We have a Linkham heating/cooling stage that appears to perform according to its specifications. We have not done a rigorous evaluation of the stage temperartures/drift etc. but with the standards we have run it has provided results close to expected values. It has a stated range range of -190 to +600 degrees C. We have used it in the -50 to +200 range with no problems. Overall a good unit but a bit pricey.
Joe Neilly Abbott Labortories Dept. of Microscopy and Microanalysis 200 Abbott Park Rd. Abbott Park, IL 60064
We have attempted to addressed the issue of sample retention in a TEM in a short technical note in Journal of Microscopy, V 174, Pt. 1, April 1994, pp. 55-58, "Enhanced retention of magnetic particles (e.g. microtomed sections) in a TEM". ( E-mail valdemar-at-fast.net with a request for a reprint. )
The technique boils down to fabricating a bi-layer, electron transparent support film on a standard TEM grid of your choice. The materials in the two-layer support film are chosen for solvent incompatibility (e.g. collodion top film soluble in amyl acetate over the bottom layer of Formvar soluble in ethylene dichloride but not in amyl acetate); so that, after placing your sample on the top film, the top film is temporarily softened and transformed into electron transparent layer of adhesive with a judicious application ( a few drops to wet filter paper near your TEM grid ) of solvent specific to that film. The bi-layer support films are a lot easier to fabricate than the description makes it appear, and the technique is amenable to any combination of support films as long as the structural layer is not affected by the solvent for the adhesive layer.
Prior to developing this method, we had a severe problem with losses of microtomed sections of magnetic steel in the field of a high excitation objective lens. Now, we hardly ever loose one; and the bi-layer support films are sufficiently robust for sample deposition with a hand-held eye lash, and sufficiently stable and clean for multi-hour acquisition of quantitative composition profiles at high resolution by EDS under UHV in a dedicated STEM. Perhaps with an exception of some EELS work, I see no reason why the technique would not work under most circumstances for your persnickety sample with a judicious selection of the support films and their solvents.
We have tried the folding grid approach and, by comparison, found it a bother.
For a method of applying an adhesive to grid bars, you might look up E. Fritz (1991), "The use of adhesive-coated grids for the X-ray microanalysis of dry-cut sections in the TEM", J. Microsc. 161, 501-504. ( Sorry, but I'm out of copies of this one. )
Valdemar Furdanowicz valdemar-at-fast.net Homer Research Labs Bethlehem Steel Co. Bethlehem, PA 18016
******************************************************** Isabel Nogueira wrote: ----------------------------------------------------------------------- I work with a TEM and I'm currently observing and performing EDS analysis on gold samples. The problem is that the samples don't stick to the grids. I need folding grids but the ones I've seen so far are Cu, Ni, Au or Ag.
I need carbon folding grids and would appreciate if anyone can tell me where I can get them. -----------------------------------------------------------------------
Nancy Buening wrote: ----------------------------------------------------------------------- I am analyzing calcite shells in the TEM. I am having difficulty gluing the calcite samples onto the grid. I have been using colloidal silver liquid, but it either dries before I can get the calcite in place or I get too much liquid and the grid "drowns". Is this just a matter of practice, or is there another gluing material that I can try that is more forgiving ? -----------------------------------------------------------------------
Does anyone have any simple solutions to reduce the amount of endogenous fluorescence that seems to be both tissue type and fixation specific? We work with fish retinas and as a rule immersion fix in 4% paraf. for 1 hr, room temp. When glut is added the problem is worse. The fluorescence seems to be worse in the inner segments of the photoreceptors-all these years the results have been satisfactory, but any improvement would be great. Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
I am trying to serial section some fungal spores for TEM, and I am running into problems with the sections wrinkling.
Details: I am using formvar and carbon coated slot grids. The specimen is embedded in epon. I am using a Reichert Ultracut E (mechanical advance)
ultramicrotome, and an Edgecraft diamond knife. I have been moving the sections onto the grids with the aid of a micromanipulator holding the grid into the water, touching the section ribbon to the formvar, and winding the grid up out of the water.
I have tried varying the section size - from about 1/2mm*1/2mm to less than 1/4mm each side, with little or no change in the degree of wrinkling. I tried varying the angle at which the grid is held into the water, from almost parallel, to perpinduclar, with no success. I tried varying the speed at whcih the grid is removed from the water, and hence the ribbon is sucked onto the formvar, also with no success. Finally, I thought it might be the formvar/carbon coat -- when held close to the boat, the humidity caused the film inside the slot to go slack. I thought maybe the slight waviness in the film might be introducing the wrinkles, so I changed my formvar recipe and procedure. I did manage to produce a couple grids with intact films which did not slacken near the boat, but the sections still wrinkled. The sections are uniform silver/gold interface, and are generally part of a very strong ribbon (can be hard to break it up into managable pieces using eyebrow brushes).
So, here I am: does anyone have any suggestions for avoiding the %&*%$$$# wrinkles, other than changing the embedding medium properties?
Also, has anyone else ever noticed the slackening of the formvar film near to water, what might cause it, and how best to avoid it? (my good grids were made with formvar that was thoroughly dessicated in dichloroethane - I was previously using chloroform without the variety of drying procedures).
Thanks in advance (from me and my supervisor whoe would appreciate it if I didn't throw the ultramicrotome through the window),
Russell
PS I have also tried picking up the sections with an empty slot grid and overlaying that grid over another, coated grid, and allowing the sections to dry. They still wrinkled. R.
I have a Durst Laborator S-45-EM and the varipoint unit that adjusts light intensity has broken. Does any one have any idea how to fix or replace this unit. I don't expect that a new replacement is available but maybe someone has a Laborator that they are no longer using can provide this part. Or maybe some one can tell me how to build a unit that will accomplish the same function so that I can continue to use the Laborator.
Also can anyone tell me how to find Steve Miller who used to be with Integrated Microsystems
Bob Schmitz Robert J. Schmitz Electron Microscope Lab Department of Biology, CNR Building University of Wisconsin Stevens Point Stevens Point, WI 54481 phone (715) 346-2420 FAX (715)346-3624
I know how you feel. Be patient. I have been doing serial sectioning for 6 years. We have an elaborate set up which allows us to section in pairs. One person sections and one person transfers the sections to the formvar coated slot grid. If you want to know about this transfer apparatus I can find the reference for you.
In the meantime you will have to work by yourself. Whenever I section by myself I get some wrinkles(nothing to upset research though). The wrinkles occur as I slide the formvar coated slot grid into the diamond knife water, and draw up the sections. The wrinkles are the result of the formvar stretching a little when it goes under water. Then, as the sections lie down on the stretched formvar, they wrinkle a bit. I never have wrinkles which are bad enough to stop science.
To minimize wrinkles I suggest: Make your block face as tiny as possible.
Immerse your grid at a 45 degree angle.
Slowly coax your sections onto the formvar, and slowly and gently pull the grid out of the water at a 45 degree angle. The key is to move slowly and deliberately.
Work on a day that you are happy, the block is happy, the sectioning room is happy. Do not attempt EM on a bad hair day.
Cleaning the ol' closets. Before I throw these in the dumpster, I thought I'd ask: JEOL 100B specimen carousel and specimen holders JEOL 100B specimen exchange mechanism (parts) 2-3 boxes of HU11E replacement parts and vacuum tubes EDAX detector and 183 preamp, from AMRAY SEM. Broken window, maybe other damage
You pay to ship it, it's yours. Contact me at the address below:
Julian P.S. Smith III Biology Winthrop University Rock Hill, SC 29733 803-323-2111 x227 (vox) 803-323-2246 (fax)
jd wrote: } } It has been a few years since I have been involved with temperature } control stages on optical microscopes. Last I remember there were } issues such as surface area temperature mapping(variations) functions, } isolation, target range overshooting, steady state vs. drift } functions, heating and cooling techniques, depth of focus and other } optical issues, maximum and minimum feasible temperatures, heating and } cooling techniques, etc. Can anyone refresh my memory , bring me up to } date ,and refer me to journal and review articles on the subject. } I would like references to current manufacturers. Also I would be } interested in personal experiences with specific current equipment } manufacturers of both microscopes and control stages for this use } (preferably any potentially negative feedback opinions sent only to my } personal email address below, so no manufacturer is embarrassed or } damaged by personal opinions in this forum/possibly an obvious point } of controversy). } Finally does anyone have any personal experience with modifications to } existing designs or references to in house construction of these } devices. } I also might be prospect for use equipment of this nature. } } JD } EMAIL1: wa5ekh-at-juno.com } please cc to EMAIL2: wa5ekh-at-cyberramp.net Dear JD,
There are several people who make very good heating stages. Yes, there are issues such as surface area, heating and cooling techniques, etc. Typically, however, the better stages have fairly enclosed heating chambers, so there is less concern about heat transfer. A few quick cautions: you probably will have to get long working distance objectives, especially if your application requires higher magnifications. Also,you have not mentioned much about the specifics of your work, so it is hard to answer in a more focused fashion, but you may not be able to look at a very extensive area at any given time.
I have had personal experience with two manufacturers: Mettler and the Koffler hot stage available through Arthur Little. Mettler's stages are well built and, depending on the options purchased, can provide extremely delicate temperature control and sophisticated heating and cooling cycles. They are also very expensive. The Koffler hot stages are much more practical and allow you to look at larger samples. As I remember it, they had an interesting large glass plate which covered the chamber and kept the temperature fairly constant. Both typically have a range from some sub-room temperature value to about 350 degrees C.
There are three other companies which I would suggest that you investigate as well: Bioptechs, PhysiTemp, and World Precision Instruments. All three are better suited to the 32-35 degree C, constant temperature conditions necessary for live cell work. I have only seen brochures for the PhysiTemp but I have had some lengthy conversations with the people at Bioptechs. If you are doing critical live cell work, they have a number of options whichhallow you to look at somewhat larger fields. Also, for things like Calcium flux test, where even the temperature of the objective lens might be critical, they have an objective heater.
Finally, on the do-it-yourself front: Walter McCrone runs courses on hot stage microscopy out of the McCrone Institute in Chicago. Part of the class revolves around using a rheostat, wires, and thermally conductive glass to build your own stage. The good news is that the stage has a very low profile so that you can look at larger areas and don't need extra long working distance objectives. Secondly, they are very inexpensive to build. However, unless you have experience with thermocouples, I don't know how you would accurately measure the tempature at the sample. Also, the sample is open to ambient conditions. I am not quite sure how Walter's design accounts for air currents, local cooling, etc. I have worked with a client who built one of these devices for pigment and polymer analysis and he seemed quite happy with it. (I will attach names and addresses for any contacts which I have in my files, below).
Microscopy/Microscopy Education tries to act as a clearinghouse for this type of information, so we would really appreciate a summary of your findings.
Hope this all helps.
Barbara Foster Consortium President MME 53 Eton Street Springfield, MA 01108 US PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
America's first consortium of expert microscopists dedicated to helping you be more productive with your microscope.
DISCLAIMER: MME does not sell or have any financial interest in any of the above mentioned equipment.
List of contacts: Bioptechs - Dan Focht (412)282-7145 Web site: www.Bioptechs.com Mettler - Mark Kelsey (800)638-8537 ext 7041 McCrone Institute - ask for any of their fine technical staff - (312)842-7100
PhysiTemp advertises in most of the common trade journals. I will have to send you both their information and Arthur Little's under separate cover.
You could try waving a Q-tip soaked in chloroform over the sections before bringing the grid anywhere near them (this is not the same as waving a dead chicken - it works). Or you could buy a heat-pen from (I think) JB EM, and probably other suppliers. In this case, you wave a hot wire over the sections and they flatten like magic. Hope this helps.
Lesley Weston.
On Mon, 1 Dec 1997, Wyeth, Russell wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } TWIMC, } } I am trying to serial section some fungal spores for TEM, and I am } running into } problems with the sections wrinkling. } } Details: I am using formvar and carbon coated slot grids. The specimen } is } embedded in epon. I am using a Reichert Ultracut E (mechanical advance) } } ultramicrotome, and an Edgecraft diamond knife. I have been moving the } sections onto the grids with the aid of a micromanipulator holding the } grid } into the water, touching the section ribbon to the formvar, and winding } the } grid up out of the water. } } I have tried varying the section size - from about 1/2mm*1/2mm to less } than } 1/4mm each side, with little or no change in the degree of wrinkling. I } tried } varying the angle at which the grid is held into the water, from almost } parallel, to perpinduclar, with no success. I tried varying the speed } at whcih } the grid is removed from the water, and hence the ribbon is sucked onto } the } formvar, also with no success. Finally, I thought it might be the } formvar/carbon coat -- when held close to the boat, the humidity caused } the } film inside the slot to go slack. I thought maybe the slight waviness in } the } film might be introducing the wrinkles, so I changed my formvar recipe } and } procedure. I did manage to produce a couple grids with intact films } which did } not slacken near the boat, but the sections still wrinkled. The } sections are } uniform silver/gold interface, and are generally part of a very strong } ribbon } (can be hard to break it up into managable pieces using eyebrow } brushes). } } So, here I am: does anyone have any suggestions for avoiding the } %&*%$$$# } wrinkles, other than changing the embedding medium properties? } } Also, has anyone else ever noticed the slackening of the formvar film } near to } water, what might cause it, and how best to avoid it? (my good grids } were made } with formvar that was thoroughly dessicated in dichloroethane - I was } previously using chloroform without the variety of drying procedures). } } Thanks in advance (from me and my supervisor whoe would appreciate it if } I } didn't throw the ultramicrotome through the window), } } Russell } } PS I have also tried picking up the sections with an empty slot grid and } overlaying that grid over another, coated grid, and allowing the } sections to dry. They still wrinkled. R. } }
Bob - Find a place through your yellow pages which rewinds electric motors. Phone them and ask if they can rewind a variable transformer. Otherwise, buy a variable transformer with similar specifications and adapt it to the equipment. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } } I have a Durst Laborator S-45-EM and the varipoint unit that adjusts } light intensity has broken. Does any one have any idea how to fix or } replace this unit. I don't expect that a new replacement is available } but maybe someone has a Laborator that they are no longer using can } provide this part. Or maybe some one can tell me how to build a unit } that will accomplish the same function so that I can continue to use the } Laborator. } } Also can anyone tell me how to find Steve Miller who used to be with } Integrated Microsystems } } Bob Schmitz } Robert J. Schmitz } Electron Microscope Lab } Department of Biology, CNR Building } University of Wisconsin Stevens Point } Stevens Point, WI 54481 } phone (715) 346-2420 } FAX (715)346-3624
} From: "jd" {wa5ekh-at-cyberramp.net} } To: {microscopy-at-sparc5.microscopy.com} } Subject: Temperature Control Stages for Optical Microscopy } Date sent: Fri, 21 Nov 1997 03:39:31 -0800
I sort of feel that people using this list should reveal at least their name, if not their affiliation. I don't want to start anything both big and trivial, but what do others think?
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
} I have a Durst Laborator S-45-EM and the varipoint unit that adjusts } light intensity has broken. Does any one have any idea how to fix or } replace this unit. I don't expect that a new replacement is available } but maybe someone has a Laborator that they are no longer using can } provide this part. Or maybe some one can tell me how to build a unit } that will accomplish the same function so that I can continue to use the } Laborator.
Bob,
I'm assuming that you're talking about the potentiometer that controls the point light source. I AM NOT an electronics whiz, so I may be completely off base, but I'm pretty sure that those units are nothing more than expensive, heavy-duty "dimmer switches", combined with a voltage transformer. Check with an electronics shop who might be able to fix it fairly easily---I can't imagine that they can be too complicated. If it can't be fixed, I'd bet that a substitute could be made and perhaps fitted with the connectors on the unit you have. It might even be that a hardware store with a good electrical section might have a transformer and dimmer switch with specifications compatible with your light source.
One caution: I would NOT---repeat, NOT---try to use just a dimmer switch hooked to a regular 110-120 outlet. I once had just finished changing from conventional tungsten light to the point light source, grabbed the wrong cord and plugged it directly into an outlet. When I turned on the switch, the bulb exploded like a pistol going off and sprayed half the darkroom with hot shards of glass (I hadn't closed the enlarger head access panel, yet). Get a good electrician's advice before rigging a substitute, but I bet it can be done.
Randy Tindall Electron Microscope Lab New Mexico State University Las Cruces, NM 88003 Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
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I recently came across an article where someone was using an "Air Stream Incubator" from Nevtek as a temperature controler. It is apparently a blower that moves heated air across the stage area heating everything in its path. I have never heard of this before. Any comments on the effectiveness of this solution? Dave Knecht
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu
1. Any glut will cause autofluorescence, in fact we use it as counterstain if we want the tissue to be fluorescent.
2. I remember that someone was using .05% Pontamine Sky Blue in PBS with 1% DMSO after the immunohistochemistry was carried out, to reduce the autofluorescence in the FITC channel.
3. Another person was using .3g eriochrome black in 100ml PBS as a counterstain for the same purpose.
Hope this helps,
Bob
On Mon, 1 Dec 1997, Linda Barthel wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Does anyone have any simple solutions to reduce the amount of endogenous } fluorescence that seems to be both tissue type and fixation specific? We } work with fish retinas and as a rule immersion fix in 4% paraf. for 1 hr, } room temp. When glut is added the problem is worse. The fluorescence } seems to be worse in the inner segments of the photoreceptors-all these } years the results have been satisfactory, but any improvement would be } great. } Linda Barthel } Research Associate II } Department of Anatomy and Cell Biology } University of Michigan } lab (313) 764-7476 } fax (313) 763-1166 } barthel-at-umich.edu } } } }
Thanks for the overwhelming response! Could this possibly a reaction to a common, and all to evil foe?
Here's a quick (sort of) summary for everyone's benefit, and I'll try to answer all the further questions I received.
First, thank you everyone for the chloroform suggestions, but that's not the problem here. I've used that method many-a-time for wrinkled sections coming off the knife blade. These sections, though, are beautifully flat -- they shouldn't require any stretching. The wrinkles are _definitelly_ introduced when the sections attach to the formvar (I carefully checked the grids at all stages). There's just 3 or 4 per section, and they appear no matter whether they are left to dry down onto the formvar from another uncoated grid, or attached directly on to the formvar. (Someone pointed out this shouldn't get in the way of the observations, and in this case - with lots of spores in each sample - the wrinkles are not a critical problem. But I'm a perfectionist, and I can certainly envisage times when I wouldn't want any wrinkles, so I figured I'd try to learn with this otherwise very cooperative sample.)
A good suggestion was made regarding refinement of the micromanipulator technique I already use -- I'll give that one a shot first, but I have a feeling my perfectionist attitude towards the wrinkles may mean I'll need to try something else. The other suggestions centered around alternative means to get the sections on the grid. Basically, the sections are picked up on a formvar film attached to another tool with a hole big enough for the entire film to be passed over onto a clean slot grid. This means the sections can be positioned much more precisely over the slot. (I had heard of this method, but we are cash strapped and I was hoping to avoid a purchase of the loops or 'domino racks'. Perhaps it is unavoidable.) I was also given a couple references which I am about to go look up. I've attached the originals of various suggestions below.
Finally, everyone advocated patience. I was going to say I'd already tried that, but I guess I'll have to go out and buy some more.
Once again, thanks everyone, and I'll report back.
Russell
} snip I use domino racks that can be purchased from the EM companies like EMS or Ted Pella. Instead of working with the formvar on the grid you put the film over the domino rack which is a piece of sheet metal with slightly larger than grid size holes. You then pick up the sections from the water with a grid and don't have to worry about the film wrinkling when you get close to the water. The grids are standard cleaned slot grids. After touch down to the water the sections should be held in the center of the hole by surface tension. You then lay the grid on the hole of the domino rack and let it dry overnight. After the sections have dried, you carefully punch out the grid by using the tips of the forceps around the outside of the grid. I find that this approach works much better for me.
Patty Jansma
} snip I have been cutting ultrathin sections for a number of years now and the most recent advance in section collection to me was the invention of the 'Perfect Loop'. With this loop one can pick up sections from the water bath and place them on a grid sitting on filter paper by the side of the ultramicrotome. The excess water is then wicked away with a wedge shape of filter paper. To date, this I find is the best way to collect sections without wrinkles. This is sold in the USA by EMS. Happy sectioning. Ian Lamswood
} snip Whenever I section by myself I get some wrinkles(nothing to upset research though). The wrinkles occur as I slide the formvar coated slot grid into the diamond knife water, and draw up the sections. The wrinkles are the result of the formvar stretching a little when it goes under water. Then, as the sections lie down on the stretched formvar, they wrinkle a bit. I never have wrinkles which are bad enough to stop science.
To minimize wrinkles I suggest: Make your block face as tiny as possible. Immerse your grid at a 45 degree angle. Slowly coax your sections onto the formvar, and slowly and gently pull the grid out of the water at a 45 degree angle. The key is to move slowly and deliberately. Sally Shrom
} snip I did serial section reconstructions of flagellar apparatuses of two algae for my dissertation work using polystyrene films (Journal of Electron Microscopy Technique 13:268-269. The technique is a modification of the following: Rowley III, J.C. and Moran, D.T. 1975. A simple procedure for mounting wrinkle-free sections on formar-coated slot grids. Ultramicroscopy 1:151-155. I highly recommend this way. It's very easy - I taught it to several persons and they all became better at it than I am. Good luck, Heather Owen
} one final snip A suggestion: I was once told that formvar is less hydrophobic if you refrigerate the grids overnight before use. I never needed to, but it was suggested. Otherwise, good luck. Serial sectioning is no fun trick. Gregg Sobocinski
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In a message dated 97-12-02 10:14:08 EST, rwyeth-at-pfc.forestry.ca writes:
{ { I am trying to serial section some fungal spores for TEM, and I am running into problems with the sections wrinkling.
Details: I am using formvar and carbon coated slot grids... } }
Russell,
I have encountered similar problems in one of my past lives as an ultramicrotomist, and here's what we came up with:
We found the major contributor was the hydrophobic nature of the carbon film. Even storing the grids under UV light and glow-discharging in a partial vacuum with an air atmosphere really didn't help that much.
We finally collected the sections onto slot grids which had a *formvar only* film, wicked away the excess fluid from the side of the grid, let everything dry well, then did our staining. Immediately before going to the microscope, we placed the grids into a bell jar and gave them a light dusting of carbon. This helped immensely.
Good luck; let us know how things work out in the end!
} } Winter Workshop on In-Situ Electron Microscopy } } A workshop on In-Situ Electron Microscopy will be held in Scottsdale, } Arizona from Jan. 7-10, 1998. The goal of this workshop is to provide a } forum for presentations and discussion of recent developments in } instrumentation, current applications and future directions of in-situ TEM } and SEM. Some areas of particular interest are: } } . In-Situ Heating Experiments: Electron Diffraction and Imaging or } Temperature Controlled Experiments. } . Ion Implantation studies and ion irradiation effects } . Environmental Cells or Gaseous Environment Controlled TEM/SEM } . Effects of Stress/Strain including fracture studies and stress-induced } phase transformation } . Magnetic Materials Studies } } The list of invited speakers includes: } } Ernst Bauer, Arizona State University } Ed Boyes, Dupont Corp. } Kazuo Furuya, National Research Institute for Metals } James Howe, University of Virginia } Robert Hull, University of Virginia } T. Kamino, Hitachi Instruments Eng. Co. } John Mansfield, University of Michigan } Ulrich Messerschmidt, Max Planck Institute of Microstructure Physics } Amanda K. Petford-Long, University of Oxford } Francis M. Ross, IBM Thomas J. Watson Labs } Robert Sinclair, Stanford University } Nubuo Tanaka, Nagoya University } } Abstracts are still being accepted and should be sent as soon as possible. } For more information contact: } } Eloise Kadri } Center for Solid State Science } Arizona State University } P.O. Box 871704 } Tempe, AZ 85287-1704, USA } } Email: Eloise.Kadri-at-asu.edu } Tel: 602 965 9004 } } You can also register via our Web Site at: } http://www.asu.edu/clas/csss/workshop/ } } } } Peter A. Crozier } } Industrial Associates Program } Center for Solid State Science } Arizona State University } Tel: 602 965 2934 } Fax: 602 965 9004 } } Website: http://www.asu.edu/clas/csss/IAP/
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
} } Winter Workshop on In-Situ Electron Microscopy } } A workshop on In-Situ Electron Microscopy will be held in Scottsdale, } Arizona from Jan. 7-10, 1998. The goal of this workshop is to provide a } forum for presentations and discussion of recent developments in } instrumentation, current applications and future directions of in-situ TEM } and SEM. Some areas of particular interest are: } } . In-Situ Heating Experiments: Electron Diffraction and Imaging or } Temperature Controlled Experiments. } . Ion Implantation studies and ion irradiation effects } . Environmental Cells or Gaseous Environment Controlled TEM/SEM } . Effects of Stress/Strain including fracture studies and stress-induced } phase transformation } . Magnetic Materials Studies } } The list of invited speakers includes: } } Ernst Bauer, Arizona State University } Ed Boyes, Dupont Corp. } Kazuo Furuya, National Research Institute for Metals } James Howe, University of Virginia } Robert Hull, University of Virginia } T. Kamino, Hitachi Instruments Eng. Co. } John Mansfield, University of Michigan } Ulrich Messerschmidt, Max Planck Institute of Microstructure Physics } Amanda K. Petford-Long, University of Oxford } Francis M. Ross, IBM Thomas J. Watson Labs } Robert Sinclair, Stanford University } Nubuo Tanaka, Nagoya University } } Abstracts are still being accepted and should be sent as soon as possible. } For more information contact: } } Eloise Kadri } Center for Solid State Science } Arizona State University } P.O. Box 871704 } Tempe, AZ 85287-1704, USA } } Email: Eloise.Kadri-at-asu.edu } Tel: 602 965 9004 } } You can also register via our Web Site at: } http://www.asu.edu/clas/csss/workshop/ } } } } Peter A. Crozier } } Industrial Associates Program } Center for Solid State Science } Arizona State University } Tel: 602 965 2934 } Fax: 602 965 9004 } } Website: http://www.asu.edu/clas/csss/IAP/
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
Rick Felten 12/02/97 03:06 PM I am looking for some Pyrope Garnet, Mg3Al2Si3O12, to use as a X-Ray Microanalysis Standard. I have tried many of the EM suppliers (Energy Beam Sciences, Polysciences, Fullam, and Ted Pella) w/o any luck. I would be amenable to another standard that has major levels of Mg, Si and O and a minor level of Al and no other elements. Any suggestions?
} Does anyone have any simple solutions to reduce the amount of endogenous } fluorescence that seems to be both tissue type and fixation specific? We } work with fish retinas and as a rule immersion fix in 4% paraf. for 1 hr, } room temp. When glut is added the problem is worse. The fluorescence } seems to be worse in the inner segments of the photoreceptors-all these } years the results have been satisfactory, but any improvement would be } great.
This problem was recently discussed here. The fluorescence is due to the aldehydes and the fix [:-)] is to reduce them with NaBH4. I am definitely not competent in specimen preparation, but somewhere in our group the procedure has been worked out. If none of the experts in this field respond, I'll try to get the recipe for you. Yours, Bill Tivol
Dear Ritchie, } } I sort of feel that people using this list should reveal at least } their name, if not their affiliation. } I don't want to start anything both big and trivial, but what do } others think? } As one who has first met many people electronically through this list and then seen them at MSA meetings, I think it's useful to put names on postings. I don't think that "jd" had anything sinister in mind, and I certainly would not want Nestor to insist that all postings meet any requirements other than those of relevance, etc. already in place. Yours, Bill Tivol
A friend of mine wants to sell two microscopes. Please contact him directly.
Lietz Dialux Microscope, trinocular head with phototube (modified Orthomat System for SLR camera. Bright field objectives 4x, 10x, 25x (npl), 40x plan, 100x. Two .9 na condensers (brightfield and darkfield). Asking $2,000.
Leitz Diavert Microscope. Mint condition foruse in tissue culture. Phase contrast 10x, 20x, 4x low power objective, 12V 100w light source. Asking $1500 ($2,000 with transformer).
For more information call 614 688 4471 and ask for Steve.
Rick Felten wrote: ================================================ I am looking for some Pyrope Garnet, Mg3Al2Si3O12, to use as a X-Ray Microanalysis Standard. I have tried many of the EM suppliers (Energy Beam Sciences, Polysciences, Fullam, and Ted Pella) w/o any luck. I would be amenable to another standard that has major levels of Mg, Si and O and a minor level of Al and no other elements. Any suggestions? ================================================ Actually you don't have to settle for anything other than what you want! The pyrope garnet, often times requested, is mineral #37 on the SPI #02753 Mineral Mount. It has excellent homogeneity and has been offered by our firm since about 1980. More details about this mount, including 52 other minerals, are available on our website given below. Contact me off line if you have any questions.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
You may have tried borohydride. The following is a brief writeup of how we have used it in the past. A. Kent Christensen, University of Michigan.
BOROHYDRIDE
If glutaraldehyde was used in the fixative, then it may be advantageous to quench extraneous aldehyde groups with 1% sodium borohydride (NaBH4) (Eldred et al., 1983, J Histochem Cytochem 31:285), which is a particularly strong reducing agent (USE WITH CAUTION). Borohydride can eliminate most tissue autofluorescence, thus reducing background in LM ICC studies involving fluorescent markers. It has been suggested that borohydride may partially restore antigenicity after glutaraldehyde fixation by reducing Schiff bases (carbon-nitrogen double bonds) that can be formed when glutaraldehyde reacts with free amino groups on proteins; the reduced bonds are less rigid, and the increased mobility may restore some of the antigenicity.
Borohydride treatment can be carried out on pieces of tissue after fixation and the overnight buffer wash (Eldred et al, 1983). Use 1% sodium borohydride in PBS for 30 min at room temperature. The solution should be freshly prepared from sodium borohydride powder that has previously been stored in a manner that has protected it from moisture. The tissues will bubble vigorously as hydrogen gas leaves them, which will worry you but doesn't seem to damage the tissues. The treatment is followed by a wash of 2 x 30 min in PBS.
To use borohydride as a quenching agent during an LM immunocytochemical run, put a drop of 1% sodium borohydride on each tissue section and leave it for about 10 minutes. Then wash in a coplin jar of PBS or PBS-G.
For convenience, 10 mg aliquots of sodium borohydride powder can be stored in microtubes at -20=A1C in a plastic box containing silica gel desiccant. When you need 1% borohydride, add 1 ml of PBS to a tube and vortex briefly (CAUTION: open the microtube promptly after vortexing, or the hyrogen gas generated as the powder goes into solution will pop it open, possibly causing a spill).
Wrick: Shop around. No problem getting that standard; for one, it is in our catalogue. Note the A$ is 40% less than the US$. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } From: Rick Feltenby way of Nestor J. Zaluzec {rfelten-at-Macdermid.com} } To: microscopy-at-sparc5.microscopy.com } Subject: Pyrope Garnet } Date: Wednesday, 3 December 1997 9:15 } Rick Felten } 12/02/97 03:06 PM } I am looking for some Pyrope Garnet, Mg3Al2Si3O12, to use as a X-Ray } Microanalysis Standard. I have tried many of the EM suppliers (Energy Beam } Sciences, Polysciences, Fullam, and Ted Pella) w/o any luck. I would be } amenable to another standard that has major levels of Mg, Si and O and a } minor level of Al and no other elements. Any suggestions? } }
Three announcements from the San Francisco Microscopical Society:
First, we have established an email list of correspondents who wish to receive periodic announcements of our activities, significant updates to our web pages, and other notes of the Society. To be placed on this list, respond to me personally (not the MSA List!) via email, or respond through the solicitation on the first of our web pages (see below).
Second, we would like to invite all friends of our Society to join us at our annual Christmas Social Hour on Thursday evening, December 11, at 7:00 PM at the Rockridge Branch of the Oakland Public Library. Further information can be obtained at our web pages (again, see below).
Third, we would like to announce the establishment of the San Francisco Microscopical Society Web Pages. While in development they will reside at:
After a shaking out period and filling in some gaps we'll probably move to a more permanent URL, but for now I've posted the pages adjacent to my personal pages.
With regard to the web pages, we are curious to know if commercial firms would be interested in a corporate membership which would support the society (we are a CA non-profit corporation) as well as providing an opportunity for exposure to our members through our regular newsletter and to the broader microscopy community through our web pages. If you're with such a firm, please contact me off list and let me know if such a membership would be of interest to your firm. I would anticipate a very modest fee for such a membership.
Fourth, and perhaps most importantly, may you all have a safe, happy, and full holiday season!
Steve Shaffer -- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
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Hi Rick.
Micro-Analysis Consultants Ltd has been in the business of supplying standards for micro analysis for 16 years now. We have over 300 different materials in stock with certificates of analysis. We are an ISO 9002 registered company for the supply of standards, advertise regularly in Microscopy and Analysis, and have a web site at:
http://www.macstandards.co.uk/town/street/yr49/
We specialise in the supply of full custom standards. In other words tell us what you want and we will quote you for it. You can have whatever block size you would like in either Stainless Steel, Brass, Aluminium or Carbon resin. The number of standards we can fit into a block is form 1 to what ever will fit in the block specified. As an example for you a single 3mm x 5mm Brass mount standard of Almandine Garnet will cost £65. Delivery will be 1 week from receipt of your order.
} Rick Felten } 12/02/97 03:06 PM } I am looking for some Pyrope Garnet, Mg3Al2Si3O12, to use as a X-Ray } Microanalysis Standard. I have tried many of the EM suppliers (Energy Beam } Sciences, Polysciences, Fullam, and Ted Pella) w/o any luck. I would be } amenable to another standard that has major levels of Mg, Si and O and a } minor level of Al and no other elements. Any suggestions?
Rick: If you just need a specimen of pyrope garnet and do not want to buy an expensive standards collection of several dozen standards, then I suggest you try one of the many Internet Rock Shops. A quick check at one: http://www.gemhut.com revealed that they have many gem quality specimens of pyrope garnet starting at $9.95. I've purchased several isolated mineral specimens like this for standards and they work very well. Stanley L. Flegler, Assistant Director Center for Electron Optics Michigan State University
At 10:10 AM 12/1/97 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I don't know if I can help but I haven't seen anyone suggest what we do. Rather than picking up the sections with the grids we traditionally pick them up with a loop before placing them on the grid. In short, we make a round loop by hand - about 3 mm in diameter although the size or shape can be dependent on your specimen - with wire intended for metal evaporation such that if lowered onto a water surface it picks up the water droplet inside the circumference. This loop is then attached to the end of an applicator stick. THe loop is lowered onto the water surface around the sections - the sections are picked up with the water droplet, and then the entire contents are lowered onto a grid which is sitting on filter paper. With a film coating on the grid you might want to have your loop slightly larger than the grid so that the water flows out around it into the fitler paper - or cut "V" shaped pieces of filter paper to hold around the loop as you lower it onto the grid. I don't know if this will solve your problem but it's cheap and might be worth a try. Good luck!
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
We are interested in getting a digital camera for light microscopy. The simpler, the better. Just want to capture a good quaility image into a PC or SGI O2 platform that could then be handled by Photoshop or the like. We are considering both color and B7W only cameras. Cost is a consideration, as usual.
I would appreciate opinions from users and information from dealers.
TIA Greg Erdos
******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Dear All, Does anyone have a better idea to detect intracellular calcium than by using the potassium pyroantimonate method as suggested in Hayat? We've tried it but we're not sure if it's precipitating calcium or the cesium that we use to induce metamorphosis in our experimental animals. Our thesis depends on it. Help!
-------------------------------------------------------- Name: Winston W Wiggins, Supervisor Vox:704/355-1267 CRC-Electron Microscopy Lab Fax:704/355-7648 Carolinas Medical Center Lab:704/355-7220 P.O. Box 32861 Charlotte, NC 28232-2861 USA Date: 12/3/97 E-mail: wwiggins-at-carolinas.org Time: 11:59:18 AM --------------------------------------------------------
a researcher wants me to prepare some thin section micrographs of red and white blood cells. Simply adding 2X fixative to a few hundred microliters of blood resulted, as expected, in a large clot.
Could someone recommend a simple protocol for such a sample--my botanical biases are showing through
thanks is advance
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
We have been using the Pixera system for a little more than a year now. It does color or B/W up to 1200 pixels across. The original cost was around $1200 for camera and interface, I think.
We started with it on a 486-66 and it was a bit slow. We now have it running on a 200 MHz Pentium thru a PCI card and it works okay for a inexpensive solution. The preview is a little slow for focusing but workable. The camera is TWAIN-compliant, sort of. The preview images to other imaging apps have the colors goofed up as far as our version of the software (1.15, I think), but the stored images are ok.
We ordered a 0.5x relay lens from Edmund Scientific for about $230 to adapt the camera to our photo-tubes. We lose some of our field of view (compared to our Polaroid camera) since the Pixera uses a 1/3" CCD, but it is adequate.
At 11:41 AM 12/3/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We also needed a digital camera with good quality image and versatile. After testing many we bought the PHotometrics Sensys. It is monochrome,chilled (10 degrees) 1400 chip. We have been very pleased with the resolution and sensitivity.
Bob
On Wed, 3 Dec 1997, Greg Erdos wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are interested in getting a digital camera for light microscopy. The } simpler, the better. Just want to capture a good quaility image into a PC } or SGI O2 platform that could then be handled by Photoshop or the like. We } are considering both color and B7W only cameras. Cost is a consideration, } as usual. } } I would appreciate opinions from users and information from dealers. } } TIA Greg Erdos } } ******************************************************* } G.W. Erdos, Ph.D. Phone: 352-392-1295 } Scientific Director, } ICBR Electron Microscopy Core Lab } PO Box 118525 Fax: 352-846-0251 } University of Florida E-mail: gwe-at-biotech.ufl.edu } Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ } } ***** } "Many shall run to and fro, and knowledge shall be increased" } Daniel 12:4 } }
Any idea is better than potassium pyroantimonate. Find the nearest friendly electron probe analyst (Peter Ingram or Ann LaFurgey?) and see whether they can help you, but it depends on Ca concentration that you seek. Good luck!
On Wed, 3 Dec 1997 wwiggins-at-carolinas.org wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear All, } Does anyone have a better idea to detect intracellular calcium } than by using the potassium pyroantimonate method as suggested } in Hayat? We've tried it but we're not sure if it's precipitating } calcium or the cesium that we use to induce metamorphosis in our } experimental animals. Our thesis depends on it. Help! } } -------------------------------------------------------- } Name: Winston W Wiggins, Supervisor Vox:704/355-1267 } CRC-Electron Microscopy Lab Fax:704/355-7648 } Carolinas Medical Center Lab:704/355-7220 } P.O. Box 32861 } Charlotte, NC 28232-2861 USA Date: 12/3/97 } E-mail: wwiggins-at-carolinas.org Time: 11:59:18 AM } -------------------------------------------------------- } }
I, and others in my situation, would appreciate hearing of contract labs that perform thin sectioning and other TEM related services. It would be helpful to hear testimonials of satisfaction (or otherwise) as well as names, telephone numbers, turnaround times, pros and cons. TIA.
Steve
Steven Samuelsson, Ph.D. Procter & Gamble Pharmaceuticals, Inc. PO Box 8006 Mason, OH. 45040-8006 (513) 622-1753 office (513) 622-1752 lab (513) 622-1196 fax samuelsson.sj-at-pg.com
You may wish to use scanning probe microscopy as it allows some friction, elasticity and conductivity data as well as molecular imaging at 37 C in bio buffers.
Zhifeng Shao of Virginia is doing some work on muscle, but there is a big difference between muscles in different parts of the body.
George
} From: Katri Vuopala {katri.vuopala-at-juniper.pp.fi} } To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} } Subject: how much is too much lipid in muscle-em? } Date: Tue, 2 Dec 1997 21:26:17 +0200
You might try high resolution in situ SPM as it can give you friction, elasticity, and conductivity data while doing molecular imaging at 37 C in biological buffers. The ease of use (simplicity of sample prep) is often a time saver over SEM.
Zhifeng Shao of U Virginia works on muscle, but there is a big difference between muscles in different parts of the body.
George
} } From: Katri Vuopala {katri.vuopala-at-juniper.pp.fi} } } Subject: how much is too much lipid in muscle-em? } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
} Dear All, } Does anyone have a better idea to detect intracellular calcium } than by using the potassium pyroantimonate method as suggested } in Hayat? We've tried it but we're not sure if it's precipitating } calcium or the cesium that we use to induce metamorphosis in our } experimental animals. Our thesis depends on it. Help! } } -------------------------------------------------------- } Name: Winston W Wiggins, Supervisor Vox:704/355-1267 } CRC-Electron Microscopy Lab Fax:704/355-7648 } Carolinas Medical Center Lab:704/355-7220 } P.O. Box 32861 } Charlotte, NC 28232-2861 USA Date: 12/3/97 } E-mail: wwiggins-at-carolinas.org Time: 11:59:18 AM } -------------------------------------------------------- } Dear Winston,
I recommend the following methods:
1. Bichromate - Probst W., Histochemistry 85, 231-239, 1986.
2. Fluoride I - Ponie and Epel, J Histochem and Cytochem, vol 35,no 9, 939-956, 1987.
3. Fluoride II - Vohringer P., Microscopy Res and Technique, vol 31, 317-325, 1995.
I have been using the bichromate method mostly and with great success. Antimonate is very close in x-ray energy to calcium so it is difficult to support the findings of precipitated calcium by using EDX. This is not a problem with bichromate.
Good luck!
=============================================================== Rune Sundset Dept. of Medical Physiology, Inst. of Medical Biology, University of Tromsoe, N-9037 Tromsoe Phone : +47 77 67 54 42 or +47 77 64 46 96 Fax : +47 77 64 54 40 http://www-users.fm.uit.no/~knutst/medfys/medfys.htm --------------------------------------------------------------- Private Tunveien 21, D-10, N-9018 Tromso,Norway Phone: +47 77 67 45 48 ===============================================================
Hello I am looking for the distributor of AUROBEADS, colloidal gold used for coupling to proteins. It is not longer available from Amersham, but they couldnt tell were to get it now. reinhard
I wanted to find out about the pros and cons of leasing an SEM. I would also like to hear from any vendors off line, who could provide me with some information as well.
