} We are currently in the last throws of setting up a confocal microscope } with all the related ancilliaries including the system required for mass } storage of data. We were very keen to include a CD read/write system for } collection and storage of the images. These appeared to be the way of the } future for storage and for transfer from the confocal workstation to the } many user sites on this campus. Much to our dismay we have found(heard) } that these systems often `crash' the Silicon Graphics system we will be } using to drive our imaging software. This has made us do an about face and } move back towards an MO drive. Has anyone else faced this dilemma ? Can we } still confidently go down the CD path. } Can anyone out there give us some confidence boosting news? } I would love to hear from you. } } Tony McKenna } NZ Dairy Research
We have a Kodak CDR225 connected to a PowerMac 8100 and use a program called Toast CD-ROM to write CDs. The files are mostly image files which we fetch from our SG Indy by Fetch 2.1. We didn't consider putting the CDR on the SG machine as we want to edit etc. the images in Photoshop on the PowerMac. We usually write the CDs in the ISO 9660 format so that they can be read by any computer. The setup works fine except that the Indy is terribly slow in mounting the CDs afterwards, especially if there are many (} 30-40) files in one folder. It may be something in the system of our machine that's a bit screwed up, in which case I hope to get it fixed, but still it is probably worth having in mind.
Via: uk.ac.bbsrc; Fri, 1 Sep 1995 13:50:33 +0100 X400-Received: by /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Fri, 1 Sep 1995 13:52:08 +0000 X400-Received: by mta arcs.bbsrc.ac.uk in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Fri, 1 Sep 1995 13:52:08 +0000 X400-Received: by mta LARS.MUAS in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Fri, 1 Sep 1995 13:49:08 +0000 X400-Received: by mta LARS in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Fri, 1 Sep 1995 13:49:09 +0000 X400-Received: by /PRMD=UK.AC/ADMD= /C=GB/; converted (ia5-text); Relayed; Fri, 1 Sep 1995 13:49:21 +0000
Dear Fellow Microscopists, I am having great difficulty in getting methacrylate-embedded sections to adhere to glass microscope slides. The slides are coated with 0.1% poly-lysine (MW c. 19000). Sections, c. 6 or 8 um thick, in a drop of water are dried down onto the slides for a few hours at c. 43C, then overnight at room temperature. The resin is removed with 100% acetone and the sections (still stuck at this stage) are hydrated in phosphate-buffered saline. Sections are then processed for indirect immunofluorescence localisation of tubulin or actin. However, a (very!!) high % os sections float off the slides after the c. 45 min pre-incubation blocking stage. The plant material used is secondary vascular tissue of roots of Aesculus hippocastanum L., ie a little bit of bark, cambium, and a large 'slab' of xylem, in radial longitudinal sections. In view of the heat-sensitivity of the tubulin antigen, I want to avoid excessive heating of the slides. Can anybody suggest a fool-proof (or, at least a tried and tested) way of ensuring that the sections stick and stay stuck? Any help or comments would be most appreciated,
Nigel Chaffey, IACR - Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, Long Ashton, Bristol BS18 9AF, UK: nigel.chaffey-at-bbsrc.ac.uk
This will not be news to many of the old hands but one or two may find it useful following the recent thread on water-free LN2. An effective way of dealing with this for small quantities is to put a couple of 'Kimwipes' in the funnel before you start pouring the LN2. They absorb/filter out even very tiny ice crystals AND ARE CHEAP AND HANDY. They allow the LN2 through quite freely and, if the flow begins to slow down half way through the fill, you can throw them out and put fresh ones in. No good for automatic or large volume systems, of course. Regards
} Dear Fellow Microscopists, } I am having great difficulty in getting methacrylate-embedded sections } to adhere to glass microscope slides. The slides are coated with 0.1% } poly-lysine (MW c. 19000). Sections, c. 6 or 8 um thick, in a drop of water } are dried down onto the slides for a few hours at c. 43C, then overnight at } room temperature. The resin is removed with 100% acetone and the sections } (still stuck at this stage) are hydrated in phosphate-buffered saline. } Sections are then processed for indirect immunofluorescence localisation of } tubulin or actin. However, a (very!!) high % os sections float off the slides } after the c. 45 min pre-incubation blocking stage. ....
We have been using this methacrylate system for localizing tubulin and actin in plant roots for some time. We do not have problems adhering our sections, before or after extraction. We prepare slides coated with 3 aminopropyltriethoxysilane. Then we collect sections in a drop (about 10 microlitres) of water and then treat the slides for 2-5 min on a hot plate at 60 C, and then air dry at room temp usually overnight. This seems to work great for us. The few minutes at 60 C seem to cause no trouble for our staining. The silane is really easy to use. If you want to try it, I can give you details of coating the slides. Hope this helps.
WORKSHOP OBJECTIVE: This course will cover all aspects of pre-thinning and focus on final thinning via Tripod Polishing. Due to the limited class size and the extensive hands-on opportunities, this course is well suited to novices as well as advanced Tripodders. The course will include sections on:
How to do it and why should I? What's really going on and what am I really seeing? How to prepare small, specific area cross-sections. The problem of widely differing materials (eg tungsten). Rapid preparation of TEM cross-sections. Preparation of a wide range of materials: semiconductors, ceramics, metals,...
HANDS ON OPPORTUNITY: This course will be unique in that it will provide a hands-on opportunity for every class participant. Tripod Polishers, Polishing Wheels, and pre-thinning equipment will be made available to all participants and actual samples will be prepared - by the students - as part of the course. This is a great opportunity to get your hands dirty and actually learn by doing. The instructor will walk you through each step of the process and then let you loose on the equipment. This course is designed to teach the Tripod Polishing technique. Silicon samples will be provided to the students and used as the basis for the course teaching.
WORKSHOP LOCATIONS AND DATES: South Bay Technology - San Clemente, CA Dates: November 10-11, 1995
INSTRUCTOR: Ron Anderson, IBM, East Fishkill Facility, Hopewell Junction, NY
CLASS SIZE: Due to the extensive hands-on aspects of this course, class size will be strictly limited to 10 participants.
REGISTRATION FEE: $795.00 (includes lunches and Friday night dinner) $ 695.00 if registration is paid by October 1, 1995
REGISTRATION DEADLINE: October 15, 1995
For additional information please contact SOUTH BAY TECHNOLOGY at 800-SBT-2233, Fax 714-492-1499 or e-mail: 73531.1344-at-CompuServe.com
Dear net, Following the tread about making slides sticky, I received several requests for the protocol for silanizing slides. So I am replying at large to the net in case others are also interested. I got this protocol from another lab on campus (Tom Phillips) and I have never modified it. So, if it doesn't work for you, I probably can't help. Anyway, here it is:
The silane compound comes as a liquid from sigma. Its name is:3, aminopropyltriethoxy silane
Dip slides in a 2% silane/acetone solution for one minute Dip slides in 100% actetone for 1 min. Dip slides in ddH2O for 1 min. Repeat this step with agitation Air dry.
The slides seem consistently sticky in our hands, although we once had trouble with really teeny weeny sections that someone here was cutting (but this was also for an aplication with lots of long rinses in running water).
Hope this helps. A citation and a little bit of further information about this can be found in:
Angerer, L.M. & Angerer, R.C. (1991) Localization of mRNAs by in situ hybridization. Functional Organization of the Nucleus: a Laboratory Guide. Methods in Cell Biology, Vol 35, (ed. by B.A. Hamalko & S.C.R. Elgin), pp. 37 - 71. Academic Press, San Diego.
Dear List Subscriber, I would be interested in obtaining phase contrast accessories for a Leitz Diavert optical microscope. Specifically, the Phase Contrast Condenser according to Zernike (513-84) would be ideal for our work. These parts are on longer manufactured by Leitz. We are also interested in Phase Contrast rings for the Diavert. The Diavert used to be the top of the line Leitz microscope, 10 to 20 yrs ago. If you have some of these units still around, but inactive, we could make use of some of the optical parts --- and even unload off you (at a reasonable price) major optical components to repair our own unit. Paul Heroux Medicine McGill University
unsubscribe bjg-at-uniwa.uwa.edu.au Brendon J. Griffin Centre for Microscopy and Microanalysis The University of Western Australia Nedlands, WA, AUSTRALIA 6907 ph 61-9-380-2739 fax 61-9-380-1087
Subject: Time: 11:54 AM OFFICE MEMO SEM mounting Date: 9/3/95
Microscopy subscribers- I understand Duco brand cement is sometimes used for mounting specimens for SEM imaging. Anyone have experience using this? I would like to know what its general properties are- Cure time, outgassing, volatility under the beam, etc. Thanks all. doug_davis-at-maillink.berkeley.edu EML, UC Berkeley
I am interested on the community input on performing in-situ analysis of solder to determine composition. We have tried to do this in the past and have not had acceptable results. We used both multiple areas and multiple points and averaged the data.
I realize that this is probably an impossible task with EDS, but would like to confirm my opinion and obtain input as to any other techniques (micro probe XRF, etc.) which may provide a non-destructive method for in-situ solder analysis.
A student wants to electropolish a 1cm square of M2 high speed steel - he says this contains Mo, W and C. I would be grateful for a solution composition, temperature and voltage suggestion. Thanks in advance. Mike
Dr MJ Witcomb Electron Microscope Unit University of the Witwatersrand Private Bag 3 WITS 2050 South Africa
I haven't used the Duco cement in a long time. Since it is a solvent based cement, it will have a long drying time and the danger of rewetting dry biological samples. If memory serves, there was no great problem with outgassing after the glue was dried thoughly. It was used for a larger sample so the beam was not on the adhesive often, but I do remember trials where some damage was evident.
There was a fantastic, comprehensive, review article by Judy Murphy that covers most mounting techniques well;
Judith A. Murphy. Considerations, Materials, and Procedures for Specimen Mounting Prior to Scanning Electron Microscopic Examination. Scanning Electron Microscopy 1982 II. SEM Inc. Chicago. 1982. 657-696.
Generally, with biological samples and impervious samples, I shy away from solvent based adhesives. The former rewet and suffer surface tension artifact on redrying, and the latter trap liquid adhesive underneath and outgas for a long time. Usually, like to stay with the coated adhesives like the transfer tabs (Avery "spot o' glue"), metal tapes, and for tiny samples Scotch 850 polyester tape, so the samples don't sink, or form ripples.
Sorry for the rant. later dlb
David Bentley Imaging Facility, Div. of Biotechnology, ARL The University of Arizona Tucson, Az 85721
We recently changed microscopes but retained our old EDS detectors. I am working on finding the best way to put some of our old EDS things on the 'new' microscope. The conversion may require changing windows and/or converting from a horizontal to a high angle configuration on a TEM.
Can anyone offer some sage advice about strategies, configurations, prices, and/or vendors who can perform the work?
