508, Fax 301-238-3361
To members interested in fixatives, formaldehyde solution in particular.
Here are some additional informations about storage of commercial form- aldehyde. The principal changes which may take place in formaldehyde on storage are as follows (listed in their order of importance from a practical standpoint): (1) Polymerisation and precipitation of polymer. (2) The Cannizzaro reaction, involving oxidation of one molecule of form- aldehyde to formic acid and reduction of another to methanol. (3) Methylal formation. (4) Oxidation to formic acid. (5) Condensation to hydroxyaldehydes and sugars. The changes are detrimental to product quality but may be avoided or kept at a minimum by maintenance of proper storage conditions. With optimum conditions of storage, commercial formaldehyde will remain unimpaired for long periods of time. In general, proper storage involves avoidance of temperature extremes and the use of storage in glass bottles, inert to corrosion by the mildly acidic solution. Low temperature favor polymer precipitation, high temperatures accelerate the reaction leading to chemical loss of formaldehyde. At improper storage temperatures, a form- aldehyde solution gradually becomes cloudy and eventually solid hydrated polymer separates as a precipitate. Much more could be said about this highly interesting fixative which possesses many unusual characteristics. ---------------------------------------------- * Sverker Enestr|m * * Department of Pathology, Link|ping, Sweden * ----------------------------------------------
Since an aqueous solution of formaldehyde (even made fresh from paraformaldehyde) exists as an equilibrium between methylene glycol and formaldehyde (skewed heavily towards the m.g.), covalent linkage of formaldehyde to tissue molecules is very slow , reaching equilibrium in 16 h -at-37 C (eg., reviewed in Fox et al. J Histo Cyto 33:845-853, 1985). This argues that BRIEF fixation in formaldehyde (as in immuocytochemistry preps) induces artifacts because of the high concentration of methylene glycol, and that little actual crosslinking occurs. Why use formaldehyde as a fixative for EM?
(((((((((((((((((((((((((((((())))))))))))))))))))))))))))) R. Howard Berg Biology Department University of Memphis, Memphis, TN, 38152
If you are really interested in reading about fixatives and how they work there is a chapter devoted to this in the book by G. Griffiths. The ref. is "Fine Structure Immunocytochemistry" 1993 published by Springer Verlag, Heidelberg.
Let me add a note for those workinf on mineralized tissues after formaldehyde fixation. Small larval bivalve shells (80 to 250 mis microns) dissiolve quite ras rapidly in even buffered formaldehyde 10% solution. If fixation is required in aldehyde, use glutaraldehyde by preference, or use formaldehyde the minimum time and replace with 70-95% ethanol. or better yet (if only mineralized material will be studied, use 95% ethanol as sole fixative. Shells last for years o in ethanol alan pooley marine sci sem lab rutgers univ
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