Thanks,
Paula
Paula Allan-Wojtas Atlantic Food and Horticulture Research Station Kentville, Nova Scotia B4N 1J5 Canada
I would appreciate if anyone could let me know of companies that do phosphor screen restoration. Please e-mail me directly at my address. Thank you in advance. Cathy Kelloes
I would appreciate if anyone could let me know of companies that do phosphor screen restoration. Please e-mail me directly at my address. Thank you in advance. Cathy Kelloes
We have been using carbon replicas of proteins sprayed or adsorbed on mica. It used to work quite well, but recently we have had problems with floating replicas off the mica support. It is the same batch of mica as before and I do not think we changed anything. Any bright ideas?
Thanks,
Michal
Michal Jarnik Lab of Structural Biology Research, NIAMS, National Institutes of Health, Bethesda, Maryland 20892. Ph: (301) 435-2587
I posted a message yesterday about the ASU Workshop. Apparently some people had problems reading the web page. I apologize. If you want information about the ASU Workshop on In-Situ Electron Microscopy and the ASU Winter School in January 1998, please see the following website: http://www.asu.edu/clas/csss/
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
Our Sorval MT 2B needs an overhaul. Are there any who you would suggest in the proximity of Boston who can service an MT 2B? Thank you. Thane Benson {thane-at-epl.meei.harvard.edu}
We are in the market for a digital image acquisition system to retrofit to our 100CX. Could you suggest vendors who supply such a system? Thank You Thane Benson {thane-at-epl.meei.harvard.edu}
I am looking for sources of LR White embedded human muscle tissue suitable for IEM. The ideal situation would be if someone has, or could prepare, unstained sections on grids. Alternately, if I could locate LR White embedded material I could section it and return the block. Any help would be greatly appreciated.
Brett M. Connolly, Ph.D. Merck Research Laboratories Human Genetics Dept. WP26A-3000 PO Box 4 West Point PA 19486
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I would be interested in your findings. Elaine
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
I too have been looking at low cost options for video/photomicroscopy. = I was ready to go for the color ccd and a Snappy. Then my advisor said = he would consider purchasing a Digital camera of moderate cost. The main problem with the mid-range digitals, as I am sure many of you = are aware, is that they are all fixed lense cameras. I noticed the Kodak = DC120 ($800) because it has fairly high resolution, it has threading on = the front of the lens to accept filters and adaptor lenses, and it also = has a macro mode. When I spoke with their tech. service, they told me = that a photomicroscopy system utilizing the DC120 system should be = released in late December (approximate cost - $2100). The system will = include the camera to C-mount adaptor, cables, power supply and software = for onscreen preview of images. For my application, a nice thing about = this system is that it uses the stock camera. The camera can then be = used for other tasks around the lab.=20 I actually bought the camera on my own to give it a try before trying = to sell the idea to my advisor. Of course, I didn't have any of the = adaptors or the luxury of the onscreen image. The camera easily = balanced atop a widefield ocular lens in the phototube. I was able to = preview pictures in the DC120's lcd display. With a litte trial and = error adjustment of the cameras lens position in macro mode I was able = to achieve simultaneous focus through the binocular and the camera.=20 With this crude system, I was able to get great quality images of = Acanthamoeba under phase contrast. I was most interested and concerned = with the cameras ability to capture fluorescent samples. I did get good = images of AO stains in the range of 1 to 8 second exposure times.=20 With very litte fuss, I was also able to take good quality gel photos = with the camera balanced on top of the polaroid gel hood.=20 My impression was that if you machined your own adaptor, the system = purchase might not be necessary but it would certainly be more = convenient.=20 My advisor was sufficiently impressed, but the verdict is still out on = whether I'll get to order the system.
Kevin Brent Smith - Masters Student University of Louisville Biology Dept. Life Science Bldg. Rm.#12 Louisville, KY 40292 Phone: (502) 852-6773 Fax: (502) 852-0725
We are research microscopists at Polaroid who have been using the Polaroid DMC Digital Microscope Camera in our laboratory since before its introduction in July of this year. We are very pleased with the high resolution digital images acquired with this camera. The DMC is an affordable (just under $6K) high resolution color CCD camera for the light microscope. The million pixel 3/4" CCD camera has a built in C-mount interface, which takes standard 1" or most 2/3" C-mounts. No special adapters are required. It is a SCSI device (no frame grabber board required) and quickly transfers images to your computer without any compression involved. It comes with a TWAIN driver, Adobe Photoshop plug-in for the Mac, and also a stand alone program (PC and Mac) to acquire and perform some image processing on images. The camera captures 24 bit color images in two sizes: a 1600 X 1200 pixel 5.5 MB image, or an 800 X 600 pixel 1.4 MB image. There is also an 8 bit B/W capture mode. The camera driver has a B/W preview mode with up to 5 frames/second capture to aid in positioning and focusing. A focus meter mathematically monitors and helps you optimize focus. Please visit Polaroid's website for more information - www.Polaroid.com.
At 11:15 AM 12/4/97 -0500, Thane Benson wrote: } Our Sorval MT 2B needs an overhaul. Are there any who you would suggest } in the proximity of Boston who can service an MT 2B?
I would recommend Bill McGee. He worked for Dupont-Sorvall when they made this microtome. His company is Microtome Service Company of Liverpool, NY. He can be reached at 315-451-1404.
Best regards, Steven E. Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
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It really depends on what you want to do with the images i.e. print them = out to photo quality printer, etc. We have a 100CX with an image system on it, but could not afford the high = end system. It is likely that this will prevent us from truly using it = for what it was intended although the manufacturer indicated that it = should do the job i.e. print most of our images directly to a Codonics = NP1660 printer. What will you be using it for?
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave. Stockton, CA 95207 209/954-5284 __________________________________________________________________________= _____
We are in the market for a digital image acquisition system to retrofit to our 100CX. Could you suggest vendors who supply such a system? Thank You Thane Benson {thane-at-epl.meei.harvard.edu}
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Hello All,
Does anyone know what has happened to Dr. Om Johari and the journal
{bold} Scanning Microscopy {/bold} . I have been unable to contact him via e-mail, phone,
or fax and have not been able to find reference to the journal in over two
A colleague has a dust problem. He has several mobile carts used to transport plastic components that he wishes to keep free of dust and very clean. The components are kept inside of a 36 cubic inch box on top of the cart. Anyone know of a way to keep the components from attracting dust when the box is opened to remove one of the plastic parts?
Since the carts are going to be moved a lot, the anti-static device should also be mobile. I suggested electrostatic precipitators, but he did not like the idea of high voltages? Likewise, he did not like my idea of using an alpha emitter (Americium, for example). Can't think of much else, however.
Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Gaylord Garroway has rendered routine maintenance service for our Sorvall microtomes (we have 9) for over 20 years. He is very competent, and we have been pleased with his service. He may be reached at 1-508-473-9579 (Milford, MA).
Vachik Hacopian
} Our Sorval MT 2B needs an overhaul. Are there any who you would suggest } in the proximity of Boston who can service an MT 2B? } Thank you. } Thane Benson {thane-at-epl.meei.harvard.edu}
We're setting up a new lab and we will be doing a whole lot of microscopy: light, confocal, EM. We are looking for recommendations on a versatile printer with which we can print publication-quality (at least 600dpi but preferably better) prints. Anyone willing to tell all about their printer? -=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= Paul Tiseo | "It's funny that pirates were always going Mayo Clinic - Jacksonville | around searching for treasure, when they Birdsall 3 | never realized that the real treasure (904) 953-8254 (pager) | was the fond memories they were creating." tiseo.paul-at-mayo.edu | http:// coming soon | - Jack Handey -=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=
No reason why you can't embed the clot to see RBCs, but you won't have many WBCs. To see lots of these, you need a buffy coat. You can either separate them with Lymphocyte Separation Media or Ficoll Hypaque, or you can just let the cells settle out (heparinized) on the benchtop or in a low speed centrifuge. Gently remove the serum and gently add glut without disturbing the pellet. Let fix for a couple of hours and take off the very top layer (either cut the plastic tube with a razor blade, or remove the cells with a Pasteur pipet. This layer will be enriched with WBCs, but will also have many reds. It will appear slightly pinker than the cells in the bottom (a creamy hue). Re-pellet and encase with agar to keep them together as a block while embedding.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
It seems to me that you are forced to go ex situ with SEM?
It would seem that you will loose versatility rather than gain. The current SPM design can give you atomic resolution with STM and AFM, Plus allow control of the environments (PH, Temperature, Electro chem Potential) and work under liquids (wide range of viscosity).
For food studies you get the atomic resolution imaging while you simulate real conditions such as digestion, cooking, freezing, reactions with drugs, alcohol, etc.
This is also the big interest from Biological SEM users moving to SPM for in situ studies on live samples at 37 C.
To my knowledge what may be missing is spectroscopy.
But you gain dramatically by moving to high resolution in situ microscopy.
Let me know about your needs, SPM may be a good alternative.
George
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Dr. Elaine Humphrey wrote: =================================================== I wanted to find out about the pros and cons of leasing an SEM. I would also like to hear from any vendors off line, who could provide me with some information as well. =================================================== As some one who has purchased outright SEMs, bank financed SEMs and leased SEMs over the past almost thirty year period, think I can answer with some amount of experience. And the answer: It is "all in the eyes of the beholder", or putting it another way, it is all a matter of trade offs between interest rates, bank loan rates vs. lease rates. The decision that is the right one at one point in time, might be the wrong decision at some other time.
And this kind of decision might be one way in the USA and it might be an entirely different one in Canada where there are different tax laws and different rules relating to the tax deductibility of lease payments as a business expense.
In general, the "best deal" is to pay cash but you also have to take into consideration the return you would other wise be getting if you left that money invested where it was and you took out a bank loan to pay for the instrument. In other worlds, it all depends on interest rates at the time the decision has to be made.
Leasing, is in general, just a more expensive form of bank financing. It is more expensive because the leasing company has to worry about what they would do if they had to foreclose, say, on someone's TEM. This risk can be partially off-set by a buy-back guarantee from the manufacturer, but it has been our own experience that a manufacturer, since they now have to cover this added "cost" or "risk" of having to undo the sale some day, tends to be a bit less competitive on the final negotiated selling price.
Even the decision whether to use an "open" or "closed ended" lease depends on who is going to carry the burden of risk of predicting fair market value at the end of the lease term.
So the real answer is that there are a number of trade offs, the sum total of which are influenced by a) your country, b) your tax status (e.g. for profit vs. nonprofit vs. government lab), and c) relative lease vs. bank loan rates. This has, for us, always been an exercise for our outside accounting firm and not for us scientists or even our in-house accounting department people.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Does anyone have any experience with analysis of particulates on Nucleopore filters? We have an ongoing project involving particle counts and EDS analysis of air sample particulates collected on Nucleopore filters. For particle counts, I have been cutting out sections of the filters and attaching them with carbon tape to aluminum stubs, then gold coating them. For EDS, I have simply attached cut sections of the filters to clean carbon stubs with carbon tape and viewed/analyzed them using the variable-pressure mode of our SEM. Since we need to image particles in detail at mags up to 20,000x, it's not feasible to do both imaging and EDS on uncoated samples, due to resolution limitations of backscatter imaging and variable pressure conditions.
On occasion, it has seemed that fewer particles are seen on the sputter-coated samples than on the uncoated ones, although they are taken from the same filters from adjacent locations. Since the particles themselves are only attached loosely to the collecting filters (i.e., no special adhesive techniques are used), is it possible that the sputter-coating process can dislodge significant numbers of particles? Needless to say, this could have serious consequences for the data....
Thanks for any feedback on this.
Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
How about slight positive pressure while the cabinet is opened? Pressure could be provided from a small compressed air or nitrogen cylinder. This should ensure that no particles from outside the cabinet would enter - if the opening is not too large. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} } A colleague has a dust problem. He has several mobile carts used to } transport plastic components that he wishes to keep free of dust and very } clean. The components are kept inside of a 36 cubic inch box on top of the } cart. Anyone know of a way to keep the components from attracting dust } when the box is opened to remove one of the plastic parts? } } Since the carts are going to be moved a lot, the anti-static device should } also be mobile. I suggested electrostatic precipitators, but he did not } like the idea of high voltages? Likewise, he did not like my idea of using } an alpha emitter (Americium, for example). Can't think of much else, } however. } } Thanks. } } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } #################################################################### } }
This was my thought also, but keep in mind that the gas in cylinders is not always "particle free" either. I would suggest that you filter the gas just as it enters the cabinet. Rating of the filter should be determined by how small a particle you are concerned with. My bias is that I am a microscopist for a filter company, but I am continually suprised at how dirty many "new" materials are as delivered.
Bob Holthausen Pall Corporation Port Washington, NY
jim-at-proscitech.com.au on 12/05/97 01:11:40 AM
To: bozzola-at-siu.edu cc: microscopy-at-Sparc5.Microscopy.Com (bcc: Bob Holthausen/SLSNY/Pall/US)
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
i do similar analyses in some undergraduate courses at the UofR. the technique we use is to collect on 0.2um pore size 13mm filters. after the particles are collected i take a 1/2" pin mount and coat it with carbon paint. while the paint is still wet i put the whole filter in it...it sticks well and the carbon does not migrate up through the pores (let it air dry in a clean area though). i then coat with either Au or C depending on the intention of EDS work...
i suspect that you may be losing particles in handling; the technique above may avoid some of these concerns (although small particles are pretty intimately attached to the polycarbonate filters)
hope this helps
b-
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
} The main problem with the mid-range digitals, as I am sure many of you are aware, } is that they are all fixed lense cameras.
That is a problem but not the main one. The main one is that most "consumer level" cameras do compression onthe image - usually JPEG - to reduce file size. This is absolutely inimical to subsequently trying to do any serious analysis on the images later - details are altered, moved, etc., brightness and color altered differently in different regions, etc. The "serious" cameras like the Kodak DCS and Polaroid DMC (I use the latter) ship the image to the computer without compression. John Russ
I HAVE A COMMERCIAL STAKE IN WHAT I AM ABOUT TO TELL YOU!
Have you seen/tried the Polaroid DMC? It connects to the microscope via standard C-mount (no lens on the camera, just C-mount thread), creates a TIFF file into your computer at 1600x1200 or 800x600 pixel resolution, and converts quickly and easily for macro work on your copy stand by adding C-mount macro lens. List price under $6K. Details on the Polaroid website at http:\\www.polaroid.com
Hoping this isn't too commercial. The product is still rather new (introduced July '97) and seems directly applicable to your question.
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I too have been looking at low cost options for video/photomicroscopy. I was ready to go for the color ccd and a Snappy. Then my advisor said he would consider purchasing a Digital camera of moderate cost. The main problem with the mid-range digitals, as I am sure many of you are aware, is that they are all fixed lense cameras. I noticed the Kodak DC120 ($800) because it has fairly high resolution, it has threading on the front of the lens to accept filters and adaptor lenses, and it also has a macro mode. When I spoke with their tech. service, they told me that a photomicroscopy system utilizing the DC120 system should be released in late December (approximate cost - $2100). The system will include the camera to C-mount adaptor, cables, power supply and software for onscreen preview of images. For my application, a nice thing about this system is that it uses the stock camera. The camera can then be used for other tasks around the lab. I actually bought the camera on my own to give it a try before trying to sell the idea to my advisor. Of course, I didn't have any of the adaptors or the luxury of the onscreen image. The camera easily balanced atop a widefield ocular lens in the phototube. I was able to preview pictures in the DC120's lcd display. With a litte trial and error adjustment of the cameras lens position in macro mode I was able to achieve simultaneous focus through the binocular and the camera. With this crude system, I was able to get great quality images of Acanthamoeba under phase contrast. I was most interested and concerned with the cameras ability to capture fluorescent samples. I did get good images of AO stains in the range of 1 to 8 second exposure times. With very litte fuss, I was also able to take good quality gel photos with the camera balanced on top of the polaroid gel hood. My impression was that if you machined your own adaptor, the system purchase might not be necessary but it would certainly be more convenient. My advisor was sufficiently impressed, but the verdict is still out on whether I'll get to order the system.
Kevin Brent Smith - Masters Student University of Louisville Biology Dept. Life Science Bldg. Rm.#12 Louisville, KY 40292 Phone: (502) 852-6773 Fax: (502) 852-0725
Paul, I would recommend looking into a Tektronix 450? color wax printer. We have an older 350 on our network that is great for color and greyscale. I believe ours is 300 dpi. 600 dpi files will take up an enourmous amount of memory. Journals will trash your 600 dpi images back to 300 dpi anyway when reducing to half-tone. Only drawback is you cannot write directly on the images Photoshop annotation as a seperate layer works well). Just my .02 worth.
ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/ Privilege does not absolve one of ecological responsibility.
At 10:20 PM 12/4/97 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I do SEM/EDX particle analysis on a fairly regular basis. I had also found that samples which I had sputter coated had fewer particles. I now use just sticky carbon tabs for most of my particles, but the ones in liquids I first pick up on a grid, and then mount on the sticky carbon. These can be sputter coated without losing the particles. If you absolutely have to collect your particles on a Nucleopore, you might try collecting as usual, and then using a conductive tape/tab to pick up the particles from the Nucleopore, then mounting the conductive tape/tab.
Stephanie Wind McCray Process Chemist Moltech Corp. 9000 S Rita Rd, Bldg 61 Tucson, AZ 85747 520-799-7631 (office) or 520-799-7535 (lab) wind-at-moltech.com
} Hi, } } Does anyone have any experience with analysis of particulates on Nucleopore } filters? We have an ongoing project involving particle counts and EDS } analysis of air sample particulates collected on Nucleopore filters. For } particle counts, I have been cutting out sections of the filters and } attaching them with carbon tape to aluminum stubs, then gold coating them. } For EDS, I have simply attached cut sections of the filters to clean carbon } stubs with carbon tape and viewed/analyzed them using the variable-pressure } mode of our SEM. Since we need to image particles in detail at mags up to } 20,000x, it's not feasible to do both imaging and EDS on uncoated samples, } due to resolution limitations of backscatter imaging and variable pressure } conditions. } } On occasion, it has seemed that fewer particles are seen on the } sputter-coated samples than on the uncoated ones, although they are taken } from the same filters from adjacent locations. Since the particles } themselves are only attached loosely to the collecting filters (i.e., no } special adhesive techniques are used), is it possible that the } sputter-coating process can dislodge significant numbers of particles? } Needless to say, this could have serious consequences for the data.... } } Thanks for any feedback on this. } } Randy Tindall } 2017 Princess Jeanne } Las Cruces, New Mexico 88001-4157
Hi Randy, A trick I have had some success with is to pre-coat the filters with a conducting metal, Au or AuPd before collecting particulates. If there is peak overlap you might even try a Cr replicate. It does a great job of making polycarbonate filters conductive, I also attach them with Ag paint. Since we're using a mass spec technique (TOF-SIMS) the carbon tape adhesive shows up. Another advantage to the AuPd pre coating is that I can use the peaks as a SIMS calibration aid.
I have used this to investigate uncoated, unfixed, freeze dried yeast cells with both LVFESEM (2kV) and TOF-SIMS. I can't say that I've tried EDS on them. The SEMs come out great up to about 9000x. I've posted some images on my web-site, follow the links to "Research Projects" then to "Sample prep for LVSEM and TOF-SIMS".
good luck ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/ Privilege does not absolve one of ecological responsibility.
I' tryng to collect all info possible about "Image Plates" devices.
I know that FUJI had in the past a sytem called FDL 5000. (I have a 1 page leaflet). Fuji, in Italy does know nothing about this produc. Can anyone help me with more info, approx. price, where to find it, etc.
Regards to everybody - Have a nice Christmas and happy New Year !
PLEASE REMOVE "_NO_SPAM" and "NO_SPAM_" before and after -at- symbol to reply me.
We recently purchased the Epson Sylus Photo printer (only costs about $500) to use as an intermediate-level printer. However, we have been pleasantly surprized at the quality we get when using the Epson Photographic Quality Glossy *Film*. A colleague recently submitted such prints of histology samples for publication.
The printer resolution goes up to 720 dpi, and there are 6 color jets vs. the usual 4. It is a little slow (takes about 5-8 minutes to do high resolution full sized prints), but we don't have a large volume so that's OK. The only problem we've had is getting the color settings to produce consistent white backgrounds at the same time as producing reasonable representation of reality. This is an issue not only of printer settings but computer software, monitor, etc. However, even when the image on the monitor is reasonable, the printout can be quite different. Takes a bit of fiddling to get desired color representation.
The usual disclaimer: no connection to Epson other than customer.
Karen
tiseo.paul-at-mayo.edu wrote: } } Hi, } } We're setting up a new lab and we will be doing a whole lot of } microscopy: light, confocal, EM. We are looking for recommendations on a } versatile printer with which we can print publication-quality (at least } 600dpi but preferably better) prints. Anyone willing to tell all about their } printer? } -=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= } Paul Tiseo | "It's funny that pirates were always going } Mayo Clinic - Jacksonville | around searching for treasure, when they } Birdsall 3 | never realized that the real treasure } (904) 953-8254 (pager) | was the fond memories they were creating." } tiseo.paul-at-mayo.edu | } http:// coming soon | - Jack Handey } -=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN 55144 "Opinions above are my own, not necessarily my employer's"
Microscopy members, I am finishing a paper on the "Evolution of microscopy in the XX century", (in hypertext) and have been unable to find pictures of Zworikin and Oatley in the locally available literature. If anyone can help me by attaching a .jpg, tif or similar picture to email, I would be very grateful.
please answer directly to
wamann2-at-metalmat.ufrj.br
Prof. Walter A. Mannheimer Dept. of Metallurgy and Materiais Eng. Federal University of Rio de Janeiro POBox 68505, 21945 Rio de Janeiro, Brazil Vox (55 21) 590-0579 Fax (55 21) 290-6626 wamann-at-metalmat.ufrj.br
} } Hi there we are looking for a very small objective aperture for our JEOL } 2000FX. Since these apertures come in strips there is a limited selection } and the "standard" set comes with a 20micorn aperture as the smallest. We } can get a 10micron from JEOL, does anyone know of a smaller one say } 5microns? } I have not called all of the Microscope Spares and Equipment suppliers yet, } I thought I would solicit commnets for the community. Many thanks. } Reply by email and I will summarize to the list if there is sufficient } interest and response. } } Cheers } } Jfm.
} Dear John,
Ladd has been producing Apertures for over forty years now. Since we produce them ourselves we can give you any hole size that you wish, within certain technical limitations. Please e-mail me with the size of all the holes you would like on the strip and the material, I suspect PT, and we will send you a quote.
Best Regards,
JD Arnott Ladd Research 13 Dorset Lane Williston, VT 05495
TEL 1-802-878-6711 worldwide 1-800-451-3406 US, Canada Fax 1-802-878-8074
Ann Fook Yang wrote: ================================================ I am planning to do immunogold on brassica pollen grains. I am looking for an adhesive that remain tacky when dried and would hold pollen through out the process of washing, fixing, immuno-treatment, postfixing, dehydrating and critical point drying.
Any suggestions will be appreciated. Thanks. ================================================ You might want to consider trying SPI's Tacky Dot(TM) Slides which can be found on our website (along with an example of their use) at URL
http://www.cccbi.chester.pa.us/spi/new/tacky.html
If the pollen is reasonably free flowing, one grain (or possibly several) will end up "sticking" per dot and since there is a build up of strength of the adhesive bond with time (as is the case for most adhesives), after about 48 hours it is at its maximum. For most particles, the bond is reasonably resistant to water, especially if not subjected to turbulent conditions. If you can do the preparation on the slides, the end result will be infinitely more easy to characterize with any kind of microscope than if the pollen grains are in some random distribution on a substrate.
The "adhesive" is a dry adhesive, there is no chance of any particle sinking into it, and there is nothing to off-gas to contaminate a vacuum system. One mounted, so long as the slides are stored long term under dry conditions as would any other SEM specimen, their life time seems to be indefinite.
Disclaimer: SPI Supplies manufactures Tacky Dot Slides under license from DuPont, the patent holder and we therefore would like to see more people using them! We are unaware of any other product of this type in the world.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
One of the cheapest and simplet "glues" for pollen grains is to dissolve the glue from about a 30cm length of Scotch tape in about 10ml of chloroform. Apply this to a clean abd shiny stub allow to d5ry and sprinkle on or place pollen grains on the surface. Dry6 overnight in a 35oC oven, loghtly coat 8-10nm Au/Pd and away you go.
Patrick Echlin Cambridge UK
On Sun, 30 Nov 1997, Garber, Charles A. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Ann Fook Yang wrote: } ================================================ } I am planning to do immunogold on brassica pollen grains. I am looking for } an adhesive that remain tacky when dried and would hold pollen through out } the process of washing, fixing, immuno-treatment, postfixing, dehydrating } and critical point drying. } } Any suggestions will be appreciated. Thanks. } ================================================ } You might want to consider trying SPI's Tacky Dot(TM) Slides which can be } found on our website (along with an example of their use) at URL } } http://www.cccbi.chester.pa.us/spi/new/tacky.html } } If the pollen is reasonably free flowing, one grain (or possibly several) } will end up "sticking" per dot and since there is a build up of strength of } the adhesive bond with time (as is the case for most adhesives), after about } 48 hours it is at its maximum. For most particles, the bond is reasonably } resistant to water, especially if not subjected to turbulent conditions. If } you can do the preparation on the slides, the end result will be infinitely } more easy to characterize with any kind of microscope than if the pollen } grains are in some random distribution on a substrate. } } The "adhesive" is a dry adhesive, there is no chance of any particle sinking } into it, and there is nothing to off-gas to contaminate a vacuum system. } One mounted, so long as the slides are stored long term under dry conditions } as would any other SEM specimen, their life time seems to be indefinite. } } Disclaimer: SPI Supplies manufactures Tacky Dot Slides under license from } DuPont, the patent holder and we therefore would like to see more people } using them! We are unaware of any other product of this type in the world. } } Chuck } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== }
I'm not usually one for "me too" posts, but all you people who are shelling out thousands of dollars for Textronics-level printers to make "publication quality" prints ought to heed Karen Zaruba's (and my) words. Today, the best bang-for-bucks is acheived with the Stylus Photo. If you find the Epson Glossy Film a bit pricey (at roughly $2.50/sheet, I do), you can go to Kodak Inkjet Photo Quality paper (it looks and feels like photo paper) for about one-third that cost and not lose much definition. The prints that come off the Epson device are publication quality - I defy anyone call them "unacceptable" especially given the previous post on what actually goes on in the transfer to print! I will submit some next week for publication and I plan not to say a word about how they were generated - let the printer complain (if they even notice). The only thing noticeably better than these prints is the digital image itself. Rob Palmer CEB/UT
Re the comment "Takes a bit of fiddling to get desired color representation." (Seems like I may have heard that once or twice before!)
One of the big benefits I hear for the Codonics printer is their "bracketing" feature, where it will print series of thumbnails of an image, varying several combinations of settings such as gamma, so that the best-appearing combination can be chosen easily without guessing. Does anyone know if such a program (maybe a RIP, Raster image processor program?) exists that can be used with other printers, e.g. Epson or HP? Any comments on price/benefit?
Thanks Richard Thrift DepoTech Corp. Richard_Thrift-at-DepoTech.com
} } } {kszaruba-at-MMM.COM} 12/05/97 09:40am } } } . . . We recently purchased the Epson Sylus Photo printer . . . . . . The only problem we've had is getting the color settings to produce consistent white backgrounds at the same time as producing reasonable representation of reality. This is an issue not only of printer settings but computer software, monitor, etc. However, even when the image on the monitor is reasonable, the printout can be quite different. Takes a bit of fiddling to get desired color representation.. . .
} Have you seen/tried the Polaroid DMC? It connects to the microscope } via standard C-mount (no lens on the camera, just C-mount thread), } creates a TIFF file into your computer at 1600x1200 or 800x600 pixel } resolution, and converts quickly and easily for macro work on your } copy stand by adding C-mount macro lens. List price under $6K. } Details on the Polaroid website at http:\\www.polaroid.com
How does this differ from the PDC-2000/T which, I believe, is much cheaper?
Article posted on Lindsay's Lab at ASU web shows force data of 7 liquid molecular layers at the solid liquid interface, molecular orientation, and molecular deformation based on position.
This combined with in situ control of environments should be the enabling technology for molecular tribology studies.
For corrosion and tribology studies you can get atomic imaging, as well as several types of force data LFM (lateral force) MAC Force (direct vertical force), MAC Phase data, and MAC Phase with fractionalized tips (chemical force)
George
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Will you be at Cell Biology show in Washington D.C., December 15 - 18?
If you are please come to talk to us about Scanning Probe Microscopy. We will be conducting training and "live" demonstrations of a technical breakthrough for in situ high resolution biological imaging.
George
At 11:02 AM 12/4/97 -0600, henk-at-vt8200.vetmed.lsu.edu wrote:
} } } }
{excerpt} ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello All,
Does anyone know what has happened to Dr. Om Johari and the journal
{bold} Scanning Microscopy {/bold} . I have been unable to contact him via e-mail, phone,
or fax and have not been able to find reference to the journal in over two
years. Have he and the journal "retired"?
Bill Henk
Dept. of Anatomy & Cell Biology
LSU School of Veterinary Medicine
Baton Rouge, LA 70803
phone -(504)346-3237
e-mail - henk-at-vt8200.vetmed.lsu.edu
{/excerpt} { { { { { { { {
F.Y.I. ASU/MI Winter Microscopy Workshop is hosting its annual AFM in Biology Hands-on Workshop. It will be held February 9th to 11th, 1998. This is a wonderful opportunity for someone to learn the fundamentals of AFM. On the second day of the workshop, participants will get a chance to image atoms and molecules. Participants are encouraged to bring their own samples for free testing.
I have used Me salicylate to clear whole (gutted, 100mm sculpins) fish. This was a pretty simple procedure, just dehydrate the fish in an ethanol series, then transfer to 100% Me salicylate through a 3:1, 1:1, 1:3 EtOH:Me sal. series. If parts stay cloudy, it is probably because they weren't completely dehydrated. I used 1 to 3 days in each step, tissue pieces would take less. When finished, the fish almost looked like they were made of glass.
Phil
} From: Ramin Rahbari 313 998-3383 {rahbarr-at-aa.wl.com} } } I am interested to hear from individuals that have used methyl salicylate (? } conc.) to clarify tissue in particular skin. }
Philip Oshel PO Box 5037 Station A Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu or poshel-at-hotmail.com ***** looking for a job *****
Disclaimer: I do represent a business that sells computers and their parts. We build and repair computers.
Where did you get the price figures for the hard drives? If you are talking about IDE, you are several years behind in prices. 6.0 Gig HDD's only retail for about $300.00 or so. Look around. The price of things may surprise you.
SCSI HDD's are sinking in price as of early summer. The price is very affordable.
The best way to back up is still via a second HDD. It is less expensive than most alturnatives and there is no media to concern yourself with. Take care of it and keep it clean and it should last for a long time.
Don't forget that we are on the horizon of 1.0 Gig floppy drives.
Things are getting better and better as well as less expensive.
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Hello All I am sorry - the message with above subject was dedicated to Microprobe list. regards Krzysztof M.Herman LabSoft Sp.C. 21 Kosciuszki Str. 05-500 Piaseczno, Poland tel/fax: (48 22) 7502024, 7502028, 7570671 fax: (48 22) 483787, mobile (48 90) 213438 E-mail: labsoft-at-labsoft.com.pl http://www.labsoft.com.pl/ ------=_NextPart_000_01BCFD25.4B37D9E0 Content-Type: text/html; charset=ISO-8859-2 Content-Transfer-Encoding: base64
If you be at Cell Biology show in Washington D.C., December 15 - 18, please come to talk to us (booth # 253) about Scanning Probe Microscopy.
We will be conducting training and "live" demonstrations of a technical breakthrough for in situ high resolution biological imaging.
George
F.Y.I. ASU/MI Winter Microscopy Workshop is hosting its annual AFM in Biology Hands-on Workshop. It will be held February 9th to 11th, 1998. This is a wonderful opportunity for someone to learn the fundamentals of AFM. On the second day of the workshop, participants will get a chance to image atoms and molecules. Participants are encouraged to bring their own samples for free testing.
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Paul, Don=92t like using their name so let=92s say Evex & similar=3D the gener= al term "Slime-x". I suspect that their product line consist of a broad, very broad but shallow array of airware, little else. No idea what their claims are. No reason to believe what I=92d see or hear. Don=92t care. I am glad to see this subject up again. During the intense frothing over the Slime-x ethics, it seemed the common denominator was frustration at the lack of punitive actions available. No cyber bullets....wrong!.... information is a wonderful thing. Well folks here is my shot. Our institution maintains a list of preferred suppliers as well as problem children. They make a reasonable effort to be fair in assessments. I supplied a sampling of the better researched messages, their unsigned responses & a memo suggesting that a company that behaved in this manor could not be trusted at any level. This is not an absolute fix but is progress. BANG I believe the we have an obligation to protect our counter parts in the field, most of which I dare to say do not subscribe to this list. For now, the low life ethical standards of Slime-x are an issue to us, probably to no one else. Many of our students are potential customers of high tech. hardware. Their decisions will reflect on & may impact us directly. Down the road, they/we may be oblivious to this companies ethical status. Any of you who have centralized purchasing or preferred vendor list can help many of our associates now & in the future by doing as I have done. Document the demon. Naturally there will be others who follow in the footsteps of Slim-x but I think a good public spanking is in order and a fairly effective deterrent to others.
from my extream side: in a loose paraphrase of George Patton in Hollywood... when you put your hand it the goo that used to be your funding's face you=92ll know how to kill the enemy, shoot them in the belly ....
Are there any Zeiss Wizards out there that can help me with a few questions concerning a Zeiss UNIVERSAL 'M' microscope ?
1-What are the electrical demands of a GLAREX viewing hood, and what does the electrical attachment do ?
2-What is the purpose of the light bulb on the 35mm camera attachment ?
PARTS WANTED:
1-Adaptor ring to attach light source for reflected light microscopy on back of microscope. ie: Adaptor ring 3.4 mm (internal thread) {-} 4.3 mm bayonet mount or Adaptor ring 3.4mm (internal thread) {-} 3.3 mm bayonet mount
2- Extension tubes (12 mm) for EPIPLAN lenses with M24mm thread
3- Low power (1 to 8x) EPIPLAN, EPIPLAN LD or EPIPLAN HD lenses
4- H-Pr-POL reflector for vertical Illuminator IIC and EPIPLAN POL lenses
ZEISS parts for sale/swap
1- Vertical Illuminator IIC (lens mount does not hold 'quick change ring') 2- Adaptor ring (brass, home made, 4.2mm thread (externior thread) {-} 4.3 mm bayonet mount.
OR, do you know the name, address, telephone number, e-mail of some one that does.
Hello!: I was wondering if one of your associates might help me. I have an older (20-30 years) microscope. It is a Spencer/American Optical, bifocal, oil immersion, serial number 155715, but with no other type number. The instruction manual was lost (prior owner) many years ago. Is there anyone out there who might have a copy of the manual that they might share with an ignorant biochemist? Please help. E Mail defilip1-at-flash.net. Thank you
I have found that floating carbon off mica, as with floating Formvar off glass slides, seems to be affected by a combination of the alignment of the planets, the phase of the moon and the day of the week :-)
Other factors appear to be: Humidity of the air in the room (if too dry, no good); Surface condition of the mica (I rinse in alcohol and acetone after I cleave the mica); Condition of the evaporation system (possible oil contamination etc.).
Mostly though I just wait a day or two and try again. Usually works.
JUST A QUICK LETTER TO SHOW YOU SOME LASERS- OPTICS AND OPTICAL TABLES SURPLUS THAT WE JUST RECEIVED.
ITEM TRIMMU12 14 WATT ARGON LASER MADE FOR HEART SURGERY, TRIMEDYNE MODEL 900 TEMOO, POLORIZED,220VAC INPUT , WATER COOLED , FIBER LAUNCH, ALL ON ROLLAROUND CART EXCELENT FOR LAB USE, THE POWER WAS MEASURED AT 13 TO 14 WATTS. PRICE $9500 12 MONTH WARRANTEE.
ITEM: COHERENT ARTICULATING ARM FROM A MODEL 451 CO2 MEDICAL LASER. ECCELLENT COND. $200
ITEM CO220A: CO2 LASER MADE BY PFIZER ,1990, FOR SURGERY, TATTOO REMOVAL ECT. 20 WATT OUTPUT , TESTED AND IN EXC. COND. 110 VAC INPUT, COST $40,000 NEW OUR PRICE 4,900. MODEL 20-C
ITEM:PDA-1U1 SPECTRA PHYSICS QUANTRA RAY PULSED DYE LASER , GOOD FOR SPARE PARTS MODEL PDA-1 $500
ITEM NEWU1 NEWPORT OPTICAL TABLE 16" BY 36" 4" THICK, 1 " HOLE SPACING, COMES WITH A RUBBER ISOLATED TABLE STAND, NOT AIR SUPPORTED, $750
ITEM: HEPSN1 HELIUM NEON POWER SUPPLY KIT OPERATES UP TO A 15 mW LASER, INCLUDES ALL COMPONENTS AND PRINTED CIRCUIT BOARD, ALL YOU HAVE TO DO IS STUFF AND SOLDER THE CIRCUIT BOARD . 4" BY 3" BY 3", PRICE $75
ITEM HENEU12 1 TO 1.5 MW HE-NE LASER 632.8 nM INCLUDES 12VDC INPUT POWER SUPPLY ALL IN A PLASTIC HOUSING 6.25 IN. BY 1.375IN BY 2.25 IN. TEMOO,RANDOM POL. ,1.7 MR DIVERGENCE. 12 MONTH WARRANTEE , PRICE $45
ITEM MELU12 1 TO 2 mW HE-NE LASER 632.8 NM , PULLS FROM MEDICAL EQUIPMENT .EACH UNIT INCLUDES HE-NE HEAD AND POWER SUPPLY[110VAC INPUT]. ALL YOU NEED TO PROVIDE IS A POWER CORD AND A FUSE TO MAKE THE UNIT OPERATIONAL. THE BEAM IS TEM00, POLORIZED WE WILL COVER EACH UNIT WITH A 12 MONTH UNLIMITED HOUR WARRANTEE, EXCELLENT FOR FOR LAB OR HOME USE. NEW THESE COST APPROX. $350 OUR PRICE $85. DIMENSIONS 9.75 BY 1.25 INCHES, P.S. 4.25 BY 3.25BY 1.25 INCHES.
ITEM RAMCNS1: RAMAN CELL OPTICS 308 nm AR/AR 4600 A 0=0 DEGREES 1000 MM FL. 2" DIA. NEW. ORIGINAL PRICE $520 OUR PRICE $175
ITEM TFPOLNS1: POLARIZERS , THIN FILM FOR 532 nm , NEW, ORIGINAL COST $590 EACH OUR PRICE $200 EACH 10 MM DIA.
ITEM CO2OCNS1: CO2 HIGH REFECTOR AND OUTPUT COUPLER 10.5 MM DIA, OC =79%R NEW. $200 A SET.
ITEM 25MNS1: DIELECTRIC BROADBAND MIRRORS 450 TO 700NM , NEW WITH PLASTIC PROTECTIVE COATINGS , 2 SIZES 25 MM SQ. AND 50 MM SQ. RECOMENDED FOR HIGHER POWER LASERS.
ITEM # BSDNS1: 50/50 DIELECTRIC COATED PLATE BEAM SPLITTER 630 TO 660 NM COMES IN A TRIANGLE SHAPE EACH SIDE APPROX. 1" PRICE $20
ITEM # 45NS1 45 DEGREE RED REFLECTOR , PASSES 488 TO 532NM , CAN BE USED TO COMBINE RED AND GREEN/BLUE LASERS TO CREATE A WHITE LIGHT LASER. 1" SQ. PRICE $15
ITEM# PCINS1 PLANO/CONVEX LENS COATED FOR YAG 1064NM , AR COATED, 10MM DIA. NEW, ORIG. COST $250 OUR PRICE $100
ITEM# INFILTER : INTERFERENCE FILTERS USED FOR PASSING A PARTICULAR SPECTRAL LINE , 11.8 MM DIA. CAREFULLY REMOVED FROM MEDICAL EQUIPMENT AND WRAPPED IN LENSE PAPER. THE FOLLOWING WAVE LENGTHS ARE AVAILABLE. 523.5, 547.4 , 572.1, 512.9, 550.6, 488, 505.7 nm price $20 each.
FOR A COMPLETE LINE OF NEW AND USED LASERS - OPTICS -ELECTRO OPTICS- LASER SHOWS ORDER A COMPLETE CATALOG AT MWKINDUSTRIES.COM
TO: ORDER GO TO OUR WEB SITE MWKINDUSTRIES.COM {SECURE ORDERING SITE}
QUESTIONS OR REMOVAL FROM MAILING LIST EMAIL: MWK-at-WORLDNET.ATT.NET
I am interested in visualising cellulose microfibrils and microtubules/microfilaments in wood-forming cells of trees in the confocal microscope. I use FITC-labelled antibodies for cytoskeleton and would like to stain the cellulose with a fluorescent dye. If I had access to a UV laser I would have no hesitation in using calcofluor/tinopal, but I don't. So, can anybody suggest a visible light-excited fluorochrome that will localise the cellulose and will permit imaging of both cellulose and cytoskeleton? (If it helps we have a Zeiss 510 with 488, 568 and 633 nm lines.)
I thank you in advance,
Yours sincerely,
Nigel Chaffey
----------------------------------------------------- Dr Nigel Chaffey, Dept Forest Genetics & Plant Physiology, Swedish University of Agricultural Sciences, S-901 83 Ume=E5, Sweden Phone: +46-90-786-6305 =46ax: +46-90-786-5901 eMail: nigel.chaffey-at-genfys.slu.se
I am interested in visualising cellulose microfibrils and microtubules/microfilaments in wood-forming cells of trees in the confocal microscope. I use FITC-labelled antibodies for cytoskeleton and would like to stain the cellulose with a fluorescent dye. If I had access to a UV laser I would have no hesitation in using calcofluor/tinopal, but I don't. So, can anybody suggest a visible light-excited fluorochrome that will localise the cellulose and will permit imaging of both cellulose and cytoskeleton? (If it helps we have a Zeiss 510 with 488, 568 and 633 nm lines.)