Thanks,
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 jmkrupp-at-cats.ucsc.edu
David- The Avery "spot o glue" is no longer made, and hasn't been available in some time. Check with your favorite consumables supplier for their recommended substitute. Steven Slap, Energy Beam Sciences
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
On Sept. 3, Doug Davis asked about "Duco" brand cement:
"Duco" cement will certainly "work", however, there are much better alternatives, "better" being defined from several perspectives:
a) Double sided adhesive conductive carbon sheets and tape will do the same job (provided the samples being mounted are not too large) but have the advantage of no outgassing. They are also "relatively" stable under the beam, and since the substrate is already conductive, to whatever degree one might have to apply gold or carbon to impart surface conductivity, in this case, one would have to apply much less. Cure time is not applicable, and the sample is instantly ready to be put into the vacuum of either a sputter or carbon coater or the SEM. Remember, Duco cement is certainly not conductive. Also, if not completely cured (who has the time for a complete "cure" in a busy SEM lab?), solvent vapors off-gas.
b) One can use products called "Leit-C-Plast" or even "Tempfix" which have the advantage that more massive samples can be held in place and the adhesive itself is conductive. Again, cure time is not an issue.
The only apparent "disadvantage" is that these other alternatives all cost more money than "Duco" cement. On a "per sample" basis, we are still talking pennies, but they do cost more.
Now for this disclosure: SPI Supplies has been offering these "Duco" cement alternatives for some number of years. They are in wide use worldwide. Other types of dry adhesive products are available from other suppliers, not necessarily the same item in a different wrapper. They differ in terms of conductivity, purity, surface smoothness, and freedom from pin-holes and of course, price. You have to try them out to see which ones work best for you. All would seem to be a better choice than "Duco" cement (or even any of the so-called "five minute epoxies"). Products like "Duco" might seem to be "cheaper" but in the long run, in many cases, they tend to be more "expensive" in terms of wasted time doing samples over again and also, in terms of more frequent column cleanings, aperture replacements,etc.
See pages 43-47 of the SPI Supplies "SourceBook" and/or contact SPI by e-mail for appropriate SPI #'s and current prices. Sorry, these products are not yet up in the SPI Supplies "electronic catalog" on our "web site" but check after Oct. 1.
Chuck
===================================================== Charles A. Garber, Ph. D. Ph: 1-(800)-2424-SPI President 1-(610)-436-5400 SPI SUPPLIES/Structure Probe, Inc. FAX: 1-(610) 436-5755 PO BOX 656 West Chester, PA 19381-0656 USA
e-mail: GVKM07A-at-prodigy.com [Direct for C. Garber] SpiSupp-at-aol.com [SPI Customer Service e-mail box]
On Sept. 6 Alwyn Eades inquired about adhesives for mounting samples for cryo SEM:
We have found that mechanical mounting in the equivalent of a tiny vise built into the appropriate specimen stub makes for rapid mounting, good thermal contact, and good stability under the beam at low temperatures (down to that of liquid nitrogen in our case. Adhesives, including "cryoglues" such as toluene failed or were awkward. Good thermal contact has been critical in our experience so we fracture or saw cut at least one or two flat faces on the sample to make contact with the jaws of the vise.
A photograph and diagram of a 12 mm holder we use can be found in "Bastacky, J., C. Goodman, and T. L. Hayes (1990). A specimen holder for low-temperature scanning electron microscopy. Journal of Electron Microscopy Technique 14(1): 83-84." A 5mm holder for the Gatan coldstage is described in "J. Bastacky, C. Lee, T. Freeman, G. Weber, A. Baeza, T. Hubbins, Y. Chen. (1995). A specimen holder for high-resolution low-temperature scanning electron microscopy. Microscopy Research and Technique, in press." The technique of sawing flat faces is illustrated in Bastacky, J., G. R. Hook, G. L. Finch, J. Goerke, and T. L. Hayes (1987). Low-temperature scanning electron microscopy of frozen hydrated mouse lung. Scanning 9: 57-70. I expect these approaches would work for liquid helium temperature samples.
Jacob
Jacob Bastacky, MD Room 116 Donner Laboratory Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
We never turn our EDS detectors off, and depend on the automatic shutdown in the rare occasions that the LN2 is depleted. The only question I have about turning them on and off routinely is whether the stability of the calibration is maintained after numerous shutdowns and re-starts. Anyone have any observations? Larry
I have just undergone heavy duty surgery on my right shoulder: tremendous arthritis, bone spurrs, etc. etc. We did the right shouder because I am right handed and because it was much worse than the left which will probably need to be done in a year or two. Someone just said to me that all electron microscopists end up with shoulder problems. I had not heard this before. I've been doing EM full time for well over twenty years so I guess I qualify as a "lifer". Have any of you EMers out there ever heard of anything like this or have you experienced similar proble pro I'm just curious; if it's true we should warn new EMers and perhaps get the manufacturers to institute design changes to ameliorate such oblems.
As a favor for a collaborator, I used PTA to negatively stain a phage prep. I found lots of example of what I think may be filamentous phages but am not certain. are there any phage morphologists out in cyber space? I am interested in knowing the size range I should be seeing. my filaments are about 12 nm wide. is this too big? can anyone suggest references with good TEM pictures with which to compare my stuff? Thanks for any help.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (314)-882-4712 (voice) (314)-882-0123 (fax)
I have questioned colleagues at this location and nobody has heard of an EM related shoulder injury (we are all 20+years). A service engineer visiting us today does relate having heard of wrist related pain following repeated exposure to freon during cleaning procedures..
Message-Id: {9509062133.AA29067-at-las2.iastate.edu} To: GVKM07A-at-prodigy.com Cc: MICROSCOPY-at-AAEM.AMC.ANL.GOV {013.00881436.GVKM07A-at-prodigy.com}
Let me second Charles Garber's comments on conductive/adhesive mounting. I have had to run samples of powders applied to double-sticky "carbon dots" on several occasions when it was impossible to coat the samples at all. I don't recommend it, but you can get it to work. If anyone has a similarly tricky SEM or EPMA problem, I'd be happy to supply more information. --- Alfred Kracher akracher-at-iastate.edu http://www.public.iastate.edu/~akracher/
This is in response to Dr. Bently's message regarding the Avery Spot-O-Glue. It is absolutely still in existence and never was unavailable as was posted on the listserver. Two years ago Avery discontinued making the product and we at Electron Microscopy Sciences bought out the equipment needed to manufacture the Spot-o-Glue and have been manufacturing it under the same name for the past 2 years. It still comes in the 2,592 tabs per pack with 72 sheets per pack. The pricing remains as it was when Avery was making it. If anyone wants further information on the product please just give me a call and I will be more than happy to assist. Sincerely,
Stacie Kirsch SGKCCK-at-aol.com ELECTRON MICROSCOPY SCIENCES TEL:215-646-1566 FAX:215-646-8931
I apologize in advance for this lengthy letter but I'm in desperate need of help & feel it is necessary to describe the whole picture.
I am performing cryoultramicrotomy of wool fibres which have been treated previously with a stain to provide sufficient contrast to observe ultrastructure & the changes which take place as the water in the wool fibre is dried. The aim is to compare ultrastructure of frozen hydrated & freeze dried cryosections of wool to determine where water is located/contained in the fibre.
A method has been established during a number of trials based on different cryogens, embedment methods & media, temperatures, knife angles, collection methods etc. To summarize, an RMC MT7 ultramicrotome fitted with CR-21 cryoattachment, is used at a temperature (both specimen & knife) of -125=B0C= , 45=B0 glass knives with cutting angle of 5=B0 and dry collection (because= frozen hydrated sections required) of cryosections with eyelash probe onto Cu sandwich grids coated with celloidin & carbon.
The embedment method uses a plastic (Gilson's 2-100=B5l) pipette tip to= clamp a bundle of fibres which protrude from the tip. It is necessary to use a bundle of fibres to provide sufficient support - wool fibres do not freeze very well & remain relatively flexible but although a bundle of fibres gives good support it does result in a rather large block face.
The bundle of fibres is soaked in water (to hydrate them), dipped in 40% PVP with 10% glycerol (this embedment medium was found to be most compatible with wool fibres in terms of hardness & ease of cutting) & frozen in a hexane/petroleum spirit slush (found to be most suitable).
The problem(s) is that the wool fibres tend to compress on cutting so that for some of the cutting cycles no wool sections are obtained & then some very thick sections are cut. Occasionally, a thinner section will be cut which is OK for STEM (but then there are other problems: folding, no cuticle on fibre...).
Secondly, & perhaps more easily solvable (?), due to the nature of the block face (as described earlier with a fibre bundle surrounded by enough PVP/glycerol to support the bundle) a large amount of "ice"/PVP + glycerol sections - ie. without wool - are produced. This, in conjunction with the compression problem, results in many small (because they are compressed) sections overlapping or on top of one another, instead of nice cryosections "peeling off" as occurs with "easier" tissues. All of these problems mean that cryotransfer of these sections to the microscope results in a grid containing alot of ice, some very thick sections & the very occasional thinner section (& the ice usually obstructs examination of the ultrastructure of these sections). These time-consuming procedures give a low yield of useable sections.
I would be most grateful if anyone could pass on any useful advice, comments, references, methods or related experiences. Many thanks
In response to Rachel's difficulties with cryosectioning wool fibres, I have a few comments which I thought I would throw to the net also to stimulate some ideas.
1. It's important that the specimen is firmly held and does not flex during sectioning, therefore it should be mounted so it doesn't project far out from the chuck. 2. A cryo diamond knife might help to give more reproducible sectioning. 3. In past experience I've found that some stubborn materials sometimes cut better with a lower angle glass knife, ie. a 40 or 35 degree knife, though the knife edge probably doesn't last as long as a 45deg one. 4. Freezing in liquid propane might be better, though it probably would not solve the primary problem that the fibres remain flexible at low temperature. 5. Try a very low sectioning speed, or section manually so you can control it. If it doesn't help, very occasionally a fast speed might work instead. 6. Is it possible to mount the fibre bundle in a thin tube made of some material which could be sectioned together with the fibres? 7. It's often difficult with non-cryoprotected specimens to get nice regular sections, and collecting sections with an eyelash probe can be fiddly, so productivity is usually fairly low anyway. If collection of the sections is an additional problem, perhaps an anti-static device might help. 8. Rather than using the sections, the frozen sectioned fibre bundle could be mounted on a cold stage in a cryo-SEM, and the cut ends examined and analysed directly. Unfortunately we do not have such a facility in Brisbane, but perhaps arrangements could be made with another lab.
Any other ideas anyone? Regards Thor ---------------------------- Dr Thor Bostrom Analytical EM Facility Queensland University of Technology (QUT) GPO Box 2434, Brisbane, QLD 4001, Australia Ph: (61) 7-3864-2351 FAX: (61) 7-3864-5100
In response to John A. I have had bursitis in both shoulder for years but I doubt that it is job related. It has been with me for to long. I do however known many EM'ers who have had sinus problems. Mine have gotten worse over the years. I have been doing EM for about 14.5 years.
} From JOHNA-at-SCI.WFEB.EDU Wed Sep 6 16:23:32 1995
} Hello folks,
} I have just undergone heavy duty surgery on my right shoulder: tremendous } arthritis, bone spurrs, etc. etc. We did the right shouder because I am } right handed and because it was much worse than the left which will } probably need to be done in a year or two. Someone just said to me that } all electron microscopists end up with shoulder problems. I had not heard } this before. I've been doing EM full time for well over twenty years so I } guess I qualify as a "lifer". Have any of you EMers out there ever heard } of anything like this or have you experienced similar proble pro I'm just } curious; if it's true we should warn new EMers and perhaps get the } manufacturers to institute design changes to ameliorate such problems.