I thank you in advance,
Yours sincerely,
Nigel Chaffey
----------------------------------------------------- Dr Nigel Chaffey, Dept Forest Genetics & Plant Physiology, Swedish University of Agricultural Sciences, S-901 83 Ume=E5, Sweden Phone: +46-90-786-6305 =46ax: +46-90-786-5901 eMail: nigel.chaffey-at-genfys.slu.se
Below is the result of your feedback form. It was submitted by (Barbara308-at-aol.com) on Saturday, December 6, 1997 at 20:24:01 ---------------------------------------------------------------------------
Email: Barbara308-at-aol.com Name: Barbara Oakley
School: Oakland University
State: Michigan
Zip: 48317
Question: Dear Madame or Sir,
My name is Barbara Oakley--I'm a grad student at Oakland University in Rochester, Michigan. I've been given a budget of $25,000 to buy a new metallograph for our material science laboratory. Can you give me any advice? Right now I'm looking at the PME3 from Leco....
Below is the result of your feedback form. It was submitted by (bhaab-at-zinc.cchem.berkeley.edu) on Friday, October 31, 1997 at 14:12:11 ---------------------------------------------------------------------------
Email: bhaab-at-zinc.cchem.berkeley.edu Name: Brian B. Haab
School: U.C. Berkeley
State: CA
Zip: 94720
Question: Hi,
My question has to do with imaging and the thickness of the cover plate used. How important is it to use a cover plate thickness for which the objective was specifically designed? For example, if I'm using an objective which says 0.17 (presumably corrected for a 170 micron cover slip thickness), would image quality be greatly distorted when using something, say, twice as thick? Also, what is the definition of "working distance?"
Louis DeFilippi (by way of Nestor J. Zaluzec) wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello!: I was wondering if one of your associates might help me. I have an } older (20-30 years) microscope. It is a Spencer/American Optical, bifocal, } oil immersion, serial number 155715, but with no other type number. The } instruction manual was lost (prior owner) many years ago. Is there anyone } out there who might have a copy of the manual that they might share with an } ignorant biochemist? Please help. E Mail defilip1-at-flash.net. Thank you } } Louis Dear Louis,
I have some older information on AO microscopes. If you can send a more complete description (size, color, any other markings on objectives, stand, etc.), I will try to see if it matches any of our literature.
Even better, if you can send a Polaroid print to our offices.
Thanks, Barbara Foster Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
I am trying to make a TEM specimen of Diamond, I have a Gatan PIPS at my disposal, but to use it my Diamond wafer has to be less than 100 microns thick. At the it is 1mm thick. Any suggestions on how to thin the Diamond wafer? The Diamond is actually poly-crystaline CVD
Thank you in advance Adam Dr Adam Papworth Dept Materials science & Engineering Ashton Building The University of Liverpool L69 3BX
Phone No 0151 794 5372 Fax No 0151 794 4675 E-Mail adamp-at-liv.ac.uk
Hello Michel...are you evaporating Carbon or a Pt-C mixture for your replicas? I can only comment on my experience using Pt-C. If your buffer has changed, this may affect the ability of the replica to release. High salt or EDTA causes difficulties. We routinely use 0.1 M ammonium bicarb or 1% acetic acid. You might try exposing the mica to the vapors of 1% acetic acid, in a closed container, following evaporation. This works wonders for releasing otherwise difficult replicas.
Good Luck,
Doug Keene Shriners Hospital Research ---------------------- Doug Keene DRK-at-shcc.org
} My question has to do with imaging and the thickness of the cover plate used. } How important is it to use a cover plate thickness for which the objective } was specifically designed? For example, if I'm using an objective which } says 0.17 (presumably corrected for a 170 micron cover slip thickness), } would image quality be greatly distorted when using something, say, twice } as thick?
The cover plate thickness is the distance between the top of the cover plate and the specimen so it includes the mounting medium AS WELL AS the thickness of the cover plate itself. The refractive index of the mounting medium can therefore be quite important. AND the depth of the mount.
If the mount/plate is twice as thick you will see severe distortion of the image with the periphery quite out of focus.
Cover plate thickness does not much affect oil immersion lenses as the oil optically bonds the front element of the lens to the plate. For this reason all 100x and some very good 63x and 40x objectives are used with oil immersion. But it is very important for "high dry" lenses where you are hoping for good images. The best high dry (63x, 40x) objectives have a correction collar which you can adjust to compensate for differing cover plate/mounting medium thickness. If you don't have one of these, you have to be careful to use a cover plate of the thickness the lens is corrected for
Also, what is the definition of "working distance?"
Its the distance between the front lens of the objective and the top of the cover plate.
} } } } } --------------------------------------------------------------------------- } } } } Mel Dickson Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
I received this directly from Dennis. It should interest anyone who is doing microscopy outreach, particularly if it's with SEM, so I'm taking the liberty of placing it on the listserver:
Dear Friends and Associates, Discovery Channel's documentary series, "Movie Magic", is airing a segment called "Far Out Creatures" that features special effects used in the making of science fiction films (Alien Resurrection and Starship Troopers). Part of the 1/2 hour segment will include a short interview with me and some of my "MicroAliens". I thought this might be of interest to you.
The segment will run: December 11, 1997 9:30 - 10:00 PM - West Coast Time December 12, 1997 1:30 - 2:00 AM - West Coast Time December 13, 1997 2:00 - 2:30 PM - West Coast Time
Please check your local listing for the time of Movie Magic on the Discovery Channel for these days.
Best Regards, Dennis Kunkel
*********************************************** * Dennis Kunkel Ph.D. * * Pacific Biomedical Research Center * * University of Hawaii * * * * email - kunkel-at-pbrc.hawaii.edu * * www - http://www.pbrc.hawaii.edu/~kunkel/ * ***********************************************
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
bhaab-at-zinc.cchem.berkeley.edu wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Below is the result of your feedback form. It was submitted by } (bhaab-at-zinc.cchem.berkeley.edu) on Friday, October 31, 1997 at 14:12:11 } --------------------------------------------------------------------------- } } Email: bhaab-at-zinc.cchem.berkeley.edu } Name: Brian B. Haab } } School: U.C. Berkeley } } State: CA } } Zip: 94720 } } Question: Hi, } } My question has to do with imaging and the thickness of the cover plate used. } How important is it to use a cover plate thickness for which the objective } was specifically designed? For example, if I'm using an objective which } says 0.17 (presumably corrected for a 170 micron cover slip thickness), } would image quality be greatly distorted when using something, say, twice } as thick? Also, what is the definition of "working distance?" } } Thank you very much, } } Brian Haab } } --------------------------------------------------------------------------- Dear Brian, Coverslips are considered part of the optical design of a microscope. If the objective carries the marking "0.17", it expects to see a coverslip of approximately that thickness on your sample. While it may tolerate a slight deviation (0.15-0.18), the image may be seriously degraded outside those limits. By the way, as you might have noticed, there is very little information in the catalogs relating this thicknessness to the ordring specifications. For 0.17mm, order a #1 1/2 coverslip.
Regarding working distance: it is open distance or space between the front lens of the objective and the top of the preparation. Condensers also have working distances": from the top element of the condenser to the bottom of the slide/preparation.
We cover all of this and much more in our book, "Optimizing Light Microscopy for Biological and Clinical Laboratories". If you would like to order one, email me and we will forward an electonic order form.
Hope this helps.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
Barbara308-at-aol.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Below is the result of your feedback form. It was submitted by } (Barbara308-at-aol.com) on Saturday, December 6, 1997 at 20:24:01 } --------------------------------------------------------------------------- } } Email: Barbara308-at-aol.com } Name: Barbara Oakley } } School: Oakland University } } State: Michigan } } Zip: 48317 } } Question: Dear Madame or Sir, } } My name is Barbara Oakley--I'm a grad student at Oakland University } in Rochester, Michigan. I've been given a budget of $25,000 to buy a new } metallograph for our material science laboratory. Can you give me any } advice? Right now I'm looking at the PME3 from Leco.... } } I'd appreciate any help you could provide. } } Barb Oakley } } --------------------------------------------------------------------------- Dear Barb,
Most of the microscope companies carry superb metallographs. What do you need in the way of specific functionality (i. e., camera ports, magnification, upright vs. inverted, ability to project reticles for visual measurement, etc.).
In your budget range, you probably would do well to consider the Olympus (sold through LECO) or a Unitron system. Another alternative is a good quality used Reichert MEF3 or MEF4. If you need contacts, phone numbers, etc., please email me.
Send me a copy of your specifications and applications and I will see who else might have what you are looking for.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
Valdre' Andrea wrote: } } } I' tryng to collect all info possible about "Image Plates" devices. } } I know that FUJI had in the past a sytem called FDL 5000. } (I have a 1 page leaflet). Fuji, in Italy does know nothing about this produc. } Can anyone help me with more info, approx. price, where to find it, etc.
Try http://home.fujifilm.com/info/products/science/ip/index.html I hope that helps.
Philip -- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
Does anyone know of any commercially available ABs that react with cytoskeletal elements in cereal plants? We wish to do immunofluorescence but have failed using commercially available alpha- and beta-tubulin monoclonal ABs (N356 and N357 ABs from Amersham). We have done Western blots and these do not appear to recognize the respective tubulins in rice. Any advice? Thanks.
Tim Bourett DuPont Experimental Station Wilmington, DE USA
I do not have any experience with the Leco line of microscopes but I have used others. We currently have a Nikon Epiphot that I think performs better than most of the other metallographs that I have used. Make sure you get demos and have plenty of samples to try test on them before you buy. You should alos get a box of film and record an image from each one. Good luck.
I don't know about wood but in human tissue we use a .01% Evans Blue as a total protien counterstain in conjunction with FITC tagged primary. The evans blue excites and emmits like rodamine or texas red.
Bob
On Sun, 7 Dec 1997, Nigel Chaffey wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } =20 } Dear Fellow Microscopists, } =20 } I am interested in visualising cellulose microfibrils and } microtubules/microfilaments in wood-forming cells of trees in the confoca= l } microscope. I use FITC-labelled antibodies for cytoskeleton and would li= ke } to stain the cellulose with a fluorescent dye. If I had access to a UV } laser I would have no hesitation in using calcofluor/tinopal, but I don't= =2E } So, can anybody suggest a visible light-excited fluorochrome that will } localise the cellulose and will permit imaging of both cellulose and } cytoskeleton? (If it helps we have a Zeiss 510 with 488, 568 and 633 nm } lines.) } =20 } I thank you in advance, } =20 } Yours sincerely, } =20 } Nigel Chaffey } =20 } ----------------------------------------------------- } Dr Nigel Chaffey, } Dept Forest Genetics & Plant Physiology, } Swedish University of Agricultural Sciences, } S-901 83 Ume=E5, } Sweden } Phone: +46-90-786-6305 } Fax: +46-90-786-5901 } eMail: nigel.chaffey-at-genfys.slu.se } =20 } Looking for another job/position/post... } =20 } =20 } =20
} PDC-2000 is a fine hand-held digital camera but does not have a way to } adapt for the microscope. Amont other things there are internal } optics that don't cooperate well with the microscope optics. It also } could do some macro work with close-up lenses, but that is not } optically the best imaging system.
OK. So, Polaroid removes the optics.
} DMC, while using the same Polaroid designed sensor chip as PDC-2000, } otherwise is designed and built specifically for microscopy. Its } C-mount thread (no optics) allows easy mounting on almost any } microscope using standard C-mount adapters, and also can accept many } "C-mount" threaded macro lenses for use on the copystand.
Adding a c-mount and a tripod socket is worth about $50. I fail to see how the price of the DMC is justified. This seems to be another example of how specialised users pay higher prices for less product. I know that 'economies of scale' do not apply to the technical market.
Can anyone supply me with a source of human skeletal muscle suitably prepared for IEM? Or.. would anyone be able to loan blocks of LR White embedded human skeletal muscle that I could cut sections from and return?
Brett M. Connolly, PhD Merck Research Laboratoriess email: brett_connolly-at-merck.com
I have a customer who is looking for a technique to examine a few small ( {1 mm) bubbles in quartz via Raman or other IR technique. The objective is to thin the quartz to within 1 micrometer of the bubble. The sample size is a few centimeters but can be reduced if necessary.
Any help will be appreciated.
Thanks,
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
} Please could somebody help me. } } I am trying to make a TEM specimen of Diamond, } I have a Gatan PIPS at my disposal, but to use it my Diamond wafer } has to be less than 100 microns thick. At the it is 1mm thick. } Any suggestions on how to thin the Diamond wafer? } The Diamond is actually poly-crystaline CVD } } Thank you in advance } Adam
Hi Adam, Well, I have looked at some single crystal diamond, and the results were great, this is how the sample was prepared;
The sample was sliced with a diamond saw as thinly as we could get, and we tried to dimple it but that was really pointless, we only got 30um reduction in thickness in seven days!
So we PIP's it from quite a thick sample, total time taken was 3080 minutes! Beam at 5KV rot speed 3.5 rpm Ion currents 25uA Gun angle 5'.
Your sample may not be quite as hard, so you many have less time in the PIPS. Also, I found that trying to punch a 3mm disk with a ultrasonic disk cutter is also pointless, much better to mount on a disk then grind the edges round. Be prepared to change the mounting ring several times as the beam will destroy this quicker relative to the diamond.
I hope this helps, btw. Tony Garratt-Reed says hello and sends his regards!
Cheers
David
Dr. David C. Bell Room 13-1018 E-Mail: dcb-at-MIT.EDU Center for Mat. Sci. and Eng. PH: (617) 253-3317 Massachusetts Institute of Technology FAX: (617) 258-6478 77 Massachusetts Ave, Cambridge, MA 02139-4307
Have any of you placed 160 mm objectives to a Zeiss Axioplan or Axiophot with good results? Carl Zeiss does sell an adapter that threads into many 160 mm objectives, but I would perfer to put a negative lens on a slider below the tube lens. I estimate that a -360 mm lens placed about 40 mm below the tube lens would do the trick. I would also welcome comments on the Axioplan II and the merits of the new optics.
Rob ---------------------------------------------------------------------------- ---------------------- Robert W. Williams Center for Neuroscience, Department of Anatomy and Neurobiology 875 Monroe Avenue, Memphis TN 38163 USA Tel: 901/448-7018 or -7050 FAX: -7193 http://mickey.utmem.edu/neuron.html rwilliam-at-nb.utmem.edu
Can anyone supply me with a source of human skeletal muscle suitably prepared for IEM? Or.. would anyone be able to loan blocks of LR White embedded human skeletal muscle that I could cut sections from and return?
Brett M. Connolly, PhD Merck Research Laboratoriess email: brett_connolly-at-merck.com
You might try ultramicrotomy to prepare cross-sections of diamond for TEM analysis. I've had a great deal of experience and luck cross-sectioning hard materials using the technique. You can review an ultramicrotomy procedure for cross-sectioning diamond and cBN in "Ultramicrotomy of Diamond Films for TEM Cross-section Analysis," P. Swab, Microscopy Research and Technique, Wiley-Liss Inc., vol. 31, pp. 308-310 (1995) and other references found in the paper. The paper describes the formation of "micro chips" that are preferentially oriented, embedded in epoxy, and then cross-sectioned with a diamond knife. These cross-sections may show mechanical artifacts, but are uniformly thick and free of beam damage and chemical artifacts.
The concoidal micro chips generated in this procedure are very thin at the edges and may be sufficiently thin for direct TEM observation. If not, the microchips may be secured to a grid and thinned using conventional ion beam techniques. Ion-thinned cross-sections show a minimum of mechanical artifacts, but are not uniformly thick and may show beam damage and associated chemical artifacts.
For assistance with ultramicrotomy in the UK, contact John Forsdyke at Oxford Brookes University (jforsdyk-at-bms.brookes.ac.uk).
Regards,
Phil Swab Advanced Coatings Division Advanced Refractory Technologies Buffalo, NY, USA Phone: 716-875-4091 E-mail: pswab-at-art-inc.com
} ---------- } From: Adam Papworth[SMTP:A.J.Papworth-at-LIVERPOOL.AC.UK] } Sent: Sunday, December 07, 1997 3:37 PM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: TEM of Diamond } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Please could somebody help me. } } I am trying to make a TEM specimen of Diamond, } I have a Gatan PIPS at my disposal, but to use it my Diamond wafer } has to be less than 100 microns thick. At the it is 1mm thick. } Any suggestions on how to thin the Diamond wafer? } The Diamond is actually polycrystalline CVD } } Thank you in advance } Adam } Dr Adam Papworth } Dept Materials science & Engineering } Ashton Building } The University of Liverpool } L69 3BX } } Phone No 0151 794 5372 } Fax No 0151 794 4675 } E-Mail adamp-at-liv.ac.uk } }
It is not appropriate to share human tissue. Each project should be justified according to the local and national regulations. Most importantly the people (or their survivors) who provided the tissue should be informed and allowed to decline participation in the project. I hope that this request does not represent the state of ethics at all commercial organizations! Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
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I am trying to make a TEM specimen of Diamond, I have a Gatan PIPS at my disposal, but to use it my Diamond wafer has to be less than 100 microns thick. At the it is 1mm thick. Any suggestions on how to thin the Diamond wafer? The Diamond is actually poly-crystaline CVD
Thank you in advance Adam Dr Adam Papworth Dept Materials science & Engineering Ashton Building The University of Liverpool L69 3BX
Phone No 0151 794 5372 Fax No 0151 794 4675 E-Mail adamp-at-liv.ac.uk
Adam:
You might try ultramicrotomy to prepare cross-sections of diamond for TEM analysis. I've had a great deal of experience and luck cross-sectioning hard materials using the technique. You can review an ultramicrotomy procedure for cross-sectioning diamond and cBN in "Ultramicrotomy of Diamond Films for TEM Cross-section Analysis," P. Swab, Microscopy Research and Technique, Wiley-Liss Inc., vol. 31, pp. 308-310 (1995) and other references found in the paper. The paper describes the formation of "micro chips" that are preferentially oriented, embedded in epoxy, and then cross-sectioned with a diamond knife. These cross-sections may show mechanical artifacts, but are uniformly thick and free of beam damage and chemical artifacts.
The concoidal micro chips generated in this procedure are very thin at the edges and may be sufficiently thin for direct TEM observation. If not, the microchips may be secured to a grid and thinned using conventional ion beam techniques. Ion-thinned cross-sections show a minimum of mechanical artifacts, but are not uniformly thick and may show beam damage and associated chemical artifacts.
For assistance with ultramicrotomy in the UK, contact John Forsdyke at Oxford Brookes University (jforsdyk-at-bms.brookes.ac.uk).
Regards,
Phil Swab Advanced Coatings Division Advanced Refractory Technologies Buffalo, NY, USA Phone: 716-875-4091 E-mail: pswab-at-art-inc.com
Hi everyone, Would anyone have the address / phone number for HITEK Cryogenic Engineering ?The last address I have is: 3190 Park Road Bay Vista Business Park Benicia, CA 94510
TIA Rosemary
#################################################### Rosemary Walsh Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:rw9-at-psu.edu ####################################################
I can only suggest that, if you are sincerely interested in a high quality digital camera for your microscope, you evaluate the DMC first-hand and compare it with similar products, most of which cost thousands of dollars more. Compare, also, some of the lower-cost solutions and adapters available and determine what image quality level you truly require. Then buy whatever meets your needs at the lowest cost to you.
There is significant investment involved when developing a product specifically for a specialized market, such as optical microscopy. Companies, Polaroid included, need to recover their investment costs so that they can continue to serve their customers.
If you'd like to discuss further, or if you'd like to arrange a product demonstration locally, please get back to me directly.
I hope this helps you in your concerns and that we are able to be of service to you. Thanks for your interest!
} PDC-2000 is a fine hand-held digital camera but does not have a way to } adapt for the microscope. Amont other things there are internal } optics that don't cooperate well with the microscope optics. It also } could do some macro work with close-up lenses, but that is not } optically the best imaging system.
OK. So, Polaroid removes the optics.
} DMC, while using the same Polaroid designed sensor chip as PDC-2000, } otherwise is designed and built specifically for microscopy. Its } C-mount thread (no optics) allows easy mounting on almost any } microscope using standard C-mount adapters, and also can accept many } "C-mount" threaded macro lenses for use on the copystand.
Adding a c-mount and a tripod socket is worth about $50. I fail to see how the price of the DMC is justified. This seems to be another example of how specialised users pay higher prices for less product. I know that 'economies of scale' do not apply to the technical market.
Can anyone tell me about how often a diamond knife needs to be sharpened? I realize it would depend on the amount of wear it gets. Our EM dept. is only open three days a week, with cutting being done about 2-4hrs a week. Our knife was sharpened six months ago and seems to be getting dull again. Is this to be expected?
I know that there are MAC and PC versions of NIH Image but has anyone used NIH Image on a Silicon graphics system? If so, would you please tell me what is needed to set it up.
Salzburg, 12/9/97 Dear Brian,=20 unfortunately I do not know wether you got my reply to your posting on = that=20 subject correctly by 11/02/97 as you can see as follows: =20
Salzburg, 11/02/97
---------- Von: azriel gorski[SMTP:azrielg-at-cc.huji.ac.il] Gesendet: Sonntag, 02. November 1997 09:24 An: David S. Wexler, Ph.D. Cc: Microscopy-at-sparc5.microscopy.com Betreff: Re: cover slip thickness
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On Fri, 31 Oct 1997, David S. Wexler, Ph.D. wrote:
} = ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of = America=20 } To Subscribe/Unsubscribe -- Send Email to = ListServer-at-MSA.Microscopy.Com } = -----------------------------------------------------------------------. } =20 } Hi, } =20 } I have a question about the importance of cover slip thickness. = Namely, } how important is it to use a cover slip thickness for which the } objective } is designed? For example, if I'm using a 40X, NA 1.3 oil immersion } objective which has the number 0.17 on it (corrected for a 170 micron } cover slip thickness), what would be the effect on image quality if I } used a cover slip with, say, a 300 micron thickness?
} Brian Haab 11/02/97 wrote: } U.C. Berkeley } bhaab-at-zinc.cchem.berkeley.edu } =20
} Good luck,
} Shalom from Jerusalem, } Azriel=20
} ******************************************************** } Azriel Gorski, Head azrielg-at-cc.huji.ac.il } Optical Microscopy Laboratory } Division of Identification and Forensic Science } Israel National Police } Jerusalem } ISRAEL
} It is important. You are using a precision optical instrument and } the cover slip is an indespensible part of the optical correction. = Since=20 } you are using an oil immersion objective at 40 X (with oil I hope) it } appears you want as much defined and clear detail as posible. Using any } lense above about 40 X you can see the difference between #1 cover = slips } (0.13 to 0.17mm thick), #1 1/2 coverslips (0.16 to 0.19mm thick), and = #2=20 } cover slips (0.17 to 0.25mm thick). Now having said that, there is an } assumption implicit in that. That is that the sample adhears directely = to } the underside of the coverslip. So don't "flood" with mounting medium.
W. MUSS writes: The cover slip for "dry" objectives with a numeric aperture N.A. below /up to 0.3/0.4 is said to be no problem in terms of optic geometry as well as image quality. You can use "dry"objectives with an N.A. {=20 0.40 with or without a coverslip. For "dry" objectives with an N.A. of=20 =3D/} 0.4 the cover slip is part of the "optical correction" system of = the=20 optical lenses. If you don=B4t use such cover slips, due to spherical=20 aberration you will get diminished contrast quality. The higher the N.A of the objective will be, the smaller will be the tolerances: for example:
Numerical Aperture (NA): effective thickness of cover slip: 0.40 0.17 +/- 0.09 mm 0.60 0.17 +/- 0.013 mm 0.75 0.17 +/- 0.004 mm
These theoretical values not necessarily don=B4t apply for the reality: They are valid assuming tissue sections adhere closest to the lower=20 side of the coverslip. In practice between upper side of the tissue=20 section and the lower side of the cover slip there is a small layer of=20 mounting medium, the actual thickness you won=B4t know. So, in=20 practice you should/can use a minimum of mounting medium, and cover slips measuring 0.15-0.16 mm. If using N.A.=B4s } 0.75 you = won=B4t be able to reach the goal "0.17" precisely. Therefore you can choose dry objectives with a "correction collar" which allows you to adjust = the optical correction between 0.12 and 0.22 mm.
With "immersion objectives" it seems to be more critical: For obtaining optimal resolution and contrast =3D optimal image quality you should be aware of the quality of the immersion oil, of the=20 coverslip, of the mounting medium as well as of the temperature. Immersion oil, and the coverslip then will be a PART of THE OBJECTIVE itself. An immersion oil should have at 23 degr. C a=20 refractive index "n lower case e" of 1.518 (the dispersion, "Abbe=B4s numeral" not considered here). Coverslips for oilimmersion-objectives mostly are obligatory.=20 Deviation/variations of thickness of the coverslips here wouldn=B4t be that critical than with the "dry objectives" (see above): compensating for that will work the drop of immersion oil between the cover slip and the objective lens. An other point worth to be considered with respect to that would be variation(s) in the refractive power. So, as a "theoretical" consequence one should use cover slips with the same refractive index of the mounting medium/immersion medium. Normally, you=20 won=B4t get the information about the refractive index of cover slips, = or do you??
But anyway, you will get optimal results only (especially if using an objective with NA of 1.40) when using additionally an immersion-oil- condensor lens (because oil immersion with its high NA needs a=20 corresponding illumination aperture: if you want to benefit from the = full=20 performance of your optical system the illumination aperture =3D condensor lens has to be brought up to the objective aperture =3D objective lens. The immersion-condensor should have a similar big aperture than the objective.)
} As an asside, I know one microscopist who measures his coverslips with = a } micrometer and only uses the ones which are actually 0.17mm.
W.MUSS writes: You can buy also coverslips which are proved to be 0.17 +/- 0.01 mm as well as other qualities which are the more expensive the less they exhibit variation from 0.17 mm. But, as Brian proposes, I too know LM-freaks measuring a package of "normal" coverslips one by one with a micrometer. This may be the cheapest way in being sure about 0.17 mm=20 thickness.
------------------------------------------------------=20 } Also, what is the definition of "working distance?" I understand it = to } be the distance between the top of the cover slip and the lens of the } objective, but I want confirmation of this definition.
} You are basically correct.... to be a "sticler" it is the nearest part = of } the lense, not any optical center of it.
W.MUSS writes: The higher the magnification number of an objective, the smaller the=20 "working distance" will be. Besides problems in positioning a high power objective carefully over a section/cover slip and thereby without damaging it (which wouldn=B4t be possible with modern objectives with=20 a "sliding/retractive" mechanism of the objective itself) you certainly =
will have or can get problems with an increased thickness of your cover slip (see above): say, working distance of your } N.A. 1.40, 40x=20 Oil Immersion Objective { will be 0.2 mm (see info-sheet of your=20 microscope's or objective's instruction manual, you *can* use a=20 coverslip 0.3 mm thickness: but then you will not be able to "optically reach" the tissue section for correct imaging. At least you will decrease resolution and contrast capacity. =09 Another suggestion: why not try a 40x or 50x objective with an N.A. of 1.00? This would be a compromise with respect to working distance as well as not necessarily the "must" of using immersion illumination/ immersion of the condensor front lens to achieve resolution/contrast=20 conditions as provided and possible by the optical system you use.
Also: if you view semithin sections with your scope, you won=B4t use = cover=20 slips at all: try viewing directly with a drop of immersion oil on your =
sections and the oil immersion objective. You will get marvellous image quality (provided you have adjusted your optical system according to KOEHLER=B4s instructions). Removal of the immersion oil is just immersion of the object slide(s) into a coplin jar containing xylene. After soaking a while you can wipe off (or blow off by use of compressed air) very easily remnants of immersion oil.
Hope this explanation helps you with your work best regards "on a lazy sunday 2nd of november from SALZBURG- Mozart=B4s birthplace"
Wolfgang MUSS the same on Tue, 9th Dec. 1997 Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: bhaab-at-zinc.cchem.berkeley.edu[SMTP:bhaab-at-zinc.cchem.berkeley.edu] Gesendet: Sonntag, 07. Dezember 1997 18:29 An: microscopy-at-sparc5.microscopy.com Betreff: LM: imaging and the thickness of the cover plate
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Below is the result of your feedback form. It was submitted by (bhaab-at-zinc.cchem.berkeley.edu) on Friday, October 31, 1997 at 14:12:11 -------------------------------------------------------------------------= --
Email: bhaab-at-zinc.cchem.berkeley.edu Name: Brian B. Haab
School: U.C. Berkeley
State: CA
Zip: 94720
Question: Hi,
My question has to do with imaging and the thickness of the cover plate = used. How important is it to use a cover plate thickness for which the = objective was specifically designed? For example, if I'm using an objective which says 0.17 (presumably corrected for a 170 micron cover slip thickness), would image quality be greatly distorted when using something, say, = twice as thick? Also, what is the definition of "working distance?"
The DMC and any other product of its kind are worth what people will pay for it. It is the nature of the free market economy. You must consider=
the product development costs, which are substantial. Also, how many wil= l they sell? The market size is not huge.
The Polaroid DMC brought was introduced at about $6000. This is FAR less=
than any other quality digital microscope camera on the market. The only=
less expensive options come from something like the Pixera, which is not realistic for anyone who wants to get true high quality images without massive pixel enlargement. It seems odd to blast Polaroid for introducin= g a camera that was $5000 less than their nearest competitor when introduce= d. The good news is that prices always go down, and quality goes up.
} The DMC and any other product of its kind are worth what people will pay } for it.
Undeniable.
} The Polaroid DMC brought was introduced at about $6000. This is FAR less } than any other quality digital microscope camera on the market. The only } less expensive options come from something like the Pixera, which is not } realistic for anyone who wants to get true high quality images without } massive pixel enlargement. It seems odd to blast Polaroid for introducing } a camera that was $5000 less than their nearest competitor when introduced.
That's not the thrust of my argument. Why is a camera with fewer features/components so much more expensive than the more complex one? It seems less of a market-driven issue than a marketing one, similar to the observation that I can buy DuMont forceps from a jeweler's supplier for half the cost as from a medical supplier.
} The good news is that prices always go down, and quality goes up. } }
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It also depends on what is being cut as well as who is doing the cutting. If it is plant material, 6 months is a LONG time. Animal tissue is = softer but again some animal tissues have hard components which are hard = on the knife edge. The experience of the cutter obviously also comes into play. The less = experience, usually the more resharpenings may be necessary. Are they = also taking thicks on the same knife? If so, you might want to get a = histo knife for thicks and perhaps your thin diamond would last longer. Good Luck Judy Murphy San Joaquin Delta College Microscopy Technology Center Stockton, CA __________________________________________________________________________= _____
Can anyone tell me about how often a diamond knife needs to be sharpened? = I realize it would depend on the amount of wear it gets. Our EM dept. is = only open three days a week, with cutting being done about 2-4hrs a week. Our knife was sharpened six months ago and seems to be getting dull again. Is this to be expected?
------------------ RFC822 Header Follows ------------------ Received: by sjdccd.cc.ca.us with ADMIN;9 Dec 1997 08:58:05 -0800 Received: from Sparc5.Microscopy.Com (206.69.208.10) by = ms.sjdccd.cc.ca.us with SMTP (Eudora Internet Mail Server 1.2); Tue, 9 Dec 1997 08:57:56 = -0800 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) id HAA12949 for dist-Microscopy; Tue, 9 Dec 1997 07:56:36 = -0600 Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by = Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA12946 for = {microscopy-at-msa.microscopy.com} ; Tue, 9 Dec 1997 07:56:32 -0600 X-Sender: zaluzec-at-microscopy.com Message-Id: {v03007803b0b301d55bd4-at-[206.69.208.21]} Mime-Version: 1.0 Content-Type: text/plain; charset=3D"us-ascii"
} Does anyone know of a quick test that would confirm pen ink (Flair, Bic, } etc.) on paper? I have plenty of sample that consists of large, black, } blue-fringed blotches on a flooring felt backing. There are no visible } pigment particles up to about 400X, and the stain is moderately soluble in } an ethanol/methanol blend. } } Thanks in advance.
David, you might find it helpful to submit this question to the forensic science list. To submit, send the message to forens-l-at-ACC.FAU.EDU. To subscribe, send a message to MailServ-at-Acc.Fau.Edu with the following in the body of the message: Subscribe Forens-L Your Real Name.
This is just the kind of thing the forensic people do all the time.
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
does anyone have a good source for transfer roles and paper for a tektronix Phaser 440 dye sub printer?
thanks
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Dear Diane, have found your posting on an issue which only can be answered very subjectively. For further elucidation one should know not only weekly/daily hours of usage but, more interestingly:
- what company product ( e.g. DIATOME, DEKKER, MICROSTAR.........) - which type of Diamond knife (e.g. HISTO-Type for semithin sections, ULTRA-Type for ultrathin sections.....) - length of cutting edge - proper working (cutting) as usually done (starting cutting from left to right edge) - max. width of specimens you are cutting - which kinds of tissues (with incrustations, artificial inclusions,...any to be expected?) - which type of, of which hardness resin(s) you use - which person will perform cutting (I understand that this will be an experienced person, familiar with cutting with diamond knife(-ives) - cutting angle, knive free angle, cutting speed: is this done correctly as implicated by the manufacturer's (guaranteed) data sheets on cutting properties - cleaning methods used (sometimes remnants of resin and other cutting debris imparts cutting quality: have a careful look on your cutting edge at least at mag. x 45, seen with back light)
If you can provide some more details according to this items, maybe there is a more objective answer. Best regards Wolfgang
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
DISCLAIMER: No commercial interest in products/product lines, company/-ies, if such names are mentioned or such are refered to.
---------- Von: Diane M. Smith[SMTP:smithde-at-valunet.com] Gesendet: Dienstag, 09. Dezember 1997 15:10 An: microscopy-at-sparc5.microscopy.com Betreff: Q: Diamond Knife-resharpening
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Can anyone tell me about how often a diamond knife needs to be sharpened? I realize it would depend on the amount of wear it gets. Our EM dept. is only open three days a week, with cutting being done about 2-4hrs a week. Our knife was sharpened six months ago and seems to be getting dull again. Is this to be expected?
In my initial response to this thread, I extolled the virtues of the = Kodak DC120 camera for photomicroscopy based on trials at our lab. = Maybe I should have given the standard disclaimer that I have no = affiliation with Kodak ... I definitely should have listed the other = cameras I considered.=20 Unless anyone would like to include another product, the three similar = choices mentioned in this thread are the Pixera, the Polaroid DMC, and = the soon to be released Kodak MDS120 system (based on the DC120). = Obviously, this list excludes more capable, more expensive cameras. The = question of interest is how capable are these cameras and are they = capable enough... beyond being just teaching and demonstration systems. Seth Grotelueschen makes a good point about not blasting Polaroid for = being the first to release a relatively low cost system that is = comparable to much more expensive systems. Hopefully, we are at a point = where prices for these systems continue to fall precipitously toward = affordability... and hopefully before Kalman Rubinson gets a chance to = retire : )
A quick comparison of the 3 systems follows with questions or comments = related to capability:
1.) PRICE Pixera $1200 Polaroid DMC $6000 Kodak MDS120 $2100 2.)CCD SIZE Pixera 1/3" Polaroid DMC 12.15mm(~1/2") Kodak MDS120 1/2" Question: Is the physical size of the CCD array important or is = resolution more important to image quality? 3.) CCD RESOLUTION Pixera - ~750k=09 Polaroid DMC ??? claimed to be "megapixel" Kodak MDS120 836K 4.) IMAGE RESOLUTION Pixera 1 meg =20 Polaroid DMC 1.9 meg (1600x1200) Kodak MDS120 1.2 meg (1280x960) All these systems use integration and averaging to increase final image = resolution. Question: Does this type of claim based on integrated = resolution overstate the capabilities of the camera? Are these claims = analogous to "false magnification"? Is resolution in the = photomicroscopy as important as other issues such as low light = sensitivity and data integrity? 5.) DATA INTEGRITY It is my understanding that all of these systems are capable of transferring images uncompressed to PC or Mac or loss-less compression = such as Tiff formats. As Dr. John Russ pointed out, many comsumer grade = digitals store images using lossy compression algorithms.=20 6.) IMAGE TRANSFER TO PC- all systems are TWAIN compliant Pixera - limited to speed of serial port =20 Polaroid DMC - faster SCSI transfer rate Kodak MDS120 - limited to speed of serial port=20 7.) FLEXIBILITY OF CAMERA Both the Polaroid and the Pixera are tethered to the PC and are = generally dedicated to the microscope. They can be used with a c-mount = video lens (sold separately) for other applications but must remain = tethered to a computer.=20 The Kodak camera can be used as a stand alone digital camera away from = the computer. No addition lenses are required. 8.) Strengths - all systems promise to be user friendly compared to a = video camera/ frame grabber solution=20 Pixera - Most affordable =20 Polaroid DMC - Highest resolution and speed of transfer Kodak MDS120 - Most Versatile, affordable, and good resolution
All the information above was taken from the respective spec. sheets for = each system.=20
For my application, I am trying to capture images allowing me to = optimize my probing protocols which will later be used on the confocal. = So rather than looking for "analyzable" images I am more concerned with = optimizing for analyzable images. That doesn't mean that these systems = are not capable of this. Hopefully, someone will take the time to = explain how/why these systems are/are not capable of generating = "analyzable" images
Kevin Brent Smith=20 Graduate Student University of Louisville Biology Dept. Life Science Bldg. Rm.#12 Louisville, KY 40292 Phone: (502) 852-6773 Fax: (502) 852-0725 =20
I remember there have been reports since many years ago regarding to the alternative chemical treatment in place of critical point drying using a critical point dryer. Can any of you tell the name(s) of the chemicals and the companies which sell them? Or your experience using such chemicals? Sorry for not being able to look up a SEM book for myself.
Thanks!
Yuhui Xu Core EM Facility Dana Farber Cancer Institute
We have just acquired an old Gatan Ion Mill and the Electronic Gas Flow Control (EGC) valve is not working. I am just wondering if anyone happen to have a spare EGC valve or a manual needle valve assembling to give away or for sale.
Thanks a lot.
Eric Yu Wang North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313) 936-3353 Fax: (313) 763-5567 eywang-at-engin.umich.edu http://emalwww.engin.umich.edu/people/eywang/eywang.html
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Subject: Time:3:40 PM OFFICE MEMO reply} test for ink Date:12/9/97
Reply to: RE} Test for Ink? ************************* You wrote, "Does anyone know of a quick test that would confirm pen ink (Flair, Bic, etc.) on paper? I have plenty of sample that consists of large, black, blue-fringed blotches on a flooring felt backing. There are no visible pigment particles up to about 400X, and the stain is moderately soluble in an ethanol/methanol blend. Thanks in advance. Dave Stadden" ************************* Dave, One technique might be to use X-ray fluorescence (XRF), exciting the specimens with secondary x rays. This could probably be done in a couple of hours, and doesn't require a lot of specimen prep work. XRF won't do the chemistry, but it could be very good for matching elemental content of inks (at least for those elements in the ink heavier than sodium) and their relative quantities, even in the presence of organic components. Using XRF, first look at a gross ink sample to determine elemental content and relative concentrations to optimize the type of x-ray excitation. Then collect an x-ray spectrum from the ink-on-paper specimen. Collect another spectrum (same geometry, same excitation, same acquire live time, etc) of paper only. Strip out (subtract) the paper spectrum from the ink-on-paper to get an ink signature from paper. Save. Repeat this process, subtracting the flooring felt backing "background") for the ink spectrum from the ink on felt . Compare the two spectra. If the Flair, Bic, ??, is the source of the stain, you should see a pretty good match. It won't be exact since the samples were taken from two quite different substrates. You might be able to minimize the mismatch if you prepared a "standard", using known inks on otherwise identical, unstained, felt backing. Is this an on-going analysis problem or a one-time test? Do you have access to an XRF system? Dennis DGCollins-at-lbl.gov
I'm David Langlais, Vice President of Marketing for Pixera Corporation. Since Pixera was introduced into this discussion and into discussions on some USENET news groups lately, I feel that a response to the Mr. Grotelueschen's remark is appropriate.
I'm posting from the email account of one of my employees. My email is ddl-at-pixera.com and I'll be subscribing to the listserver today so anyone can reach me with comments......
And so...
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On Tuesday, 9 Dec 1997, Seth J. Grotelueschen wrote:
} The DMC and any other product of its kind are worth what people will pay } for it. It is the nature of the free market economy. You must consider } the product development costs, which are substantial. Also, how many will } they sell? The market size is not huge.
} The Polaroid DMC brought was introduced at about $6000. This is FAR less } than any other quality digital microscope camera on the market. The only } less expensive options come from something like the Pixera, which is not } realistic for anyone who wants to get true high quality images without } massive pixel enlargement.
Mr. Gruteleuschen and others are certainly entitled to their opinions regarding the quality of Pixera's cameras in microscopy. I just wish that he and others would take the time to be technically accurate. He and they are not. There is a common misconception and disbelief that Pixera can really achieve 1260x960 pixels in our catpured images without some nefarious trick or (gasp) 'massive pixel enlargement".
Mr. Gruteleuschen evaluated our Pixera Professional some months ago and decided not to purchase. If he chose not to purchase due to the technical reason he stated in this message, then he made his decision for the wrong re asons.... Because he either doesn't understand how our camera system works or he doesn't believe it. If the pictures he captured didn't satisfy his requirements, I would have no problem with him saying so. But his remark is technically erroneous and misleading.
Pixera's high resolution cameras - Professional and Personal, can capture 24-bit RGB color images in resolutions up to 1260x960 pixels using only a 250K CCD. Unless you understand our technology (some of which is proprietary and trade secret) it is easy to disbelieve that this can really be accomplished. After all, if it could be done, Kodak, Polaroid, Sony and others would have done so, right? Wrong.
I'll add here that additional details about Pixera and its technology are available in the January and September issues of Advanced Imaging Magazine in articles written by Mr. Charles Reis...
The Pixera Professional (the camera we sell in microscopy) achieves 4x the resolution of our CCD this way. The key to our high resolution technology is three related pieces. 1.) Our camera head contains an electro-mechanical light refractor (patented in the US and Japan) in front of the CCD. We use this light refractor to shift the incident light from the subject onto the pixel elements. This mechanism is extremely accurate down to the sub-micron level. 2.) We use a proprietary (and patented) color filter pattern for our CCD. It's custom made for Pixera to work with our light refractor. 3.) We have proprietary software image processing software that works in conjunction with the light refractor and CCD filter pattern to achieve the image quality and resolution of our images. Pixera uses 100% software image processing to generate its images, the only image processing done in the camera is AGC and a little Gamma.