The following message was sent to the WWW Microscopy & Microanalysis site can anyone help this person?
If you have the information reply to the address at the end of the message (Email: pj8v-at-nih.gov) . Do not use Email reply as the message will go to me not the originator.
I would like to know, if by any chance you might have the information, the address/phone of MacArthur or McArthur Microscores of Cambridge, England. The company used to make a small hand held light microscope that was used in the field, especially by scientists conducting work on parasitic diseases. Would you happen to know the address/phone or someone who might? I am trying to get the info to a missionary trying to do malaria diagnosis in Hati under primative conditions. No electricity so a self contained high power (oil immersion to high dry) scope is needed.
Thanks,
Peter Jackson, Ph.D. fax 301-402-2638 Ph 301-496-8426 Email: PJ8V-at-NIH.GOV
Dear Rachel, What thickness sections are you cutting? I have no real experience with cryosectioning (I was at a demo, and we have a Reichert), but very thin sections are difficult in general, and your specimen seems particularly patho- logical. Could you get away with thicker sections? Do you have access to an IVEM? If so, that would be a possible way out. Good luck. Yours, Bill Tivol
} I am interested on the community input on performing in-situ analysis of solder } to determine composition. We have tried to do this in the past and have not } had acceptable results. We used both multiple areas and multiple points and } averaged the data. } } I realize that this is probably an impossible task with EDS, but would like to } confirm my opinion and obtain input as to any other techniques (micro probe } XRF, etc.) which may provide a non-destructive method for in-situ solder } analysis.
Dear John, What size specimen do you need to analyse, and what size is the solder blob? Is your problem related to the volatility of the solder components? How accurate do you need the quantitation? If your specimen fits in a SEM, you could try EDS at relatively high voltage (necessary for Pb) and low beam current to minimize volatility problems. Possibly proton-induced x-ray emis- sion (PIXE) will do the job. If you are interested in this technique, I'd contact Dr. Tom Cahill at UC Davis. He has examined everything from air pollution to a page from a Guttenberg Bible. I don't have his email address, but his phone # is (916) 752-1460. Good luck. Yours, Bill Tivol
} OFFICE MEMO EDS detector HV stability Date: 9/6/95 } } We never turn our EDS detectors off, and depend on the automatic shutdown in } the rare occasions that the LN2 is depleted. The only question I have about } turning them on and off routinely is whether the stability of the calibration } is maintained after numerous shutdowns and re-starts. Anyone have any } observations? } Larry } Dear Larry, We have checked the calibration on our TN2000 (an oldie), and we find that it is maintained pretty well--to one or two channels, or 10 or 20 ev. Noran has a somewhat tedious procedure for energy calibration which works well for us with aluminum on a copper grid. This procedure takes less than 1 hr and is accurate to better than one channel. I don't know what components are in the circuits, but I'd worry more about drift due to decay of one of them than due to restarts. BTW, we use our EDS only infrequently; if you use yours daily, it's probably right to leave it on. Yours, Bill Tivol
For the first time ever, I have been asked to section some material prepared in UV polymerized Lowicryl resin. The sample material appears to refuse to be cut while the plastic cuts routinely. I have tried some heat cured blocks from the same materials at the same time; these cut without any problem, but the antigenicity is comprimised in the sample. Because these samples are from an outside source, I do not have any control over the preparation. Another contributing factor is ,without enormous expense and difficulty, the material may not be replaceable.
Questions: Is there a specific alternative flotation fluid used with the UV versus heat cured Lowicryl?
Will the antigenicity be comprimised if the samples are heat cured after they were UV cured?
Has anyone else had and solved this problem?
Please send any and all suggestions / recommendations gained through experience with this resin. melsen-at-MICROBIO.emory.edu L.R. Melsen Emory University Microbiology and Immunology 3029 Rollin Research Center Atlanta, Ga 30322 404 727 3508 FAX 404 727 3659
Hi, Any chance the persons that supplied you with the tissue used OsO4? Methacrylate resins (Lowacryl is in this "family") do not like to polymerize in the presence of OsO4. If they used OsO4, I'm afraid you may be out of luck.
Just a guess re: your problems with sectioning. Doug ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: dcromey-at-ccit.arizona.edu) : :...................................................................:
I am sending this for the EM lab in Marine Biology at Bodega Bay, CA (in the UC, Davis system) Please contact them for further information. Thanks
Two EMs are available, Zeiss EM 902 TEM and a Hitachi S570 SEM. The listed prices through UC, Davis Central Stores and Receiving are $79,000 for the Zeiss and $12,500 for the Hitachi. Bids at lower prices will be considered if necessary. The sale will be processed by Central Stores and Receiving at UC, Davis, but persons interested in submitting bids may contact me at the Bodega Marine Lab. Fred Griffin, PhD Telephone 707/875-2045 FAX 707/875-2009 e mail: fjgriffin-at-ucdavis.edu ______________________________________________________________________________ San Joaquin Delta College World Wide Web:http://www.sjdccd.cc.ca.us 5151 Pacific Avenue email:webmaster-at-ms.sjdccd.cc.ca.us Stockton, CA 95207 general information:(209) 474-5151 or FAX-at-(209)474-5600
Fortunately, the designer of the Lowicryl family (K4 M, HM20, K11, HM23) Eric Carlemalm can be consulted directly. I think we all will learn a lot from his reply so I am forwarding your question to Eric.
} For the first time ever, I have been asked to section some material prepared } in UV polymerized Lowicryl resin. The sample material appears to refuse } to be cut while the plastic cuts routinely. I have tried some heat cured } blocks from the same materials at the same time; these cut without any } problem, but the antigenicity is comprimised in the sample. Because these } samples are from an outside source, I do not have any control over the } preparation. Another contributing factor is ,without enormous expense and } difficulty, the material may not be replaceable. } } Questions: } Is there a specific alternative flotation fluid used with the UV versus heat } cured Lowicryl? } } Will the antigenicity be comprimised if the samples are heat cured after } they were UV cured? } } Has anyone else had and solved this problem? } } Please send any and all suggestions / recommendations gained through } experience with this resin. } melsen-at-MICROBIO.emory.edu } L.R. Melsen } Emory University } Microbiology and Immunology } 3029 Rollin Research Center } Atlanta, Ga 30322 } 404 727 3508 FAX 404 727 3659
********************************************************* Sverker Enestrom M.D., Ph.D. Department of Pathology University of Linkoping, Sweden Phone: +46 13 22 15 20 Fax: +46 13 13 22 57 *********************************************************
Fortunately, the designer of the Lowicryl family (K4 M, HM20, K11, HM23) Eric Carlemalm can be consulted directly. I think we all will learn a lot from his reply so I am forwarding your question to Eric.
} For the first time ever, I have been asked to section some material prepared } in UV polymerized Lowicryl resin. The sample material appears to refuse } to be cut while the plastic cuts routinely. I have tried some heat cured } blocks from the same materials at the same time; these cut without any } problem, but the antigenicity is comprimised in the sample. Because these } samples are from an outside source, I do not have any control over the } preparation. Another contributing factor is ,without enormous expense and } difficulty, the material may not be replaceable. } } Questions: } Is there a specific alternative flotation fluid used with the UV versus heat } cured Lowicryl? } } Will the antigenicity be comprimised if the samples are heat cured after } they were UV cured? } } Has anyone else had and solved this problem? } } Please send any and all suggestions / recommendations gained through } experience with this resin. } melsen-at-MICROBIO.emory.edu } L.R. Melsen } Emory University } Microbiology and Immunology } 3029 Rollin Research Center } Atlanta, Ga 30322 } 404 727 3508 FAX 404 727 3659
********************************************************* Sverker Enestrom M.D., Ph.D. Department of Pathology University of Linkoping, Sweden Phone: +46 13 22 15 20 Fax: +46 13 13 22 57 *********************************************************
I'm puzzled by the idea that methacrylate resins don't polymerize properly in the presence of OsO4. When I started EM back in the 1950s, everyone fixed tissues in OsO4 and embedded in methacrylate resins. That was before glutaraldehyde and Epon arrived on the scene in the early 1960s.
A. Kent Christensen, University of Michigan, {akc-at-umich.edu}
-------------------------------------
On Thu, 7 Sep 1995 dcromey-at-CCIT.ARIZONA.EDU wrote:
} Hi, } Any chance the persons that supplied you with the tissue used OsO4? } Methacrylate resins (Lowacryl is in this "family") do not like to } polymerize in the presence of OsO4. If they used OsO4, I'm afraid you } may be out of luck. } } Just a guess re: your problems with sectioning. } Doug } ..................................................................... } : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : } : Sr. Research Specialist University of Arizona : } : (office: AHSC 4212A) P.O. Box 245044 : } : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : } : (FAX: 520-626-2097) (email: dcromey-at-ccit.arizona.edu) : } :...................................................................: } }
We ordered Mowiol powder for mounting coverslips and can't seem to find a recipe for making it up and storing it Please share your lab instructions Many thanks
Sandra K. Masur Box 1183 fax: 212-289-5945 Mount Sinai School of Medicine phone: 212-241-6544 or 0089 NY NY 10029-6574 e-mail:masur-at-msvax.mssm.edu
Subject: Time: 4:21 PM OFFICE MEMO Freeze Slammer for sale Date: 9/5/95
FOR SALE: One Hitek (RMC) copper block freeze slammer, can be used with LN2 or liquid helium sources; appropriate transfer lines for each included. Extras, please inquire. Original purchase price $5860, all offers will be considered. Doug Davis doug_davis-at-maillink.berkeley.edu EML, 26 Giannini Hall University of California Berkeley, CA 94720
I, too, have had trouble with complete*UV* initiated of polymerization of some samples that had been embedded in Lowacryl resins. Since I had no problems with heat initiated polymerizations in the same tissue, I assumed the the problem was with the reduced Os absorbing enough of the UV to inhibit polymerization in the tissues.
A phenomenon I find more perplexing is the un (intentionally) initiated polymerization of just some of a batch of plant samples (that were OsO4 fixed) after final infiltration in LR white, in a lab that should have no stray UV light in it . Any ideas on this?
John Heckman Center for Electron Microscopy Michigan State University
A collegue (Jill Verlander) of mine needs the address of Dr. Apkarian. If you could e-mail it to her directly as I will be on vacation she would be grateful. Thanks
Most of the problems (and solutions) to the acrylic resins, as Dr. Christensen alluded to, goes back to, and have been solved in the the early days of EM with methyl/butyl methacrylate. Although we haven't discovered all the problems with the acrylic resins and still expect to see more, at this point, consider it to be the resin of choice in many cases for sample embedment. I do have several suggestions in dealing with LR white embedding problems. The curing of all acrylic resins in an exothermic process where heat is released. In the case of osmicated tissues, heat is absorbed by the dark osmicated tissue, which raises the temperature, which is further absorbed, which raises the temperature further, ect. Ultimately, this will result in full polymerization of the plastic in the bottle prior to embedding (you may be able to "rescue" some important tissues even after gelling by cutting the soft plastic with an applicator stick and embedding in fresh resin). The best solution seems to store the samples in the refrigerator during infiltration and keep the samples in the dark until just ready to embed and polmerize them. I have heard rumors that a fair amount of UV light does escape the flourescent lamps, although I have nothing substantial to back such a rumor up. Two other tricks that seem to help with LR White: One is to purchase it from your supplier uncatalysed, and add the catalyst when you are ready. We divide the bottle, after adding the catalyst, into 10 ml plastic scintillation vials, and store at -80C, taking it out as we need it. Check with your EM supplier for availability. This idea comes from Newman, G.R. and J. A. Hobot. Resin Microscopy and On-section Immunocytochemistry. 1993. Springer Verlag. New York., and a suggestion a while back on this listserver. The other trick comes from V. Lindley in: Microscopy Research and Technique. A new Procedure for Handling Impervious Biological Specimens. 21:355-360 (1992). After the osmicated tissue is placed in gelatin capsules filled with resin, leave the specimens at room temperature overnight prior to putting in the oven for cure. This has eliminated the gas bubble formation around the samples as they are cured. (This article has many other tricks and is well worth reading.) I agree with your analysis that dark or highly colored samples don't polymerize well with UV and may need subsequent heat treatment to fully cure. 50C for 24-48 hours has seemed ok thus far, but the antigens we look at may not be that heat sensitive. We have been collecting the ways to mess up the acrylic resins. We are somewhere between 50 and 100, and we have only scratched the surface. Sorry about the length of this message. later dlb
David Bentley Imaging Facility, Div. of Biotechnology, ARL The University of Arizona Tucson, Az 85721
With reference to your request for information about heating cooling stages. You might like to look in my book "Low Temperature Microscopy and Analaysis" Plenum Press New York 1992. There are many primary and secodary references to heating cooling stages.
Patrick Echlin Director, Multi-Imaging Centre School of Biological Sciences University of Cambridge UKOn Fri, 8 Sep 1995, Michael Mallamaci wrote:
} Dear Microscopists, } } I thought I would open this question up to a more diverse microscopy audience, } since I received only limited responses from the SPM list. } } We are interested in purchasing a controlled temperature stage for an AFM. It } doesn't necessarily have to be made for the AFM, but only needs to be } retrofitted to operate on an existing scope. The ideal stage would cool to LN2 } temperatures, heat to around 100-200 C, and offer temperature control +/- a } couple of degrees. We realize that this may require a couple of different } stages, and that such heating/cooling may be hazardous to the scope (a risk we } are willing to take). We also know that a Peltier device operates within this } temperature range, although not to the extremes, and we would like to start our } studies with a Peltier stage if possible. } } Does anyone know of any vendors which supply Peltier or any other types of } heating/cooling stages (perhaps for an SEM) which could fit our needs? } } We plan to build these devices ourselves if nothing is available commercially. } We are limited to sample stages since it will be impossible for us to put the } entire microscope in a freezer/oven due to its size. I would appreciate hearing } from anyone who has some practical experience with the hazards and pitfalls of } building such sample stages. } } Thanks to all in advance, } } Mike } } _________________________________________________________ } Michael P. Mallamaci mallamaci-at-goodyear.com } Goodyear Tire & Rubber Company Tel: (216) 796-7436 } D/415A Analytical Sciences Fax: (216) 796-3304 } 142 Goodyear Blvd } Akron, OH 44305 USA } }
I'm still puzzled. I received two direct E-mail messages about problems in UV curing of Lowicryl for OsO4 specimens before this additional thoughtful message arrived. I wonder if Lowicryl/LR White are more vulnerable to UV with OsO4 specimens than ordinary methacrylate was. When I began EM as a graduate student in 1956, we always cured methacrylate with heat, but during the time I was a postdoc with Don Fawcett we switched to UV curing of methacrylate because it gave better results (the tissues in those days were fixed only with OsO4).
A. Kent Christensen, University of Michigan, {akc-at-umich.edu}
--------------------------------------
On Fri, 8 Sep 1995, David Bentley wrote:
} Most of the problems (and solutions) to the acrylic resins, as Dr. } Christensen alluded to, goes back to, and have been solved in the the early } days of EM with methyl/butyl methacrylate. Although we haven't discovered } all the problems with the acrylic resins and still expect to see more, at } this point, consider it to be the resin of choice in many cases for sample } embedment. } I do have several suggestions in dealing with LR white embedding } problems. The curing of all acrylic resins in an exothermic process where } heat is released. In the case of osmicated tissues, heat is absorbed by the } dark osmicated tissue, which raises the temperature, which is further } absorbed, which raises the temperature further, ect. Ultimately, this will } result in full polymerization of the plastic in the bottle prior to } embedding (you may be able to "rescue" some important tissues even after } gelling by cutting the soft plastic with an applicator stick and embedding } in fresh resin). The best solution seems to store the samples in the } refrigerator during infiltration and keep the samples in the dark until just } ready to embed and polmerize them. I have heard rumors that a fair amount } of UV light does escape the flourescent lamps, although I have nothing } substantial to back such a rumor up. } Two other tricks that seem to help with LR White: } One is to purchase it from your supplier uncatalysed, and add the } catalyst when you are ready. We divide the bottle, after adding the } catalyst, into 10 ml plastic scintillation vials, and store at -80C, taking } it out as we need it. Check with your EM supplier for availability. This } idea comes from Newman, G.R. and J. A. Hobot. Resin Microscopy and } On-section Immunocytochemistry. 1993. Springer Verlag. New York., and a } suggestion a while back on this listserver. } The other trick comes from V. Lindley in: Microscopy Research and } Technique. A new Procedure for Handling Impervious Biological Specimens. } 21:355-360 (1992). After the osmicated tissue is placed in gelatin capsules } filled with resin, leave the specimens at room temperature overnight prior } to putting in the oven for cure. This has eliminated the gas bubble } formation around the samples as they are cured. (This article has many } other tricks and is well worth reading.) } I agree with your analysis that dark or highly colored samples don't } polymerize well with UV and may need subsequent heat treatment to fully } cure. 50C for 24-48 hours has seemed ok thus far, but the antigens we look } at may not be that heat sensitive. } We have been collecting the ways to mess up the acrylic resins. We } are somewhere between 50 and 100, and we have only scratched the surface. } Sorry about the length of this message. } later dlb } } David Bentley } Imaging Facility, Div. of Biotechnology, ARL } The University of Arizona } Tucson, Az 85721 } }
} Subject: Time: 11:54 AM } OFFICE MEMO SEM mounting Date: 9/3/95 } } Microscopy subscribers- } I understand Duco brand cement is sometimes used for mounting specimens for } SEM imaging. } Anyone have experience using this? I would like to know what its general } properties are- Cure time, outgassing, volatility under the beam, etc. Thanks } all. } doug_davis-at-maillink.berkeley.edu } EML, UC Berkeley } I have used Duco cement for mounting large specimens:
- place a small drop of Duco cement on the stub - place a small drop of Tube Koat (or any other conductive glue) next to it on the stub - mix well with a toothpick - apply the sample - dry at least overnight before coating or putting in scope
- I have not attempted to determine outgassing times, etc but this seems to work for me - I just put the sample in a fume hood and cover partially -
- adding the tube koat makes the adhesive layer conductive - this works really well for large samples, - if the surface in contact with the stub is very uneven you can use a little more cement to fill in the gaps -
I do not have any reference for this but it is my impression that this is a commonly usd technique
{Pine.SOL.3.91.950909161447.11231B-100000-at-breakout.rs.itd.umich.edu} To: "A. Kent Christensen" {akc-at-umich.edu} Cc: David Bentley {dlb-at-u.Arizona.EDU} , Microscopy-at-aaem.amc.anl.gov Message-id: {Pine.PMDF.3.91.950911074544.541071992B-100000-at-CCIT.ARIZONA.EDU} X-Envelope-to: Microscopy-at-aaem.amc.anl.gov MIME-version: 1.0 Content-type: TEXT/PLAIN; charset=US-ASCII Content-transfer-encoding: 7BIT
Kent, I don't know what to say about your OsO4 experience with Lowacryl and Methacrylates. I doubt it's because Dave Bentley and I are both from Arizona (we can cure epoxy outdoors this time of year, "but it's a dry heat" ;-)). Seriously, my experience with Methacrylates is limited, but here's what I remember: JB-4 refused to heat polymerize around pieces of glut/OsO4 fixed nerve tissue (a real mess) and I believe the instructions hinted that this could be a problem. Lowacryl (I wish I still had a copy of the package insert) mentioned the OsO4 problem, particularly with UV polymerization (poorly polymerized tissue in the center of the block, this can be a problem with too large pieces of pigmented tissues eg liver that have never seen OsO4). LR White, I have a package insert from EM Corp that states "Osmium Tetroxide reacted tissues should not be 'cold cured' with the accelerator. This process is strongly exothermic and the dark colour of the tissue leads to focal heat accumulation which can cause local problems in and around the tissue."
This is not to cast doubt on the veracity of your experiences. We all know that there is as much "art" as there is "science" in doing EM.
Have a good week. Doug
..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: dcromey-at-ccit.arizona.edu) : :...................................................................:
Recently I sent a message to the listserver offering a EM300 for sale. The instrument has been sold. Thanks to those who requested donations; there were some interesting prospects for the donation list. Bob Roberts Arizona State University Center for Solid State Science
Hey folks -- Is anybody out there looking to sell a (hopefully) low-milage Reichert FC4E cryo attachment? Our FC4 (a museum piece, but used on a Reichert Ultracut E) is on it's last legs and we would like to avoid having to buy BOTH a new microtome and cryo unit. Please reply to me personally at gmartin-at-welchlink.welch.jhu.edu Thanks in advance,
Greg Martin Dept. of Cell Biology and Anatomy Johns Hopkins School of Medicine
We are looking for a 2nd hand CAMERA for a JSM-T330A. The JEOL name for this unit is "CSI-5". This is the camera unit for polaroid photos. If you are not using this unit anymore, we are interested in buying it from you.
Please, let us know the price.
Thanks in advance.