When taking a high-res picture (800x600 or above), our camera takes 4 exposures of the subject. This takes a little under a second. For each exposure, our light refractor shifts the incident light in a particular direction for a particular distance in order to get four separate and distinct samples. We do not pixel double or enlarge. To understand this in more depth, look up the patent. Mr. Yuji Ide, founder of Pixera is the principal author. This process is analgous to shifting the CCD for each exposure like some other high priced, high-resolution cameras do.
After each exposure, the raw analog pixel data from the CCD is sent to our interface card where all we do is convert the analog raw pixel data into digital raw pixel data. The raw digital pixel data is then sent into the computer (Winows 95, NT, or Mac) for processing. We do not compress any of the information for transfer into the computer. Another benefit of this technique and technology is that we get two color samples for each pixel. So our camera has the color information and reproduction of a 2-CCD camera. So even though we do have to interpolate to get the 24-bits of color (as do all single CCD cameras) we at least start with twice the information. Our software image processing of course understands our color filter patter, image shifting, etc., and processes the raw pixel information into a image that is approx. 3.6 million pixels.
Our camera also functions as a "video" camera although no formatted video signal is generated. We simply stream the raw pixels (without using the light refractor) into the computer and build the motion frames on the fly. The faster the computer, the faster the video...Today we are hardware limited by our card architecture to 10 fps, next year we won't have this limit.
This is it in a nutshell. We don't pixel double, we don't interpolate other than for color and a little to adjust for aspect ratio, and we don't do "massive pixel enlargement". If anyone wants or need additional information I'd be glad to discuss it with you. If you still don't believe it, we have a couple of thousand customers that did, and we'd like the opportunity to turn you into a believer as well...
} It seems odd to blast Polaroid for introducing } a camera that was $5000 less than their nearest competitor when introduced. } The good news is that prices always go down, and quality goes up.
Gee, I guess then it's really odd to blast Pixera for introducing a camera that is several thousands less than anyone (including Polaroid) simply because you don't believe what the camera is doing and even worse because you don't take the time to understand......
One place where I do agree with Mr. Gruteleuschen is - We at Pixera believe that prices SHOULD always go down and quality go up. That's why we're working hard on new cameras of 2 and 4 million pixels based on our technology. And you can bet that they'll be priced and have quality that some people, at least, will find hard to believe....
I'm not trying to make this personal at all. I'm just tired of "people of science" on various news groups and listservers taking unwarranted and inaccurate potshots at Pixera's products without really knowing the facts.
I appolgize for my unwarranted swipe at commercial institutions. Dr. Connolly has provided an appropriate response--see below. However, his request did not state clearly the conditions and considerations for an exchange of human tissue and since there is considerable controversy regarding these issues amongst pathologists these days I believe this community (Microscopy) should be careful with language as well as any exchange of tissues.
I believe you're thinking of HMDS, hexamethyldisilizane. Most EM supply houses sell it, I think. Procedures for use were covered in a note in the May 97 issue of Microscopy Today, and Scott Whitaker (sp? correct me if I'm wrong) as information on this on the U Florida web pages at: http://www.biotech.ufl.edu/~emcl/tips.html
I've used it successfully. Email me with specifics about what you're up to, and maybe I can give you some ideas.
Phil
} Dear Colleagues: } } I remember there have been reports since many years ago regarding to the } alternative chemical treatment in place of critical point drying using a } critical point dryer. Can any of you tell the name(s) of the chemicals and } the companies which sell them? Or your experience using such chemicals? Sorry } for not being able to look up a SEM book for myself. } } Thanks! } } Yuhui Xu } Core EM Facility } Dana Farber Cancer Institute }
Philip Oshel PO Box 5037 Station A Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu or poshel-at-hotmail.com ***** looking for a job *****
If anyone is looking into this topic or knows of any relevant references or SEM methods, I would greatly appreciate the information. I have scoured our meager library and the current contents database with virtually no luck. My kingdom for access to the science citation index.
Thank you so much for any aid you can provide.
Jill Craig University of Northern British Columbia
I am still using an old ETEC Autoscan (SEM, vintage 1973). It is still working well after more than 12 000 hrs of usage and usually we get the results that we want.
However, a calcium containing deposit has been forming in the cooling water supply (in Cu tubes, in cooling coils around diff.pump, in heat sinks, ect). The microscope was donated to us several years ago but was not connected for the last 18 months to the recirculating water system in our laboratory ( we are using filtered tap water). The water flow has now been reduced drastically over the last few weeks and I fear that the 'pipes' may eventually totally block up.
What is the best (and safe) way to reduce or remove this deposit. Back-flashing was only partially successful.
Any suggestion ?
Thank You
Hans Brinkies SWINBURNE, University of Technology School of Engineering and Science Electron Microscopy Laboratory HAWTHORN, 3122, Australia
Sorry folks; the Discovery channel has CHANGED the air times of Dennis Kunkel's microscopy, described on the listserver yesterday. Here's the new message:
} Dear Friends and Associates, } Sorry for the incovenience but Discovery Channel has preempted } Movie Magic for this week. It will air the following week (see below). } Discovery Channel's documentary series, "Movie Magic", is airing a } segment called "Far Out Creatures" that features special effects used in } the making of science fiction films (Alien Resurrection and Starship } Troopers). Part of the 1/2 hour segment will include a short interview } with me and some of my "MicroAliens". I thought this might be of } interest to you. } } The segment will run: } December 18, 1997 9:30 - 10:00 PM - West Coast Time (PST) } December 19, 1997 1:30 - 2:00 AM - West Coast Time (PST) } December 20, 1997 2:00 - 2:30 PM - West Coast Time (PST) } } Please check your local listing for the time of Movie Magic on the } Discovery Channel (or check their website schedule at } www.discovery.com/diginets/discovery/discovery.html). } } Best Regards, Dennis Kunkel } } *********************************************** } * Dennis Kunkel Ph.D. * } * Pacific Biomedical Research Center * } * University of Hawaii * } * * } * email - kunkel-at-pbrc.hawaii.edu * } * www - http://www.pbrc.hawaii.edu/~kunkel/ * } **********************************************
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
I have just learned that there has been a delay in the distribution of the
Call for Papers and Advanced Registration for the Microscopy & Microanalysis '98 Meeting in Atlanta, Ga July 12-16
due to a delay at the printers.
It is expected that they will be mailed during the week of Dec. 15th.
In order to avoid having to reply to repeated questions which have started to flow in my direction I have taken the liberty of posting this Email message to both the MSA Membership as well as the Microscopy Listserver Databases.
As additional information becomes available I will make sure that the M&M'98 WWW pages are updated. These WWW pages are directly accessible from the URL
I would like to hear comments or suggestions about sample preparation of biofilms builded up by methanogenic and acidogenic facultative anaerobic bacteria deposited on sand grains for SEM. So far, we have done fixation using glut araldehide and further dehydration with alcohol.
Does anyone have any experience in sampling sand grains from a bioreactor in order to preserve the biofilm for SEM observations?
Thanks in advance,
Nora Pratta Centro Regional de Investigacion y Desarrollo Santa Fe - Argentina
Hi everyone, I would like to hear comments or suggestions about sample preparation of biofilms builded up by methanogenic and acidogenic facultative anaerobic bacteria deposited on sand grains for SEM. So far, we have done fixation using glutaraldehide and further dehydration with alcohol. Does anyone have any experience in sampling sand grains from a bioreactor in order to preserve the biofilm for SEM observations? Thanks in advance, Nora Pratta Centro Regional de Investigacion y Desarrollo Santa Fe - Argentina
You might want to repost this to the biofilms listserver: biofilms-at-net.bio.net. "Conventional" EM (TEM, SEM) has limited utility in study of most bacterial biofilms because they suffer much artifact introduction (spatial distortion) in the drying process. I would encourage you to supplement your SEM studies with ESEM (environmental scanning electron microscopy; what I call "hydrated" SEM), laser confocal microscopy, or cryosectioning light microscopy. I can help you with confocal and can refer you to people who can assist with the rest. Rob Palmer Director - Biofilm Imaging Facility CEB/UT
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This is the text of Stephen Kohn's message. It was attached as a Word document which makes it a little harder to open.
I wonder if it would be worthwhile to provide some guidance or reminders to the list members as to what makes for good message practice. In addition to these attachments, I have been seeing a number of messages packed with HTML coming across. Most of those appear to come from the Microsoft Outlook mail program. I would rather we stick to plain text so that the most can read the message with the least difficulty.
Regarding the comments made during the past week about the use of inexpensive high-resolution digital cameras for microscopy purposes, please note the following update information about The Pixera "Professional" camera which clarifies any possible misunderstandings about the product that subscribers may have:
Pixera's most recent software upgrade - Version 1.2, contains a 10 frame per second "fast viewfinder" window. This is equivalent to the speed of what many digital camera video conferencing systems run at.
TWAIN compatibility has been improved - Pixera recently tested more than 10 programs from image analysis vendors/providers and found the Pixera Professional camera to be TWAIN compliant with all of them. Feel free to contact me If you'd like a copy of the list.
RGB Color mix in pre-image capture mode now allows for viewing a color balanced image which is equivalent to the final image captured.
With reference to other comments made, the variable lens Pixera Professional "can ship" an uncompressed 3.7 megabyte image to the computer for viewing and or image analysis purposes.
"While I have a vested interest in Pixera as an employee, the purpose of this reply as stated above is to clarify details regarding the system's capability and is presented as a non-commercial message for informational purposes only.
Mike Saulnier is no longer with IVS. Please remove this address from your mailing-list:
msaulnier-at-ivsinc.com
Thanks, Dave Shipman Network Administration
} ---------- } From: Dave Shipman } Sent: Wednesday, December 10, 1997 5:51 AM } To: Dave Shipman } Subject: Notification: Inbound Mail Failure } } The following recipients did not receive the attached mail. Reasons } are listed with each recipient: } } {msaulnier-at-ivsinc.com} msaulnier-at-ivsinc.com } MSEXCH:IMS:IVS, Inc.:SW_DOMAIN:WINNT_1 0 (000C05A6) Unknown } Recipient } } The message that caused this notification was: } } { {Message: Microscopy & Microanalysis '9...} } }
In short, I would like to inquire as to the methods and chemicals that people are using for cleaning their filament assemblys on their electron microscopes. Especially the final step.
When changing our filament for our TEMs, we have in the past cleaned any tarnish or deposits from the filament assembly pieces using a mild polish, then sonicated the parts in a detergergent solution, followed by water rinses, sonication in a few rinses of ETOH, then finally sonication in a freon-based ultra-precision cleaning solution. (The used freon solution is dumped into its own waste bottle.)
We are considering changing this process (especially in consideration of the availability and disposal of the freon solution) and are wondering what other labs use for cleaning their filament assemblys thoroughly. We are especially interested in the final cleaning step (removing any leftover residues).
Thank you for your input. Don't be afraid to be brief.
Gregg Sobocinski Parke-Davis Pharmaceutical Research Ann Arbor, Michigan USA Sobocig-at-aa.wl.com
A related question is: Does anyone know if the proceedings of the 15th Pfefferkorn conference held in May 1996 are ever going to published by Johari et al.
Matt Libera Stevens Institute of Technology
henk-at-vt8200.vetmed.lsu.edu wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello All, } Does anyone know what has happened to Dr. Om Johari and the journal } Scanning Microscopy. I have been unable to contact him via e-mail, } phone, } or fax and have not been able to find reference to the journal in over } two } years. Have he and the journal "retired"? } } Bill Henk } Dept. of Anatomy & Cell Biology } LSU School of Veterinary Medicine } Baton Rouge, LA 70803 } phone -(504)346-3237 } e-mail - henk-at-vt8200.vetmed.lsu.edu
Anyone using HF: Where do you buy your acid neutralizing chemicals? We've purchased the whole HF safety kit in the past, but somehow the chemicals never come out right - I currently have 2 bottles of the HF inactivator (#2) and part of a bottle of the acid neutralizer (#4). The neutralizer is just sodium carbonate (right?) - any reason why I shouldn't just use that from any source? I've spoken with chemistry types from several vendors and nobody is sure. And to buy just the Mallinckrodt stuff packaged for the safety kit is insane - they can't sell the regular jar; I'd have to buy a tub of it. So, I'm hoping someone out there has a wonderful answer - I'm afraid to assume anything with handling this particular chemical.
I'm sure that everyone will tell you that the lifetime of your diamond knife depends on what you cut, who uses it, and how you take care of the knife - it's all true. The lifetime of a knife can be measured in minutes or years - it's up to you. For best results, treat your diamond knife as you would your toothbrush, don't share it with anyone. In truth, a knife needs to be sharpened when the sections it cuts no longer meet your needs. I map the use of my knife edge and systematically work my way across the knife as it degrades with use. For particularly tough samples I may use a previously degraded region of the knife for the initial cuts. With regard to cleaning, minimize cleaning and never allow sections to dry on the knife.
My first diamond knife was a 4 mm knife that lasted 8 years, cutting 4 to 8 hours a week on glass, metals, semiconductors, and dielectrics. So much for predicting lifetimes. Typically there was only myself and a trained tech using the knife. My blocks are made with Spurrs, the blockfaces are prepared on the microtome using a glass knife, and are only 50 to 100 um across. I am meticulous about the preparation of the blockface, the orientation of the sample in the blockface, and strictly minimize the size of the embedded sample that the diamond knife cuts.
Phil Swab Advanced Coatings Division Advanced Refractory Technologies Inc. Buffalo, NY
} ---------- } From: smithde-at-valunet.com[SMTP:smithde-at-valunet.com] } Sent: Tuesday, December 09, 1997 9:10 AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Diamond Knife } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Can anyone tell me about how often a diamond knife needs to be } sharpened? I } realize it would depend on the amount of wear it gets. Our EM dept. is } only } open three days a week, with cutting being done about 2-4hrs a week. } Our } knife was sharpened six months ago and seems to be getting dull again. } Is } this to be expected? } }
I am also beginning to evaluate hand-held digital cameras for an upcoming image analysis project. The recent discussion has been very helpful for me - thank you to everyone for posting to the general list.
One product I haven't seen mentioned is the Minolta RD-175. Does anyone have experience with this camera, or know how it fits into the spectrum? Does it use the JPEG compression that Dr. John Russ spoke about? Does it provide good color representation? I should mention that I want a camera that is not "tethered to the computer".
Thanks again for your help,
Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN 55144 "Opinions above are my own, not necessarily my employer's"
} ... Anyone using HF: } Where do you buy your acid neutralizing chemicals? We've purchased } the whole HF safety kit in the past, but somehow the chemicals never come } out right - I currently have 2 bottles of the HF inactivator (#2) and part } of a bottle of the acid neutralizer (#4). The neutralizer is just sodium } carbonate (right?) - ...
I might suggest only that the neutralizer be Ca carbonate which would allow production of the inert by-product CaF.
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
ASTIMEX has a set of 53 minerals - standards for microanalysis, in a typical mount of 1 inch diameter. This includes two garnets - pyrope Mg3Al2Si3O12 and almandine Fe3Al2Si3O12.
The address:
ASTIMEX Scientific Ltd. 351 Wellesley Str. East Toronto Canada M4X 1H2 fax: (416) 961-2402 e-mail: jcr-at-quartz.geology.utoronto.ca (Prof. J.C. Rucklidge)
Best regards,
Wojtek xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz National Accelerator Centre PO Box 72 Faure 7131 South Africa E-mail: PRZYBYLOWICZ-at-nac.ac.za Fax: +27-21-8433543 Phone: +27-21-8433820 xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
Please can you give me the necessary information/instructions how to realize publication of positions of Philips Electron Optics on the MSA's Listserver.
I am Jacques Weterings personnel manager of Philips Electron Optics in Eindhoven the Netherlands.
What are you cutting? Steel? My diamond knives are sharp after 10 years of daily use. We are cutting neural tissue embedded in epon(very soft) We have never resharpened them.
Perhaps your knife was not sharpened well. Or perhaps your knife was beyond repair in the first place.
Sally
On Tue, 9 Dec 1997, Diane M. Smith wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Can anyone tell me about how often a diamond knife needs to be sharpened? I } realize it would depend on the amount of wear it gets. Our EM dept. is only } open three days a week, with cutting being done about 2-4hrs a week. Our } knife was sharpened six months ago and seems to be getting dull again. Is } this to be expected? } } }
At 08:55 AM 12/10/97 +0100, Heinz Fehrenbach wrote: } can anyone of this newsgroup give me information about and subscription } procedure to any related newsgroup ? In particular, I would appreciate } to hear whether there is a special newsgroup discussing immuno-histology, } -fluorescence, -blotting, and pathology subjects.
There is an immunohistochemistry list, called ipox-l
To subscribe, send a message to majordomo-at-pathology.stanford.edu and write subscribe ipox-l in the body of the message. I believe that there is also a pathology list, and instructions for this are available at "The Histotech's Home Page" (http://www.histology.to)
Best regards, Steven E. Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Om Johari sent out an announcement either late las year, or earlier this year, that he was retiring. He also said that he hadn't found anyone who was willing to take over the job of editing the Scanning Microscopy journal or organizing the annual meeting. I don't have a current email or regular mail address to reach him. It's probably not likely that anything not currently in press will be published by him.
Call Prof. Wil Bigelow in Ann Arbor, 764-3321. I would be surprised if he did not recommend a final rinse/sonication in isopropanol, if freon is a no-no for you. He will certainly have some suggestions for the overall cleaning process also.
Larry
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
} I wonder if it would be worthwhile to provide some guidance or reminders to } the list members as to what makes for good message practice. In addition to } these attachments, I have been seeing a number of messages packed with HTML } coming across. Most of those appear to come from the Microsoft Outlook mail } program. I would rather we stick to plain text so that the most can read the } message with the least difficulty.
I agree. While I have the capability of downloading and viewing such messages, the bother is such that I will not do so. Let's keep to plain text.
Colleagues... we have old Hitachi S-650 SEM with KEVEX ANALYST 8000 microanalyser with old architecture ( on LST-11 minicomputer). Can anyone give a advice about cheap and easy ways to change this system on modern system on PC platform? What we need to this? Thanks, Yury Shipilov AME Department UofA
I'm in the market for a new inverted microscope that will form the basis of a Fura 2, Ca++ imaging/photometry system. I am considering either a Leica DMIRB or a Nikon TE200/300. Has anyone compared these microscopes and their long working distance Fluor objectives (20, 40 and 63 or 60X)? Any help would be appreciated.
--
William F. Jackson, Ph.D. Professor Department of Biological Sciences 5380 McCracken Hall Western Michigan University Kalamazoo, MI 49008
I have used a diamond knife without any need for resharpening for about 9 years, only cutting plant material. Although I did not cut with it every day, I was the full time technologist in this lab doing most of the ultrathin sections for a variety of researchers. I NEVER cut any thick sections with that knife, and I was careful to always have very small block faces and handle my knife very carefully. If I ever had any sections stick and dry on the knife edge, I would wipe them off with alcohol soaked balsa wood. (first I soaked that balsa in acetone for several weeks to remove all the resin in the wood) Although nobody really recommends wiping the knife edge with anything, I find that it is essential to really have a clean edge. I wipe it under the microscope, and keep wiping until I can see that it is truly clean, but of course I'm careful to avoid using much pressure, or any force perpendicular to the knife edge. BEING GENTLE is the key. If one isn't careful, a lot of garbage can dry on the back of the knife edge without you knowing that it is there. That is why I inspect it carefully under a dissecting microscope, looking especially at the back of the knife edge. This garbage can make you think that your knife needs resharpening, when in fact it is only dirty. In order to loosen up any dirt on the knife edge, I soak it overnight or over the weekend in alcohol. This will not harm the knife, in fact I know a researcher who STORES his diamond knife under alcohol. Of course acetone will ruin it almost instantly, so DON'T EVER MAKE THAT MISTAKE, because the acetone will quite readily dissolve the glue that holds the diamond in place. But the alcohol is perfectly fine.
In a human pathology lab now, I notice that our knives get sharpened more frequently, perhaps after 1 or 2 years, depending on the size of the knife. We cut sections absolutely every day, and very big sections, where a single section might cover a significant portion of the grid. That way the pathologists don't need to waste a lot of time scanning around looking for the section!!
Hope this helps, Garry
} ---------- } From: Murphy, Judy[SMTP:murphy-at-sjdccd.cc.ca.us] } Sent: 9 December, 1997 11:24 } To: Diane M. Smith; MSA } Subject: RE: Diamond Knife } } It also depends on what is being cut as well as who is doing the cutting. } If it is plant material, 6 months is a LONG time. Animal tissue is softer } but again some animal tissues have hard components which are hard on the } knife edge. } The experience of the cutter obviously also comes into play. The less } experience, usually the more resharpenings may be necessary. Are they also } taking thicks on the same knife?
It is also wise to have some Calcium Gluconate 2.5% gel handy to apply liberally to the affected area and cover loosely with gauze wrap. It is available by prescription only I believe.
Honey Nut Cheerios Harry ---------- ----------------------------------------------------------------------- .
Tamara Howard wrote:
} ... Anyone using HF: } Where do you buy your acid neutralizing chemicals? We've purchased } the whole HF safety kit in the past, but somehow the chemicals never come } out right - I currently have 2 bottles of the HF inactivator (#2) and part } of a bottle of the acid neutralizer (#4). The neutralizer is just sodium } carbonate (right?) - ...
I might suggest only that the neutralizer be Ca carbonate which would allow production of the inert by-product CaF.
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
I would like to sincerely apologize to The Pixera Corp. for the = misinformation in my post to the listserv. My intention was not to = disinform. I am not an expert ... just a curious student trying to = learn a little by generating discussion with the previously posted = comparison.
IMAGE RESOLUTION=20 Pixera Their web site states:=20 Resolution in still mode 1.2 meg (1260x960) Image size - 1.3 meg(1280x1024) =20 Polaroid DMC 1.9 meg (1600x1200) Kodak MDS120 1.2 meg (1280x960)
A number of private messages send to me expressed a concern with the = image resolution claims these systems make given the effective = resolutions of the CCDs used.=20
IMAGE TRANSFER I mistakenly stated that Pixera's transfer rate is limited to the = serial port rate. The truth is they have a PCI or PMCIA connection. In = general, is slower image capture is a limitation of these sytems? I am = currently trying to reach tech service for each company to learn more = about how the images are transfered and what the realistic transfer = rates are (basically how long does it take to capture 2 images = consecutively).
Kevin Brent Smith Graduate Student University of Louisville Biology Dept. Life Science Bldg. Rm.#12 Louisville, KY 40292 Phone: (502) 852-6773 Fax: (502) 852-0725
Looking to identify MELANIN in fungal walls. There are no antibodies for it (Its not a protein- I don't think), and I can't find any lectins for it. It is diagnostic in urine samples, and skin, hair etc.
o.k., surely someone somewhere - one of those great old Histologist / microscopists has some eye-of-newt & pinch of bat-wing technique for staining it - possitively would be even better - but we have non-melanized mutants whcih can be used as a control (putatively we can turn melanization On/Off and this is what we are trying to verify, eh?).
Any hints or techniques would be greatly appreciated. Applicable microscopies include SEM (SEI & BEI), TEM, LM, Confocal, and fluorescence. Rather NOT attempt any autoradiography techniques though.
Thank you in advanced for any help!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
Harry - Thanks for pointing this out! Our safety department gets us a fresh tube every year; it is kept in a special pouch on the wall next to the hood where we use and store the HF. We also have an "Instructions for physicians" handout with the gluconate, it deals with treatment following HF exposure.
On Wed, 10 Dec 1997, Ekstrom, Harry wrote:
} It is also wise to have some Calcium Gluconate 2.5% gel handy to apply } liberally to the affected area and cover loosely with gauze wrap. It is } available by prescription only I believe. } } Honey Nut Cheerios } Harry } ----------
We are looking for some Teflon tweezers, to dip a TEM sample in HF. Anyone know of a source?
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60208-3108 tel: (847) 491-3996 fax: (847) 491-7820 email: l-marks-at-nwu.edu http: //www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
I am seeking the advice of any member who wishes to make this a truly joyful season.
I am working with some skin samples, (human, mouse, rat) that are between 100=B5m to 150=B5m thick. They have been labeled with various 1=B0s and three 2=B0s. The labeling is specific and strong. The problem is the clarification of the tissue and the proper mounting in a suitable fashion for confocal. Most of the problems stem from the fact the tissue is so thick; and before you ask, yes it has to be that thick. I am looking for the relationship between matrix, nerve and blood structures. I have performed the 3D analysis successfully on smaller, 40=B5m sections, but clearing the tissue becomes important in the thicker sections for proper imaging.
I have tried dehydration w/ EtOH 70%-100% and Methyl salicylate, but the crystals left behind by the Me-S and washing them displaced the tissue.
I have tried 70-30 v/v glycerol but only moderately improved my image.
I dislike using anything which might leave an organic residue. That includes old favourite cleaners like "Brasso" and "Wenol". AND even the organic solvents which YOU HOPE remove that greasy residue can leave their own residues. Steve Chapman (whom God preserve) advises that "Wenol plus K" in particular leaves behind an organic film intended to retard oxidation.
So my preference is for an aqueous polish which rinses off with copious volumes of our glass distilled water.
Microgrit alumina (say grade 1600) on a damp cotton bud is fine. Or you can buy polishing alumina ready made up (you may have it already) from say Buehler.
But my preference is for diamond paste on a cotton bud. ProScitech sold me our existing tube which was 5g. Its 0.25 micron grit, quality of diamond 100k (whatever that matters). One uses so little it seems to last forever.
Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
I dislike using anything which might leave an organic residue. That includes old favourite cleaners like "Brasso" and "Wenol". AND even the organic solvents which YOU HOPE remove that greasy residue can leave their own residues. Steve Chapman (whom God preserve) advises that "Wenol plus K" in particular leaves behind an organic film intended to retard oxidation.
So my preference is for an aqueous polish which rinses off with copious volumes of our glass distilled water.
Microgrit alumina (say grade 1600) on a damp cotton bud is fine. Or you can buy polishing alumina ready made up (you may have it already) from say Buehler.
But my preference is for diamond paste on a cotton bud. ProScitech sold me our existing tube which was 5g. Its 0.25 micron grit, quality of diamond 100k (whatever that matters). One uses so little it seems to last forever.
Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Hi Richard, Our lab studies frog melanophores which contain hundreds of melanosomes (melanin-filled pigment granules). I have had a great deal of success visualizing melanosome distribution on the confocal by imaging backscattered (reflected) light. Using this technique allows me to double stain cells with rhodamine & fluorescein and superimpose reflected light to create very nice 3 color images. On our old Zeiss LSM210 this is possible by scanning with the 543 HeNe laser line (usually used to excite rhodamine) and detecting for green fluorescence (usually used to detect emitted light from fluorescein). You can accomplish the same effect by removing the fluorescent filter block altogether, but images acquired this way are misaligned with respect to fluorescence collected thru the dichroic. The only difficulty I've had using this technique is that the samples must be sufficiently below the plane of the coverslip so that reflection from the glass doesn't contribute to the backscatter image.
Hope you find this input useful. Best,
Steve Rogers Dept. of Cell & Structural Biology & The Beckman Institute - Optical Visualization Facility University of Illinois -at- C/U srogers-at-delphi.beckman.uiuc.edu
} Looking to identify MELANIN in fungal walls. There are no } antibodies for it (Its not a protein- I don't think), and I can't } find any lectins for it. It is diagnostic in urine samples, and } skin, hair etc. } } o.k., surely someone somewhere - one of those great old Histologist } / microscopists has some eye-of-newt & pinch of bat-wing technique } for staining it - possitively would be even better - but we have } non-melanized mutants whcih can be used as a control (putatively we } can turn melanization On/Off and this is what we are trying to } verify, eh?). } } Any hints or techniques would be greatly appreciated. Applicable } microscopies include SEM (SEI & BEI), TEM, LM, Confocal, and } fluorescence. Rather NOT attempt any autoradiography techniques } though. } } Thank you in advanced for any help! } } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } } "640K ought to be enough for anybody." } -- Bill Gates, 1981
Its fair and reasonable for Steven Slap to provide this information, but why does he not indicate that he is the editor of the suggested histology site?
Sure his name appears on that site and his company is shown as a sponsor (a cheap form of advertising), but no connection is made and most visitors would not realise that Steven Slap is the manager of the sponsoring firm and thus has a commercial interest: In essence it is a commercial site. Predictable his company is the only consumable supplier company shown. Nothing wrong with his sponsoring or editing such a page but visitors to the page and readers of this listserver are entitled to know not just who is sponsoring what but what commercial interests the editor has. Its known as propriety! Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } From: Energy Beam Sciences, Inc. {ebs-at-ebsciences.com} } Dear fellow microscopists, } } At 08:55 AM 12/10/97 +0100, Heinz Fehrenbach wrote: } } can anyone of this newsgroup give me information about and subscription } } procedure to any related newsgroup ? In particular, I would appreciate } } to hear whether there is a special newsgroup discussing immuno-histology, } } -fluorescence, -blotting, and pathology subjects. } } There is an immunohistochemistry list, called ipox-l } } To subscribe, send a message to majordomo-at-pathology.stanford.edu and write } subscribe ipox-l in the body of the message. I believe that there is also a } pathology list, and instructions for this are available at "The Histotech's } Home Page" (http://www.histology.to) } } Best regards, } Steven E. Slap } ******************************** } Energy Beam Sciences, Inc. } Adding Brilliance To Your Vision } ebs-at-ebsciences.com } http://www.ebsciences.com/ } ******************************** }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: Time:5:39 PM OFFICE MEMO RE} Cooling water problems Date:12/10/97
Hans Brinkies wrote, "I am still using an old ETEC Autoscan (SEM, vintage 1973). It is still working well after more than 12 000 hrs of usage and usually we get the results that we want.
"However, a calcium containing deposit has been forming in the cooling water supply (in Cu tubes, in cooling coils around diff.pump, in heat sinks, ect). The microscope was donated to us several years ago but was not connected for the last 18 months to the recirculating water system in our laboratory ( we are using filtered tap water). The water flow has now been reduced drastically over the last few weeks and I fear that the 'pipes' may eventually totally block up.
"What is the best (and safe) way to reduce or remove this deposit. Back-flashing was only partially successful.
"Any suggestion ?
"Thank You
"Hans Brinkies SWINBURNE, University of Technology School of Engineering and Science Electron Microscopy Laboratory HAWTHORN, 3122, Australia" **************************************** Hans, The same problem occurs in lots of water-cooled, high power equipment such as vacuum tube amplifiers, x-ray tubes, and particle accelerators. For components with copper cooling water passages we use only low conductivity water (min of 750K-ohm-cm: 1 Megohm-cm is better) in a closed-loop cooling system. Even then deposits will accumulate. We use water flow switches to ensure at least a minimum water flow in sensitive equipment, and try to keep an eye on the pressure drop across each water cooling circuit, back-flushing with a dilute solution of Sulfamic acid when we see a clear rise in the pressure drop at the required flow. Hope this helps keep your ETEC Autoscan running. Dennis Collins DGCollins-at-lbl.gov
Dear Yuri, I have been researching the same thing to replace my two old Kevex 8000s. So far I have found iXRF, (www.ixrfsystems.com) who specialize in putting new computers on old Kevex's, ANS, (www.qtmsys.com) who have a very inexpensive product, and, of course, Kevex has an upgrade that is a little more. All are worth talking to. You wrote:
} Colleagues... } we have old Hitachi S-650 SEM with KEVEX ANALYST 8000 microanalyser with } old architecture ( on LST-11 minicomputer). Can anyone give a advice } about cheap and easy ways to change this system on modern system on PC } platform? What we need to this? } Thanks, } Yury Shipilov } AME Department } UofA } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Sara, We have been running the VCR Ion Mill for about eight years, now. We have found the instrument to be solid, reliable and easy to use and clean. The people at VCR are very helpful and easy to deal with. Ours is an older model, now they have one run totally by computer. You wrote: } Hi All } } We need to buy a new ion mill for our lab. Any suggestions, warnings, } tips would be appreciated. } } Thanx in advance } } Sara Prins
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Gregg, Everyone has their favorite technique and all will work. I use a good scrub with Wenol, a rinse in acetone, which removes any Wenol residue, a check under 20X dissceting scope to be sure I got it all clean, a rinse in denatured ehtanol, then a final rinse in pure ethanol and dry with hot air. The hot air dry is important to prevent drying residue. Each time I say "rinse", I mean immersing the assembly in the solvent and sonicating for at least a minute. Another technique that helps is to sonicate the piece first in a 10% oxalic acid solution, then rinse in several distilled water changes. Dry with clean alcohol as above. The oxalic acid loosens the residue and saves hard scrubbing. Good luck. You wrote: } } In short, I would like to inquire as to the methods and chemicals that people } are using for cleaning their filament assemblys on their electron microscopes. } Especially the final step. } } When changing our filament for our TEMs, we have in the past cleaned any } tarnish or deposits from the filament assembly pieces using a mild polish, then } sonicated the parts in a detergergent solution, followed by water rinses, } sonication in a few rinses of ETOH, then finally sonication in a freon-based } ultra-precision cleaning solution. (The used freon solution is dumped into its } own waste bottle.) } } We are considering changing this process (especially in consideration of the } availability and disposal of the freon solution) and are wondering what other } labs use for cleaning their filament assemblys thoroughly. We are especially } interested in the final cleaning step (removing any leftover residues). } } Thank you for your input. Don't be afraid to be brief. } } Gregg Sobocinski
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Laurence, A forceps supplier such as Fine Science Tools may have them, but a cheaper source is the Optician that handles soft contact lenses. That's where I got mine. Some drup stores have them with contact lens supplies. They may just be plastic, but should stand HF. You wrote: } } We are looking for some Teflon tweezers, to } dip a TEM sample in HF. Anyone know of a source? } } ++++++++++++++++++++++++++++++++++++++++++++++++ } Laurence Marks } Department of Materials Science and Engineering
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
At 03:44 PM 12/10/97 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Dear Colleague,
I think that one should consider the following products:
1) VCR Group XLA 2000 Ion/Atom Mill (USA) Suite 31, 250 East Grand Avenue, South San Francisco, CA 84080
2) Gatan Model 691 Precision Ion Polishing System PIPS (USA) Gatan Inc. 6678 Owens Drive, Pleasanton, CA 94588-3334, USA
3) Bal-Tec RES 100 (Lichenstein) BAL-TEC AG, F=F6hrenweg 16, Postfach 62, FL-9496 Balzers, F=FCrstent= um Lichenstein
4) Technoorg-Linda IV3 or IV3/H/L (Hungary) H-1077 Budapest, R=F3zsa u. 24, Hungary, phone: 36-1-3428-713, fax:= 36-1-3224-089
5) Fishione Instruments Model 3000 Ion Mill (USA) E. A. Fishione Instruments Inc. 9003 Corporate Circle, Export, PA 15632= USA
I think that IonTec (UK) is not present on this market now, but I am not sure. I can give you further details separately if you want. Best wishes, Bela Pecz 11th Dec. 1997 ----------------------------------------- Dr. Bela Pecz Research Institute for Technical Physics H-1325 Budapest, POBox 76 Hungary phone: 36-1-2332 865 fax: 36-1-2332-794 E-Mail: pecz-at-mufi.hu -----------------------------------------
Laurence Marks wrote: ======================================================= We are looking for some Teflon tweezers, to dip a TEM sample in HF. Anyone know of a source? ======================================================= If what you want to dip is something more substantial in size than a TEM grid, then the molded Teflon (R) tweezers found on our website will hold up extremely well. They were originally developed for use in the electronics industry where HF dipping is a pretty frequent and routine operation. But don't try to pick up with them a TEM grid, it won't work.
If it is a TEM grid or grid sized sample you want to pick up, then you will need Teflon coated high precision tweezers which can also be found on our website. It is a constant battle of trade offs: You want the tips to retain their sharpness, therefore the coating must be extremely thin, but at that coating thickness, there is a good chance for some small population of pin-holes to form at the tips. So the protection is not perfect, but it is lightyears better than using unprotected tweezers. Also, Teflon is soft and lacks abrasion resistance and it will, therefore, wear off, some might say too quickly. But the good news is that the tweezers can still be used as ordinary tweezers for other purposes.
Now while the molded Teflon tweezers are a pretty commonly found item, high precision Teflon coated tweezers are not so easily found. My comments relative to pin-holes and the coating being worn off were directed specifically to the SPI brand of this kind of product since we have not made comparisons between our product and anyone else's.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ =================================================
Om's last Scanning Microscopy International meeting was billed as just that last May in Chicago, I was tthere. He has oganised them for 30 years and I guess has done it enough. Some of the groups were talking of continuing their own little specialist gatherings elsewhere in future, who knows?
There is a Pfeffercorn meeting being organised currently:
_______________________________________________- Scanning Microscopy International Sixteenth Pfefferkorn Conference Optimising Scanning Electron Microscopy April 5 - 8 1999 Aberystwyth, Wales, UK
Full details of the conference, updated regularly, can be viewed on the World Wide Web page: http://www.aber.ac.uk/~dbswww/pfeffer16/home.html
Dr Iolo ap Gwynn, (iag-at-aber.ac.uk) Institute of Biological Sciences, The University of Wales, Aberystwyth, Wales, SY23 3DA, UK Fax: +44 1970 62 23 50 ________________________________________________
I am trusting that the journal is continuing because I received a manuscript back this week, along with the reviewers' comments. More work!
Just to join in on a discussion that involves a large part of our daily = tasks, I would like to point out the dangers in cleaning in solvents. If you clean your own E.M., then usually you are also aware of the = "freshness" and source of the chemicals/solvents in your lab. In our = travels we often find that we cannot get the solvents we would like or = need and have to make do with a lot of strange variations. We have also = found contaminated solvents which can cause all sorts of problems. = Usually you can visibly see the residue. It appears as white stains. = It's the invisible residues that catch us some times. The basic rules we follow is as follows: Clean only if necessary. If you find that the test specimen is proving = that the E.M. is not up to spec, only then clean the column bits.=20 If the gun assembly is still fairly clean, do not polish. You will only = add dirt. Use a polishing paste that you know works and what it takes to remove = it. This can only be tested by experience unfortunatly. Pastes we use = vary from Wenol, Pol metal polish, pikal metal polish( supplied with = most Japanese E.M.'s) and Hyprez diamond pastes. All of which work in = different circumstances. As we are normally charging per hour, the = customers don't appreciate us trying to polish away for hours on end. So = we find the diamod pastes work better on the gun parts as they remove = the tungsten very quickly. We then commonly use Ethanol as a solvent for these pastes, as acetone = is not always available. One of the better solvents we do use is = Arklone. But it is also not always available and is very expensive.
The secrets we have learnt are simple and work well. Polish the parts well with cotton buds and the paste. Once it is clean, = immerse them immediately in the ethanol/solvent. This ensures that the = paste does not harden on the parts. Ultrasonically clean the parts for at least 10 minutes a time. Remove = the solvent and fill the beaker again with new solvent and ultrasonic = again. If Arklone is available, then use this as the final solvent. When taking the parts out of the solvent, "polish" again with clean = "solvent dunked" cotton buds. Normally you find that there is still = quite a lot of "dirt" that comes off. Use clean solvent to dunk the buds = into. Beware of fibres that can be left after polishing. Check the parts under a light microscope for any particles, residues or = fibres. Once the E.M. is re-assembled allow it to pump down over night, at = least, before checking for a beam.
With the gun assembly, we would again suggest you do not clean if it is = not necessary, as a light coating of tungsten does not affect = performance that badly. For the higher KV. TEMs obviously this would = have to be monitored very closely.=20 We have had good success in simply ultrasonically cleaning with the = Protrain formula of 10% Silvo, 10% ammonia and water. Rinsing thoroughly = afterwards in water then some alcohol. Again I will point out that we donot always have all the fancy chemicals = most labs have available to them, but if this method works for us, I am = sure the added steps and specialised chemicals listed can only improve = cleaning by a smaller margin.
Good luck and happy polishing.
Luc Harmsen Anaspec, South Africa Technical support for E.M. clients, world wide. Anaspec-at-icon.co.za Tel:+27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290
Can somebody please send a copy of a method how to make holey carbon to me? I know it was on the listserver a while ago but somehow I can't find the e-mail or my printout. Thanx
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Dehydrate with graded ethanol 50 through 100 % then clear in 2 changes of Xylene 5-10 minutes each, that shoul render the tissues translucent.
-- Begin original message --
} Hello: } } I am seeking the advice of any member who wishes to make this a truly } joyful season. } } I am working with some skin samples, (human, mouse, rat) that are } between 100=B5m to 150=B5m thick. They have been labeled with various 1=B0s } and three 2=B0s. The labeling is specific and strong. The problem is the } clarification of the tissue and the proper mounting in a suitable } fashion for confocal. Most of the problems stem from the fact the } tissue is so thick; and before you ask, yes it has to be that thick. I } am looking for the relationship between matrix, nerve and blood } structures. I have performed the 3D analysis successfully on smaller, } 40=B5m sections, but clearing the tissue becomes important in the thicker } sections for proper imaging. } } I have tried dehydration w/ EtOH 70%-100% and Methyl salicylate, but the } crystals left behind by the Me-S and washing them displaced the tissue. } } I have tried 70-30 v/v glycerol but only moderately improved my image. } } Please help } } Ramin Rahbari } 313-998-3383 -- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
I have not seen any cheap conversions, unless perhaps you do the whole thing yourself (and if your "rates" are very, very cheap). There are a number of
vendors who offer pc conversion packages, but the cost is from about $8000 to $20,000 (US).
Woody White Mcdermott Technology, Inc.
Work http://www.mtiresearch.com
Me http://www.geocities.com/capecanaveral/3722 ______________________________ Reply Separator _________________________________
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Colleagues... we have old Hitachi S-650 SEM with KEVEX ANALYST 8000 microanalyser with old architecture ( on LST-11 minicomputer). Can anyone give a advice about cheap and easy ways to change this system on modern system on PC platform? What we need to this? Thanks, Yury Shipilov AME Department UofA
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We are looking for some Teflon tweezers, to dip a TEM sample in HF. Anyone know of a source?