Our address :
JEOL (EUROPE) BV Service Department Ikaroslaan 7A - 1930 Zaventem BELGIUM TEL : (32) 02 720.05.60 FAX : (32) 02.720.61.34
} } Post-doc on TEM available - The NSF-Materials Research Group at the } School of Electrical and Computer Engineering of Purdue University has an } immediate opening for a post-doctoral research associate. The position } requires hands-on experience on transmission electron microscopy (TEM) } including electron diffraction pattern analysis, bright-field, dark-field, } weak-beam, and high-resolution TEM, as well as specimen preparation for } plan-view and cross-sectional TEM. The employee will be responsible for } the characterization of molecular-beam epitaxy (MBE) grown II-VI, III-V } and/or nitrides wide bandgap semiconductor heterostructure devices, } including analysis of the failure mechanism of II-VI lasers and light- } emitting diodes. Duties will also include maintenance of sample } preparation facilities including a Gatan Dual ion mill and a Jeol-2000 EX } high-resolution transmission electron microscope which is under service } contract. Rate of pay will be commensurate with qualifications and } experience. All applications should send resumes to Professor Robert L. } Gunshor, School of Electrical and Computer Engineering, Purdue University, } West Lafayette, IN 47907-1285; Fax: (317) 494-2706. Purdue University } is an AA/EEO/ADA employer. }
Please do not reply to me directly, send your inquiries to Albert Yee (afyee-at-engin.umich.edu)
ADJUNCT LECTURER POSITION
THE UNIVERSITY OF MICHIGAN
The Department of Materials Science and Engineering at The University of Michigan, Ann Arbor, is seeking candidates for a non-tenure track lecturer. Responsibilities include: assisting or leading in the development of new undergraduate teaching materials and methods that place a strong emphasis on multimedia presentation techniques; the development of self-paced, interactive learning modules; participation in the development and teaching of new undergraduate laboratory modules that place a strong emphasis on processing and physical and mechanical property characterization. The department currently enrolls approximately 140 undergraduate students and is an integrated materials program encompassing metals, ceramics, polymers and EMO materials. Applicants must have a Ph.D. in materials science and a demonstrated interest in undergraduate education. Minimum one year appointment, renewable to three years based on funding and performance. Send resume and list of references to:
Professor Albert F. Yee, Chairman Department of Materials Science and Engineering The University of Michigan 2300 Hayward Street Ann Arbor, MI 48109-2136 or afyee-at-engin.umich.edu
An Equal Opportunity Affirmative Action Employer
John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu, John.F.Mansfield-at-umich.edu or jfmjfm-at-umich.edu they all reach me! URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
Integrated Microscopy Resource (IMR) a Biomedical Research Resource
at:
The Wisconsin Center 702 Langdon Street Madison, WI 53706
Progran Schedule and Abstracts can be found on the IMR Web page:
http://www.bocklabs.wisc.edu/imr/imr.html
#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#* Presentations will focus on biological problems for which a combination of microscopies [i.e. integrated microscopy] has been used. The speakers will demonstrate by example the power, potential and limitations of various microscopical techniques. The techniques which will be discussed include: DIC, Confocal, 2-Photon Excitation Imaging, SEM, TEM, Cryo-specimen Preparation, and AFM. #*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*
FEES: General Registration $100.00 (On Site: $130.00) (Includes: Opening Reception, Social and Buffet Dinner, Coffee Breaks and Materials)
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TO RECEIVE A BROCHURE AND REGISTRATION FORM SEND COMPLETE MAILING INFORMATION TO:
IMR Univ. Wisconsin-Madison 1675 Observatory Drive Madison, WI 53706
There is an opening at the Stevens Institute of Technology for a post doctoral researcher with interest and expertise in the area of electron microscopy of polymers. The optimal applicant will be fluent in the techniques of cryo-ultramicrotomy, heavy-element staining, and traditional diffraction-contrast TEM imaging techniques. He/she must have excellent communication skills. Experience with energy-loss and x-ray spectroscopies will be helpful. The successful applicant will work collaboratively with both graduate students and industrial researchers using a 300keV TEM, a 200keV FEG TEM/STEM, and a digital FEG SEM among other tools. The microstructural problems of interest will be related primarily though not exclusively to structure/processing relations in compatibilized homopolymer blends. A one-year renewable appointment is anticipated to begin in January 1996. Applicants should submit their resume, publication list, copies of selected publications, and the names of three references to:
Prof. M. Libera Dept. of Materials Science and Engineering Stevens Institute of Technology Hoboken, New Jersey 07030
We digitise some of our electron micrographs using a flat bed scanner. We are looking for a possible site for downloading Ted Pella letraset type fonts. The type we looking for should preferably be black characters on a white background. Information concerning any sources will be welcome.
Thanks ******************************** Keith Williams Experimental Biology Programme Medical Research Council Tygerberg, SA
--Boundary (ID sL9J3/G6+sQD75uBz4KiOg) Content-type: TEXT/PLAIN
EM related injuries? Well, there's failing eyesight from too many day's spent straining to see in that dark room (Mom always warned me), and this curious dent in my forehead from leaning it against the column whilst trying to peer in the window. And then there's this nasty pain in my neck which goes away as soon as my clients do.
Sorry to make light of this, actually my physio therapist tells me that the classic bent-over-arms-out TEM position is a likely contributor to my own neck and shoulder problems (that and age). Maybe the mouse driven scopes of the future will spare microscopists to come.
Why not use any of the desktop or graphics software (e.g., Corel) to make your own set of fonts, symbols or arrows for labelling digitized copy? Almost all of the various packages today allow importing a wide variety of image formats and would provide the maximum in flexibility as to size and font.
You can do this in PhotoShop. Place a white or black letter on your image using the standard type tool. Leave it selected so that the marching ants still outline it. Switch the foreground/background colors so now you have black instead of white or vice versa. Under the Edit pull down menu there is a selection called Stroke. Select stroke and a dialog box comes up. We typically are working with text being placed on the image at 324 dpi so we set the pixel option to 3 and select the outside option. this will place a black border around a white letter similar to the old letraset method. good luck.
} Hi there fellow microscopists } } We digitise some of our electron micrographs using a flat bed } scanner. We are looking for a possible site for downloading Ted Pella } letraset type fonts. The type we looking for should preferably be } black characters on a white background. Information concerning any } sources will be welcome. } } Thanks } ******************************** } Keith Williams } Experimental Biology Programme } Medical Research Council } Tygerberg, SA } } email: kwilliam-at-eagle.mrc.ac.za } *********************************
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (314)-882-4712 (voice) (314)-882-0123 (fax)
Dear Keith- I'm probably going to take a lot of flak for this, but it is important that our colleagues understand what goes on in the commercial world. The black-on-white dry lettering sheets offered by Ted Pella, or Energy Beam Sciences or anyone other EM supplier are not "Letraset type fonts." That is, they are not sheets that Letraset or any other graphic arts company has designed to sell in art supply shops, with an EM suppliers name on it. They are custom sheets, designed and paid for by the EM supplier. In the case of black-on-white sheets, this involves two printing plates. Most of the existing dry lettering sheets were developed by EM supply companies at the request of customers. In some cases, the demand for that particular sheet never paid back the cost of the artwork and plates, but we do these things because our customers are our best resource for new ideas. Ted Pella uses a modified Helvetica font for his sheets; we use a modified Futura font. But the artwork involved in the creation of these sheets is our intellectual property, just as if it were copyrighted. The notion that someone would consider "downloading" this art without a license from the company that owns the design makes me sad. Thanks for letting me vent! Steven Slap
Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" To: MICROSCOPY-at-AAEM.AMC.ANL.GOV
Greetings, Steve Slap is correct. Downloading or copying fonts without a license is inappropriate. However, what you want is achievable at little cost and some effort. Font disks are fairly inexpensive to buy. That gives you the license to use a font for the intended purpose, i.e. typing (better yet, just use whatever fonts you have, Helvetica is very common). The only thing you really want is a white bacground so the alphanumerics will stand out on a micrograph. Make the text bold and shadow your font with white (this will provide a form fitting look) or put the alphanumerics on a white background. I know you can do this in Photoshop and suspect that any number of other graphics programs can do the same. One can always create an acceptable substitute to dry transfer lettering by using a laser printer. I just used MS Word with a bold, outlined and a bold shadowed script to print white text/lines on a black background on Labelon Laserdraft Film. What you do is create whatever alphanumerics you need. Print it on this clear, sticky-backed material through a standard laser printer, cut it to size, pull off the backing and put it on any micrograph you want. Or you can always buy what you need from our vendors.....
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
At the risk of offending virtually all of you, I feel the need to wade into this discussion.
I work for Structure Probe, Inc./SPI Supplies, which is a major supplier of customized transfer sheets for the benefit of microscopists, specifically electron microscopists and other laboratory types, among many other things.
My daughter works for an outfit called Font Haus, which is in the business of selling fonts to people who use them AND LICENSING THEIR USE!
The use of this bulletin board to request the assistance of fellow microscopists in stealing the intellectual property of my esteemed competitor, Ted Pella, is something that I find personally offensive.
If you need any of the (literally) tens of thousands of fonts that Font Haus sells, along with an appropriate license for your intended use, please call them at 1 800 942 9110. They will be happy to lead you through the process of selecting and licensing a font. They do not, however, sell transfer sheets.
I don't normally comment much on this BB, but the use of it to propose to commit the theft of intellectual property is an indication of the extent to which the moral climate of our community has deteriorated, and it sickens me.
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. 63 Unquowa Road Fairfield, CT 06430-5015 Ph: 1 203 254 0000 FAX: 1 203 254 0000 e-mail: AWBlackwoo-at-aol.com WWW: http://mail.cccbi.chester.pa.us/spi/spihome.html
I think the vendors are getting in a snit for nothing concerning these fonts. I read the original request to simply mean the individual was trying to achieve digitally a font which was black with a white border or white with a black border similar to what Ted Pella sells. Obviously scanning in Pella's fonts would be a copyright violation but the individual obviously wasn't interested in this since he owns a scanner and wasn't doing it. Even if one wanted to violate Pella's copyrights, it wouldn't be worth it. The resulting images would be lower quality compared to the original, not Postscript scalable and be filled in and not just outlines. Surely Pella and the other vendors aren't suggesting Pella owns the copyright on the concept of placing contrasting borders of fonts. One can buy such fonts from dealers as has been suggested but a simpler and cheaper and fully legal way is to create them in Photoshop or an equivalent program. To do it in Photoshop: Place a white or black letter on your image using the standard type tool. Leave it selected so that the marching ants still outline it. Switch the foreground/background colors so now you have black instead of white or vice versa. Under the Edit pull down menu there is a selection called Stroke. Select stroke and a dialog box comes up. We typically are working with text being placed on the image at 324 dpi so we set the pixel option to 3 and select the outside option. this will place a black border around a white letter similar to the old letraset method. Does any vendor think this is an infringement of Pella or Letraset? If so, they should talk to Adobe and a lot of other graphic programs.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (314)-882-4712 (voice) (314)-882-0123 (fax)
This question was forwarded from the Center for Reproduction of Endangered species here in San Diego. Please respond to sbarlow-at-sunstroke.sdsu.edu:
We are trying to differentiate between healthy and atretic follicles in dog ovaries and have run into problems when we shift from whole tissue to isolated preparations. In paraffin embedded tissue sections, follicles can be unequivocally differentiated using the Shorr stain (contains Biebrich Scarlet, Orange G, Fast Green, PTA, Phosphomolybdic acid, acetic acid in 50% alcohol). In healthy follicles, zona pellucida stain green , whereas atretic zona pellucida stain orange. In isolated follicles, this differentiation is not observed between known healthy vs atretic follicles. To approximate the treatment the paraffin embeded tissue undergoes, isolated follicle preparations were fixed, then treated to a dehydration series similar to paraffin embedding then hydrated and stained.
Can anyone offer 1) any explanation for the apparent lack of differentiation in isolated vs embedded, deparaffinized material and/or 2) an alternative protocol or stain that would enable us to discriminate between healthy and atretic follicles?
Thanks in advance for your assistance
B. Durrant ( please respond to sbarlow-at-sunstroke.sdsu.edu) ---------------------------------------------------------- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University San Diego CA 92182-0057 phone: (619) 594-4523 fax: (619) 594-5676 email to sbarlow-at-sunstroke.sdsu.edu
} A student wants to electropolish a 1cm square of M2 high speed steel - } he says this contains Mo, W and C. I would be grateful for a } solution composition, temperature and voltage suggestion. } Thanks in advance. } Mike } } } Dr MJ Witcomb } Electron Microscope Unit } University of the Witwatersrand
I don't know if you've recieved a response or not, but Bernie Kestel's book on electropolishing (still available from Argonne National Labs, I think - at least in the US) suggests the use of 10% HClO4 in ethanol at 5 degrees C and a current of 500 mA (voltage as required). This etch is for a Mo-Si stainless steel but it may be worth a try.
-Kirk Rogers
Kirk A. Rogers krogers-at-materials.ecn.purdue.edu 317-494-8751 office http://materials.ecn.purdue.edu/~krogers/ (coming soon) 317-494-1204 fax Purdue University, School of Materials Engineering, 1289 MSEE building, W. Lafayette, IN 47907-1289
New York Society of Experimental Microscopists Presidential Symposium Friday, September 22, 1995 Stern Auditorium Annenberg Building, 100th Street between Madison and 5th Ave The Mount Sinai School of Medicine
in honor of the memory of Eric Holtzman CELLS AND ORGANELLES
9:00 REGISTRATION
9:30 WELCOME Dr. David R. Colman, NYSEM president 1994-1995 Dr. Sandra K. Masur, NYSEM president 1995-1996
9:40 Dr. Ursula W. Goodenough, Washington University The evolution of sex at a cellular level
10:30 Dr. Pamela Cowin, New York University School of Medicine Cadherin-mediated cell adhesion
11:20 Dr. James E. Rothman, Memorial Sloan Kettering Cancer Center Machinery and mechanisms of intracellular protein transport
12:10 LUNCH
2:00 Dr. Kathryn E. Howell, University of Colorado School of Medicine New insights into the structure and function of the trans-Golgi network
2:50 Dr. David D. Sabatini, New York University School of Medicine The generation of transport vesicles in the trans-Golgi region
3:40 Dr. Paul B. Lazarow, Mount Sinai School of Medicine Peroxisomes in yeast and man
4:30 Dr. Lorna Role, Columbia University College of Physicians & Surgeons Development of synaptic transmission between neurons
5:20 RECEPTION
Co-sponsored by The Mount Sinai School of Medicine
For additional information:
Best regards,
Sandra K. Masur Box 1183 fax: 212-289-5945 Mount Sinai School of Medicine phone: 212-241-6544 or 0089 NY NY 10029-6574 e-mail:masur-at-msvax.mssm.edu
My name is David Kristensen, and I am trying to do a project for the Westinghouse Talent Search. I am trying to find the structure of a virus by looking at an electron micrograph. I will do it using Fourier space, so I only need one of them. I would need a small virus (preferably a polio-virus or one of its relatives) in order to be able to find out the structure (a computer might not be able to handle a large virus without parallel processing). So what I'm asking is, can anyone give me/tell me where to find an electron micrograph of a small virus?
My name is David Kristensen: e-mail: dkristen-at-hobbs.leesummit.k12.mo.us The MIND Conquers ALL.
I was wondering if anyone has used or is using an x-ray detector by Amptek. The physics dept. here at UMBC wants to put a model number XR-100T x-ray detector onto my JEOL JSM-35CF SEM. What I need to know is, what kind of problems could I encounter having this unit installed on my SEM. It is basically a low res detector (200eV) so I don't think it would be too good for biological purposes. It is also of the dry type of detector system that doesn't need liquid nitrogen. Any info about this system would be greatly appreciated.
You can not obtain a viruse's structure with just any electron microscope image of a virus using fourier transform processing --- If a transmission electron microscopy image is taken of a virus in vitreous ice and a number of tilt views of this virus are also taken you may be able to reconstruct the virus to about ~30A resolution. You could not reconstruct the viruses internal structure with a surface replicated virus with any of the various metals. It is also unlikely that the internal viral structure is preserved in a negatively stained preparation of viruses for transmission electron microscopy. By taking just a single image the sample is heated in the electron beam to 100-200deg.C and many of the structural proteins are incinerated in the vacuum of the microscope. This does not happen as readily when a sample is embedded in vitreous ice at -196deg.C.
Good luck on your project, George C. Ruben Dept Biological Sciences Dartmouth College Hanover, NH
Hello there. Someone in my lab is having a problem which I believe has been covered before, but am not sure. Any help would be appreciated. The person is forcing neutrophils through a polycarbonate filter, fixing the filter, embedding the filter in agarose and then into paraffin so that are cutting the filter on end to see the cells passing through the filter. The cells look fine when an H&E is done but when they are attempt to do an immunolabel for Brdu, the cells and membrane are gone. We routinely use silane coated slides for immunno without any problems until now. The staining procedure requires a pepsin pretreatment (pH 2.5) followed by an incubation in 2N HCl. Is there something else we could use to attach the cells to the slides??? Would poly-L-lysine be affected by these conditions?? We feel that the use of albumin would not work because of the pepsin treatment. Any information/suggestions would be greatly appreciated. Thanx
Mark Elliott, PhD UBC-Pulmonary Research Lab Vancouver BC Canada acidic incubation as well. Is there something else wee could use
Mark, How are you cleaning your slides? Chromic acid or 1% HCl in 95% ethanol are the two methods that work the best for us, regardless of the subbing agent used on the slides. Poly-L-lysine will survive proteases and the 2N HCl pretreatments for BrdU, as will chrome alum-gelatin.
Regards, Glen MacDonald Hearing Development Laboratories University of Washington Seattle, WA 98195-6515 (206)543-8360 glenmac-at-u.washington.edu
I have a customer who wants TEM surface characterization of polyester filters which are "clean and dry". Pore sizes are 0.05 - 0.03 microns. They are interested in the pore size, shape and distribution on both top and bottom surfaces.
1) do I need to wash prior to embedding to "clean" off any manufacturing debris?
2) will embedding in an epoxy such as EMbed-812 or Spurr's be adequate or should I use a polyester resin?
3) should I osmicate to impart contrast and/or are there any other contrast enhancing techniques I should use? I'm anticipating a contrast problem since there will be no tissue (filter only).
***these filters are non elastic and are very stable on a substrated grid
This is for Fred Hayes regarding TEM methods for polyester filters.
Hi, Hayes:
I have been working with ultrafiltration membrane materials on TEM and SEM several years and here are some methods I have used and hopefull it can help you.
1. Metal shadow for topography of polyester menbranes on TEM : Just follow regular Pt/Pd metal shadow method with 10-30 degree angle then disolve membrane material and you will get a metal replica of the sample. This replica can be observed up to X20000 and possible to count the holes.
2. GMA embeding and ultrasection for TEM: Epon 812 (or its components) will disolve or damage sample. GMA is a water soluble resin and no many components needed. I use GMA (with 1-2% PTA) for embeding and it works well. PTA will react with active layer of the sample during embeding and show the active layer clearly. No staining and fixation are needed. The result looks like dark field image on TEM. Vacuum filtration with GMA embedding resin and cut membrane into small pieces before embeding will show pores more clear since resin fill out the holes. The pore sizing up to 100 A can be counted and measured easily up to X50000 on film.
Hopefull it helps.
Zhiyu Wang Department of Biosystem Engineering University of Hawaii Honolulu HI 96822 zhiyu-at-hawaii.edu
On Thu, 14 Sep 1995, Fred Hayes wrote:
} I have a customer who wants TEM surface characterization of polyester } filters which are "clean and dry". Pore sizes are 0.05 - 0.03 microns. } They are interested in the pore size, shape and distribution on both top } and bottom surfaces. } } 1) do I need to wash prior to embedding to "clean" off any manufacturing } debris? } } 2) will embedding in an epoxy such as EMbed-812 or Spurr's be adequate or } should I use a polyester resin? } } 3) should I osmicate to impart contrast and/or are there any other } contrast enhancing techniques I should use? I'm anticipating a contrast } problem since there will be no tissue (filter only). } } ***these filters are non elastic and are very stable on a substrated grid } } all suggestions will be appreciated, many thanks. } } Fred A. Hayes } Dixon, CA } 916-678-6280 } }
I am planning to analyze coal macerals for cabon, sulfur, and oxygen contents using an electron microprobe. Does anyone know if there are any microprobe standards (coal or synthetic coke) available somewhere ?
I am putting together a class on advanced imaging methods and one of the subjects I want to cover is tomography. Does anyone know of any freeware, shareware, or even a commercial program, that can demonstrate the principles of tomographic reconstruction using any of the standard algorithms (and appropriate input data of course). Any guidance would be much appreciated.
David Joy "joy-at-utkvx.utk.edu"
********************************************************************** * F239 Walters Life Sciences Bldg. NOTE NEW AREA CODE! * * University of Tennessee Phone (423)-974-3638 * * Knoxville, TN 37996-0810 FAX (423)-974-3642 * **********************************************************************
Does anyone have or know of a set of database routines suitable for cataloging microprobe standard collections? I thought I would check before I designed my own. What I want is to be able to search the database on different fields such as mineral type or chemistry. Any info would be appreciated.
Glenn ********************************************************************** * Glenn Poirier glenn_p-at-geosci.lan.mcgill.ca * * Electron Microprobe Lab Phone: (514) 398 6774 * * Earth and Planetary Sciences Fax: (514) 398 4680 * * McGill University THERE ARE THREE SIDES TO EVERY STORY; * * Montreal, Quebec YOUR SIDE MY SIDE AND THE TRUTH * **********************************************************************
The best way of imaging the surface of polyester filter is to look at them by high-resolution SEM (low-voltage FESEM). It can view surface features directly without complicated specimen preparation. I looked at some kind of filter 4 years ago in the Hitachi S-900 LVSEM. The filter surface can be imaged up to 100,000x in stereo. The 3-dimensional 5-10nm surface details are clearly seen at this magnification. Hope this information can help you.
Ya Chen
========================================================================= \ / Assistant Researcher-Cryo/SEM Coordinator TEL : 608-263-8481 \ / __ Integrated Microscopy Resource (IMR) FAX : 608-265-4076 \/ / / University of Wisconsin-Madison / / / 1675 Observatory Drive #167 Email:YChen-at-macc.wisc.edu / /__/_ Madison, WI 53706 Email:chen-at-calshp.cals.wisc.edu
IMR WWW Home Pages: http://www.bocklabs.wisc.edu/imr/imr.html =========================================================================
} } I have a customer who wants TEM surface characterization of polyester } filters which are "clean and dry". Pore sizes are 0.05 - 0.03 microns. } They are interested in the pore size, shape and distribution on both top } and bottom surfaces. } } 1) do I need to wash prior to embedding to "clean" off any manufacturing } debris? } } 2) will embedding in an epoxy such as EMbed-812 or Spurr's be adequate or } should I use a polyester resin? } } 3) should I osmicate to impart contrast and/or are there any other } contrast enhancing techniques I should use? I'm anticipating a contrast } problem since there will be no tissue (filter only). } } ***these filters are non elastic and are very stable on a substrated grid } } all suggestions will be appreciated, many thanks. } } Fred A. Hayes } Dixon, CA } 916-678-6280
In article {Pine.SOL.3.91.950914232209.26887A-100000-at-chip.ucdavis.edu} Fred Hayes {fahayes-at-ucdavis.edu} writes: } Date: Thu, 14 Sep 1995 23:40:13 -0700 (PDT) } From: Fred Hayes {fahayes-at-ucdavis.edu} } Subject: Help re: TEM of polyester filter
} I have a customer who wants TEM surface characterization of polyester } filters which are "clean and dry". Pore sizes are 0.05 - 0.03 microns. } They are interested in the pore size, shape and distribution on both top } and bottom surfaces.