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60208-3108 tel: (847) 491-3996 fax: (847) 491-7820 email: l-marks-at-nwu.edu http: //www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Jim Darley wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Its fair and reasonable for Steven Slap to provide this information, but } why } does he not indicate that he is the editor of the suggested histology site? } } Sure his name appears on that site and his company is shown as a sponsor (a } cheap form of advertising), but no connection is made and most visitors } would not realise that Steven Slap is the manager of the sponsoring firm } and } thus has a commercial interest: In essence it is a commercial site. } Predictable his company is the only consumable supplier company shown. } Nothing wrong with his sponsoring or editing such a page but visitors to } the page and readers of this listserver are entitled to know not just who } is sponsoring what but what commercial interests the editor has. Its known } as propriety! } Jim Darley } } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 77 740 370 Fax: +61 77 892 313 } Great microscopy catalogue, 500 Links, MSDS, User Notes } ************************ http://www.proscitech.com.au } } ---------- } } From: Energy Beam Sciences, Inc. {ebs-at-ebsciences.com} } } Dear fellow microscopists, } } } } At 08:55 AM 12/10/97 +0100, Heinz Fehrenbach wrote: } } } can anyone of this newsgroup give me information about and subscription } } } procedure to any related newsgroup ? In particular, I would appreciate } } } to hear whether there is a special newsgroup discussing } immuno-histology, } } } -fluorescence, -blotting, and pathology subjects. } } } } There is an immunohistochemistry list, called ipox-l } } } } To subscribe, send a message to majordomo-at-pathology.stanford.edu and } write } } subscribe ipox-l in the body of the message. I believe that there is } also a } } pathology list, and instructions for this are available at "The } Histotech's } } Home Page" (http://www.histology.to) } } } } Best regards, } } Steven E. Slap } } ******************************** } } Energy Beam Sciences, Inc. } } Adding Brilliance To Your Vision } } ebs-at-ebsciences.com } } http://www.ebsciences.com/ } } ******************************** } }
Jim, Don't be so childish. If your company had the gumption and interest to support the profession you serve, perhaps you would have had the insight to establish such a site as Steven has. Ronnie Houston Dallas, TX
} } Does anybody out there } } Looking to identify MELANIN in fungal walls. There are no } antibodies for it (Its not a protein- I don't think), and I can't } find any lectins for it. It is diagnostic in urine samples, and } skin, hair etc. } } o.k., surely someone somewhere - one of those great old Histologist } / microscopists has some eye-of-newt & pinch of bat-wing technique } for staining it - possitively would be even better - but we have } non-melanized mutants whcih can be used as a control (putatively we } can turn melanization On/Off and this is what we are trying to } verify, eh?). } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu -- End original message -- Richard,
While not quite eye of newt and bat wing, though it doesn't miss that era by much, try the following procedure by Masson (1928) which is still in use today.
Masson-Fontana method for melanin:
Fontana Silver Solution:
10 percent silver nitrate 20 ml Ammonia Distilled water 20 ml
To the 10% silver nitrate add the ammonia a drop at a time untill only a faint opalescence remains. Add the distilled water and filter, allow to stand overnight prior to use. Store in the dark and the solution will be good for about 30 days. Do not reuse.
Method:
1-hydrate the tissue sections 2-wash well in distilled water 3-transfer to Fontana silver solution for 12-16 hours in the dark in a covered container 4-Wash well in 2-3 changes of distilled water 5- (optional step)tone in 1% gold chloride 6-stabelize in 5% sodium thiosulfate for 5 minutes 7-wash in tap water for 5 minutes 8-counterstain wit 1% neutral red for 2 minutes 9-dehydrate, clear and coverslip
NOTE!!!!!Ammoniacal silver decomposes into an explosive compound after time (months)
From Histopathological Stains and their Diagnostic Uses, J.D. Bancroft & A. Stevens
If I can be of further assistance please feel free to call me.
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
A coworker needs to prepare plan view and cross sectional TEM samples of metal thin films (30-500 angstroms) on GaAs. Any suggestions on the best sample prep for this type of material? TIA
Dick Fonda
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 _____________________________________________________________
The journal Pigment Cell Research has much to say about melanin in it's different forms. Also, work done in our lab some time ago used a whole list of nonradioactive methods for detecting melanin. Check out "Pigmented cells of the stria vascualris and spiral ligament of the chinchilla" by C.G. Wright and D.H. Lee, Acta Otolaryngologica (Stockholm) 108:190-200, 1989 or "Pigmented epithelial cells of the mebranous cassular wall of the chinchilla" by C.G. Wright and D.H. Lee, Acta Otolaryngologica (Stockholm) 102:438-449. There is even a way to see if your nonpigmented cells might actually be producing the precursors of melanin without making the final product. I didn't do any of the work myself- so I can't give you much detail. Hope this info is helpfull.
Karen Pawlowski Sr. Res. Assoc. Dept. of Otolaryngology UT Southwestern Med. Ctr. Dallas, TX
On Wed, 10 Dec 1997 edelmare-at-casmail.muohio.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Does anybody out there } } Looking to identify MELANIN in fungal walls. There are no } antibodies for it (Its not a protein- I don't think), and I can't } find any lectins for it. It is diagnostic in urine samples, and } skin, hair etc. } } o.k., surely someone somewhere - one of those great old Histologist } / microscopists has some eye-of-newt & pinch of bat-wing technique } for staining it - possitively would be even better - but we have } non-melanized mutants whcih can be used as a control (putatively we } can turn melanization On/Off and this is what we are trying to } verify, eh?). } } Any hints or techniques would be greatly appreciated. Applicable } microscopies include SEM (SEI & BEI), TEM, LM, Confocal, and } fluorescence. Rather NOT attempt any autoradiography techniques } though. } } Thank you in advanced for any help! } } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } } "640K ought to be enough for anybody." } -- Bill Gates, 1981 }
For the record, the Polaroid PDC-2000 is available for use either "tethered" or "untethered", the latter with onboard storage for 40 or 60 images depending on the model. You will then need to connect to the computer to download your images, though. Price of the PDC-2000 is now under $2K for all models.
Hope this helps your information gathering process. For more, get back to me or check www.polaroid.com
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I am also beginning to evaluate hand-held digital cameras for an upcoming image analysis project. The recent discussion has been very helpful for me - thank you to everyone for posting to the general list.
One product I haven't seen mentioned is the Minolta RD-175. Does anyone have experience with this camera, or know how it fits into the spectrum? Does it use the JPEG compression that Dr. John Russ spoke about? Does it provide good color representation? I should mention that I want a camera that is not "tethered to the computer".
Thanks again for your help,
Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN 55144 "Opinions above are my own, not necessarily my employer's"
For the record, the Polaroid PDC-2000 is available for use either "tethered" or "untethered", the latter with onboard storage for 40 or 60 images depending on the model. You will then need to connect to the computer to download your images, though. Price of the PDC-2000 is now under $2K for all models.
Hope this helps your information gathering process. For more, get back to me or check www.polaroid.com
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I am also beginning to evaluate hand-held digital cameras for an upcoming image analysis project. The recent discussion has been very helpful for me - thank you to everyone for posting to the general list.
One product I haven't seen mentioned is the Minolta RD-175. Does anyone have experience with this camera, or know how it fits into the spectrum? Does it use the JPEG compression that Dr. John Russ spoke about? Does it provide good color representation? I should mention that I want a camera that is not "tethered to the computer".
Thanks again for your help,
Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN 55144 "Opinions above are my own, not necessarily my employer's"
The absolute best, fastest, and cheapest way to prepare non-site specific cross section samples with this type and thickness of metallization on GaAs is with the small angle cleavage technique. There is a detailed pictorial outline of the technique in the MRS Specimen Prep for TEM of Materials IV, Vol 480 that just came out. The authors are myself and John McCaffrey who developed the technique and has several pubs out on it. The technique requires very little in terms of equipment that is not usually found in a lab. Southbay Technology is selling a starter kit that has all the required supplies. I can typically prepare about 9 samples in about an hour and exchange and examine a sample within about 5 minutes to see if it is good. A big advantage to this method is that there is no need to worry about contamination of the sample. I've used this on GaAs and other materials and examined the samples in field emission microscopes with no problems. I do use electronic grade acetone and double rinse them.
For the plan view samples, you will have to dimple and ion mill because of the metallization. Don't forget to protect the good side from ion sputtered material.
If you didn't have the coating and just wanted to make plan view samples from the GaAs, I'd recommend chemically polishing the GaAs from the backside after mechanically thinning to about 100 um to make the plan view samples. Peter Goodhew showed me an inexpensive way to do this. I think that the solution used was 5% bromine in ethyl alcohol (it might have been methanol) which was dripped onto the sample from a burette. The sample was low temperature waxed to a coverslip slide that was put onto a Teflon pedestal with a small amount of vacuum grease mounted in a plastic cup. The grease was not exposed to the solution because it was in the middle of the coverslip and far from the edges. The cup was tilted at an angle of about 30 degrees and was rotating with the use of a small DC motor. A stereomicroscope was used to help determine when the sample was perforated.
You could use this method instead of dimpling if you stopped the process before perforation and then continued the thinning process with ion milling. The disadvantage is that you will not know the thickness left to ion mill.
I hope this helps.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
A coworker needs to prepare plan view and cross sectional TEM samples of metal thin films (30-500 angstroms) on GaAs. Any suggestions on the best sample prep for this type of material? TIA
Dick Fonda
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 _____________________________________________________________
Sorry, folks; the schedule that I posted last week has been changed. CS } ************************************************************************ } Dear Friends and Associates, } Sorry for the incovenience but Discovery Channel has preempted } Movie Magic for this week. It will air the following week (see below). } Discovery Channel's documentary series, "Movie Magic", is airing a } segment called "Far Out Creatures" that features special effects used in } the making of science fiction films (Alien Resurrection and Starship } Troopers). Part of the 1/2 hour segment will include a short interview } with me and some of my "MicroAliens". I thought this might be of } interest to you. } } The segment will run: } December 18, 1997 9:30 - 10:00 PM - West Coast Time (PST) } December 19, 1997 1:30 - 2:00 AM - West Coast Time (PST) } December 20, 1997 2:00 - 2:30 PM - West Coast Time (PST) } } Please check your local listing for the time of Movie Magic on the } Discovery Channel (or check their website schedule at } www.discovery.com/diginets/discovery/discovery.html). } } Best Regards, Dennis Kunkel } } *********************************************** } * Dennis Kunkel Ph.D. * } * Pacific Biomedical Research Center * } * University of Hawaii * } * * } * email - kunkel-at-pbrc.hawaii.edu * } * www - http://www.pbrc.hawaii.edu/~kunkel/ * } ***********************************************
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
} Dear all, } } A coworker needs to prepare plan view and cross sectional TEM samples of } metal thin films (30-500 angstroms) on GaAs. Any suggestions on the best } sample prep for this type of material? TIA } } Dick Fonda
Ion milling of course. Bela Pecz 11th Dec. 1997 ----------------------------------------- Dr. Bela Pecz Research Institute for Technical Physics H-1325 Budapest, POBox 76 Hungary phone: 36-1-2332 865 fax: 36-1-2332-794 E-Mail: pecz-at-mufi.hu -----------------------------------------
Right, prices seem to be dropping. I recently visited one web site = advertising the PDC-2000 for ~$2500 only to visit again a few weeks = later to see it reduced to ~$1600. I haven't seen a comensurate price = drop in the DMC 2000 but that is probably because I haven't recieved a = recent quote. =20 Kevin Brent Smith University of Louisville Biology Dept.
-----Original Message-----
The PDC 2000/40 has a new(45 days old) list price of $1699 and the PDC/2000/60 has a list price of $1999. There has not been a price reduction on the DMC as of yet.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation "See What Develops" 4525 Leonard Parkway Richmond, Virginia 23221-1809 804 254 1011 804 254 1013 Fax warrenj1-at-polaroid.com
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Right, prices seem to be dropping. I recently visited one web site advertising the PDC-2000 for ~$2500 only to visit again a few weeks later to see it reduced to ~$1600. I haven't seen a comensurate price drop in the DMC 2000 but that is probably because I haven't recieved a recent quote. Kevin Brent Smith University of Louisville Biology Dept.
-----Original Message-----
For the record, the Polaroid PDC-2000 is available for use either "tethered" or "untethered", the latter with onboard storage for 40 or 60 images depending on the model. You will then need to connect to the computer to download your images, though. Price of the PDC-2000 is now under $2K for all models.
Hope this helps your information gathering process. For more, get back to me or check www.polaroid.com
} Jim, } Don't be so childish. If your company had the gumption and interest to } support the profession you serve, perhaps you would have had the insight } to establish such a site as Steven has. } Ronnie Houston } Dallas, TX
Fair comment, but as one who visits it occasionally, I'd like to point out that Jim does host an interesting and useful website.
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Tom Phillips wrote, I am having a problem with my Bio-Rad MRC-600 and I was wondering if any other users have had a similar one. On the Normal scan speed, I have a very consistent "vibration" in the image in which each horizontal line on the monitor are slightly zig-zagged.
This problem can be caused by 2 problems, and I have experienced both. One is laser instability, and the second is dirty contacts between the detector and the digitizer. I suspect that your problem is the later from what you describe. To fix it remove the front panel of the scan head and disconnect the 9 pin connect coming from the detectors. They are located on the left side of the box, if the scan head is on an upright scope. They are the same as the cable running from the scan head to the computer scan card. Remove them and clean the pins with an eraser to remove the carbon build up that occurs, and then blow out the dust. Electrical contact cleaner or small amount of methanol or acetone should work as well. I do this on a semi-annual basis. If this appears at the 1 second scan speed it will eventually show up in the slow, 4 second scan speed.
Joe Goodhouse Confocal Core Facility Molecular Biology Princeton University
We have a Gatan wide-angle CCD camera with digital processor for EM201 and have been asked to set up an image capture, processing and archiving computer system. I know there are several expensive packages out there but feel that since this is a standard TV signal, 640x480, there should be some effective products,(i.e. All-In-Wonder from ATI) that work together and are available from computer shops.
Does anyone have a similar system set up already (not necessarily for TEM) that uses universal file formats and hardware connections?
I would appreciate any feedback - Thanks!
Karen Rethoret York University, Toronto 416-736-2100 x33289
I have access to a LARGE number (ie several cases) of microscope cover glasses that I would send to anyone willing to pay the cost of shipping.
These cover glasses are #1 and are 24 mm X 60 mm (not what I call a standard size). They are "Bioloid" Brand from Will Corp. Rochester, N. Y. The packaging says "uniform quality non-corrosive".
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There is an excellent paper that I would be pleased to send you a copy of=
which deals with ion milling of hard coatings. The paper is:
"Transmission Electron Microscopy Characterization of Hard Coatings and Films; Sample Preparation Aspects and Results" G. Radnoczi, Arpad Barna Research Institute of Technical Physics, Hungarian Academy of Sciences. =
Surface and Coatings Technology, vol 80 (1996) pp 89-95.
This paper deals with several materials such as: =
10u diamond film on silicon mechanically alloyed Al-Cu Powder SiC fibers in plastic SiC/Si TiN/Si
All of the work was done with an IV3 Research Grade Ion Milling System which is offered by South Bay Technology. Therefore, I do have a financi= al interest in this posting. Nonetheless, it is a very good paper. I also have several other papers by Dr. Barna et al which deal specifically with=
ion milling difficult materials. I think they would be valuable refernce=
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Adam Papworth } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Please could somebody help me.
I am trying to make a TEM specimen of Diamond, I have a Gatan PIPS at my disposal, but to use it my Diamond wafer has to be less than 100 microns thick. At the it is 1mm thick. Any suggestions on how to thin the Diamond wafer? The Diamond is actually poly-crystaline CVD
Thank you in advance Adam Dr Adam Papworth Dept Materials science & Engineering Ashton Building The University of Liverpool L69 3BX
Phone No 0151 794 5372 Fax No 0151 794 4675 E-Mail adamp-at-liv.ac.uk {
SOBOCIG wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } In short, I would like to inquire as to the methods and chemicals that people } are using for cleaning their filament assemblys on their electron microscopes. } Especially the final step. } } When changing our filament for our TEMs, we have in the past cleaned any } tarnish or deposits from the filament assembly pieces using a mild polish, then } sonicated the parts in a detergergent solution, followed by water rinses, } sonication in a few rinses of ETOH, then finally sonication in a freon-based } ultra-precision cleaning solution. (The used freon solution is dumped into its } own waste bottle.) } } We are considering changing this process (especially in consideration of the } availability and disposal of the freon solution) and are wondering what other } labs use for cleaning their filament assemblys thoroughly. We are especially } interested in the final cleaning step (removing any leftover residues). } } Thank you for your input. Don't be afraid to be brief. } } Gregg Sobocinski } Parke-Davis Pharmaceutical Research } Ann Arbor, Michigan } USA } Sobocig-at-aa.wl.com
Gregg I use this procedure on SEM gun parts, apertures, aperture strips and any other critical parts: 1.) Clean with 1u diamond paste. Chuck Garber tells me that his has no silicones, unlike the metalographic pastes. I don't know for certain, but it works fine and he has the best price. I do third party service and an 18 gram syringe lasts me for years.
2.) Clean (ultrasonic) with Joy dishwashiing liquid and hot water. I understand that the Proctor & Gamble labs clean their critical AAU parts in this and find no detectable residue (-at- ppm or better levels).
3.) Rinse under hot tap water to remove all detergent residue (cold tap water has also always worked for me when that is all that is available).)
4.) If available, rinse with distilled water or DI water. If not avaliable, don't worry because I"ve never had any trouble finishing with tap water.
5.) Blow dry to avoid drying residue, especially in critical areas (wehnelt opening, anode opening, etc.). This IS important!
I'm going to get spammed on the use of water, but the reasoning is as follows: All organic solvents come in contact with plastics of various types, most of which contain plasticizers. Plasticizers will and definitely do contaminate e-beam columns. They're a lot like Apiezon grease. Keep them out of your vacuum system. Water will not polymerize, will not contaminate and will not stay in your vacuum system. It may take some time to pump it out, but when it's gone, it's gone and there is no trace of it left on your apertures, baffles and other critical parts. It is non-toxic (at least so far), it is non-flammable, it is readily available and it is cheap. The only negative effect that I've seen is the temporary increase in total pressure in the vacuum system. Before you anti-water people get wound up, have you ever put a resudual gas analyzer on your microscope? Try it and I guarantee, even with a turbo pump, you'll have at least 90% water. If you're using a diffusion pump you will have 97% or better water in your vacuum system (assuming that it's not leaking). That's why I don't worry about water. For those who are using cryopumps, I'll cry "uncle" (but how do you put up with the vibrations?).
Ken Converse owner Quality Images 3rd party SEM service
Dennis Collins wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Subject: Time:5:39 PM } OFFICE MEMO RE} Cooling water problems Date:12/10/97 } } Hans Brinkies wrote, } "I am still using an old ETEC Autoscan (SEM, vintage 1973). It is } still working well after more than 12 000 hrs of usage and usually we } get the results that we want. } } "However, a calcium containing deposit has been forming } in the cooling water supply (in Cu tubes, in cooling coils around } diff.pump, in heat sinks, ect). The microscope was donated to us } several years ago but was not connected for the last 18 months } to the recirculating water system in our laboratory ( we are using } filtered tap water). The water flow has now been reduced drastically } over the last few weeks and I fear that the 'pipes' may eventually } totally block up. } } "What is the best (and safe) way to reduce or remove this deposit. } Back-flashing was only partially successful. } } "Any suggestion ? } } "Thank You } } "Hans Brinkies } SWINBURNE, University of Technology } School of Engineering and Science } Electron Microscopy Laboratory } HAWTHORN, 3122, Australia" } **************************************** } Hans, } The same problem occurs in lots of water-cooled, high power equipment such } as vacuum tube amplifiers, x-ray tubes, and particle accelerators. } For components with copper cooling water passages we use only low } conductivity water (min of 750K-ohm-cm: 1 Megohm-cm is better) in a } closed-loop cooling system. Even then deposits will accumulate. } We use water flow switches to ensure at least a minimum water flow in } sensitive equipment, and try to keep an eye on the pressure drop across each } water cooling circuit, back-flushing with a dilute solution of Sulfamic acid } when we see a clear rise in the pressure drop at the required flow. } Hope this helps keep your ETEC Autoscan running. } Dennis Collins } DGCollins-at-lbl.gov
Hans
Some how I didn't get the original message and don't have your e-mail address and so can't reply directly, but here goes:
How much flow can you get? The system only needs 4 to 6 gallons/hour but you should be able to get more than 10 gph at 20 psi and a good deal more at higher pressures. You may be able to do an acid flush on the whole thing but I would only flush the whole thing if the problem isn't localized. The ETEC is fairly easy to trouble-shoot in the plumbing area and you should find where your blockages are, first. They can be most anywhere, but start with the DP, the nylon "J" tube following and the 1/4" copper tubing from there to "water out". Also, sometimes it is only the fittings, not the tubing, so just replace them. Most likely the DP is plugging up. Remove the DP from the system. Remove the 1/4" PolyFlo x 1/8 mpt elbow from the outlet end and scoop out as much muck as you can. Try water or air (at 60 - 100 psi) from the top. If the flow remains restricted, pass some HCl (10%-30%) through under low pressure, about a 1 to 2 foot head, and be careful of the splatters. Follow with water and repeat if needed. If the nylon "J" tube is the problem, you can try and clean it or just replace it. Polyethylene tubing is not recommended here due to the high temperatures, but can be used in a pinch. Either type of tubing can be bent in boiling water, then cooled to form a permanent "J". If the copper tubing between the nylon tubing and the outlet is plugged, replace it with 1/4" refrigeration tubing. Once you get your system cleaned out, periodically flush it. The easiest way is to turn off the water, disconnect the "water in", and attach the "air" to the "water in" to blow out the lines. Reconnect the lines correctly and run the water at as high a pressure as you can. Repeat this until the lines run clear. AFter you get familiar with this, you won't even bother to turn the DP off because it can be done so quickly.
Ken Converse owner Quality Images 3rd party SEM service (ETECs are our specialty)
Just an update from an electron microscopist who as a service engineer has probably cleaned more electron guns round the world than very many others.
As many will know I have been involved with the maintenance of electron microscopes for 33 years. Firstly as a service engineer and then through my own training organisation where we train both operators and service engineers. We run regular maintenance courses around the world when we get a good idea of which cleaning materials and solvents are available.
Once again I have been watching the discussions with interest to see if there were many holes in the cleaning explanations. That said may I toss in my few pennies (cents) worth?
TUNGSTEN Gun Systems
The cathode assembly should be cleaned every filament change, the anode every other change and the electron gun at least once a year.
Materials - Almost any metal polish of less than 1 micron may be used to clean electron gun components however it must not be LONG LIFE. Long life additives coat the cleaned item with a polymer that causes chaos in the electron gun. Look out for any indication on the bottle or tube that the manufacturer is claiming that you will not need to clean the metalwork so often after using their product!
Method - Almost more important than the cleaning efficiency is our ability to completely remove the polishing media. So many service call outs are due to problems caused through inefficient removal of the media. For this reason it makes sense to use a metal polish that is easily removed by a solvent for tungsten. In this way we not only remove the metal polish but also clean the areas that are difficult to approach with the polish, nooks and crannies! Also very important is the need to clean without damaging the component, scratching it or placing cotton hairs within the "traps" that the manufacturers seem to put in our way. The best cleaning technique is a wet clean, that is to use solutions and an ultrasonic cleaner. In this way the damage that mechanical forces apply to the components are minimised. Sure the cathode aperture may need a little more encouragement to give up its deposit but only do this if the wet cleaning procedure falls short. We like "Silvo" or "Bluebell" or "Brasso", liquid metal polishes that will mix with a dilute ammonia solution to form a cleaning media, but a solution that may be removed with further washes in dilute ammonia. The mix - 10% metal polish in 90% ammonia solution - where the solution is 10% ammonium hydroxide in water. Place the components, one at a time, in the solution with their least important face down wards. Never put gun components together in the solution as they will damage each other. Do not put an aluminium cathode in ammonia as it will go black, oxide! After 20 minutes in an ultrasonic the component should be clean, wash off in running water and run for another 5 minutes in straight 10% ammonium hydroxide in water. Swill off with running water and then wash in alcohol and dry. NEVER throw away your solutions until you have reassembled the cathode as it is quite possible for the small screws to have fallen out and to reside in the debris at the base of one of the cleaning containers. If you do have a deposit remaining in the aperture area of the cathode a little mechanical effort with the cleaning media may be required,
The gun chamber IS important and this should be cleaned through disassembly once a year, particularly with a TEM. Dirty guns hold gas and induce micro discharge which spoils images. Clean the gun chamber with metal polish, remove the metal polish with dilute ammonia and buff up the walls with a clean chamois or dear skin leather. To retain the cleanlyness of the chamber, each time you change a filament buff up the walls with the leather. If the chamber smells, oily-ozone smell, but is not visibly stained, this is the result of discharge and all traces of the smell should be removed with dilute ammonia.
Look after your gun, it is probably the dirtiest area of the microscope, other than the specimen area in a SEM or the camera chamber in a TEM, its state will determine the ultimate performance of the instrument (high voltage stability) and your filament life.
LANTHANOM HEXABORIDE
Technique developed by ANU Electron Microscopy Unit Canberra
Clean the cathode with 25% hydrochloric acid in water by immersing for 60 seconds and then cleaning with a weak alkaline (ammonia or sodium hydroxide). Wash with water and then alcohol before drying.
LaB6 sources should last a long time (1000 hours plus) but they do need an intermediate cleaning session about every 250 to 350 hours. Some people amaze us by getting away with 1100 hours without cleaning but this is the exception not the rule.
Good luck!
Steve Chapman, Senior Consultant, Protrain, Oxford ,UK Tel & Fax +44 (0) 1844 353 161 web page - http://ourworld.compuserve.com/homepages/protrain
I about to purchase a double tilt heating stage (~1000=83C) and would like any suggestions or critics of Oxford vs Gatan (or any others). This would be for use in a JEOL 2010 with +_ 30=83 of tilt in both the x and y. In particular I would like info regarding temperature stability and accuracy, TEM sample stability, extra EDS signals, "smoothness" of tilting, durability of the holder, sample thickness limitations of the holder, etc. I know I could get info from the vendors but they sem to think they are perfect and the others are substandard. Thanks in advance
Brad Storey Materials Scientist Argonne National Lab - West P.O. Box 2528 Idaho Falls, ID 83403 Ph. 208-533-7685 (office) Ph. 208-533-7439 (lab) =46ax 208-533-7683 =05brad.storey-at-anl.gov
We're looking for a "highest-resolution at low-cost" laser printer for SEM micrographs. LaserMaster has an 1800 dpi printer at about 5-10 cents per page. Experience/suggestions appreciated.
JB
Jacob Bastacky, MD Room 116 Donner Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Telephone: 510.486.4606 Berkeley, California 94720 FAX: 510.486.4750
Dear sir, I'm read your home page and known your address on it.My name is Ms.Vimonwan from Faculty of Public Health,Burapha University.I'm Anatomist and lecturer.I'm beginning study about skin Thai frog and I hope to know about processed some skin frog for TEM,SEM? Because skin frog have thinkness of mucous layer.I don't know" How to wash -out ?" What is the best fixative to keep it? How long! If you don't mine,please introduce me about that and name of text book to improove my work.Area of skin on thumb in male frog and behind axilla in female frog. I will to comparative in this area in during breeding season and non-breeding. Thank you very much for your kindness. Finally,I'm looking forward to hearing from your soon. Yours sincerely, Ms.Vimonwan
My address: Ms.Vimonwan Nakiem Faculty of Public Health, Burapha University, Chonburi 20131 THAILAND
Hello All I would appreciate suggestions to following matter: in } 5 years old TEM microscope (particularly CM-20 Philips, with Riber IGP) the ion getter pump is getting old and the pumping times increase(specially after admitting air) - for many reasons I am aware of, but one of them is erosion of Ti electrodes inside IGP. 1) When would You decide to exchange the pump body - what level of decrease in performance ? 2) Did You exchanged allready by Yr machine the IGP pump ? - if yes - after what period of operation, and what were the symptoms ? Thanks in advance for any info from experience and practice.
regards Krzysztof M. Herman LabSoft S.c. 05-500 Piaseczno, ul. Kosciuszki 21, Polska tel/fx: (48 22) 7502024, 7502028, 757067, mobile: (48 90) 213438, (48 90)299748 fax: (48 22) 483787, Email: labsoft-at-labsoft.com.pl zapraszamy do http://www.labsoft.com.pl/ ------=_NextPart_000_01BD06D9.1534E5A0 Content-Type: text/html; charset=ISO-8859-2 Content-Transfer-Encoding: base64
Ronnie: Are you indifferent to know who wrote and sponsored a research project? Do you thing that in our society it is unimportant to know who recommends a product? Do you think its o.k. for a person with obvious commercial interests to edit a "Trade journal" without at least advising readers of those commercial interests?
These things matter a good deal, otherwise the big buck will rule all and not just most things. "Childish"? I have not been called that for some decades! I would, however, advise that ad hominem (against person rather then the argument) should not be used in a professional forum. "Had gumption"? Well, our online amounts to 11 megabytes. Over half of the "pages" are services like: MSDS, User Notes and Links. Tell me when you find a better catalogue, I love to learn.
Our catalogue is obviously a commercial site and its not masquerading as a society's site. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au ---------- } Jim, } Don't be so childish. If your company had the gumption and interest to } support the profession you serve, perhaps you would have had the insight } to establish such a site as Steven has. } Ronnie Houston } Dallas, TX
} Date: Friday, 12 December 1997 3:26 } Jim Darley wrote: } } } } Its fair and reasonable for Steven Slap to provide this information, but } } why does he not indicate that he is the editor of the suggested histology site?
} } Sure his name appears on that site and his company is shown as a sponsor (a } } cheap form of advertising), but no connection is made and most visitors } } would not realise that Steven Slap is the manager of the sponsoring firm and } } thus has a commercial interest: In essence it is a commercial site. } } Predictable his company is the only consumable supplier company shown. } } Nothing wrong with his sponsoring or editing such a page but visitors to } } the page and readers of this listserver are entitled to know not just who } } is sponsoring what but what commercial interests the editor has. Its known } } as propriety! } } Jim Darley } } } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 77 740 370 Fax: +61 77 892 313 } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } ************************ http://www.proscitech.com.au } } } } ---------- } } } From: Energy Beam Sciences, Inc. {ebs-at-ebsciences.com} } } } Dear fellow microscopists, } } } } } } At 08:55 AM 12/10/97 +0100, Heinz Fehrenbach wrote: } } } } can anyone of this newsgroup give me information about and subscription } } } } procedure to any related newsgroup ? In particular, I would appreciate } } } } to hear whether there is a special newsgroup discussing } } immuno-histology, } } } } -fluorescence, -blotting, and pathology subjects. } } } } } } There is an immunohistochemistry list, called ipox-l } } } } } } To subscribe, send a message to majordomo-at-pathology.stanford.edu and } } write } } } subscribe ipox-l in the body of the message. I believe that there is } } also a } } } pathology list, and instructions for this are available at "The } } Histotech's } } } Home Page" (http://www.histology.to) } } } } } } Best regards, } } } Steven E. Slap } } } ******************************** } } } Energy Beam Sciences, Inc. } } } Adding Brilliance To Your Vision } } } ebs-at-ebsciences.com } } } http://www.ebsciences.com/ } } } ********************************
Krzysztof M. Herman =20 LabSoft S.c. 05-500 Piaseczno, ul. Kosciuszki 21, Polska tel/fx: (48 22) 7502024, 7502028, 757067,=20 mobile: (48 90) 213438, (48 90)299748 fax: (48 22) 483787, Email: labsoft-at-labsoft.com.pl zapraszamy do http://www.labsoft.com.pl/
Dear Krzysztof, I don't know about your special type of Riber IPG; I have in my lab a = LEYBOLD-HERAEUS IPG, which has been changed as a whole (Ti-netting) 2 = times now (within 17 years of use). In addition, 2 times we could turn = the Ti-nettings 180 degrees round (after cleaning). In the manuals = provided by LEYBOLD for the IG-pump there was indicated a life-span of = about 47.000 working hours, provided vacuum better than 1.0x 10 to the = minus 6 mbar, and it was suggested that it was more economic to change = the whole pump after those 47.000 hours of pumping. So I am convinced, = you should have an instruction manual on the RIBER IG-pump too, provided = by the TEM-dealer (Philips, or any else). If you don#t get to a solution, try the following Compnay/+address for = more information on re-building your particular IG-Pump:
DUNIWAY STOCKROOM CORP. 13?5 Space Park Way MOUNTAIN VIEW, CA. 94043 USA Phone: USA/650/969-8811 Fax: USA/650/965-0764 or visit their www-Site:
No commercial interest in products/product lines, company/-ies, if such = names are mentioned or such are refered to. In this case I am only a = catalogue holder.
Best regards and Season's greetings, Merry Christmas and calm holidays Best wishes for a HAPPY, HEALTHY, SUCCESSFUL and PROSPEROUS NEW YEAR TO = ALL of YOU
Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: labsoft[SMTP:labsoft-at-labsoft.com.pl] Gesendet: Freitag, 12. Dezember 1997 08:36 An: MSA Microscopy Betreff: Old Ion Getter Pumps...
At 07:57 AM 12/12/97 +0200, you wrote: } yes. i meant carbon coated holey films. } Sara }
Sara,
Here is a method I have used in the past. It's a hassle, believe me, but it usually works. It also makes you realize why purchasing holey films commercially costs so much---they're not easy to make.
1) If you can find it, purchase a product called "Victawet" (I believe Electron Microscopy Sciences carries it). This comes in very small vials and it lasts forever. It serves as a lubricant to release plastic films from glass slides in the following steps.
2) Get some high quality glass microscope slides. We have found that Esco brand slides work more reliably than any other kind. Don't know why. The ones with one frosted end are useful for keeping orientation.
3)Polish these slides until they gleam and show NO contamination upon close examination. Do this even if they are "precleaned".
4)Take a piece of Victawet about the size of a cooked grain of rice and put it in a tungsten basket in a vacuum evaporator. Arrange as many CLEAN glass slides around it facing the basket. A distance of several to many centimeters is fine----i.e., distance is not too critical. It may be possible to make a custom rack for this purpose out of plastic or metal---the material used should not outgas too much.
5) Pump down the evaporator and when high vacuum has been reached, gently increase current to the basket. At a certain point the victawet will start to melt and you will see a fog developing on the slides. (If you use too much current, the piece of victawet may jump out of the basket and you have to start over, so be patient.) This "fog" is what you want. No need to overdo it, a little bit will work fine.
6) Repeat Step 5 until you have as many slides as you need. Use a new piece of victawet for each batch. Store these slides in a container for later use, since they keep for a few weeks.
7) Get some formvar (some people use butvar) in 1,2 dichloroethane (ethylene dichloride). 0.5% or 0.25% usually works. You can order 1% and dilute it, too. Keep this solution in a dessicator. Water in the solution causes problems, so only open it when necessary and for as brief a time as possible. Pour this in a tube wide enough to hold a slide, to a depth just short of the frosted end of the slide. (You can conserve formvar by using a special tube with a constricted lower end. One product is called "Dip Miser" and I think it's sold by Ted Pella. We had a special dipping tube made by a glassblower from a regular glass cylinder fused with a very wide constricted bottom.)
8) Cover the top of the tube with the formvar with something so that the atmosphere inside becomes saturated with the evaporating dichloroethane.
9) Get a container big enough to hold water to a depth of several inches. It should be 8-10 inches in diameter for comfortable working. Fill it to the top with, preferably, double-distilled water.
10) Now you're ready. Take the victawet coated slides and polish them again. They should feel slippery. Clean them until they gleam. Clip the frosted end of the slide with something to hold it, like a paper clamp attached to a piece of wire, and put it in the formvar solution in the tube. Keep it there for about 5-10 seconds, then pull it up and let it drain INSIDE THE TUBE. Only leave the top of the tube uncovered while putting the slide in and taking it out, in order to keep the atmosphere saturated. While the slide is draining, keep something over the tube opening, allowing only enough opening for the wire. The length of time you let the slide drain determines the final thickness of the formvar film. Start at about 10 seconds. If you desire a thinner film, increase the draining time up to about 15-20 seconds. (This is why the atmosphere must remain saturated with dichloroethane inside the tube. If it is not, the formvar just dries with draining properly.)
11) To make the holes, expose the slide to moisture IMMEDIATELY upon removing the wet slide from the dipping tube. We would breathe on it, but passing it quickly over a bath of steaming water might do the same thing. The microdroplets of water in the steam or in your breath make the holes. To repeat, this must be done IMMEDIATELY before the slide has time to begin drying. This is also a good time for prayer, invocations to the ancestors, luck rabbit's feet, and anything else you might find effective in appeasing the gods of holey films.
12) Lean the coated slide up against something in a dust-free area to dry for at least two minutes.
13) Back to the basin of water---take a clean laboratory tissue paper and drag the surface of the water to rid it of oil and dust. You can also put a drop of collodion in amyl acetate on the water, let it form a film, then pick this up and discard it to clean the surface.
14) Take the slide and cut around the outside perimeter of the film with a clean razor blade or scalpel. Then take the slide and, holding the frosted end, SLOWLY dip it into the water. If all goes well, the victawet was good, and you have been virtuous recently, the film will separate from the slide and float on the surface of the water. (HINT: a friend of mine---thanks, Steve---discovered that heating the water really helps in releasing the film. Don't boil it, just warm it up.)
15) You can now take your clean TEM grids and place them carefully on the floating film. When you have enough, take another CLEAN glass slide (no need for victawet this time) and scoop the film up. This is tricky and hard to describe: the idea is to catch the end of the slide on the end of the floating film so that when the slide is pushed down into the water the film will be pulled down with it and against it. When you finish, the grids should be between the formvar film and the glass slide. Put it somewhere dust free to dry. When dry, you place the slides in a vacuum evaporator and coat them with 100-300 angstroms of carbon. Then, you can GENTLY AND CAREFULLY push on the edge of the grid to pop it loose from the film. At this point, you will hopefully have a carbon-coated holey film on a grid, ready for use.
Alternatively, you can use a "domino rack", a piece of metal with small holes in it and two bent ends that serve as legs (kind of like a little bench with holes a few millimeters larger than TEM grids). These can be purchased commercially, or made yourself if you can find the right kind of metal with holes. When the film is floating on the water, before putting the grids on it, slowly lower the CLEAN domino rack onto the film, which will adhere to it by surface tension. Lift it out and put it in a dust free place to dry. The result is formvar film covering a bunch of little holes. The grids are later set upon these films by placing a drop of double-distilled water on the film, placing the grid on the drop, and letting it dry down. The film with grids can then be carbon-coated, or you can coat the individual grids later.
I warned you. This is a good starting procedure and variations can be done at several points, depending upon your circumstances. Things can go wrong at any stage, affected by humidity especially. It's a frustrating procedure and I would welcome any suggestions for making it easier, but it does work if you're patient and willing to experiment a little. If you do get a batch to work, I suggest making a whole bunch at once while luck is with you. You may not be so fortunate next week.
Hope this helps. Let me know if you have any questions, and I'll try to help more.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
I've had a request from someone for information on the wee mites - I think they're mites! - that lurk around our eyelashes. They especially wanted to see a picture. Someone suggested that Discover Magazine had published such a micrograph. I went through a stack of back issues but missed any such article. Does anyone know of a handy source of this information/photo? Thanks.
Carolyn J. Emerson email: cemerson-at-plato.ucs.mun.ca
Biology Department Memorial University St. John's, NF A1B 3X9 Tel: (709) 737-7515 Fax: (709) 737-3018
Fellow Histoneters, I know that this has been discussed but obviously I need more imput. What is everyone doing about using controls for immunofluorescent studies. My understanding is that CAP dictates that a control must be run on each antigen being used. Has CAP provided the source where we can "purchase" these controls? If a Positive Patient is used as a control, are we legally allowed to do this without informing the Positive patient? Please help! Teresa
Brad- both holders are probably very well made, I have experience with the=20 Gatan double tilt, temp. controlled specimen holder, the temp control unit= =20 was very accurate, the stability is somewhat compromised (compared to=20 standard double tilt holders), using the Be locknut and Hex ring reduces th= e=20 chances for any unwanted signal, the vacuum on the liquid nitrogen vessel= =20 needs "recharged" every year or so, the only problem I found (which has=20 probably been corrected by now) is with the precision of the tilt axis,=20 there was a very coarse tilt mechanism for the secondary tilt axis. Ask yourself what you need from your tools, then look at each company's=20 spec.s, and haggle price! -M. Rock
On Thu, 11 Dec 1997, Brad Storey wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } =20 } I about to purchase a double tilt heating stage (~1000=83C) and would lik= e } any suggestions or critics of Oxford vs Gatan (or any others). This } would be for use in a JEOL 2010 with +_ 30=83 of tilt in both the x and y= . } In particular I would like info regarding temperature stability and } accuracy, TEM sample stability, extra EDS signals, "smoothness" of } tilting, durability of the holder, sample thickness limitations of the } holder, etc. I know I could get info from the vendors but they sem to } think they are perfect and the others are substandard. } Thanks in advance } =20 } Brad Storey } Materials Scientist } Argonne National Lab - West } P.O. Box 2528 } Idaho Falls, ID 83403 } Ph. 208-533-7685 (office) } Ph. 208-533-7439 (lab) } Fax 208-533-7683 } =05brad.storey-at-anl.gov } =20 } =20 } =20
I'm trying to find out if there are going to be any good short courses or workshops on molecular biology techniques such as in situ hybridization, apoptosis detection and so forth in the next few months. I know that some companies run these courses as do societies and educational institutions. Any information would be greatly appreciated!
To all those who wrote to me before with suggestions for curing my wrinkle problems with semi-thin sections, I thank you, but the problem still persists, after trying those suggestions.
But I only get the problem really with nerve longitudinal sections. Could any technologists or scientists, or techno-scientists suggest any other plastic other than Epon/Araldite or Spurr that might yield better results, such as Epon or Araldite itself???
Ray, We recently retired our Delta system. Over the years we had troubles with the monitor and tried finding anything else that would work with their video signals. The scan rates are just unusual enough that other monitors can't quite synchronize on the signal, and we tried a few.
I would suggest taking the image from the Kevex and transporting it to another computer of your choice. Kevex had a package for converting images. I wrote similar programs on my own which I would be happy to share.
One program converts IMS and FXM files already collected. A second program grabs the graphic screen image and saves it to a file. It works best if you have TSX+ available for running multiple programs at once.
Then there is the matter of shipping the files to a PC. The Kermit program does okay and the Kevex can be attached to the ethernet, but that is not so cheap a solution. Feel free to ask for more details.
At 08:43 AM 12/11/97 -0500, you wrote:
} Has anybody successfully hooked up the color display output from a Kevex } } } Delta EDX to a personal computer running Windows; if so how did you do } it? Our system uses an old DEC computer and an Electrohome monitor with } } } individual RGB and separate horizontal and vertical sync connections. The } } } only printer output is to a serial connected to an old OKI dot matrix } printer. } } Thanks in advance. } } Ray Haythornthwaite ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
Both web pages are very nice, I just subscribed to the IHC newsgroup, and am looking forward to improving my IHC techniques. Everybody has some kind of interest, don't get so paranoid, thanks for the info., drop the argument.
-Mike Rock no disclaimer, no affiliation, no stress
On Fri, 12 Dec 1997, Jim Darley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Ronnie: } Are you indifferent to know who wrote and sponsored a research project? } Do you thing that in our society it is unimportant to know who recommends a } product? } Do you think its o.k. for a person with obvious commercial interests to } edit a } "Trade journal" without at least advising readers of those commercial } interests? } } These things matter a good deal, otherwise the big buck will rule all and } not just most things. } "Childish"? I have not been called that for some decades! I would, however, } advise that } ad hominem (against person rather then the argument) should not be used in } a professional forum. } "Had gumption"? Well, our online amounts to 11 megabytes. Over half of the } "pages" } are services like: MSDS, User Notes and Links. Tell me when you find a } better } catalogue, I love to learn. } } Our catalogue is obviously a commercial site and its not masquerading as a } society's site. } Jim Darley } } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 77 740 370 Fax: +61 77 892 313 } Great microscopy catalogue, 500 Links, MSDS, User Notes } ************************ http://www.proscitech.com.au } ---------- } } Jim, } } Don't be so childish. If your company had the gumption and interest to } } support the profession you serve, perhaps you would have had the insight } } to establish such a site as Steven has. } } Ronnie Houston } } Dallas, TX } } } Date: Friday, 12 December 1997 3:26 } } Jim Darley wrote: } } } } } } Its fair and reasonable for Steven Slap to provide this information, } but } } } why does he not indicate that he is the editor of the suggested } histology site? } } } } Sure his name appears on that site and his company is shown as a } sponsor (a } } } cheap form of advertising), but no connection is made and most visitors } } } would not realise that Steven Slap is the manager of the sponsoring } firm and } } } thus has a commercial interest: In essence it is a commercial site. } } } Predictable his company is the only consumable supplier company shown. } } } Nothing wrong with his sponsoring or editing such a page but visitors } to } } } the page and readers of this listserver are entitled to know not just } who } } } is sponsoring what but what commercial interests the editor has. Its } known } } } as propriety! } } } Jim Darley } } } } } } ProSciTech Microscopy PLUS } } } PO Box 111, Thuringowa QLD 4817 Australia } } } Phone +61 77 740 370 Fax: +61 77 892 313 } } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } } ************************ http://www.proscitech.com.au } } } } } } ---------- } } } } From: Energy Beam Sciences, Inc. {ebs-at-ebsciences.com} } } } } Dear fellow microscopists, } } } } } } } } At 08:55 AM 12/10/97 +0100, Heinz Fehrenbach wrote: } } } } } can anyone of this newsgroup give me information about and } subscription } } } } } procedure to any related newsgroup ? In particular, I would } appreciate } } } } } to hear whether there is a special newsgroup discussing } } } immuno-histology, } } } } } -fluorescence, -blotting, and pathology subjects. } } } } } } } } There is an immunohistochemistry list, called ipox-l } } } } } } } } To subscribe, send a message to majordomo-at-pathology.stanford.edu and } } } write } } } } subscribe ipox-l in the body of the message. I believe that there is } } } also a } } } } pathology list, and instructions for this are available at "The } } } Histotech's } } } } Home Page" (http://www.histology.to) } } } } } } } } Best regards, } } } } Steven E. Slap } } } } ******************************** } } } } Energy Beam Sciences, Inc. } } } } Adding Brilliance To Your Vision } } } } ebs-at-ebsciences.com } } } } http://www.ebsciences.com/ } } } } ******************************** } } }
This might be off-topic, but the news of a cure for my particularly intense and troublesome headaches was so exciting for me, that I thought I might as well share this with you folks, just in case there are some of you in the lab there suffering needlessly.
Basically I just take 2 Aspirin, and 2 Extra strength Tylenol AT THE SAME TIME. A doctor advised me that there was no problem in doing this, (ie: no adverse effects) because the 2 drugs work differently. So, whereas taking 4 aspirin would not be advised, you can fill in the extra cracks of pain relief with the tylenol. I double checked this with my pharmacist and she says that this is perfectly acceptable, and I'm not doing anything dangerous here.
Anyway, this combination of pain kill completely - blowing your headache into hyperspace, leaving you feel very comfortable.
I need to calibrate the magnification in of a TEM for magnifications above x100k, a range where the usual gratings standards are not so good. The resolution of the TEM is not great, but sufficient to use lattice images of asbestos fibers as a standard. However, I would appreciate to hear suggestions on other suitable magnification standards that can be used under diffraction contrast above x100k. Thank you for your help
Augusto _________________________________________________________________________ Augusto A. Morrone 107D-MEL, P.O. Box 116400 MAIC Materials Science and Engineering University of Florida Gainesville, FL 32611 (352) 392-1497 or 6985 Fax: (352) 392-0390 amorr-at-mse.ufl.edu
Augusto, You need to get a hold of a MaG-I-Cal sample. This was recently written up in the latest Microscopy Today by John McCaffrey. You can find them at SouthBay Technology. It not only allows you to do a mag calibration, but a rotation calibration and camera constant as well. It provides a mag calibration from the lowest to the highest mag on the TEM. I've just recently calibrated my TEM from 10kX to 500kX. -Scott Walck
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I need to calibrate the magnification in of a TEM for magnifications above x100k, a range where the usual gratings standards are not so good. The resolution of the TEM is not great, but sufficient to use lattice images of asbestos fibers as a standard. However, I would appreciate to hear suggestions on other suitable magnification standards that can be used under diffraction contrast above x100k. Thank you for your help
Augusto _________________________________________________________________________ Augusto A. Morrone 107D-MEL, P.O. Box 116400 MAIC Materials Science and Engineering University of Florida Gainesville, FL 32611 (352) 392-1497 or 6985 Fax: (352) 392-0390 amorr-at-mse.ufl.edu
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Question I need to calibrate the magnification in of a TEM for magnifications = above x100k, a range where the usual gratings standards are not so good. = The resolution of the TEM is not great, but sufficient to use lattice images = of asbestos fibers as a standard. However, I would appreciate to hear suggestions on other suitable magnification standards that can be used = under diffraction contrast above x100k. Augusto A. Morrone 107D-MEL, P.O. Box 116400 MAIC Materials Science and Engineering University of Florida Gainesville, FL 32611 (352) 392-1497 or 6985 Fax: (352) 392-0390 amorr-at-mse.ufl.edu
Response There is now a TEM calibration standard that supposedly is good the = entire range of mags on a TEM. I just got one but haven't had a chance = to try it. We got ours from Ted Pella but other supply houses may have = it
Judy Murphy San Joaquin Delta College Microscopy Technology Center Stockton, CA
some of us in the lab here are thanking you endlessly for this combination of pain kill completely - blowing our headache into hyperspace, leaving us feel very comfortable, but what do you thinking to mixingly paracetamol with acetyl salicylic acid ? Or evenly taking more paracetamol/tylenol: does your pharmacist thinking this might hit you liverishly, or grammatically?
Ray
ps for research purposes only, at which pint (sic) in your letter did you take the combination?
At 14:58 -0600 12/12/97, Garry Burgess wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ray Hicks ________________________________________________________________________ |University of Cambridge |Tel 01223 330149 | |Department of Medicine |Fax 01223 336846 | |Level 5, Addenbrookes Hospital |e-mail {rh208-at-cus.cam.ac.uk} | |Hills Road Cambridge |Web http://facsmac.med.cam.ac.uk | |CB2 |ftp server ftp://131.111.80.78 | |UK | | |_________________________________|_____________________________________|
Just a note to clarify a recent posting: I learned the technique I posted for making holey films from Steve Schmitt at the Center for Electron Microscopy at Southern Illinois University at Carbondale, which is directed by Dr. John Bozzola. I don't know where it came from originally, but I wasn't the one who developed it. Don't want to create the wrong impression.
Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
The standard you need is the MAG*I*CAL TEM Calibration Standard.
The MAG*I*CAL is a TEM calibration standard that performs all of the thr= ee major instrument calibrations for a TEM: image magnification; camera constant for indexing diffraction patterns; and image/diffraction patter= n rotation for relating crystal directions to features in the image. =
MAG*I*CAL consists of an electron transparent cross-sectional TEM sample made from a MBE grown, single-crystal semiconductor wafer. When the calibration structure is viewed in a TEM, it appears as a series of light=
and dark layers where the layer thicknesses are accurately known. The calibrated thickness measurements of these light (silicon) and dark (SiGe=
alloy) layers are based on careful TEM measurements of the {111} lattice=
spacing of silicon which is visible on the calibration sample itself, and=
are supported by x-ray diffraction measurements. The layer spacings are designed so that the sample can be used to calibrate the entire magnification range in a TEM - from 1,000X to 1,000,000X. As the sample = is also a single crystal of silicon, the calibrations requiring electron diffraction information such as the camera constant and image/diffraction=
pattern rotation can also be performed easily and unambiguously. One single calibration sample can therefore be used to provide all three of t= he major TEM instrument calibrations at all magnifications and all camera lengths.
South Bay Technology, Inc. supplies the MAG*I*CAL and so I have a defini= te financial interest in promoting its use. I also have copies of other research papers that have been written by the developer, John Mccaffrey, which will provide you with much greater detail. If you have an interest= , please let me know and I'll forward the information to you.
As a matter of additional interest, we can provide batches of the MAG*I*C= AL which are all made from the same wafer which provides the ultimate in calibration uniformity. This has proven to be an ideal solution to large=
organizations who are looking for a "company standard" calibration technique. Please inquire for more information on this service.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
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Dear Netters: =
I need to calibrate the magnification in of a TEM for magnifications abov= e x100k, a range where the usual gratings standards are not so good. The resolution of the TEM is not great, but sufficient to use lattice images = of asbestos fibers as a standard. However, I would appreciate to hear suggestions on other suitable magnification standards that can be used under diffraction contrast above x100k. Thank you for your help
Augusto _________________________________________________________________________=
Augusto A. Morrone 107D-MEL, P.O. Box 116400 MAIC Materials Science and Engineering=
University of Florida Gainesville, FL 32611 (352) 392-1497 or 6985 Fax: (352) 392-0390 amorr-at-mse.ufl.edu
Jill Craig wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi all, } } If anyone is looking into this topic or knows of any relevant } references or SEM methods, I would greatly appreciate the } information. I have scoured our meager library and the current } contents database with virtually no luck. My kingdom for } access to the science citation index. } } Thank you so much for any aid you can provide. } } Jill Craig } University of Northern British Columbia Jill,
I would suggest you contact John Delly through the McCrone Institute in Chicago. John has a wealth of info on wood.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
} Jill, } } I would suggest you contact John Delly through the McCrone Institute in } Chicago. John has a wealth of info on wood. }
The full company name (for directory assistance, etc.) is McCrone Research Institute. They are at 2820 S. Michigan Avenue, Chicago, IL 60616. 1-312-842-7100 (voice) 1-312-842-1078 (fax). http://www.mcri.org
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
Ray, lost your address but my client provides a software conversion package, KV2WIN whick will handle all your conversion problems for both images and spectra. For more info;
Bob Hessler Hessler Technical Services PHONE/FAX: 203/358-0266
Would it be possible to give me the address/phone/email of Lipshaw (Detroit) - I am interested in using a permenant mount for confocal specimens and I can't seem to find a European source for Permafluor.
thanks
Mark Auty
Dairy Products Centre Fermoy Co. Cork Ireland mauty-at-dpc.teagasc.ie
Salzburg, 14th of December 1997, 11.00 p.m. local time
CONCERN: CHEMICAL ELOXATION OF CORRODED ALUMINIUM-SURFACES
Dear all out there, I greatly would appreciate any suggestion or idea for solving the = following problem: in our Histology-Lab we use a "fixator" apparatus (for rapid fixation of = small tissue specimens contained in baskets in a cylindrical container, = made from chrome-nickel-steel, by means of alternating application of = pressure and vacuum in formaldehyde solution). The chrome-nickel-steel cylinder is durably fixed to a "collar" made of = eloxated aluminium (they cannot be separated for what reason ever), = which on its top serves as base for a sealing lid (made from plastic) = with an O-ring. Due to working with the specimen baskets (which turn out to be almost as = wide in diameter as the narrow cylinder's diameter) the eloxation of the = Aluminium-collar has been distorted and corrosion has been initiated.=20
The manufacturing company told us that there is no chance to overcome = that corrosion, the only way for stopping it could/would be to overlay = the corroded areas (after thorough cleaning: with what, if not alcohols = or acetone??) with silicone paste. Corrosion should be stopped then due = to exclusion of air/O2. The container-combination (chrome-nickel-steel - Aluminium) described = and used in the fixator now has been replaced by an other construct = which doesn't fit into the old apparatus. Also, one cannot buy the old = container as a sparepart, because it is not available any more. Since corrosion maybe goes on despite pasting silicone (as a byproduct = of condensation acetic or another acid will be formed) over the corroded = areas I am not sure about risks of a breaking of the Al-collar due to = pressure later on. Last chance would be to buy a new "fixator" which amounts appr. US$ = 3000.-
So my question:=20 is there ANYBODY WHO KNOWS OR HAS EXPERIENCE how to SEAL corroded = ALUMINIUM SURFACES, most elegantly by "artificial, CHEMICALLY INDUCED = ALUMINUM-ELOXATION" (procedure??)
Thanking you very much for your considerations best regards with my and our best wishes for a MERRY CHRISTMAS and A HAPPY, HEALTHY, PROSPEROUS and SUCCESSFUL NEW YEAR To you all and especially to you, and you
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
Dear colleagues…. Let me present you with a challenging scenario…. I am trying to Golgi stain some fixed mouse brains that have been sent to my lab…thus far I have tried using a Rapid Golgi and a Golgi Kopsch variant with no success….it turns out that although the brains were supposed to have been fixed in 10% neutral buffered formalin, instead they accidently used 10% formaldehdye (a 10% formalin solution contains only 4% formaldehyde). I am convinced that the excessive concentration of formaldehyde has contributed to the difficulty in obtaining successful Golgi staining. So, my question is --- Is there any way that the brains can still be saved for Golgi impregnation?? Might it help if the brains were rinsed in running water for a lengthy period and then put into the correct formalin solution?? Any thoughts or suggestions would be welcomed…. Thanks in advance… Ron Mervis ~~~~~~~~~~~~~~~~~~ Ronald F. Mervis, Ph.D. Neuro-Cognitive Research Labs RonMervis-at-aol.com
Have you thought of finding out the reason of your headache and fix that rather than stuffing yourself with drugs? And yes, it is really off- topic. ----------
This might be off-topic, but the news of a cure for my particularly intense and troublesome headaches was so exciting for me, that I thought I might as well share this with you folks, just in case there are some of you in the lab there suffering needlessly.
Basically I just take 2 Aspirin, and 2 Extra strength Tylenol AT THE SAME TIME. A doctor advised me that there was no problem in doing this, (ie: no adverse effects) because the 2 drugs work differently. So, whereas taking 4 aspirin would not be advised, you can fill in the extra cracks of pain relief with the tylenol. I double checked this with my pharmacist and she says that this is perfectly acceptable, and I'm not doing anything dangerous here.
Anyway, this combination of pain kill completely - blowing your headache into hyperspace, leaving you feel very comfortable.
Thanks for all who responded to my query. Especially those who were brief, but I learned a little bit from all.
For those of you interested in results of the survey, here's my abbreviated, paraphrased summary. I have focussed on the solvents involved, since that was the focus of my question. (My apologies if I misrepresent any procedures due to my brevity.) --- --- --- --- --- --- I received lots of information on polishes, and it seems that matching the solvents to the type of polish that needs to be removed is a good approach. (Some polishes clean up very easily with 10% ammonia solutions.)
It was suggested to use a 10% polish (Silvo, Bluebell, or Brasso) with 90% ammonia solution. This will turn aluminum pieces black, though.
Potassium Hydroxide (KOH), followed by dH2O and ethanol is a popular way to remove oxidation residues without the need for polish. (Cautions with this technique are that brass may discolor a bit, but it doesn't seem to cause any ill effects to the operation of the gun.)
It seems that drying the assemblys with heat lamps or hair dryers after rising in acetone, isopropanol or ethanol prevents the residue that sometimes forms. It appears that this residue may be from water condensing out of the air onto the assembly, which is cooled from the solvent evaporating.
In at least one case, someone uses dH2O as the final 'solvent' and dries with a hair dryer. The reasoning is that water vapor is the kindest of impurities to have in a column, compared to plastics and other chemicals that may dissolve in other solvents and may become deposited on the assembly during drying.
10% oxalic acid was also recommended as an added cleaning step.
--- --- --- Any more details about suggested procedures can be forwarded to you if you contact me before the end of January. After that, the information will be deleted.
Gregg Sobocinski Parke-Davis Pharmaceutical Research Ann Arbor, Michigan USA Sobocig-at-aa.wl.com
IN SITU HYBRIDIZATION RT-PCR WORKSHOP INTERDISCIPLINARY CENTER FOR BIOTECHNOLOGY RESEARCH UNIVERSITY OF FLORIDA FEBRUARY 5 & 6, 1998
THE CONTENT: In situ Hybridization (ISH) is a powerful method for the localization and quantitation of specific messenger RNA in single cells and within the natural tissue geometry. It has become the method of choice in the histopathological study of gene expression. When the technique is combined with PCR, mRNA can be detected at very low levels. ISH is also used for cellular detection of viral DNA and as a tool to obtain cytological information on the location and alteration of genomic sequences in chromosomes (FISH, PRINS).
This two-day workshop provides participants with hands on experience on the methodology and principles for the in situ RT-PCR detection of low abundance target sequences. The potential applications of the technique will be illustrated by the detection of adrenomedullin mRNA from rat brain tissue sections. In an additional experiment, participants will utilize ISH hybridization to detect viral DNA in infected cells.
Workshop includes lectures on a variety of ancillary techniques. Some of the topics include: Nucleic acid probe technology (antisense RNA probes), non-isotopic labeling and detection, tissue preparation and fixation for in situ hybridization, optimization of in situ methods (silane-coated slides, formalin fixation, protease digestion, DNase digestion and 4.5 mM MgCI2), fluorescent in situ hybridization (FISH, PRINS) mapping, cellular detection of viral DNA, etc.
THE LOCATION: Communicore Health Science Center, University of Florida, Gainesville, FL
FOR INFORMATION & REGISTATION: education-at-biotech.ufl.edu phone: (352) 392-8408 fax: (352) 392-8598
FEE: non-student: $200.00 FL Graduate students: $100.00 Registration Deadline: January 15, 1998 $25 Late Registration fee applies after deadline.
I hate to burst your bubble, but you have just re-discovered Exedrin! :-)
At 02:58 PM 12/12/97 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone have experience with TEM JEOL 1010 in dark field mode with SHP 10 OL pole piece. Im not satisfied by the results obtained with a SAP 10B OL pole piece of the 1010 JEOL. What is your point of view ? Does SHP pole piece could change something in quality of these images in dark field mode ?
Thanks for help.
P-Yves
E-Mail:sizaret-at-med.univ-tours.fr
Pierre-Yves Sizaret laboratoire de Microscopie Electronique UFR Medecine 2 bis Bd Tonnelle 37032 Tours cedex FRANCE
Dear Scott, With reference to the enquiry from Richard Fonda on the thinning of GaAs, here are some points from our experience. Br in methanol is good for the preparation of plan view specimens of GaAs, but I consider 5% is too strong a solution and likely to cause roughening of the surface. 2% is adequate and slow thinning can be achieved at a concentration of 0.5%. Be careful when adding Br to methanol as concentrated solution can explode.It is also best to use a fresh bottle of methanol as it should be dry. An alternative, which we actually use, is Cl in methanol. This can be used by bubbling the Cl into the methanol. Great care must be taken as at too strong a concentration it will set itself alight. We judge the concentration by the colour which goes from pale yellow to a stronger yellow/green as it increases. Both Br and Cl are hazardous materials and should be used in an adequate fume cupboard.
Pete Augustus Tel +1327 356362 Caswell Analytical Services Fax +1327 356775 GMMT Caswell Towcester Northants NN12 8EQ UK
} The absolute best, fastest, and cheapest way to prepare non-site specific } cross section samples with this type and thickness of metallization on GaAs } is with the small angle cleavage technique. There is a detailed pictorial } outline of the technique in the MRS Specimen Prep for TEM of Materials IV, } Vol 480 that just came out. The authors are myself and John McCaffrey who } developed the technique and has several pubs out on it. The technique } requires very little in terms of equipment that is not usually found in a } lab. Southbay Technology is selling a starter kit that has all the required } supplies. I can typically prepare about 9 samples in about an hour and } exchange and examine a sample within about 5 minutes to see if it is good. } A big advantage to this method is that there is no need to worry about } contamination of the sample. I've used this on GaAs and other materials and } examined the samples in field emission microscopes with no problems. I do } use electronic grade acetone and double rinse them. } } For the plan view samples, you will have to dimple and ion mill because of } the metallization. Don't forget to protect the good side from ion sputtered } material. } } } If you didn't have the coating and just wanted to make plan view samples } from the GaAs, I'd recommend chemically polishing the GaAs from the backside } after mechanically thinning to about 100 um to make the plan view samples. } Peter Goodhew showed me an inexpensive way to do this. I think that the } solution used was 5% bromine in ethyl alcohol (it might have been methanol) } which was dripped onto the sample from a burette. The sample was low } temperature waxed to a coverslip slide that was put onto a Teflon pedestal } with a small amount of vacuum grease mounted in a plastic cup. The grease } was not exposed to the solution because it was in the middle of the } coverslip and far from the edges. The cup was tilted at an angle of about } 30 degrees and was rotating with the use of a small DC motor. A } stereomicroscope was used to help determine when the sample was perforated. } } } You could use this method instead of dimpling if you stopped the process } before perforation and then continued the thinning process with ion milling. } The disadvantage is that you will not know the thickness left to ion mill. } } I hope this helps. } } -Scott Walck } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Guys Run Rd. (packages) } P.O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } } "The opinions expressed are those of S.D. Walck and not of PPG Industries, } Inc. nor of any PPG-associated companies." } } } ---------- } From: Richard Fonda } To: Microscopy } Subject: TEM GaAs prep } Date: Thursday, December 11, 1997 9:46AM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
INTERNATIONAL COURSES OF LIGHT MICROSCOPY, PHOTOMICROGRAPHY AND LASER SCANNING CONFOCAL MICROSCOPY GARGNANO (Lake of Garda) October 1998
The Course is a post-graduated theoretical/practical course, with propedeutical lectures and practical stages on microscopy, photomicrography and confocal microscopy. The course will take place in Gargnano (Lake of Garda) in October 1998.
Further information and registration details will be found at the following Web address.
http://imiucca.csi.unimi.it/endomi/micro.html
Thank you Paolo Castano
_____________________________________________________ Prof. Paolo Castano UNIVERSITY OF MILAN INSTITUTE OF HUMAN ANATOMY - CHAIR OF HUMAN ANATOMY FOR PHARMACY Via Mangiagalli, 31 - 20133 Milan (Italy)
Anybody has an idea how to stain starch in Chlamydomonas?
I tried (suggested in the list recently) without succes: Nile Blue, Safranin, ConA-FITC all for fluorescence. (Nile Blue was in water, sometimes after bleachingwith Na-hypochloride. Safranin coming from EtOH or also in water. All after fixation with glutaraldehyde, sometimes also after Triton. ConA stained many things but not starch).
Something else to try? Polarisation Microscopy doesn't work, J2KJ also not (I don't know really why ...)
Arthur Dr. Arthur Schuessler University of Heidelberg Zellenlehre Im Neuenheimer Feld 230 D-69120 Heidelberg Germany
The starch staining for LM on algae samples is generally done by KI+I2. Under LM the starch particals appear as dark-blue color and nothing alse can appear as this color. Acturally the usefull reagent is I, KI is used to increase the soluability of I2.
****************************************** Zhiyu Wang Electron Microscope Lab and Imaging Center Western Kentucky University(WKU) Bowling Green KY 42101
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Anybody has an idea how to stain starch in Chlamydomonas? } } I tried (suggested in the list recently) without succes: Nile Blue, } Safranin, ConA-FITC all for fluorescence. } (Nile Blue was in water, sometimes after bleachingwith Na-hypochloride. } Safranin coming from EtOH or also in water. All after fixation with } glutaraldehyde, sometimes also after Triton. ConA stained many things but } not starch). } } Something else to try? Polarisation Microscopy doesn't work, J2KJ also not } (I don't know really why ...) } } Arthur } Dr. Arthur Schuessler } University of Heidelberg } Zellenlehre } Im Neuenheimer Feld 230 } D-69120 Heidelberg } Germany }
Anytime I even got a whiff of Amyl Acetate (even when working in a fume hood) I would instantly get a very blinding headache. I used to use it as a solvent for Colloidon, a kind of support film plastic. But fortunately I have a better fume hood now, and I also rarely use this now. But I can also get headaches from fumes of epon/araldite and xylol. Even though we keep our polymerization oven in a fumehood, when we open the door of the oven to remove the plastic, some of the fumes escape. Of course safety conditions have improved a lot since days gone by, but sometimes I wonder what residual effects exposure to these chemicals might have on a person.
Garry } ---------- } From: Peggy Brannigan[SMTP:brannign-at-asrr.arsusda.gov] } Sent: 15 December, 1997 12:12 } To: Garry Burgess } Subject: Re: Blinding Headache } } Gerry, } } The subject of headaches may not necessarily be as off topic as one would } think. I had headaches almost daily for years until two things happened: } 1. I stopped using acrolein in my fixative and 2. I moved to a new } building where a.) my office was no longer in the lab itself and b). the } lab met OSHA requirements for # of air exchanges per hour. } } The headaches did not go away entirely but there was a dramatic change-I've } always wondered how many other microscopists suffer headaches. Since we } use our eyes so intensely staring at flickering screens and monitors and } expose ourselves to nasty chemicals it wouldn't surprise me if headaches } are, to some extent, an occupational concern. } } With respect to your tylenol-aspirin discovery , your're right on target } there. I go to a neurologist who basically prescribes the same thing, as } well as a few other medications. Headaches are complex, involving several } different pathways and success often requires taking several different } medications to target different components of the headache. } } Obviously it is best if you can find an environmental or dietary source for } the problem but that's not as easy as it sounds, simply because headaches } are so complex. There are numerous medications available that will prevent } headaches but you have to take them on a daily basis and should be under a } neurologist's care. } } One word of warning...if you take tylenol every day you may experience a } rebound effect where you get a headache when the tylenol leaves your } system. I don't think aspirin does this, but I''m not sure. } } Other things that have helped me: acupuncture, aerobic exercise, muscle } relaxation through biofeedback, and the herb feverfew. } } Anyway, good luck. } } Peggy } } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have made a list of what I consider to be the more common problems involved in Diagnostic Electron Microscopy as relates to specimen collection, preparation of plastic, sectioning, staining, and photography, for the benefit of outsiders to our lab, so that they might avoid some of these pitfalls. I would be interested in getting feedback from other people who might have other problems in mind that I have not considered.
My guess is you can indeed salvage the brains and still use them. Have they already been processed and embedded in paraffin? If so, you'll have to deparaffinize and rehydrate them, then rinse in running water or soak overnight in a large volume of (preferably) buffer.
Hint: Depends on what kind of tissue processor you have, but it is sometimes possible to run the processor in reverse order to deparaffinize and rehydrate...or, it is also possible to run the specimens through the "purge" or "retort clean" cycle, as well. This does a great job of deparaffinizing the specimens and leaving them in 100% alcohol; from there you can step down to water or buffer.
If you are convinced the extra formalin is the problem, this should do the trick. Unlike glutaraldehyde fixation (which is largely non-reversible), formalin fixation is to a large extent "reversible," in that large amounts can be coaxed out of the tissue by prolonged washing.
Good luck! Let us know how things work out.
Best regards,
Bob ********************************* Robert (Bob) Chiovetti E. Licht Company / 1-800-865-4248 rchiovetti-at-aol.com
********************************* Cryostats / Microtomes / Tissue Processors / Embedding Centers / Slide Stainers / Glass Coverslippers / Microscopes (Representing Leica since 1967) / Fiber Optic Systems / Linear Measuring / Micromanipulation (Linear Encoded, Video) / Image Analysis, Archiving, Capture / Video / Video Printers (Cooled CCD, Digital, RGB, Super VHS, 3-chip) / Vibration Isolation Systems / Programmable Stages / Heating & Cooling Stages
Rick Felten 12/15/97 12:52 PM Does anyone know a way to print several image files in an automatic fashion that would include the file name on the image. I am using windows 95.
Bob Schoonhoven recommended the Fontana-Masson stain for melanin pigment. We have a very fast microwave version of the Fontana-Masson stain. I would be happy to fax it to any interested parties.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
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RE} Safety using microscopes 12/16/97 11:48 AM Dear Keith,
While at ARCO Alaska we too were tasked to find a more ergonomic workstat= ion for palynologists and foram workers alike. We basically looked at the= level of their microscopes, computer screens, keyboards, chairs and heig= ht of desk(s).
Only a couple of people were experiencing problems at the time and the re= st did not care about the workstations because they had no problems. This= means that some people tolerate less than optimum ergonomics than others= .
SOLUTION: We changed the chairs to multifunctional settings type and sugg= ested differents heights of keyboards and monitors, microscopes. Most of = the problems went away except for the one person who had an existing aggr= avated lower back problem. His problem was not fixed by any of these solu= tions, nor could it be.
I personally did not have any problems, but when I changed my chair I not= iced a huge difference in comfort.
Finally, most light microscope users are "slumpers". Notice that they slu= mp down with poor posture even when not at the microscope. This presents = a problem when trying to fix their posture at the microscope and while th= ey are just sitting at their desks.
My personal opinion is that the chair is the single most important adjust= ment to make for workers at the desk and/or microscope.
Best of Luck
John Shane McCrone Research Institute
--------------------------------------
Dear Microscopy Listers
I have been tasked with coming up with something about using light micros= copes without harming yourself! Not so funny, according to two of our now-retired staff who counted plankton etc. for 20+ years. They both suff= ered neck/shoulder/back pain which eased after retirement.
In the UK we have a specific, detailed set of Regulations concerning comp= uter use, another unspecific set covers all workstations e.g. supermarket=
checkouts. A lot of the computer Regs. might be transferred to microscope= set-ups.
I'd appreciate any info, anything to avoid re-inventing the wheel!
} I have been tasked with coming up with something about using light micr= oscopes without harming yourself! Not so funny, according to two of our } now-retired staff who counted plankton etc. for 20+ years. They both su= ffered neck/shoulder/back pain which eased after retirement.
Keith, I have a handout that I use in my teaching to guide students in th= e proper positioning of the microscope and their bodies as they use the instrument. This is particularly important for those who may use the ins= trument for several hours at a time. Perhaps you will find the text of t= he hand-out helpful, though it may not reproduce particulary well in an emai= l message. Here goes:
Good Posture is Essential to Good Microscopy
Let=92s face it. Looking through a microscope is not what our bodies wer= e built for. And, looking through a microscope for an extended period of= time requires an unnatural rigidity of the body. You=92re not moving around, = even a bit, as you normally would. The result can be cramped muscles, especially in the neck and shoulders, and a whole body fatigue that can m= ake 10:00 am feel like 5:00 pm. The solution is correct posture and a pr= oper arrangement of microscope, chair, and body.
=B7 If you=92re going to be doing microscopy for any length of time, insi= st of a good, adjustable, ergonomically-designed chair!
=B7 Adjust the height of the chair so that your feet can rest comfortably= on the floor with an even pressure along the back of the thighs. It may= be advantageous to tilt the seat of the chair slightly down in front to elim= inate any pinch at the back of the knees.
=B7 Note: Adjust the chair height without regard to the microscope height= !
=B7 Adjust the position of the microscope so that you can comfortably gaz= e into the eyepieces without leaning significantly forward. This require= s two adjustments:
=A8 Set the lateral position of the microscope so that it is clos= e to the front edge of the bench with the eyepieces no further away from = you than the front edge of the bench.
=A8 Set the vertical position of the microscope so that it is a b= it high for comfortable viewing. This will normally require elevating th= e microscope on some type of stand on top of the bench. The idea is to for= ce yourself to straighten your back as you draw up to the microscope so t= hat with extended viewing your back is straight and your head in an upright p= osition. Look down into the eyepieces by letting your eyes view at a downward angle. Do not bend the neck to look =93straight ahead=94 into t= he microscope.
=B7 Obtain some type of arm rest for the arm used to focus the microscope= so that you don=92t need to be constantly raising the arm off the bench = surface and so that you are not tempted to say, =93Oh, the focus is good enough,=94= to yourself while you work. Constant, continuous focusing of the micros= cope is essential to minimize eye strain. (As an aside, proper setting of int= er-pupilary distance and eyepiece parfocalization are also essential to minimizing eye strain.)
=B7 After you=92ve established the optimal height for your microscope by = propping it up on phone books, this mornings paper, or whatever, you may = wish to have a permanent stand fabricated to the right height. Consider incorpor= ating one or two arm rests into the design. Go on, it won=92t cost you m= uch and it will make a world of difference in your comfort as you work. No p= oint growing old and crooked before your time!
=B7 Take a break from time to time. Get up, walk around, stretch arms, b= ack, neck, and legs to remove any =93kinks=94 that may be forming before = they become a bother.
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
Keith Ryan wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopy Listers } } I have been tasked with coming up with something about using light microscopes without harming yourself! Not so funny, according to two of our } now-retired staff who counted plankton etc. for 20+ years. They both suffered neck/shoulder/back pain which eased after retirement. } } In the UK we have a specific, detailed set of Regulations concerning computer use, another unspecific set covers all workstations e.g. supermarket } checkouts. A lot of the computer Regs. might be transferred to microscope set-ups. } } I'd appreciate any info, anything to avoid re-inventing the wheel! } } Season's Greetings } } Keith Ryan } Plymouth Marine Lab., UK Dear Keith,
Part of the problem is that few microscopists "align themselves", when they align the microscope. An adjustable chair, with secondary adjustments for the backrest, plus "elbow pads" or armrests on the microscope will solve most of these problems. The microscope should be addressed at eye level to avoid hunching over or stretching to reach the eyepieces.
If you decide to draft formal regs, send me a note.
Best regards, Barbara Foster (longtime FRMS) Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
Adriana P. M Rodriguez wrote: } } Dear Dr. Foster: } I just saw a message you posted on the microscopy list, and the phrase } below your address caught my attention. } I just wondered if our institution offers short courses in different } microcopy techniques, or only on-site training. } Thanks in advance for your time. } } Adriana Rodriguez } } } Best regards, } } Barbara Foster } } Consortium President } } Microscopy/Microscopy Education } } 53 Eton Street } } Springfield, MA 01108-2838 USA } } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } } **************************************************** } } Microscopy/Microscopy Education } } America's first consortium of microscopy experts offering } } customized on-site training & applications solutions } } } } } Adriana P. M. Rodriguez } Biotecnologia Vegetal, CENA/USP } Av. Centenario 303, Cx. Postal 96 } 13400-970, Piracicaba, SP, Brasil } phone: +55-19- 429-4694 } fax: +55-19- 429-4610 Dear Adriana,
We will be offering several courses during first quarter 1998: a. SPIE/Photonics West - "Modern Microscopy & Applications" - lecture deomstrationwhich reviews fundamentals of microscopy, including a range of contrast enhancement techniques, plus a discussion of advanced methods including integration of image processing technis, automated measurement, and feed-back systems. (Thursday, 1/29/98, San Jose, CA)
b. American Chemical Society - "Applied Optical Microscopy for Chemists" - a three day, hands-on, total immersion course in light microscopy which covers imaging theory, contrast enhancement, and basic measurement techniques. Emphasizes Polatized Light Microscopy. (Friday, Saturday, Sunday, 2/27, 2/28, 3/1, New Orleans, LA)
c. The MME Traveling Road Show - "Optimizing Light Microscopy" - a lively, one-day lecture demonstration on alignment, operation, optics, and contrast enhancement. (Monday, 3/16, New York City; Wednesday, 3/18, Springfield, MA/Hartford CT; Friday, 3/18, Boston, MA).
For further information, please email me directly.
Thanks for your interest. Barbara Foster Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
I just tried printing from ThumbsPlus 3.0. You can print in batch mode, you are not restricted to just a catalog and you can print a bunch of other information including:
name (with or without directory path) file size date dimensions pixels and physical size resolution keywords (that you added to it in the database) annotation (that you added)
All of these are optional in the output.
Agreeing with others, it is a great program. You can also scan images into it and the thumbnails are automatically made. We use the network version and have a LAN database that we can all select and we can have local databases for our hard drives. We open the images by double clicking on the image, copy it, and paste it into a window application such as a wordprocessor or PowerPoint. It can also be used to open a variety of formats including Photoshop and convert them.
-Scott
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Rick Felten 12/15/97 12:52 PM Does anyone know a way to print several image files in an automatic fashion that would include the file name on the image. I am using windows 95.
Please unsubscribe nsmith-at-csuhayward.edu Nancy R. Smith Director of Operations Microscope And Graphic Imaging Center California State University, Hayward Hayward, CA 94542 http://www.csuhayward.edu/SCI/sem
Has anyone use Softwindows 95 for a Silicon Graphics system with the Scion Image PC (PC version of NIH Image)? Also what has been your experiences with Softwindows 95?
Six months ago I said goodbye to the List and my job managing the Australian Museum's SEM lab. I'm now in Brussels starting a Ph.D. in Archaeology, but it's funny how things move in circles. I'm looking at 3D modelling of excavations, including Image Analysis in 3D, which is why I'm starting by asking those of you in confocal microscopy the following;
1) Can anyone help with an email address for H.J.G. Gundersen at the Stereological Research Laboratory in Denmark? I've tried Stereo-at-svcd.aau.dk but the message bounces.
2) Does anyone know of commercially available 3D IA programs? (I want to measure object sizes and object orientations)
3) Does anyone know of any Lists dedicated to 3D modelling?
Many thanks, Merry Christmas and may your New Year's "resolution" be small and stable.
Is anyone out there doing any EDX work with marine sediments - especially those in the vicinity of pipeline outfalls. if so, I'd like to communicate with you.
Please reply to my e-mail address rather than the listserver. Thanks
Mike Gregory
EM Unit University of Durban-Westville South Africa
} Does anyone have experience using the Polaroid Sprint Scan 45 for TEM } negatives? I would appreciate any opinions on this scanner.
My response:
We have been using the SprintScan 45 for 6 months. We are very pleased with the quality of images produced by this scanner. Initially, an inconvenience of the scanner was the lack of a holder for TEM negatives. Polaroid has subsequently developed one that uses anti-Newton glass that works fine. I was very skeptical about the use of glass but it has proven to work nicely. The design of the holder is such that it can be easily modified with frames to make a glassless holder if you must have it. I suspect that these frames are in the works.
I have just completed (last night, in fact) a side by side comparison of images generated conventionally by darkroom endeavors versus scanning of the same negative and printing on a Codonics 1660 dye sub printer. Even though I had to use a color paper on the Codonics (their thermal line paper is not yet ready for prime time), the results were amazingly good. I compared enlargements of 1x, 4x, 8x and 16x using a 10MB image scanned at 2000 ppi. (I am now looking at smaller sized files as well.) I must say that as a "classically trained microscopist" with considerable dark room experience, I was finally convinced that a reasonable, high quality image can be generated in this way. I spent a couple hours in the darkroom using various focal length lenses, condensor lenses and adjustments on the enlarger to achieve the results that could have been obtained in 20 minutes on the scanner and dye sub printer. Although I think that a darkroom may be needed in some cases (large central facilities, for example), digital darkrooms will work for the majority of users. So fast, so convenient, so much less environmentally damaging.....
Disclaimer: I have no commercial interests in either Polaroid or Codonics. I can be just as critical of these companies when they show shortcomings.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Hoping someone can help me. We used to buy PBS tablets from a company called OXOID. Need to order more but can't find address. Are they still around, or has someone bought them out. Any help greatly appreciated.
Thanks Mark Elliott UBC-Pulmonary Research Lab St. Paul's Hospital Vancouver, BC Canada V6Z 1Y6 604-631-5351 (fAX)
This is that magical time of the year when the thoughts of those of us in the tropics turn to... cryofixation!
We have a "cryogen freezing bath" device from Balzers, which consists of a small container on a tube which fits into a Dewar of LN2. The small container is filled with e.g., a freezing slush of Freon or propane, into which (cryoprotected) samples are plunged. Someone else on campus would like to have one of these devices, and the machine shops are really backed up. Do any of you have an idea where this kind of thing can be purchased? If you can't picture this, a schematic can be found in Bozzola and Russell's Electron Microscopy on p. 312.
Actually, while I'm at it, the intended purpose is to freeze mouse heart/aorta and liver for cryosections, and I'd be glad to pass on any tips any of you have. This group was just plunging the tissues into straight LN2 and wondering why they had big holes...! Pre-fixation is not a problem; in fact it is desirable. I think it's intended for in situ hybridization/autoradiography at the light level. It's those pesky photons, again. I just don't understand how they work, and so I'm not much help when it comes to LM!
It's about 76 degrees F and sunny in Honolulu today. Surf is coming up on the North Shore. Hope you all are having a fine holiday season!
Mele Kalkimaka, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I am also very interesting in 3D image analysis and reconstruction, especially to the biological subjects. I send you an address in web site which contains a great number of software. Hopefully you could find some of them are suitable to your current project.
http://biocomp.arc.nasa.gov/3dreconstruction.
Good luck!