} 1) do I need to wash prior to embedding to "clean" off any manufacturing } debris?
} 2) will embedding in an epoxy such as EMbed-812 or Spurr's be adequate or } should I use a polyester resin?
Don't use replication or sectioning if you can help it. They CAN be used, but they don't give an image of the real surface. For that, use the method of low voltage field emission SEM we describe in our papers:
Kim, KJ, Dickson, M. et al (1991) Electron Microscopy in Synthetic Polymer Membrane Research. Journal of Microscopy vol 162 pp 403-413
Kim, Dickson et al. (1992) Applications of FESEM to polymer membrane research. Micron & Microscopica Acta. vol 23 pp 259-271.
Find someone who has an in-lens FESEM and buy their services. (We can help you !) That will give you an immediate result much cheaper than using the other methods you suggest.
How do I resubscribe? I haven't received mail in 2 months. I must have been omitted. May I be readmitted? Sally Shrom sally-at-retina.anatomy.upenn.edu
Dear microscopists, We have an Akashi EM-002A 120kV TEM which uses freon as the insulator in the HT tank and in the gun. I am going to need to recharge the tank soon (the pressure in it drops slowly over a year or two). As I have used up the freon supplied with TEM I will replace it with SF6 as recommended by Topcon/Akashi but I have heard a rumour that its not as straightforward as simply pumping out the freon and refilling with SF6. Has anyone out there done this type of changeover or know anything about any potential problems?
Thanks in advance for any comments.
Richard Easingwood
Please reply to: richard.easingwood-at-stonebow.otago.ac.nz
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
does anyone out there have any experience in cutting wood for transmission electron microscopy? Any sugestions for cryo or epon sectioning of wood?
Andreas Brech Electron Microscopical Unit for Biological Sciences Department of Biology, University of Oslo. P.O.Box 1062 Blindern N-0316 Oslo 3 Norway Tel.: + 43-22 85 61 89 (work) + 43-22 43 83 23 (privat) Fax.: + 43-22 85 47 26 e-mail.: abrech-at-bio.uio.no
We have just purchased a Leica confocal system which will be used by the faculty and grad students here at UMBC. I would like to build a good library of reference books for their use and was wondering if anybody had good suggestions as to what books would be a necessity and what books would be nice to have. Also, does anybody know of a good security system for the microscope so non-trained personnel won't have access to using the scope. It would be nice if the system could be incorporated into the software of the confocal system. Any suggestions would be greatly appreciated.
Thanks,
Phil Rutledge, Director Center for Electron Microscopy University of Maryland Baltimore County 5401 Wilkens Ave. Catonsville, MD 21228
Hi! I am interested in locating a dye that will stain collagen and/or a monolayer (huve cells) prior to embedding in LRW for postembed immunogold labeling. This is simply to aid location of the monolayer while facing. I have found a reference using 1%glut, 0.2% picric acid and 6mM eserine (physostigmine). Does the eserine function a dye? I intend on using .5% glut with paraformaldehyde. Any ideas? Shelley Landon Kaurin , kaurin-at-rmslab.rockefeller.edu
Phil, Congratulations on your Leica confocal, its a nice instrument. We had concerns here about security and accounting for the microscope. We are currently using Direct Access for Windows (Symantec, which bought out 5th Generation Systems). DAW is not perfect, but there is so little out there. It does provide a measure of security and it collects the hours that users are on the instrument for billing later. Another possiblity I've been considering lately is a program called Full Armor (I've only seen it in the TigerSoftware catalog, 800-888-4437). It seems to do most of the same things as DAW, but it is a newer product (DAW has been in version 1.04 for over 2 years) and it "says" it can lock file manager sub-directories (something DAW can't do if you allow access to file manager).
Write me directly for more information. DAW was about $50 (educational price) and Full Armor is $69.99 in the catalog. (I have no financial interest in either program).
Doug
At 10:04 AM 9/19/95 -0400, you wrote: } } We have just purchased a Leica confocal system which will be used by the } faculty and grad students here at UMBC. I would like to build a good } library of reference books for their use and was wondering if anybody had } good suggestions as to what books would be a necessity and what books would } be nice to have. Also, does anybody know of a good security system for } the microscope so non-trained personnel won't have access to using the } scope. It would be nice if the system could be incorporated into the } software of the confocal system. Any suggestions would be greatly } appreciated. } } Thanks, } } Phil Rutledge, Director } Center for Electron Microscopy } University of Maryland Baltimore County } 5401 Wilkens Ave. } Catonsville, MD 21228 } ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: dcromey-at-ccit.arizona.edu) : :...................................................................:
Via: uk.ac.bbsrc; Tue, 19 Sep 1995 17:00:49 +0100 X400-Received: by /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Tue, 19 Sep 1995 17:02:30 +0000 X400-Received: by mta arcs.bbsrc.ac.uk in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Tue, 19 Sep 1995 17:02:30 +0000 X400-Received: by mta LARS.MUAS in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Tue, 19 Sep 1995 16:59:31 +0000 X400-Received: by mta LARS in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Tue, 19 Sep 1995 16:59:33 +0000 X400-Received: by /PRMD=UK.AC/ADMD= /C=GB/; converted (ia5-text); Relayed; Tue, 19 Sep 1995 16:59:38 +0000
I am having trouble getting the right colour balance when taking slides with a Wild M400 photomacroscope. I am illuminating specimens with light from a halogen bulb via a ring light or two swan necks.There is no brightness control on the transformer, so presumably it runs at opyimum voltage ay all times. Daylight film (Kodak ektachrome 100) gives a yellow cast with a definite khaki/yellow to what was a white background. Tungsten film (Ektacrome 160T) gives a green cast with a pale green background or what should be white. I have bracketed the exposures for both films several stops above and below the given rating. I thought that halogen light would be approximately daylight, but it is obviously not, nor is it tungsten - which is not surprising. Any ideas on a suitable transparency film or fiters I can use to get correct balance?
Many thanks, Richard
Richard.Pring-at-BBSRC.ac.uk
Long Ashton Research Station, Long Ashton, Bristol, U.K.
To: MICROSCO:CENTRAL cc: DRStadden:R_D:Armstrong Subj: Need Microtome Chuck ------------------------------------------------------------------
Attention Sorvall rotary microtome users:
I have a need to make routine, thick sections of polymer films for cross- sectional study by the FTIR microscope. Where can I get a chuck with vise jaws instead of the beam capsule clamps? I'd like to simply sandwich the three mil film of interest between polyethylene pads and not have to embed in epoxy the way our IR guy has been doing. The unit is of unknown vintage, but is probably at least 20 or 30 years old, small, gray, rather unassuming. The collet itself threads onto a 5/8 mail threaded piece with a ball fitting on opposite end. Sound familiar? You can tell I'm not a full-time microtomist. Thanks in advance for your help.
Dave Stadden DRStadde+aCENTRAL%ARMSTRONG-at-MCIMAIL.COM Lancaster, PA
Dear Sir: I would appreciate if you agree to let me join your microscopy network. Could you please tell me how to subscribe and other requirements from you? Thanks a lot for your kindly consideration. Sincerely yours
Message-Id: {m0sv7Sc-0007C8C-at-stjohns.ohsu.edu} Message-Version: 2 } To: microscopy-at-aaem.amc.anl.gov
Phil, Congratulations on your Leica confocal, its a nice instrument. We had concerns here about security and accounting for the microscope. We are currently using Direct Access for Windows (Symantec, which bought out 5th Generation Systems). DAW is not perfect, but there is so little out there. It does provide a measure of security and it collects the hours that users are on the instrument for billing later. Another possiblity I've been considering lately is a program called Full Armor (I've only seen it in the TigerSoftware catalog, 800-888-4437). It seems to do most of the same things as DAW, but it is a newer product (DAW has been in version 1.04 for over 2 years) and it "says" it can lock file manager sub-directories (something DAW can't do if you allow access to file manager).
Write me directly for more information. DAW was about $50 (educational price) and Full Armor is $69.99 in the catalog. (I have no financial interest in either program).
Doug
At 10:04 AM 9/19/95 -0400, you wrote: } } We have just purchased a Leica confocal system which will be used by the } faculty and grad students here at UMBC. I would like to build a good } library of reference books for their use and was wondering if anybody had } good suggestions as to what books would be a necessity and what books would } be nice to have. Also, does anybody know of a good security system for } the microscope so non-trained personnel won't have access to using the } scope. It would be nice if the system could be incorporated into the } software of the confocal system. Any suggestions would be greatly } appreciated. } } Thanks, } } Phil Rutledge, Director } Center for Electron Microscopy } University of Maryland Baltimore County } 5401 Wilkens Ave. } Catonsville, MD 21228 } ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: dcromey-at-ccit.arizona.edu) : :...................................................................:
Message-Id: {m0sv7ea-0007C5C-at-stjohns.ohsu.edu} Message-Version: 2 } To: microscopy-at-aaem.amc.anl.gov (!microscopy)
We have just purchased a Leica confocal system which will be used by the faculty and grad students here at UMBC. I would like to build a good library of reference books for their use and was wondering if anybody had good suggestions as to what books would be a necessity and what books would be nice to have. Also, does anybody know of a good security system for the microscope so non-trained personnel won't have access to using the scope. It would be nice if the system could be incorporated into the software of the confocal system. Any suggestions would be greatly appreciated.
Thanks,
Phil Rutledge, Director Center for Electron Microscopy University of Maryland Baltimore County 5401 Wilkens Ave. Catonsville, MD 21228
Message-Id: {m0sv7jD-0007C8C-at-stjohns.ohsu.edu} Message-Version: 2 } To: microscopy-at-aaem.amc.anl.gov
The McCrone Research Institute in Chicago is now unloading some antique Cambridge Stereoscan II parts by November 1 1995. Console, column, and chamber are being retained for museum but everything else related must go. If you are interested contact ndaerr-at-mcri.org. Thanks.
Message-Id: {m0sv7jF-0007C9C-at-stjohns.ohsu.edu} Message-Version: 2 } To: microscopy-at-aaem.amc.anl.gov
Hello
does anyone out there have any experience in cutting wood for transmission electron microscopy? Any sugestions for cryo or epon sectioning of wood?