Peiyi Wang Research Fellow Department of Engineering Materials University of Southampton Southampton SO17 1BJ UK Tel: 01703 595101 Fax: 01703 593016 E-mail: pw2-at-soton.ac.uk
There is no real need to purchase the quench cooling device you describe. All you need is a copper cylinder which will sit in a Dewer of LN2. Have a little plastic lid on the cyliner, When the cyliner is cool, slowly condense some propane into the cold cylinder (do this in a spark proof hood). Once the propane is cooled put the lid on. The purpose of this little lid is to stop LN2 bubbling into the cylinder with the propane. When you are ready to quench your sample, get a warm rid and make a little pool in the solid propane, watch the propane pool and as the surface just begins to frreeze over, plunge in little bits of your sample. Let then stay for 30 secs then quickly bring them out and put them into a dewer of LN2. Variations of this are endlezs. You can use ethane instead of propane (bit dangerous) and you can have the whole dewer-cylinder sitting on a magnetic stirrer with a magnetic flea in the secondary cryogen. If you are really serios, read my book "Low temperature Microscopy and Analysis"
Your weather sounds great. Here in Cambridge we are just recovering from the Siberian Express which gave a wind chill factor of -10C. Now it hovers around freezing with rain. But we English argue that you have to suffer cold and wet to appreciAte warm
Best wishes for the Christmas Season
Patrick Echlin Multi-Imaging Centre School of Biological Science University of Cambridge United Kingom.
On Wed, 17 Dec 1997, Tina Carvalho wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } This is that magical time of the year when the thoughts of those of us in } the tropics turn to... cryofixation! } } We have a "cryogen freezing bath" device from Balzers, which consists of a } small container on a tube which fits into a Dewar of LN2. The small } container is filled with e.g., a freezing slush of Freon or propane, into } which (cryoprotected) samples are plunged. Someone else on campus would } like to have one of these devices, and the machine shops are really backed } up. Do any of you have an idea where this kind of thing can be purchased? } If you can't picture this, a schematic can be found in Bozzola and } Russell's Electron Microscopy on p. 312. } } Actually, while I'm at it, the intended purpose is to freeze mouse } heart/aorta and liver for cryosections, and I'd be glad to pass on any } tips any of you have. This group was just plunging the tissues into } straight LN2 and wondering why they had big holes...! Pre-fixation is not } a problem; in fact it is desirable. I think it's intended for in situ } hybridization/autoradiography at the light level. It's those pesky } photons, again. I just don't understand how they work, and so I'm not } much help when it comes to LM! } } It's about 76 degrees F and sunny in Honolulu today. Surf is coming up on } the North Shore. Hope you all are having a fine holiday season! } } Mele Kalkimaka, } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** } }
Keith I assume that COSHH assessments will have taken care of fixatives, stains and immersion oil and that you aren't talking the risks of microtomes, knives and cryo work.
My feeling is that any assessment must look at levels of use:
If microscopes are used once a week for half an hour, keep them covered or in a cupboard and use them however you wish.
If microscopes are used regularly (like designated DSE user work) then a clear bench area in an a quiet area will be less stressful and ideally the user should be able to adjust the ambient light without disturbing others. Good seating should be provided which is adjustable or at least a variety of stools and chairs to suit all types of users also necessary basics like a stable bench surface, appropriate and well-placed power outlets (ie no trailing wires), lens tissues (keep those lenses clean), and a manual if necessary. Binocular eyepieces are fairly universal now and most users find these less stressful than monocular for prolonged use but they must know how to adjust them properly and should know how to set up the rest of the microscope for optimum use. Some spectacle users prefer to have high-viewpoint eyepieces so they can wear their glasses and some people prefer eyepiece eye cups (removable types are generally available). Adjustments such as X-Y stage controls, focus and lights should work properly (ie regularly maintained to avoid long term stress to the user). It may well be sensible to encourage people to take activity breaks as for DSE users. I would assume that if monitors are used to view the image they should be as suitable as modern computer screens ie sharp, no flicker and so on (if they are connected to computers then the DSE regulations should apply anyway).
Malcolm Haswell e.m. unit University of Sunderland UK
PS DSE is Display Screen Equipment, VDUs, or computer screens.
DISCLAIMER - these are my own personal opinions based on practical experience, even if I can't always achieve them.
----------
Dear Microscopy Listers
I have been tasked with coming up with something about using light microscopes without harming yourself! Not so funny, according to two of our now-retired staff who counted plankton etc. for 20+ years. They both suffered neck/shoulder/back pain which eased after retirement.
In the UK we have a specific, detailed set of Regulations concerning computer use, another unspecific set covers all workstations e.g. supermarket checkouts. A lot of the computer Regs. might be transferred to microscope set-ups.
I'd appreciate any info, anything to avoid re-inventing the wheel!
I am certain that this has been discussed before but can anyone give me guidance on the availability of large format black and white negative scanners for e.m. cut film (up to about 8.3 cm x 10.2 cm)?
I know that some flat-bed print scanners can take adaptors for film but it is not always obvious what format they can accommodate. At present the resolution of print scanners appears to be improving (600 dpi true resolution is not uncommon) and I suspect that for normal enlargement of 2 to 3x these would be just about adequate.
Any comments?
thanks Malcolm Haswell e.m. unit University of Sunderland UK
Oh and Merry Christmas to all my readers. ----------
Text item:
Does anyone have experience using the Polaroid Sprint Scan 45 for TEM negatives? I would appreciate any opinions on this scanner.
Disclaimer: The CSIR exercises no editorial control over E-mail messages originating in the organisation and the views in this message are therefore not necessarily those of the CSIR and/or its employees. Message-Id: {s4993452.076-at-csir.co.za} X-Mailer: Novell GroupWise 4.1
Hi We have a Jeol JSEM 200 STEM (200 kV) available at a nominal cost for anybody interested. If you are interested in the machine, or can want a part (high voltage transformer, lenses, scanning system, specimen holders, gun, etc), please let me know as soon as possible as the machine is heading for the scrapyard... All shipping must be paid by yourself.
Sara Prins
Surface and Structure Analytical Services Division for Materials Science and Technology CSIR PO Box 395 Pretoria South Africa
} There is no real need to purchase the quench cooling device you } describe.... a little pool in the solid propane..... Variations of this } are endless.
ISO-pentane freezes at about -160^C and comes in a bottle.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
We are interested in imaging, (non-destructively) the internal morphology of 6 - 8 micron polymeric particles. The SEM and TEM have already been used for this study and we have determined the size of internal voids to be between 0.1 and 0.4 microns. Does anyone know the resolution limits of acoustic microscopy and the location of a facility where I might get such work done?
Gerroir,Paul wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are interested in imaging, (non-destructively) the internal } morphology of 6 - 8 micron polymeric particles. The SEM and TEM have } already been used for this study and we have determined the size of } internal voids to be between 0.1 and 0.4 microns. Does anyone know the } resolution limits of acoustic microscopy and the location of a } facility where I might get such work done? } } Paul Gerroir Dear Paul,
I'd suggest to talk to the people at SONOSCAN and Leica. Both have acoustical microscopes in their product lines and may be able to suggest a facility close to yours.
My contact at Sonoscan is Don Commare (708/766-7088). I'm not sure who the Leica product manager is but you can reach Leica at (847)405-0123; they can put you in touch with the correct person.
Best of luck.
Barbara Foster Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
Am I missing something ? I never mentioned Isopentane.Of the two I mentioned ETHANE melting point 90K gives a mean cooling rate of 13-15 kKsec-1, PROPANE melting point 84K gives a mean cooling rate of 10-12 kKsec-1. Your stuff, ISOPENTANE melting point 113K does not cool as fast as the other two although I do not have the figures to hand. If you want to read more see Chapter 3 in "Low Temperature Microscopy and Analysis" by Patrick Echlin, Plenum Press, New York 1992. When choosing a cryogen there are many factors to consider including specific heat, thermal conductivity, relative cooling efficiency, viscosity at the crogen melting point andthermal inertia.
To all cryomicroscopists the Seasons Greetings.
Patrick Echlin Cambridge
Thu, 18 Dec 1997, Robert H. Olley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } On Thu, 18 Dec 1997, Dr P. Echlin wrote: } } } There is no real need to purchase the quench cooling device you } } describe.... a little pool in the solid propane..... Variations of this } } are endless. } } ISO-pentane freezes at about -160^C and comes in a bottle. } } +------------------------------------------------------------------------+ } | Robert H.Olley Phone: | } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } | University of Reading {University internal extension 7867 | } | Whiteknights Fax +44 (0) 118 9750203 | } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } | England URL: http://www.reading.ac.uk/~spsolley | } +------------------------------------------------------------------------+ } }
We have small and portable "Hanna HI8424 microcomputer pH meter" with an electrode that now reads about ph=8.1 instead of pH=7.0 the true value. When I push the calibrate button though, it just reads error (E4), and there is no way that I can calibrate this meter.
The electrode only says HI 1332 on it, and it appears to be a Ag/AgCl electrode. My problem is that I don't see "Hanna" listed in any catalogs, so I'm not sure what electrode I should buy to replace this electrode. It connects to the main control unit with a BNC connector. I have already tried rejuvinating this electrode by soaking it in various solutions, but so far nothing has changed in the way it reads.
Are there any Hanna pH meter types out there that might be able to advise me on what electrode I should get, and where I can get this electrode?
Paul Gerroir wrote: ========================================= We are interested in imaging, (non-destructively) the internal morphology of 6 - 8 micron polymeric particles. The SEM and TEM have already been used for this study and we have determined the size of internal voids to be between 0.1 and 0.4 microns. Does anyone know the resolution limits of acoustic microscopy and the location of a facility where I might get such work done? ============================================ You might want to contact a firm in suburban Chicago called Sonoscan, Inc. as follows:
Sonoscan, Inc. 530 E. Green St. Bensenville, IL 60106 USA
Tel: 630-766-7088 Fax: 630-766-4603
You would want to talk to Dr. Larry Kessler who is the founder and, so far as I know, still the President. He has helped me out of any number of tricky problems that were solvable with or only with accoustic microscopy. He is very knowledge about material science applications of accoustic microscopy. But to my understand, which might not be state-of-the-art, the resolution you are seeking I don't think is possible even with accoustic microscopy.
Chuck
Disclaimer: I have no connection with Sonoscan, financial or other wise.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Dear Judy, Scott, and David: Thank you for your response to my question on TEM calibration. I am following up on your suggestions.
Augusto _________________________________________________________________________ Augusto A. Morrone 107D-MEL, P.O. Box 116400 MAIC Materials Science and Engineering University of Florida Gainesville, FL 32611 (352) 392-1497 or 6985 Fax: (352) 392-0390 amorr-at-mse.ufl.edu
See our Web Page for detailed information about our current opening for an engineering manager. web site:kovexcorp.com or e-mail me your resume : kovex-at-spacestar.net, or telephone 612-486-9830 ext 113.
I have some investigators that are interested in doing shadow casting of DNA and large protien structures, but we do not have a carbon evaporator. They would like me to get some sources and prices for a equipment grant. Other than Electron Microscopy Sciences who else sells them? Are there any opinions on this matter? Thanks.
Rick Vaughn
RLVAUGHN-at-MAIL.UNMC.EDU EM Research Facility Dept. Cell Biology & Anatomy UNMC Omaha, NE
I have been asked to find a source for Fast Blue B.
According to the person making the request, Sigma, Fisher, and Acros have all discontinued all Fast Blue B Salts and precursors.
Does anyone know of a source or have a stockpile on hand?
According to the requester, Fast Blue B and its salts are carcinogenic and this is the reason they are no longer available. Don't ask me why he wants to fool around with something like this, I am just the messenger.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Dear Friends, I had the opportunity to share someone's: YOUR collegial friendship in = communicating more or less frequently via the MSA-Listserver or on else = opportunity. I received most valuable informations and help regarding my = requests (some were urgent) and will forward as intensely my knowledge, = if appropriate. For all those I promised informations, reprints, methods, silicone = rubber molds and instructions and else material: please apologize at = that moment for my being late with a lot of things. The month before = Christmas time every year is a strong one regarding daily stress of work = (all specimen preparations in our Lab. on my own, lectures, official and = administrative work, etc....and singing in a choir, the SALZBURG = MOZART-CHOIR with 4 different concert performances this month). I didn't forget (hope so) any of you and of my promises to you. Please allow for another some weeks, hopefully after holidays things = will go on faster than now. If one feels that something was not received = or mailed which should have been in the mail for a long time, please = feel free to urge or claim.
To all of you and yours greetings and my best wishes=20 for A Merry Christmas, at least some calm, familial and quiet days of = relaxation, as well as all the best for a HEALTHY, HAPPY, PROSPEROUS and = SUCCESSFUL NEW YEAR.
yours truly, *Wolfgang*
(Out of the Lab from 23rd Dec. to 7th of Jan, '98; but as I know me, I = will be in at some days in between for "work-up" of unsettled items)
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
SALZBURG, 19th of Dec. 1997, 01.35 a.m. local time
Dear Garry, dear all, surprising doublette:=20 the same happens/-ed with my pH-meter (BECKMAN ZEROMATIC IV) or better = the connected combination electrode (Ag/AgCl). I confess, the meter/electrode was not in use for three or four months = now, but: electrode was soaked all the time in a so called "storing = solution" originally supplied from BECKMAN (solution adjusted to pH = 4.0). I am watching now about 4 weeks what happens: despite "stand = by-function", the analog meter showed initially slowly increasing values = starting pH 7.0 to 8.5, now, after 4 weeks, it "displays" (without being = "in action") pH 11.0. If changing function from "Stand By"-mode to = "measuring" it displays pH 8.0. Now idea what goes on. Maybe a malfunction of an electrode too old = aged?? (about 5 years of age). I have not tried to compensate or to = refresh the electrode anyway, but there is white crystal growth on the = upper outside of the electrode tube, far away from the solutions in the = storage baker. Somebody out there with an idea? (BECKMAN as "last chance", I know)
Thanks in advance, best regards,
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
=20 GARRY BURGESS WROTE: ---------- Von: Garry Burgess[SMTP:GBurgess-at-exchange.hsc.mb.ca] Gesendet: Donnerstag, 18. Dezember 1997 20:49 An: 'Microscopy Society of America - Mailing List' Betreff: Hanna HI8424 pH Meter
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
We have small and portable "Hanna HI8424 microcomputer pH meter" with an electrode that now reads about ph=3D8.1 instead of pH=3D7.0 the true = value. When I push the calibrate button though, it just reads error (E4), and there is no way that I can calibrate this meter.
The electrode only says HI 1332 on it, and it appears to be a Ag/AgCl electrode. My problem is that I don't see "Hanna" listed in any catalogs, so I'm not sure what electrode I should buy to replace this electrode. It connects to the main control unit with a BNC connector. I have already tried rejuvinating this electrode by soaking it in various solutions, but so far nothing has changed in the way it reads.
Are there any Hanna pH meter types out there that might be able to advise me on what electrode I should get, and where I can get this electrode?
Dr Echlin is quite right. Isopentane had been used as a freezing compound a long time ago, but I sure was glad when propane was advocated as a better alternative. I started using propane about 15 years ago. Isopentane behaves like a "gone off" rubber solution near its freezing point and freezes well before propane. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} On Thu, 18 Dec 1997, Dr P. Echlin wrote: } } } There is no real need to purchase the quench cooling device you } } describe.... a little pool in the solid propane..... Variations of this } } are endless. } } ISO-pentane freezes at about -160^C and comes in a bottle. } } +------------------------------------------------------------------------+ } | Robert H.Olley Phone: |
Malcolm, our Links page has two sites that deal with "scanners". Use control F to find these sites. The site at Basel Uni lists numerous scanners and gives a good deal of technical details. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} } I am certain that this has been discussed before but can anyone give me } guidance on the availability of large format black and white negative } scanners for e.m. cut film (up to about 8.3 cm x 10.2 cm)? } } I know that some flat-bed print scanners can take adaptors for film but it } is not always obvious what format they can accommodate. At present the } resolution of print scanners appears to be improving (600 dpi true } resolution is not uncommon) and I suspect that for normal enlargement of 2 } to 3x these would be just about adequate. } } Any comments? } } thanks } Malcolm Haswell } e.m. unit } University of Sunderland } UK } } Oh and Merry Christmas to all my readers. } ---------- } From: Leah L Dobbs } To: Microscopy-request; microscopy } Subject: Negative Scanners } Date: 17 December 1997 17:51 } } Text item: } } Does anyone have experience using the Polaroid Sprint Scan 45 for TEM } negatives? I would appreciate any opinions on this scanner. } } Thanks, } } Leah L Dobbs } TEM Analyst } Intel Corporation }
} Dr Echlin is quite right. Isopentane had been used as a freezing compound a } long time ago, but I sure was glad when propane was advocated as a better } alternative. I started using propane about 15 years ago. Isopentane behaves } like a "gone off" rubber solution near its freezing point and freezes well } before propane.
Jim Darley is also right. I had overestimated iso-pentane, partly as a result of getting my Celsius and Kelvin mixed up! But for less critical work, like rapid quenching of polymer melts, it is sometimes helpful where working with a bottle of gas might not be allowed.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I've got a Hanna HI 8520 with autocalibrate at 4.01, 7.0 and 10.0. It is new and works fine at the moment with it's supplied electrode but will not autocalibrate with an TRIS resistant electrode that we bought at the same time. I still need to contact my suppliers. I will let you know if I get anywhere.
Malcolm Haswell University of Sunderland UK ----------
Ricky L Vaughn wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I have some investigators that are interested in doing shadow casting of } DNA and large protien structures, but we do not have a carbon } evaporator. They would like me to get some sources and prices for a } equipment grant. Other than Electron Microscopy Sciences who else } sells them? Are there any opinions on this matter? Thanks. } } Rick Vaughn } } RLVAUGHN-at-MAIL.UNMC.EDU } EM Research Facility } Dept. Cell Biology & Anatomy } UNMC } Omaha, NE
Rick,
We at Ladd Research are among those who manufracture and sell carbon evaporation systems. Please e-mail or call 1-800-451-3406 and we can talk options and pricing.
John Arnott Ladd Research 13 Dorset Lane Williston, VT 05495
TEL (US) 1-800-451-6406 1-802-878-6711 FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net
Hi Fellow Microscopists, We have just installed a "windowless" Si(Li) EDS detector for our ESEM. We would like to establish the spatial resolution of the detector in the system as a function of our typical ESEM conditions: 15KV, short working distance using the extended bullet, about 1 torr pressure, gas: air, at room and cryogenic temperatures. We are curious if fellow ESEMer's have undertaken a similar study and have some numbers to share. Are there better conditions to optimize performance? Any suggestions for good standards to test the spatial resolution?
HOPE YOUR HOLIDAYS ARE MAGNIFI-CENT!!
Lynne Garone Robert Baron Polaroid Corp. GaroneL-at-Polaroid.com
Can anyone please tell me the "a" spacing of 302 St. Steel, using the JEOL 100CX TEM/SCAN viewed at 100kv.....or how i could find that out?........Please email me ASAP Thank You.....and have a Merry Christmas Brandon Hernandez (EM student)
O.k., here's a list to get you started. I know I've left some off - no offense was indended, and hopefully the vendors themselves contact you.
NOTE: One strong recommendation. Apparently most vacuum evaporator companies supply for the optical and elctronics industries, NOT MICROSCOPY, make sure when you order that if the system has an Oil diffusion pump you specify that it NOT be filled with Silicon oil (Silicon + EM = VERY BAD), but a non-Si based oil (i.e. Santovac, or alternative).
Carbon Evaporator Vendors: (Alphbetically)
Balzers
[North American Rep.] Technotrade International 7 Perimeter Road Manchester, NH 03103-3343 Tel (603)622-5011 Fax (603)622-5211
=} Balzers is primarily involved with vacuum coating systems for the optical industry, but decades ago (1960's?) expanded their systems into the microscopy world as well. Tend to be (1) expensive, (2) finicky, (3) Techno Trade is very helpful to deal with though.
Denton Vacuum Inc. 1259 North Church Street Moorestown, NJ 08057
PH: 609.439.9100 FAX: 609.439.9111
=} I presently have two different models and they work quite nicely. The design / layout of electrical connectors, mechanical connetor (rotator) could be better for ease (Need two wrenches) and keeping clean. but not enough to swear off of them.
Edwards High Vacuum International 301 Ballardvale Street Wilmington, MA 01887
Ladd Research Industries, Inc. P.O. Box 1005 Burlington, VT 05402 PH: 802.878.6711 FAX: 802.878.8074
=} Bill Ladd designed one of the best, easiest to configure, easiest to clean, most flexible stage areas I have ever worked with (My Opinion). But Bill left Ladd a few years ago, and things were ... fuzy (?) for a while. Haven't dealt with them in three years.
Ted Pella, Inc. 4595 Mountain Lakes Blvd. Redding, CA 96003
Albert Prebus, one of the pioneers of transmission electron microscopy, died at his home in Columbus (Ohio) last Tuesday. The funeral will take place today (Friday).
After receiving B.Sc and MSc. degrees from the University of Alberta, he constructed the first North-American electron microscope at the University of Toronto in 1938, for which he earned the first Ph.D granted in electron microscopy. His achievments were honored in 1978 with an honorary Doctor of Science degree from the University of Toronto, presented at the 9th International Congress on Electron Microscopy.
In 1940, Dr. Prebus joined the Department of Physics at Ohio State University and retired as Professor Emeritus in 1978.
Ray Egerton, Physics Dept, University of Alberta, Edmonton, Canada T6G 2J1 Phone: 403-492-5095, FAX: 403-492-0714, e-mail: egerton-at-phys.ualberta.ca ------------------------------------------------------------------------
Dear Rick Vaughn: Two worthy carbon evaporator manufacturers come to mind: Edwards High Vacuum at 1-800-848-9800 and Denton Vacuum at 609-439-9100. Our Denton 502A evaporator is in its 13th year and has provided very reliable service and when technical information was needed they are very helpful people. I suppose this is a shameless plug for a company with which I have no financial interest. Good luck in your search. Don Gantz Biophysics Department Boston Univ Medical School
O.k., all you digital imaging folks: Looking for comments of any kind comparing the Condonics NP-1600 and the Tektronix Phaser 450. We're presently in the market to purchase a full page "publication quality" dye-sublimation printer, and based on vendor lit., listserver comments, and industry reviews. (Where as the Fuji Pictography 3000 looks really nice its just a little out of our price range, scarily slow).
Image Sources:
- Need to print via a networked solution, perferably a directly networked printer.
- Printing variety of images, but primarily Greyscale EM, color Confocal, and full color LM images. Ranging from 256 x 256, on up to over 5,000 x 5,000. (Fully realizing that at 300dpi 1:1 isn't going to happen)
- Printing from Intel PC's (Primarily Win NT) and Some Mac's, and a variety of software.
Codonics:
Whereas this printer comes highly regommended by the Microscopy community (Even recommened by a vendor who sold the Tektronix and NOT the Condonics) I have some grave concerns about it.
(1) Considering the highly computer oriented nature of digital imaging, the Condonics web site(http:\\www.codonics.com\) is pathetic, and last updated Aug, 6, 1997 with the NP-1600 page updated Dec, 5, 1995!). No online tech support, no on-line drivers, no FAQ's, no even a downloadable PDF manual. Whereas the "built in floppy drive for easy upgradability" maybe great but where do the "upgrades" come from? Snail mail communications? Any feeling for longer term support? (Last Codonics I worked with, '90-'94, worked great, but 1.5 years after purchase Tech support didn't really want to help at all with drivers for Windows 3.1 world, they had moved on to better things I guess)
(2) What, if any, options are there?
(3) Web search for "codonics" only results in 50-60 different sites for "Instructions for printing to the Codonics printer".
(4) No OS specific drivers, simply allows straight transfer of most image formats to the printer (this is nice) or relies on Post-Script printing (which is an option? How is it implemented?), but does require "loging on" and some rather criptic numerical "print-like-this" mode commands. I take it you are stuck with 1:1 printing and thus have to scale your image sizes prior to printing.
(5) From the user end (since most users aren't computer techno geeks like some of the rest of us) how user friendly is it really?
(6) Very fast, to Fastest on market - great. Any comments, particularly compared to Tektronix?
(7) Handles multiple jobs simultaneously - again great! Any comments?
TEKTRONIX
(1) Solid support for a variety of printing environs but primarily graphics industry, not sci. imaging.
(2) Solid on-line tech support.
(3) Requires OS specific drivers - how easily installed?
(4) Requires memory upgrade for larger image printing.
Any requested confidentiality for candid comments will be strongly protected. Vendors should feel free to reply as well.
Thank you.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
} We would like to establish the spatial resolution of the } detector in the system as a function of our typical ESEM conditions: } 15KV, short working distance using the extended bullet, about 1 torr } pressure, gas: air, at room and cryogenic temperatures.
David Joy has a Monte Carlo program to calculate the emission volume for x-rays. I don't know whether he has anything which will account for the E in the ESEM, but I'd ask him (the beam spread and x-ray absorption might be negligable anyway).
} Any suggestions for good standards to test the spatial resolution? } That depends on what you are going to examine. The SR depends on the average Z of the matrix among other things, and it can also depend on the element of interest. If you will be looking at biological specimens, perhaps something like magnetotactic bacteria would be useful--they have ~1 mu-m particles of magnetite. If you have some small particles of the element of interest, you could disperse them in a sucrose solution. (I calculated the % sucrose which gives about the right density and compo- sition for cells, but I don't have it with me.) I wonder if MAG*I*CAL can be useful for mineral specimens. You might be able to detect the Ge in the Si matrix, but I don't remember whether the Ge-containing bits are separated by enough distance. I have measured SR (crudely) by taking spectra on particles and in the matrix near them. This could be done correctly to get some function similar to a point spread function (of course, there is not enough signal from a point, so you'll have to examine an area { { your probe). Good luck. Yours, Bill Tivol
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
O.k., all you digital imaging folks: Looking for comments of any kind comparing the Condonics NP-1600 and the Tektronix Phaser 450. We're presently in the market to purchase a full page "publication quality" dye-sublimation printer, and based on vendor lit., listserver comments, and industry reviews. (Where as the Fuji Pictography 3000 looks really nice its just a little out of our price range, scarily slow).
Image Sources:
- Need to print via a networked solution, perferably a directly networked printer.
- Printing variety of images, but primarily Greyscale EM, color Confocal, and full color LM images. Ranging from 256 x 256, on up to over 5,000 x 5,000. (Fully realizing that at 300dpi 1:1 isn't going to happen)
- Printing from Intel PC's (Primarily Win NT) and Some Mac's, and a variety of software.
Codonics:
Whereas this printer comes highly regommended by the Microscopy community (Even recommened by a vendor who sold the Tektronix and NOT the Condonics) I have some grave concerns about it.
(1) Considering the highly computer oriented nature of digital imaging, the Condonics web site(http:\\www.codonics.com\) is pathetic, and last updated Aug, 6, 1997 with the NP-1600 page updated Dec, 5, 1995!). No online tech support, no on-line drivers, no FAQ's, no even a downloadable PDF manual. Whereas the "built in floppy drive for easy upgradability" maybe great but where do the "upgrades" come from? Snail mail communications? Any feeling for longer term support? (Last Codonics I worked with, '90-'94, worked great, but 1.5 years after purchase Tech support didn't really want to help at all with drivers for Windows 3.1 world, they had moved on to better things I guess)
(2) What, if any, options are there?
(3) Web search for "codonics" only results in 50-60 different sites for "Instructions for printing to the Codonics printer".
(4) No OS specific drivers, simply allows straight transfer of most image formats to the printer (this is nice) or relies on Post-Script printing (which is an option? How is it implemented?), but does require "loging on" and some rather criptic numerical "print-like-this" mode commands. I take it you are stuck with 1:1 printing and thus have to scale your image sizes prior to printing.
(5) From the user end (since most users aren't computer techno geeks like some of the rest of us) how user friendly is it really?
(6) Very fast, to Fastest on market - great. Any comments, particularly compared to Tektronix?
(7) Handles multiple jobs simultaneously - again great! Any comments?
TEKTRONIX
(1) Solid support for a variety of printing environs but primarily graphics industry, not sci. imaging.
(2) Solid on-line tech support.
(3) Requires OS specific drivers - how easily installed?
(4) Requires memory upgrade for larger image printing.
Any requested confidentiality for candid comments will be strongly protected. Vendors should feel free to reply as well.
Thank you.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
FEI Company in Hillsboro, OR has the following position open:
JOB TITLE: Particle Optics Scientist DEPARTMENT: Systems R&D REPORTS TO: R&D Manager
1) GENERAL PURPOSE:
Perform particle optics experiments and provide assistance for development of new particle optics devices.
2) ESSENTIAL RESPONSIBILITIES:
A) Perform particle optics calculations on electron / ion columns or analyzers using Munro and / or similar programs. These can include magnetic and electrostatic fields, deflection systems, as well as paraxial, 2D or full 3D modeling. Beam interactions can also be involved. B) Perform experiments on new particle optical devices. C) Keep up with the particle optics community through the literature and conferences. D) Write internal specifications, proposals and reports. E) Interact with customers regarding requirements for new particle optics. F) Serve on new product development teams as scientific advisor and for testing.
3) EDUCATION AND/OR EXPERIENCE:
A) Ph.D. is science or engineering. B) 4 years experience using Munro or similar particle optics programs. C) Particle optics experimental experience. D) Experience installing and maintaining particle optic programs on a PC. E) Demonstrated creativity. F) Ability to modify existing or write special particle optics programs.
Interested applicants should submit their resume to Lisa Olivia either by FAX at 503-640-7509 or email at lolivia-at-feico.com FEI Company, 7451 N.W. Evergreen Parkway, Hillsboro, OR 97124 ****
FEI Company in Hillsboro, OR has the following position available:
JOB TITLE: DualBeam Applications Development Engineer DEPARTMENT: Applications Development REPORTS TO: Applications Development Manager
SUPERVISORY RESPONSIBILITY:
Generally none, but may give general direction to various laboratory assistants and technicians.
JOB SUMMARY:
Develop applications for the DualBeam market, including demonstration and sales support as necessary.
ESSENTIAL RESPONSIBILITIES:
1. Conceive, plan and conduct experimental research to advance applications knowledge as applied to use of DualBeam in-line and laboratory FIB/SEM tools. 2. Acquire information about customer requirements through observation of FEI customer FIB demos, direct contact with customers and from other available marketing input. 3. Aid specification of software and hardware development or modification as necessary to implement applications consistent with safety considerations. 4. Recommend and advise on direction for potential DualBeam applications consistent with customer requirements and technical feasibility. 5. As necessary, demonstrate system operation and applications in support of Technical Marketing efforts. 6. Develop and conduct customer training programs in DualBeam applications either on site at FEI or at customer facility. 7. Develop and conduct technology transfer documents and training programs in DualBeam applications for FEI technical staff. 8. Develop scientific and technical information at conferences and meetings in support of sales and marketing efforts. 9. Disseminate general information regarding technologies relevant to advancement of DualBeam applications.
OTHER DUTIES:
1. Provide scientific and technical support to R & D, Manufacturing, Customer Service and other departments as required. 2. Assist with writing manuals and reviews. 3. Other duties as temporarily assigned by supervisor.
MINIMUM EDUCATION, EXPERIENCE AND QUALIFICATIONS:
1. MS in Physics, Chemistry or related discipline. 2. 2+ years experience in operating FIB, DualBeam=99, SEM or similar analytical systems in a lab or FAB environment. 3. Experience in defect review, inspection and other semiconductor process applications is highly desired. 4. Able to interact effectively with various employees, customers and groups at FEI and customer sites. 5. Familiar with UHV 6. Willing and able to travel up to 20% of time. 7.Eligible for passport.
Interested candidates should submit their resume to Lisa Olivia, preferably by email: lolivia-at-feico.com or FAX: 503-640-7509 FEI Company, 7451 N.W. Evergreen Parkway, Hillsboro, OR 97124
Atomic Structure / Property Relations of Interfaces in TiAl
An opening exists for a postdoctoral fellow in my research group at the University of Pennsylvania. This position is funded by a grant from the National Science Foundation on the study of interface atomic structure and diffusion in a model material system. The work will be primarily experimental with heavy emphasis on the use of state-of-the-art electron microscopy facilities at Penn. Characterization of the atomic structure and composition profile of interfaces will be correlated with measurements of interface diffusion by in-situ experiments. The in-situ experiments will be conducted at Penn and at Oak Ridge National Laboratory through an ongoing formalized collaboration. The post-doctoral fellow will likely make multiple trips to ORNL in connection with this work. The project will also include continuous collaboration with another Penn group conducting an atomistic modeling study of interface diffusion. Experimental facilities at Penn critical to this effort are a new JEOL 2010F field emission gun transmission electron microscope with several unique capabilities, a JEOL 4000 high-resolution transmission electron microscope, a Phi scanning Auger microprobe, and an optical float zone furnace. Materials Science at Penn has a wide range of other experimental facilities, excellent on-site computational hardware, an active and highly regarded faculty and a vibrant research atmosphere. If you are interested in learning more about this challenging opportunity, contact me at luzzi-at-lrsm.upenn.edu.
David E. Luzzi Professor Department of Materials Science and Engineering University of Pennsylvania Philadelphia, PA 19104-6272
The former Botany Department at the University of California at Davis has two instruments available to interested folks.
The first instrument is a JEOL 100S TEM, vintage about 1981. If you are interested, contact Gary Zamzow (530-752-8865;gwzamzow-at-ucdavis.edu).
The second instrument is an Hitachi S-800 SEM, vintage about 1981. This is one of Hitachi's early FEG instruments and has had some modifications, it may also have a Kevex x-ray system. If you are interested, contact Rick Falk (rhfalk-at-ucdavis.edu).
I am a former graduate student in the former department and would be happy to tell you what I know about the instruments, but you must contact those listed above for information regarding price or other terms.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Thanks for all of the suggestions, I've passed on the information to Matt, and it looks like the Eppendorf finder slip is the best for his application. -- Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
Merry Christmas as well as all the best for Healthy, Happy, Prosperous and Successful New Year yours truly J o z e f Stankovic p.s. tento subor je nutne odkodovat programom MIME64 resp. BASE64 a az potom spustit, lebo inac nefunguje.
This message is in MIME format. Since your mail reader does not understand this format, some or all of this message may not be legible.
I remember seeing a reference to an Immunocytochemistry Network but did not save the address. Could someone please make that available. Thankyou. Don Gantz Biophysics Dept. Boston Univ Medical School gantz-at-med-biophd.bu.edu
Lynne You may want to reconsider your windowless detector for the ESEM. Windowless detectors on a high vacuum system are difficult to maintain because all the water, hydrocarbons, and junk in the vacuum ends up on the crystal because it is at liquid nitrogen temperature. The only way that I could see this working was if you only went windowless when in an inert atmosphere such as dry nitrogen, argon, or the like. You mention air at 1 torr at room temperature which seems incompatable with windowless EDS operation. I do have some data on operating conditions for analysis and how to test your xray resolution - please contact me off the list. Happy holidays, Scott Wight
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------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
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FYI, Small World has a ESEM version of the electron flight simulator which is based on David Joys code. I have no financial interest in Small World, I am just a beta tester of the ESEM version. Scott Wight
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
edelmare-at-casmail.muohio.edu wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } } } } Ladd Research Industries, Inc. } P.O. Box 1005 } Burlington, VT 05402 } PH: 802.878.6711 } FAX: 802.878.8074 } } =} Bill Ladd designed one of the best, easiest to configure, easiest } to clean, most flexible stage areas I have ever worked with (My } Opinion).
To: Richard Edelman, Ph.D. Miami University
Thank you for your kind words concerning the Ladd evaporator. There are many users in the microscope world that would agree with you that Bill Ladd's evaporator was "one of the best, easiest to configure, easiest to clean, most flexible stage areas" etc. Ladd continues not only to produce that evaporator but we have even improved it.
Both Margaret and Bill passed on a few years ago but their long-time employees continue their tradition of quallity today after more than 40 years. It is always difficult to follow pioneers such as Margaret and Bill, but we learned from them and know they'd be proud that we continue to supply the microscopy world with the same quality of products that they did for oh so many years.
Thank you for your kind words concerning the Ladd Evaporator. There are many users in the microscope world that would agree with you that Bill Ladd's evaporator was "one of the best, easiest to configure, easiest to clean, most flexible stage areas" etc. Ladd continues not only to produce that evaporator but we have even improved it.
Both Margaret and Bill passed on a few years ago but their long-time employees continue their tradition of quality today after more than 40 years. It is always difficult to follow pioneers such as Margaret and Bill, but we learned from them and know they'd be proud that we continue to supply the microscopy world with the same quality products they did for oh so many years.
It looks as if we are about to get a bunch of 'unsubscribe' for the holiday messages (sent to the list rather than the listserver for some reason). I'm sure there is a 'postpone' command that has a complementary 'unpostpone' command. These can be sent to the listserver any time you want cessation of mail delivery in the short term.
i) Am I correct? and ii) If so, can someone in the know tell us all the proper command phrase and the address to send it to (listserv-at-msa.microscopy.com?)
Ho-Ho-Ho, John
================= C. John Runions, Ph.D Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
I work for VayTek. We sell a volume rendering product called VoxBlast.
I'm responding to Geoff Avern's question on December 17th concerning 3D. } 2) Does anyone know of commercially available 3D IA programs? (I want to } measure object sizes and object orientations) } VoxBlast is a 3D volume visualisation and measurement software that runs on UNIX, Windows 95/NT, and Mac. VoxBlast creates a volume from a stack of 2D images. The 2D images can be from many sources such as bright field, fluorescence, confocal, MRI, PET, CT, geological, etc. VoxBlast also has measuremement tools to give you object size and coordinates for selected points in 3D space. Our Windows version currently has a 3D Object Counting function that gives you the XYZ coordinates, volume, average density of each object found fitting the specified cluster size limits, and gray level limits.
If you have any interest in VoxBlast, please go to our web site at WWW.VAYTEK.COM . You can take a look at comparative rendering times for many machines on all three platforms. You can also download demo versions for each of the platforms.
Best regards
Patrick Guerin Customer Technical Support Engineer VayTek, Inc. 305 West Lowe Avenue, Suite 109 PO Box 732 Fairfield Iowa 52556-0732 Tel : 1-515-472-2227 Fax : 1-515-472-8131 E-mail : pguerin-at-vaytek.com _______________________________________
I would need information on solutions used in order to dilute antibodies in immunohistochemistry
THANK YOU
---------------------------------------------------------- Dr. Hugo H. Ortega Laboratorio de Investigaciones Histologicas Aplicadas Catedra de Histologia y Embriologia Facultad de Agronomia y Veterinaria Universidad Nacional del Litoral
There is no postpone/unpostpone command, please review your instructions which you all received when you first subscribed. Or review them on the MSA WWW Site.
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Simple send an UNSUBSCRIBE Email message to
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To avoid the mass of annual seansonal messages allow me to post a generic message:
*********************************************** Everyone on the Listserver wishes Everyone else a healthy and happy holiday season! ***********************************************
Cheers.... Nestor Your Friendly Neighborhood SysOp.
Our department is trying to find a way of making 3D reconstruction of plants under development. I am aware of something called "3D digitizer", but I am not sure how it works.
For the moment we are just making movies of plants and taking 2D images around the plant (36 images) such that we can make a QTVR film.
Maybe this is a subject for a 3D reconstruction mailing list, does somebody know of such a list?.
can anybody tell me where in Germany or Europe I can buy collodion (celloidin, parlodion), as a powder or as a solution in amylacetate. I found it in an old catalog from Electron Microscopy Sciences but do not have their e-mail or European adress. The 25% solution from Polysciences seems to me to be to concentrated for coating grids. Thanks in advance
Birgit
Dr. Birgit Neubohn Institut fuer Pflanzengenetik und Kulturpflanzenforschung (IPK) Corrensstr. 3 D-06466 Gatersleben-Deutschland
Does anyone have any experience with the fluorescently-tagged nanogolds? I'm starting to use one and have hit a snag; I'd like some practical input from anyone using these. The people at Nanoprobes are being very helpful, but I would like to talk to other "real-life" users, offline.
I am looking for the address of the company Technics, which produces the Hummer sputters. I know, its in Virginia. Could you mail me the whole adress (city...).
Thanks a lot!
CP Luftensteiner
I'm afraid the address I have is too old to be of any use ... if anyone on the Microscopy list can help then please reply directly to:
Vickie Allison, Group Leader, MOS 6 SEM Lab, Motorola, Inc.
--------------------------------------
Hello!
I am looking for the address of the company Technics, which produces the Hummer sputters. I know, its in Virginia. Could you mail me the whole adress (city...).
Thanks a lot!
CP Luftensteiner
I'm afraid the address I have is too old to be of any use ... if anyone on the Microscopy list can help then please reply directly to:
For those of us left working this close to Christmas, we are looking at a contaminant that appears to be mercury but are concerned about the potential hazard to both man (and woman) and machine. Are there any brave souls that have looked at mercury using a cryo system and is it relatively safe and non-destructive?? We would also like to get sample particle sizes and other potential elements present.
} Birgit Neubohn wrote: } } } } } Dear microscopists, } } } } can anybody tell me where in Germany or Europe I can buy collodion } } (celloidin, parlodion), as a powder or as a solution in amylacetate. } } I found it in an old catalog from Electron Microscopy Sciences but do not } } have their e-mail or European adress. } } The 25% solution from Polysciences seems to me to be to concentrated for } } coating grids. } } Thanks in advance } } } } Birgit } } } } Dr. Birgit Neubohn } } Institut fuer Pflanzengenetik } } und Kulturpflanzenforschung (IPK) } } Corrensstr. 3 } } D-06466 Gatersleben-Deutschland } } } } Tel.: (+49) 039482 5447 } } Fax: (+49) 039482 5139 } } e-mail: neubohn-at-ipk-gatersleben.de } } Dr. Neubohn, } } Ladd Research can supply your needs, as can many of the other supply } houses. Please contact me directly with your exact needs (i.e. } concentration). We can sell direct to you or through one of our agents } in Europe. } } Rita Arnott } International Sales } Ladd Research } 13 Dorset Lane } Williston, VT 05495 USA } tel 1-802-878-6711 } fax 1-802-878-8074 } e-mail ladres-at-worldnet.att.net
} Birgit Neubohn wrote: } } } } } Dear microscopists, } } } } can anybody tell me where in Germany or Europe I can buy collodion } } (celloidin, parlodion), as a powder or as a solution in amylacetate. } } I found it in an old catalog from Electron Microscopy Sciences but do not } } have their e-mail or European adress. } } The 25% solution from Polysciences seems to me to be to concentrated for } } coating grids. } } Thanks in advance } } } } Birgit } } } } Dr. Birgit Neubohn } } Institut fuer Pflanzengenetik } } und Kulturpflanzenforschung (IPK) } } Corrensstr. 3 } } D-06466 Gatersleben-Deutschland } } } } Tel.: (+49) 039482 5447 } } Fax: (+49) 039482 5139 } } e-mail: neubohn-at-ipk-gatersleben.de } } Dr. Neubohn, } } Ladd Research can supply your needs, as can many of the other supply } houses. Please contact me directly with your exact needs (i.e. } concentration). We can sell direct to you or through one of our agents } in Europe. } } Rita Arnott } International Sales } Ladd Research } 13 Dorset Lane } Williston, VT 05495 USA } tel 1-802-878-6711 } fax 1-802-878-8074 } e-mail ladres-at-worldnet.att.net
`Only a customer.' Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
Lynne Garone wrote: } Hi Fellow Microscopists, } We have just installed a "windowless" Si(Li) EDS detector for our } ESEM. We would like to establish the spatial resolution of the } detector in the system as a function of our typical ESEM conditions: } 15KV, short working distance using the extended bullet, about 1 torr } pressure, gas: air, at room and cryogenic temperatures. We are curious } if fellow ESEMer's have undertaken a similar study and have some } numbers to share. Are there better conditions to optimize performance? } Any suggestions for good standards to test the spatial resolution? }
Hi Lynne,
It is possible to restore spatial resolution for EDS in the ESEM by correcting for the beam skirt effects. The methods are described in a paper which can be found at the web-address:
http://www.risoe.dk/afm/news1new.htm
best regards, Joergen.
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
I am continuing on the subject below in seeking someones opinion on the difference betweeen the ordinary Struers Tenupol3 and a less known equipment called 550D from South Bay Technology. Are those two comparable and can 550D be used for routine work.....
Old message I am interested in your opinion about electrolyhtic thinning apparatues and their performance. I am planning to buy a jet polishing machine to be used for making thin foils from 3mm disks. The material I am interested in is High strength Aluminium alloys, steels, stainless steels.
Best wishes=20 Sten Johansson
Link=F6ping University Department of Mechanical Engineering
I just wanted to put the word out that Integrated Device Technology (IDT) has an immediate opening for a Sem Technician in our 8" Fab in Portland Oregon. The actual position is listed below along with my contact information. This is a well timed opportunity to get into a position and facility with very good growth potential. Thank you all for your time and have a great holiday season.
Dave Saiki IDT Staffing
SEM Technician
Description: Analytical SEM support for manufacturing, technology transfer, and yield enhancement. Responsible for surface and cross section sample preparation, sustaining, and imaging - Dimensional measurements (CD, film thickness) - EDX analysis. Will be working in a fully automated 8" wafer fab. New facility with a lot of growth coming.
A 2 year degree with 3+ years of experience doing Integrated Circuit construction / Failure Analysis are required for this position.
Please Respond: IDT Oregon 3131 NE Brookwood Pkwy. Hillsboro, OR 97124 503-681-6376 fax e-mail {a href="mailto:dsaiki-at-oregon.idt.com"} dsaiki-at-oregon.idt.com {/a}
We are currently looking for other options in Service Contracts for our JEOL JSM T330-A SEM. Our instrument is about 8 years old, has performed very well over this time and has been under manufacturer's Service Contract.
Because our usage has been reduced over the last couple of years, we would like to know if there are other options for maintaining and/or repairing this scope than the very expensive manufacturer's Service Contract. We are located in Philadelphia, Pennsylvania and would be interested in considering any provider who works on SEMs of this type in this region.
Any help, advice or provider recommendations would be greatly appreciated.
I beg your pardon but I failed to include my email and fax# on the preceding message.
Dave Saiki IDT Staffing
SEM Technician
Description: Analytical SEM support for manufacturing, technology transfer, and yield enhancement. Responsible for surface and cross section sample preparation, sustaining, and imaging - Dimensional measurements (CD, film thickness) - EDX analysis. Will be working in a fully automated 8" wafer fab. New facility with a lot of growth coming.
A 2 year degree with 3+ years of experience doing Integrated Circuit construction / Failure Analysis are required for this position.
Please Respond: IDT Oregon 3131 NE Brookwood Pkwy. Hillsboro, OR 97124 fax: 503-681-6376 e-mail dsaiki-at-oregon.idt.com IDT's Web Page: www.idt.com
I have owned and used both instruments as well as another instrument made by EAF Fischione that you should also consider before making you mind up. The tenepol and the Fischione model have dual polishing capability whereas the SouthBay unit only has one sided. However, the SouthBay unit can be easily converted to a chemical polishing unit for other types of materials, such as semiconductors. It can handle some pretty nasty chemicals. Another advantage of the SouthBay unit is that Bernie Kestel from Argonne National Lab has published a lot of articles on how to use this instrument for some innovative sample preparation. You can get these publications from SouthBay Technology. You should probably get a hold of them anyway since they really are definitive works on how to electropolish. The Fischione unit has a well designed sample holder that makes terminating and rinsing the sample very nice and easy. The Fischione and the SouthBay units have a reasonably sized power supply for TEM electropolishing. The Tenepol unit is huge. I modified the Tenepol unit to take the Fischione sample holder and it worked very nicely. I believe that they have modified their sample holder since I used it. If you plan to do any other type of electropolishing of large sample, the large power supply of the Tenepol unit is a plus. I used it to electroplate and electropolish parts that went into a UHV system.
Contact Dave Henriks or Shane Roberts at SouthBay Technology at "sbt-at-southbaytech.com" and Paul Fischione at EAF Fischione Instruments at "Paul.Fischione-at-internetMCI.COM"
I hope that these ramblings have helped more than they have confused you.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I am continuing on the subject below in seeking someones opinion on the difference betweeen the ordinary Struers Tenupol3 and a less known equipment called 550D from South Bay Technology. Are those two comparable and can 550D be used for routine work.....
Old message I am interested in your opinion about electrolyhtic thinning apparatues and their performance. I am planning to buy a jet polishing machine to be used for making thin foils from 3mm disks. The material I am interested in is High strength Aluminium alloys, steels, stainless steels.
Best wishes Sten Johansson
Linkvping University Department of Mechanical Engineering
Rick Felten 12/23/97 03:13 PM We need to have the stage motorized on our Hitachi S2400 SEM. The quote from Hitachi is $16,000 for X and Y w/ installation. I am sure that they have chosen a reputable sub-contractor for the job. Before we commit does anyone know of a significantly cheaper option that would still give us high quality performance?
Gerald Harrison wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello fellow microscopists, } } We are currently looking for other options in Service Contracts for } our JEOL JSM T330-A SEM. Our instrument is about 8 years old, has performed } very well over this time and has been under manufacturer's Service Contract. } } Because our usage has been reduced over the last couple of years, we } would like to know if there are other options for maintaining and/or } repairing this scope than the very expensive manufacturer's Service } Contract. We are located in Philadelphia, Pennsylvania and would be } interested in considering any provider who works on SEMs of this type in } this region. } } Any help, advice or provider recommendations would be greatly } appreciated. } } Happy year-end Holidays to all -- Gerald Harrison
Dear Gerald: Most independent service organizations are to some degree local. So it would be very helpful to know where you are located. Merry Christmas and a Happy New Year. Peter Jordan, EMSI, an independend TEM service company servicing Southern California
You could just add some solvent (probably iso amyl acetate) to your present supply. You'll have to guess the amount by adding some and watching the color of your resulting film. Start with about 10% (e.g., add 0.1 ml solvent into 1 ml commercial solution. Your film should be a light silver grey when viewed by reflected fluorescent light (like very thin silver sections). Concentration occurs through evaporation, and you can always dilute it to your specifications by experimentation. Sara
On Mon, 22 Dec 1997, Birgit Neubohn wrote:
} Date: Mon, 22 Dec 1997 14:02:02 +0100 } From: Birgit Neubohn {neubohn-at-IPK-Gatersleben.de} } To: microscopy-at-sparc5.microscopy.com } Subject: Supplier for collodion } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear microscopists, } } can anybody tell me where in Germany or Europe I can buy collodion } (celloidin, parlodion), as a powder or as a solution in amylacetate. } I found it in an old catalog from Electron Microscopy Sciences but do not } have their e-mail or European adress. } The 25% solution from Polysciences seems to me to be to concentrated for } coating grids. } Thanks in advance } } Birgit } } } Dr. Birgit Neubohn } Institut fuer Pflanzengenetik } und Kulturpflanzenforschung (IPK) } Corrensstr. 3 } D-06466 Gatersleben-Deutschland } } Tel.: (+49) 039482 5447 } Fax: (+49) 039482 5139 } e-mail: neubohn-at-ipk-gatersleben.de } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
They have a web site, www.cerious.com. Email is pcrews-at-cerious.com, phone is (704) 529-0200, fax is (704) 529-0497. You can download a shareware version from their web site to try it. The licensed version has a few more features. If you use a network, I recommend the network version. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
----------
Text item:
Does anyone have experience using the Polaroid Sprint Scan 45 for TEM negatives? I would appreciate any opinions on this scanner.
Thanks,
Leah L Dobbs TEM Analyst Intel Corporation
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There was a listing regarding an In Situ Workshop to be held in Florida. Somehow I misplaced the annoucement. If someone has the annoucement or even the phone number of the contact person, I would appreciate receiving this information. My email is 00lganion-at-BSU.edu. Thanks.
} Rick Felten } 12/15/97 12:52 PM } Does anyone know a way to print several image files in an automatic fashion } that would include the file name on the image. I am using windows 95. } } Thanks in advance.
We also use Thumbs plus. networked, multilple printers over win95/NT4 Great software!
Chris Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
Opps, I forgot to enclose my list yesterday of common problems associated with specimen collection, and tissue processing. I was hoping to get a complete list, and was hoping that other people could mention problems that they might have encountered in these areas.
Difficulties faced in the Collection of Tissue
1. Lack of the proper information on the requisitions. eg: time of fixation. Since the tissue must sit in fixative for minimum of 1 hour, if we don't know this time, then we must wait at least an hour before processing the tissue further. -name or pathology number missing, or no number on the vial, or no requisition, or a requisition with no sample. -if the requistion is not marked as a "priority", then we don't proceed to cut blue sections on the tissue, and this might result in needless delays. -illegible handwriting.
2. Insufficient sample -we often get tubes of "sample" that are apparently empty. Unless we can physically see some sample, we have trouble processing the sample because of the solution changes that we must put the sample through. If we can't see it, how do we know that we aren't "throwing the baby out with the bath water".
3. With kidney biopsy samples, the biopsy misses the glomeruli.
4. Too large of a specimen. Not only can the fixative not penetrate such a large specimen larger than a millimeter cubed, but it's extremely difficult to dehydate such tissue in the time frame that we are allowed. With incomplete dehydration, it's not possible to get the plastic to fully infiltrate the specimen, making it almost impossible to cut ultrathin, or even semithin for that matter.
Another reason why it is difficult to cut an incompletely dehyrdrated specimen block is because we cannot get complete polymerization of the plastic, so it is too soft, and does not give us enough support to cut to the very thin thicknesses required.
5. Vials with lids not tight enough.
6. Wrong specimen for the patient, or 2 specimens in the same vial, from different patients, or multiple specimens from the same patient with different tissue types in the same vial.
7. Specimen stuck to the side or lid of vial, instead of being immersed in the fixative. This might result in the specimen drying out, which of course totally destroys the ultrastructure. During the cutting of the block, it's possible to tell if the specimen has suffered this fate, because of "smoothing" problems when trying to smooth out the block surface.
8. Fresh blood or tissue sent to us directly, instead of being sent in the fixative, or sent in fixative that was poorly prepared from the referral lab. (or method of preparation was questionable.)
9. Pathologist might refer to a case by the diagnosis or tissue type, rather than the patient name or pathology number, leaving the technologist a bit baffled until they determine what case the pathologist has in mind.
10. Though it has not been a problem for us, some people might wonder what ratio of specimen to fixative should be used. In general, at least 9 parts fixative to 1 part specimen would be the largest possible ratio that one might choose.
11. Wrong container. Sperm samples have been sent in condoms, fecal samples have been sent in Cheeze Whiz jars etc, and this is not acceptable. Usually the vials supplied by our lab are adequate for most preparations, unless the sample must be centrifuged, in which case it would be preferable to sent the sample in a Falcon plastic centrifuge tube, with an adequate amount of fixative.
Problems in Polymerization
1. High humidity that we sometimes get in the summer months poses a problem for us in that the water in the air will not give us a plastic that polymerizes hard enough, consequently we have an impossible time trying to cut the specimen ultrathin. Even tbough the plastic would be hard enough for us to cut semi-thin sections, is is not hard enough to cut ultrathin.
We try to compensate through the use of dessicants and rotary vacuum pumps, to try to make our polymerization over as dry as possible, but even so, we have noticed that on the extremely humid days, the problem persists.
2. In some cases where orientation of the specimen is important, it may be difficult for us to determine the specimen orientation after fixation in Osmium Tetroxide, because the uniform black color makes it difficult to determine which side is which. In these cases reorientation after polymerization may be the only way to correct the problem.
3. Fine Needle Aspirate samples tend to give more problems in polymerization because they seem to generate more bubbles, which will often occur right at the point of interest in the specimen. We have to break all of these bubbles manually to correct the problem.
4. Use of Epon-Araldite resins, especially when combined with routine strong fixatives such as Glutaraldehyde and Osmium Tetroxide tend to destroy sensitive antigens which might have had some significance in Immunocytochemistry. This plastic also effectively renders most water soluble stains ineffective for semithin sections, essentially forcing us to adopt a monochromatic stain, rather than the preferable H+E stain which would show us contrast between the nuclei and the cytoplasm.
5. A balance must be struck between rush infiltration versus incomplete infiltration. After the sample has been polymerized though, it becomes quite useless if it has only been partially infiltrated, because at that point, it even becomes impossible to cut semi-thin sections.
Difficulties in Sectioning
1. Some tissues, because of their consistency, especially longitudinal sections of nervers, wrinkle excessively, and it is an extremely tiresome problem to deal with.
2. Plastics that have not polymerized hard enough are usually very difficult to section, and if it is possible to section, the sections break up easily of their own accord, or sometimes when under the electron beam. Usually water is to blame for this problem, especially incomplete dehydration or high humidity.
3. Plastic that is too brittle may shatter when it is being trimmed to make a useable block face. This makes it difficult to cut as well. Making plastic very quickly at high temperatures tends to result in this form of brittle plastic.
Difficulties in Staining.
1. Tissue that has stayed for too long in Glutaraldehyde my lose staining sites, and may only stain very pale, making it difficult to discern ultrastructure under the microscope.
Photography under the microscope.
1. Micrographs that are too dark result in excessive contrast. Conversely, micrographs that are too light result in prints with inadequate contrast. Micrographs that vary widely in density are difficult to print, because exposure time in the darkroom has to be changed frequently, and this results in many time wasting tests.
2. Care should be given not to shake the microscope during an exposure, or a blurred negative will result.
3. Care should be taken not to exceed the number of negatives in the electron microscope, or missing images will result.
4. With some of the older machines, one should be careful about potential camera jams, and note any unusual sounds that may occur during operation of the camera.
5. One might also be careful to note any sudden drops in vacuum after the introduction of new film. It could be that that film emulsion has not been sufficiently dried before being placed in the electron microscope.
6. Charged specimen holders or apertures can cause a slow shaking in the image that is due to charge building up and discharging. But this motion or drift can cause blurry micrographs. This can be corrected by cleaning the specimen holder in acetone, or changing the objective aperture. Increased contrast can be achieved by choosing a smaller objective aperture (of about 20 microns), and conversely, less contrast will be seen with a larger objective aperture.
7. Exact magnifications can be measured by using a calibration grid during the same session, and taking a few micrographs at the desired mag. with the calibration grid.
8. The biggest problem encountered is out of focus micrographs. This can be determined by inspection of the negative. Careful use of the wobbler focusing aid, and awareness of loss of contrast at the focused point can help reduce this problem. [some people forget to turn it off though, after using it!!] Beginners might find it useful to focus with the aid of a "hole" in the plastic, since the fresnel fringe will be clearly visible, and a large bright fresnel fringe on the inside of the hold indicates underfocus, whereas if the fringe is on the outside of the hole, it indicates overfocus.
9. The EM user should work quickly, without rushing themselves, because leaving the beam on a single area of the section burns the section. The burning might not be noticeable in the microscope, but will be readily apparent on the micrograph, and will show up as a dark circle. Excessive burning may even result in a complete penetration of the plastic by melting, resulting in total destruction of that area of the section.
10. The user should also be careful that the area of interest of the section is included on the micrograph. This is indicated on the screen by small angle marks incribed on the screen. The user should also be aware that the final image will be enlarged another 2.7 times or so in the enlarger to make the final print, and this needs to be taken into account when determining target magnification.
I will greatly appreciate if you would be able to give me a detailed protocol on how to prepare red blood cells ( I am working on fish rbc) for viewing under an SEM. Also, would it be possible to analyze for Fe using EDX.
Thank you very much. May I wish you all a Very Happy New Year.
Thanit Pewnim
%----------------------------------------------------------------------% Thanit Pewnim, Department of Chemistry, Silpakorn University Nakornpathom 73000, THAILAND } } } } } Phone +66 34 255797 Fax +66 34 255820, Internet thanit-at-kanate.su.ac.th %----------------------------------------------------------------------%
I have been tasked with coming up with something about using light microscopes without harming yourself! Not so funny, according to two of our now-retired staff who counted plankton etc. for 20+ years. They both suffered neck/shoulder/back pain which eased after retirement.
In the UK we have a specific, detailed set of Regulations concerning computer use, another unspecific set covers all workstations e.g. supermarket checkouts. A lot of the computer Regs. might be transferred to microscope set-ups.
I'd appreciate any info, anything to avoid re-inventing the wheel!
Thank you all for the replies to my query re co-visualisation of cellulose and cytoskeleton. I hope to be able to report back on the success (or otherwise) of my experiments in this area to those who requested feedback some time next year (microscope down time/holiday/trip to UK/getting hold of reagents-permitting).
In the meantime God Jul to you all,
With best wishes,
Nigel Chaffey
----------------------------------------------------- Dr Nigel Chaffey, Dept Forest Genetics & Plant Physiology, Swedish University of Agricultural Sciences, S-901 83 Ume=E5, Sweden Phone: +46-90-786-6305 =46ax: +46-90-786-5901 eMail: nigel.chaffey-at-genfys.slu.se
I have just learned that there has been a delay in the distribution of the
Call for Papers and Advanced Registration for the Microscopy & Microanalysis '98 Meeting in Atlanta, Ga July 12-16
due to a delay at the printers.
It is expected that they will be mailed during the week of Dec. 15th.
In order to avoid having to reply to repeated questions which have started to flow in my direction I have taken the liberty of posting this Email message to both the MSA Membership as well as the Microscopy Listserver Databases.
As additional information becomes available I will make sure that the M&M'98 WWW pages are updated. These WWW pages are directly accessible from the URL
Does anyone know whether the Minolta RD-175 digital camera is suitable for capturing images of biological specimens viewed with a fluorescence microscope? As I understood, this camera has an equivalent film speed sensitivity of ISO 800, which supposingly should be fast enough to capture dim fluorescent images.
Eric Cho Dept Anatomy The Chinese University of Hong Kong
can anyone of this newsgroup give me information about and subscription procedure to any related newsgroup ? In particular, I would appreciate to hear whether there is a special newsgroup discussing immuno-histology, -fluorescence, -blotting, and pathology subjects.
Thank you very much.
With kind regards,
Heinz
*********************************************************************** Dr. Heinz Fehrenbach Institute of Pathology University Clinics "Carl Gustav Carus" Technical University of Dresden
Hi Hans: Pump through the system a vinegar solution for an hour or two. If its hot its much more effective but its a bit pungent on the lungs. A bucket can serve to insert the suction and return lines a small pump is obviously required. Call me if you have any related troubles. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } From: H.BRINKIES {hbrinkies-at-lucy.cc.swin.edu.au} } To: microscopy-at-sparc5.microscopy.com } Subject: Cooling Water Problems } Date: Wednesday, 10 December 1997 22:01 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello out there. } } I am still using an old ETEC Autoscan (SEM, vintage 1973). It is } still working well after more than 12 000 hrs of usage and usually we } get the results that we want. } } However, a calcium containing deposit has been forming } in the cooling water supply (in Cu tubes, in cooling coils around } diff.pump, in heat sinks, ect). The microscope was donated to us } several years ago but was not connected for the last 18 months } to the recirculating water system in our laboratory ( we are using } filtered tap water). The water flow has now been reduced drastically } over the last few weeks and I fear that the 'pipes' may eventually } totally block up. } } What is the best (and safe) way to reduce or remove this deposit. } Back-flashing was only partially successful. } } Any suggestion ? } } Thank You } } Hans Brinkies } SWINBURNE, University of Technology } School of Engineering and Science } Electron Microscopy Laboratory } HAWTHORN, 3122, Australia
PDC-2000 is a fine hand-held digital camera but does not have a way to adapt for the microscope. Amont other things there are internal optics that don't cooperate well with the microscope optics. It also could do some macro work with close-up lenses, but that is not optically the best imaging system.
DMC, while using the same Polaroid designed sensor chip as PDC-2000, otherwise is designed and built specifically for microscopy. Its C-mount thread (no optics) allows easy mounting on almost any microscope using standard C-mount adapters, and also can accept many "C-mount" threaded macro lenses for use on the copystand.
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On Fri, 5 Dec 1997, R-Brooks Corl wrote:
} Have you seen/tried the Polaroid DMC? It connects to the microscope } via standard C-mount (no lens on the camera, just C-mount thread), } creates a TIFF file into your computer at 1600x1200 or 800x600 pixel } resolution, and converts quickly and easily for macro work on your } copy stand by adding C-mount macro lens. List price under $6K. } Details on the Polaroid website at http:\\www.polaroid.com
How does this differ from the PDC-2000/T which, I believe, is much cheaper?
Message-Id: {199712100541.AAA28126-at-ns1.axs2000.net} To: MICROSCOPY BB {Microscopy-at-Sparc5.Microscopy.Com}
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Diane M. Smith wrote: ====================================================== Can anyone tell me about how often a diamond knife needs to be sharpened? I realize it would depend on the amount of wear it gets. Our EM dept. is only open three days a week, with cutting being done about 2-4hrs a week. Our knife was sharpened six months ago and seems to be getting dull again. Is this to be expected? ====================================================== The answer to this question is about as elusive as predicting which way the Dow Jones average will close tomorrow!
But seriously, there are the "ten commandments" for a diamond knife to enjoy a long life, the most important ones being as follows:
1] Because of the extreme sharpness of a diamond knife edge, it should not be touched even for cleaning with any solid object. This is "controversial" since some manufacturers actually "recommend" that the edges be cleaned with sticks of varying types. We ourselves believe such treatment accellerates the wearing out of a diamond knife.
2] Don't let sections or the remains of sections or other debris end up drying down onto the knife edge. Keep the knife edge "wet" until it is ready for cleaning before being put to bed for the night.
3] Use a diamond knife cleaner sold by several firms (including ours) specifically for this purpose. Some typical laboratory ultrasonic cleaners can have enough power to be damaging to a knife.
4] Wash the knife edge one last time with distilled water and then dry with some kind of "blast" such as from a clean "duster".
5] Avoid conditions of "chatter" at all times. Reduce chatter by varying the clearance angle or slowing the cutting speed. Other common causes of chatter are insufficient tightening of the boat in the microtome, an insufficiently tightened block, or an incompletely cured block.
6] Final block trimming with a razor blade can leave metal particles from the blade which are of course damaging to the knife edge. This can be minimized by using a fresh razor blade each time. Then washing the end of the freshly cut block face with distilled water, followed by a drying with a "duster" blast is the final step before the first cut with the knife. This is a final chance to wash away metal particles that could damage the knife edge.
OK, there are other considerations but these are the most important. They are independent on the knife manufacturer, the type of diamond knife, length of cutting edge, nature of the samples being cut, even the price paid.
Diamond knives in an EM lab have lifetimes that are predictable like a set of tires. It depends on what kind of road you drive on, how you do your driving, not to mention the beginning quality of the product itself. We run some samples in our own laboratory that wear out a new materials science diamond knife in a week, and we run others, e.g. soft tissue samples, that are cut with a life science diamond knife that will last, in comparision, almost forever. You can't do anything about the deck of cards you have been dealt in terms of the kinds of samples you have to cut, but once having determined that, you surely can do things, under your control that can make a big difference in terms of how long your own knife will or will not last in your own environment.
Disclaimer: SPI Supplies offers a full line of diamond knives for EM and LM . Actually we have a vested interest in having knives wear out faster rather than slower. Our favorite customers are those who mistreat their diamond knives and come back sooner for resharpenings or replacements.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Meeting announcement - Focus on Microscopy 1998 Full details below - best viewed in a monospaced font.
For a classier view see our web page created by Pal Fekete http://www.physics.usyd.edu.au/physopt/fm98
Online registration will be available on the web page soon but you can also get a form (and any further details) by email from focus98-at-emu.usyd.edu.au
Focus on Microscopy 1998 11th International Conference on 3D Image Processing in Microscopy 10th International Conference on Confocal Microscopy
April 14th-17th, 1998
University of Sydney, New South Wales, Australia
Australian Key Centre for Microscopy and Microanalysis Royal Microscopical Society (UK) Image Analysis Society of Australia
International Committee Prof. Colin Sheppard, University of Sydney Prof. G.J. Brakenhoff, University of Amsterdam Prof. Tony Wilson, University of Oxford Dr. Vyvyan Howard, University of Liverpool Dr. Andres Kriete, Liebig University, Giessen Prof. P-C. Cheng, SUNY at Buffalo Prof. Alan Boyde, University of London Dr. Guy Cox, University of Sydney Prof. S. Kawata, Osaka University
Organising Committee Prof. Colin Sheppard, University of Sydney Dr. Guy Cox, University of Sydney Ms. Carol Cogswell, University of Sydney Dr. Pal Fekete, University of Sydney Dr. Min Gu, Victoria University Dr. Allan Jones, University of Sydney Ms. Eleanor Kable, University of Sydney
Introducing Sydney
Sydney, Australia's largest city, is also one of the world's most beautiful cities, built around the spectacular natural harbour which provided the site for the first European settlement of the Australian continent. It prides itself on its cultural diversity, offering a rich mix of European, Asian and indigenous Australian experiences alongside the uniquely Australian culture which has developed in the 200 years since the First Fleet landed in Sydney Cove.
The University of Sydney is the oldest in the country, established as part of the great English 19th century tradition of liberal enlightenment but unashamedly modelled architecturally on the Oxbridge pattern. It has retained both its prestige and its central location although its site has expanded greatly over the years, now accommodating more than 20,000 students. The Australian Key Centre for Microscopy and Microanalysis (AKCMM) at the University of Sydney, which is hosting the Conference this year, is the largest centre for microscopy in the Southern Hemisphere, offering a wide range of optical and electron microscope facilities both to the University and the wider community.
April is autumn (fall) in Sydney. The weather will be mild average temperature for the month is 19 C (68 F). Sydney has both a lot of sun and a high rainfall - rain can fall in any month so bring a waterproof. The ocean will be warm and very pleasant for swimming and surfing.
Scientific Programme - 15-17 April
The scientific programme will consist of poster and spoken sessions. Posters (1m x 1m) and contributed talks (15min) are invited on any of the Conference topics:
* Advances in confocal microscopy * Applications of confocal microscopy * 3-dimensional optical imaging * 3-D techniques in electron microscopy * Other 3D imaging techniques * Novel techniques in microscopy * Near-field microscopy * Multiple-photon microscopy * Multiple-dimensional image processing * Applications of image analysis
Short Courses & Workshops - 14th April
The following half-day short courses and workshops will be offered, subject to both maximum and minimum numbers of participants. All are led by internationally recognized experts in their respective fields. The cost is $75 (Aust) per half-day course.
Morning * Multiphoton microscopy * Introductory confocal microscopy * Deconvolution of 3D images * Stereology
Afternoon * Introduction to image processing * Advanced confocal microscopy * Introduction to digital imaging * 3D image processing & visualization
Social Programme
These events are included in the cost of full and accompanying member registration. A limited number of additional tickets will be available.
* Tuesday 14th April, 6pm. Welcome reception, exhibition area. Drinks and simple snacks - an opportunity to meet old friends and to get to know fellow delegates before the start of the formal business of the conference. * Thursday 16th April, 7pm. Conference Barbecue Dinner. An informal evening on an island in one of the most beautiful parts of Sydney Harbour.
Morning coffee, a light lunch, and afternoon tea are provided each day. (For workshop registrants only on the 14th).
Abstracts
Extended abstracts, up to one A4 page in length, will be published in a conference volume issued free to all delegates. Additional copies will be available for sale. Micrographs and other illustrations (monochrome only) are welcomed but must fit within the one page.
Manuscripts will be edited for format only. To simplify the editors' task please follow these simple guidelines: * Title in upper and lower case. * Authors' names with initials first, presenting author in upper case. * Full address and affiliation of all authors. * Text in a 12pt font. * References cited by name and date, not number. * References at end - do not include titles. All authors (initials first), then year, then journal citation.
All text must be submitted electronically, either as plain text, RTF or in the format of a PC or Macintosh word processor. MS Word, Word Perfect, Wordstar, MS Works, Claris Works, Write are all on site and most other formats can be converted. Postscript files and TeX are not acceptable. Illustrations should not be included within the word processor file; they should be submitted either as separate files in any common format (not postscript or eps) or as hard copy.
Send files : * by email to focus98-at-emu.usyd.edu.au * by anonymous ftp to ftp.emu.usyd.edu.au (directory \focus98) ftp://ftp.emu.usyd.edu.au/focus98) * on a floppy disk to the conference address, below.
Abstracts must be received by 31st January 1998 if they are to appear in the published volume.
Conference Details
Conference sessions and a comprehensive manufacturers' exhibition will take place in the Wentworth Building, levels 4 & 5. Some workshops will be held in the AKCMM, Madsen Building. A footbridge across City Road provides quick access between these buildings.
A discounted early registration fee applies to all registrations received and paid by 31st January 1997. All prices given are in Australian dollars - one Australian dollar is approximately 70c US.
Early Regular Full registration $ 425 $ 475 Student registration $ 250 $ 300 Day registration $ 175 $ 175 Accompanying person $ 175 $ 175
Full registration covers conference volume, admission to all scientific sessions, welcome reception, conference dinner, morning and afternoon tea, and lunch.
Student registration includes all the above except the conference dinner.
Day registration covers conference volume, admission to scientific sessions, morning and afternoon tea, and lunch on any one day.
Accompanying person's registration includes welcome reception, two half-day tours and the conference dinner
Accommodation is available at St John's College, on the University campus, for $60 per night including breakfast (single room, shared bathroom). For those who prefer a hotel, Camperdown Travelodge is $125 per night (single occupancy) or $135 (dual occupancy) including breakfast. These are specially discounted rates - to obtain them you must book through the conference. The taxi fare from the airport to Wentworth Building, St. John's College or the Travelodge is about $10.
Focus on Microscopy '98, Australian Key Centre for Microscopy and Microanalysis, F09, University of Sydney, NSW 2006, Australia.
What are some alternatives to Absolute EtOH for drying specimens for embeddment? Absolute alcohol is hard to come by. Can someone recommend a more common substance?
I have been thinking of Acetone. My specimens will be fragile: coral tissues, sponges, worms, etc. I'd like to find something fast and good, cheap and easy.
Alan Davis --
"I consider that the golden rule requires Alan E. Davis that if I like a program I must share it adavis-at-netpci.com with other people who like it" Marianas High School AAA196, Box 10001 ---Richard Stallman Saipan, MP 96950 Northern Mariana Islands GMT+10
I would like to introduce myself as a new participant at this mailing list. I am a veterinarian (*1967) working at the Institute of Veterinary Pathology in Munich, Germany. My predominant field of research is ultrastructural enzyme- and immuno-histochemistry.
Therefore, I would like to arouse a discussion on this very topic- TEM-histochemistry, preferably post-embedding. Is there anybody out there still doing enzyme histochemistry?
Looking forward to a vivid exchange of opinions...
I would like to introduce myself as a new participant at this mailing list. I am a veterinarian (*1967) working at the Institute of Veterinary Pathology in Munich, Germany. My predominant field of research is ultrastructural enzyme- and immuno-histochemistry.
Therefore, I would like to arouse a discussion on this very topic- TEM-histochemistry, preferably post-embedding. Is there anybody out there still doing enzyme histochemistry?
Looking forward to a vivid exchange of opinions...
} } What are some alternatives to Absolute EtOH for drying specimens for } embeddment? Absolute alcohol is hard to come by. Can someone recommend a } more common substance? }
Isopropanol (or Propan-2-ol, as we are told to call it these days) in many ways behaves similarly to ethanol. It is somewhat more viscous and less volatile, but not overwhelmingly so. Moreover, it is miscible with water and should be usable for dehydrating in stages of increasing alcohol concentration.
Acteone is somewhat fiercer, might go for lipids, and when it evaporates tends to chill the specimen and pull condensation out of the air.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Have not personally "looked" at metallic mercury, but have heard tales of woe from some who have tried. Mercury vapor in the sem is very bad. Will set-up a nice mercury vapor discharge lamp, disrupting everything. I have, however, examined small specimens of old amalgam without problems. You must keep the Hg from vaporizing!!!
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Seasons Greetings to all!!
For those of us left working this close to Christmas, we are looking at a contaminant that appears to be mercury but are concerned about the potential
hazard to both man (and woman) and machine. Are there any brave souls that have looked at mercury using a cryo system and is it relatively safe and non-destructive?? We would also like to get sample particle sizes and other
As a new subscriber to this group, I offer the following as a brief introduction.
My interest lie in the general area of forensic science - specifically forensic document examination. While most of my work involves optical microscopy in one form or another, I have also used SEM and CLSM to examine physical evidence in the form of "documents".
for dehydration we use either a graded series of ethanol OR acetone. This works very well, even on delicate tissues like e.g. bone marrow. You can also use propylene-glycol as the last step of dehydration, after absolute ethanol or acetone.
Yet, I'm not sure how this will work on worms. There you have the thick rigid outer shell which might impair fixation and dehydration. Have a try...
Hello, Does anyone know of a company that makes a simple micromanipulator (for making yeast tetrads) thats fitted to a Zeiss and Nikon microscopes. The original makers-Allan Benjamin Co. can not be found. Thanks.
Two vendors come to mind for micromanipulators: Leica makes excellent mechanical micromanipulators (www.leica.com), and Narishige manufactures hydraulic micromanipulators (www.narishige.co.jp). Most of these can either be placed on the microscope or put on self-supporting riser blocks that allow the needles to be brought onto the stage of the microscope.
Best regards, Bob Chiovetti ********************************* Robert (Bob) Chiovetti E. Licht Company / 1-800-865-4248 rchiovetti-at-aol.com
********************************* Leica (Wild, Leitz, Bausch&Lomb, Cambridge, AO, Reichert-Jung) / Technical Instrument Company / American Volpi / Fostek / Stocker and Yale / AEI North America / OptiQuip / Dolan-Jenner / Osram / G.E. / Philips / Ushio / Boeckler Instruments / Heidenhain / Narishige / Colorado Video / Kinetic Systems / Pryor Scientific / Compumotor / Sutter Instrument Co. / Advanced Database Systems / Cohu / Javeline Electronics / Optronics / Diagnostic Instruments, Inc. / Dage MTI / Hitachi / Panasonic / Polaroid / Kodak / Mitsubishi / Sony
My interests is in observing organic crystal growth via an optical microscope with a video/photo attachment . I would also like to study the crystallization on-line via an X-ray diffractometer. I am not after single crystals ; but a `population of crystals'.
Can anyone provide information on: -1- LINCAM - UK (manufacturer of thermomicroscopes) ; -2- in-situ crystal growth `cells' for microscopes/XRD ??
Season's Greetings! I have been asked to try to see differences in lignification in cell walls of grasses and legumes to see if it correlates with differences in expression of peroxidase. The peroxidase differences are a piece of cake to spot, but I've been having trouble with the lignin... I've tried several different protocols using phloroglucinol, but can't seem to get very intense or very consistent staining. Does anyone out there have a tried-and-true method that they are willing to share? Is there something better than phloroglucinol that I should be trying (I know it's an oldie, but seems to be standard??) thanks in advance for your help and all the best in 98!
cheers shea
Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
How about a discussion on the two most important aspects of confocal microscopy as they pertain to BIOLOGICAL Raman microspectroscopy: lenses/pinholes and specimen prep. All turnkey systems I've heard about seem to be built around high-dry lenses (e.g., 100x, NA 0.9) and incorporate fixed-diameter pinholes (if they even have more than one size!). Company scientists by-and-large start to stutter and hand-wave when I mention water-immersion lenses (either "dipping" lenses or true water-as-immersion-medium types). I've seen only one BIOLOGICAL Raman paper in which the lens is mentioned; it was reported to be a Zeiss 100x, 1.2 NA water-immersion. Is anyone familiar with this lens? My sense would dictate that, if I wanted to examine hydrated biological preps of any thickness whatsoever, I would be doing myself a favor by sticking with a water-immersion lens of some type, or am I missing something here? The above comments are complicated by sample prep. To coverslip or not to coverslip, that is the question. If I would like to use the true high res water-immersion lenses, then a coverslip is required - oui, non? But doesn't this create problems with RI mismatch and thereby throw off what is already a pretty finicky measurement? Dipping lenses would seem to be the way to go here, but we need high NA and also high mag.... And what about the pinhole - I think only one turnkey device has a variable pinhole. Others have fixed pinhole(s) that, as far as I can ascertain from speaking with company scientists, have little or no relationship to the lens (?). I'd like to hear from people doing this sort of work. Rob Palmer CEB/UT
I have a theory question about Critical Point Drying that has been bothering me. I know that the specimen is placed in a "bomb", and then transition fluid replaces the dehydrating fluid in the specimen, and the temperature is raised to the critical point, which in turn raises the critical pressure in the bomb, so that the specimen is in a sense immersed in a dense vapor phase devoid of liquid air/interface, and the vapor is slowly released until the vessel is at atmospheric pressure. But with the drop in pressure, even though it is slow, below the critical pressure for that transition fluid, why doesn't this precipitate a condensation of the vapor back to a liquid????
Is the temperature slowly increased beyond the critical temperature, to correspond to a new critical temp. for the lower pressure?
For the veterans, I'm sorry to bug them with elementary questions like this, but I can't find the answer in any book.
FOR SALE: Brand New 6.2 mm Dupont diamond knife. Never used. Current retail for a knife of this length is approximately $6000+. I would like to sell the knife for $2500 or best offer. Ron Kalil
} I have a theory question about Critical Point Drying that has been } bothering me. I know that the specimen is placed in a "bomb", and then } transition fluid replaces the dehydrating fluid in the specimen, and the } temperature is raised to the critical point, which in turn raises the } critical pressure in the bomb, so that the specimen is in a sense } immersed in a dense vapor phase devoid of liquid air/interface, and the } vapor is slowly released until the vessel is at atmospheric pressure. } But with the drop in pressure, even though it is slow, below the } critical pressure for that transition fluid, why doesn't this } precipitate a condensation of the vapor back to a liquid???? } } Is the temperature slowly increased beyond the critical temperature, to } correspond to a new critical temp. for the lower pressure? } Garry,
Basically, yes. The pressure and temperature are both raised (as you say, the pressure is increased by raising the temperature). After the critical point is passed, the pressure is released *while maintaining the elevated temperature*. This means that as the pressure is lowered the CO2 is maintained in the vapor phase. Pressure *must* be released slowly both to prevent the specimen from exploding (in quotes, if you like), and to prevent the pressure drop from lowering the temperature and thus precipitating vapor condensation back to a liquid. Usually this is around 35-40 deg. C. Higher than needed to provide a safety margin.
The CO2 could be moved through the liquid-vapor transition simply by raising the temperature above its critical value, but this would mean surface tension at the liquid-vapor interface, with all the problems that implies. So the CO2 is moved through the critical point at which there is no distinction between liquid and vapor, and so there's no interface. The vapor phase is retained by keeping the temperature elevated above the critical temperture. There is no "new" critical temperature, just the same one, but CPD makes an end run around the interface problem by raising the pressure at the same time as the temperature, keeping the CO2 liquid until the critical point is reached.
My biggest problem with CPD is what exactly happens to the specimen with the increase and decrease in pressure? Supposedly nothing, if this is done slowly enough, but I don't believe it. Things may look OK, but what really happens?
Phil
} For the veterans, I'm sorry to bug them with elementary questions like } this, but I can't find the answer in any book. } } Garry
Bother us. I find it very useful to think about things I supposedly learned (mumbletymumble) years ago. Besides, someone else likely has a better explanation, and this gives me a chance to read it.
Philip Oshel PO Box 5037 Station A Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu or poshel-at-hotmail.com ***** looking for a job *****
Does anyone have experience with preparation of CoPt(20%Pt) for SEM and EBSD. Brand new material for me and aqua regia so far doesnt seem to be the right approach.Any suggestions would be appreciated. Thanks Allen Miller
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% %%%%%%%%% Patty Miller Stained Panes momiller-at-ccia.com
} I have a theory question about Critical Point Drying that has been } bothering me. I know that the specimen is placed in a "bomb", and then } transition fluid replaces the dehydrating fluid in the specimen, and the } temperature is raised to the critical point, which in turn raises the } critical pressure in the bomb, so that the specimen is in a sense } immersed in a dense vapor phase devoid of liquid air/interface, and the } vapor is slowly released until the vessel is at atmospheric pressure. } But with the drop in pressure, even though it is slow, below the } critical pressure for that transition fluid, why doesn't this } precipitate a condensation of the vapor back to a liquid???? } } Is the temperature slowly increased beyond the critical temperature, to } correspond to a new critical temp. for the lower pressure?
} For the veterans, I'm sorry to bug them with elementary questions like } this, but I can't find the answer in any book. } } Garry
Yes - the temperature of the whole system is kept sufficiently high so that as the pressure drops, it doesn't pass through the vapour/liquid transition again.
So the system is taken in a loop around the critical point, as illustrated:) This also highlights a minor problem that you may run into in some labs - if the ambient lab temperature is too high, the CO2 is always a vapour, so the bomb may need cooling to start with, just to get liquid CO2. This caused me some difficulties when installing a system in Jakarta at the beginning of the year!
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
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