Andreas Brech Electron Microscopical Unit for Biological Sciences Department of Biology, University of Oslo. P.O.Box 1062 Blindern N-0316 Oslo 3 Norway Tel.: + 43-22 85 61 89 (work) + 43-22 43 83 23 (privat) Fax.: + 43-22 85 47 26 e-mail.: abrech-at-bio.uio.no
Message-Id: {m0sv7jH-0007CTC-at-stjohns.ohsu.edu} Message-Version: 2 } To: MICROSCOPY-at-AAEM.AMC.ANL.GOV
Greetings, The green cast with tungsten film (and you should be using tungsten film) may come from a couple of sources. Remove any polarizing lenses that may be in the light path. Thick heat resistant glass, like that found in Olympus fluorescent add-ons to the BH-2, can also give a green cast. If removing such things doesn't work, then try bracketing using 10cc, 20cc or 30 cc magenta filters. BTW, assuming the brightness is correct is not wise.
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
Message-Id: {m0sv7hS-0007C5C-at-stjohns.ohsu.edu} Message-Version: 2 } To: MICROSCOPY-at-aaem.amc.anl.gov (!MICROSCOPY)
To: MICROSCO:CENTRAL cc: DRStadden:R_D:Armstrong Subj: Need Microtome Chuck ------------------------------------------------------------------
Attention Sorvall rotary microtome users:
I have a need to make routine, thick sections of polymer films for cross- sectional study by the FTIR microscope. Where can I get a chuck with vise jaws instead of the beam capsule clamps? I'd like to simply sandwich the three mil film of interest between polyethylene pads and not have to embed in epoxy the way our IR guy has been doing. The unit is of unknown vintage, but is probably at least 20 or 30 years old, small, gray, rather unassuming. The collet itself threads onto a 5/8 mail threaded piece with a ball fitting on opposite end. Sound familiar? You can tell I'm not a full-time microtomist. Thanks in advance for your help.
Dave Stadden DRStadde+aCENTRAL%ARMSTRONG-at-MCIMAIL.COM Lancaster, PA
I am having trouble getting the right colour balance when taking slides with a Wild M400 photomacroscope. I am illuminating specimens with light from a halogen bulb via a ring light or two swan necks.There is no brightness control on the transformer, so presumably it runs at opyimum voltage ay all times. Daylight film (Kodak ektachrome 100) gives a yellow cast with a definite khaki/yellow to what was a white background. Tungsten film (Ektacrome 160T) gives a green cast with a pale green background or what should be white. I have bracketed the exposures for both films several stops above and below the given rating. I thought that halogen light would be approximately daylight, but it is obviously not, nor is it tungsten - which is not surprising. Any ideas on a suitable transparency film or fiters I can use to get correct balance?
Many thanks, Richard
Richard.Pring-at-BBSRC.ac.uk
Long Ashton Research Station, Long Ashton, Bristol, U.K.
Just ran across an ad in the back of Science. The Wheaton (IL) Biology Dept wants someone with experience in two or more of the following areas: botany, zoology, electron microscopy, etc. See Science vol 269 (15 September) page 1620 for details. And good luck.
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
I thought someone on this list with an interest in aquariums might be willing to give some advice to the following individual. Please respond directly to
Steve Murrison {toolbox-at-intertrek.com}
or to this list and I will forward him your responses.
} From: Steve Murrison {toolbox-at-intertrek.com} } X-To: editor } Subject: (no subject) } Organization: Living Colours - The Aquarium Shop - } Mime-version: 1.0 } Date: Tue, 19 Sep 95 16:55:43 -0700 } } dear editor, } I own and operate an aquarium shop in victoria b.c. } I have been searching for a microscope for some time now.I know very } little about them, but I have been holding out for something fairly } decent. I know that sounds alittle vague sorry, am interested in veiwing } various bacterium and would also like to photograph them can you help } me find something thanks steve.
Thanks!
Paul R. Perkes Webmaster Microscopy Online webmaster-at-microscopy-online.com Microscopy Online - http://www.microscopy-online.com/
Fellow microscopists, I need to digitise some tem negatives with a view to carrying out 3D reconstruction. If anyone is currently using a digitising device capable of high resolution I would like to hear about it. I would be intersted in minimum spot size speed of scanning optical density range grey levels cost If there is interest I will summarise the replies and post it to the group. Please reply to me in the first instance
Fellow microscopists, I need to digitise some tem negatives with a view to carrying out 3D reconstruction. If anyone is currently using a digitising device capable of high resolution I would like to hear about it. I would be intersted in minimum spot size speed of scanning optical density range grey levels cost If there is interest I will summarise the replies and post it to the group. Please reply to me in the first instance
Many thanks
Chris
Chris Gilpin Biological Sciences E.M. Unit Manchester University U.K. 0161 2755170 phone 0161 2755171 fax
The "Bible" of Confocal Microscopy is as follows..."Handbook of Biological Confocal Microscopy", Second Edition, Edited by James B. Pawley, Plenum Press, NY & London.
There are numerous software products (both commercial and shareware) available for Windows based software, as I believe the Leica Control Program is that will password protect any volume. As for the hardware, your Laser Box should have a key for the ignition. If you have any further questions, please do not hesitate to contact me. I am sorry we did not have a Confocal Microscope available at the time for your evaluation, however we have two new products coming for your future consideration. Good Luck, Lawrence Kordon Confocal Specialist Nikon, Inc.
This is a second posting of the original message as I have gotten many messages of returned mail that could not reach me at my personal address please respond to "microscopy..."I look forward to your comments.
I am interested in locating a dye that will stain collagen and or a monolayer (huve cells) prior to embedding in LRW for post embd immuno- labeling. This is simply to aid location of the monolayer while facing. I have found a reference using 1% glut, 0.2% picric acid and 6mM eserine (physostigmine). Does the eserine function as a dye? I intend on using 0.5% glut, with paraformaldehyde. The monolayers will be enclosed in resin then mounted for the z axis. Any ideas? Shelley Landon Kaurin
We are considering purchasing an image analysis package called "Micro-Tome" from Vaytek, Inc. in Fairfield, IA. This package porports to do digital confocal microscopy using any conventional white light microscope and digital deconvolution of the acquired images. Does anyone use this product and have an opinion regarding its usefulness or, more generally, does anyone have an opinion regarding this approach to confocal microscopy compared to the conventional laser based approach. All responses will be kept confidential.
Norman Elliott | E-mail: nee-at-lanl.gov Los Alamos National Lab | Fax: 505-665-2104 MST-7 MS E549 | Voice: 505-667-1587 Los Alamos, NM 87545 |
Message-Id: {9509201632.AA13629-at-unlinfo.unl.edu} X-Sender: tvoiles-at-129.93.1.11 X-Mailer: Windows Eudora Version 1.4.4 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
A new WWW site has just been put on-line
The Center for Materials Research and Analysis' Central Facility for Electron Microscopy now has a site detailing:
What it is What it can do Where it is How you can come to use it Current events and research projects Description of equipment with pictures Sample output of Microscopes Lots of documentation on equipment use and policies
Highlights include
A multimedia (mainly images) Standard Operating Procedure for our JEOL JSM-840A SEM
An electronic scheduling form allowing users to set up times on microscopes and submit them by e-mail
It can be accessed at the following
{http://www.unl.edu/tvoiles/}
Please visit the site and tell us what you think
Please DO NOT direct the replies to me (Todd Voiles tvoiles-at-unlinfo.unl.edu) since I am leaving as of the 21st
PLEASE DIRECT your replies to the new webmaster Gary Krichau (gkrichau-at-unlgrad1.unl.edu)
By the way, I'm leaving the list as well as my position here so Bye, Bye...its been fun.
Todd Voiles Central Facility Specialist Central Facility for Electron Microscopy Center for Materials Research and Analysis
We have a user that is looking for a quality cryo-stage (freezing stage) that will freeze down to -80 to -120 C if possible, and will fit onto a Nikon Diaphot.
Has anyone collected a list of vendors lately? How about helpful hints, or possible pitfalls to look out for, for both purchase and use.
TIA,
C. Michael Stanley, Ph.D. Coordinator, Associate Director Molecular Cytology Core Facility Molecular Biology Program 2 Tucker Hall University of Missouri-Columbia Columbia, MO. 65211 (314) 882-4895 fax= 314-882-0123
If you Email your address to us, we will send you a catalog of imported microscopes, as well as advice on video and photographic setups. Our scopes are very high quality and inexpensive. We have a nice little book called "The Microscope and How to Use It", which is an excellent and practical introduction to the whole subject.
I am a beginner in the techniques of immunocytochemistry and I need some theoretical "background" concerning the labelling with antibody and PAG staining. Would you be so kind to advice me the title of some interesting book?
Thanks in advance
Flati Silvio
Laboratory of Molecular Neurobiology Mario Negri Sud Institute (Italy)
For a good, comprehensive text on immunocytochemical methods try "Fine Structure Immunocytochemistry" by G. Griffiths, 1993, Springer Verlag.
Another way to get quickly up to speed is to attend one of the annual methods courses offered by the European Molecular Biology Organization. I can give you details if you contact me directly.
Also, why not check out the new WWW site which has a methods book foccussing on immunocytochemical methods. The URL is {http://info.med.yale.edu/cellimg} .
Paul Webster Center for Cell Imaging Yale School of Medicine New Haven, CT 06520
We are putting together a request for $ for a new ultramicrotome to replace our 25-year-old LKB. My favorite instrument of all time is the Reichert Ultracut E and I have had no expeience with the Ultracut S or later versions. Also I have not familiar with ultramicrotomes by other manufacturers. Does anyone out there have any insights, opinions, praise, etc for any one particular instrument? In other words, given the money, which one would you buy? Are used or rebuilt units available? Please reply directly to me as all responses will be kept confidential.
Thanks in advance
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
Last week I replied your massage to the microscopy net. Here I will provide your more information of the usage of LVSEM to look at filter. You can check the article of "LVSEM for high resolution topographic and density contrast imaging", by Jim Pawley, in "Advances in electronics and eletron physics", Vol. 83, pp203-274, (1992). This article discuss the priciple and application of LVSEM in details. For your project, you can pay special attension to the stereo pair of high resolution images of filter (Fig. 22 at p.250).
If you have any question or need help for using the LVSEM, please contact me off line.
Ya Chen
========================================================================= \ / Assistant Researcher-Cryo/SEM Coordinator TEL : 608-263-8481 \ / __ Integrated Microscopy Resource (IMR) FAX : 608-265-4076 \/ / / University of Wisconsin-Madison / / / 1675 Observatory Drive #167 Email:YChen-at-macc.wisc.edu / /__/_ Madison, WI 53706 Email:chen-at-calshp.cals.wisc.edu
IMR WWW Home Pages: http://www.bocklabs.wisc.edu/imr/imr.html =========================================================================
} Hayes, } } The best way of imaging the surface of polyester filter is to look at them } by high-resolution SEM (low-voltage FESEM). It can view surface features } directly without complicated specimen preparation. I looked at some kind } of filter 4 years ago in the Hitachi S-900 LVSEM. The filter surface can } be imaged up to 100,000x in stereo. The 3-dimensional 5-10nm surface } details are clearly seen at this magnification. Hope this information can } help you. } } Ya Chen }
} } } } I have a customer who wants TEM surface characterization of polyester } } filters which are "clean and dry". Pore sizes are 0.05 - 0.03 microns. } } They are interested in the pore size, shape and distribution on both top } } and bottom surfaces. } } ...... } } } } Fred A. Hayes
} } We have just purchased a Leica confocal system which will be used by the } faculty and grad students here at UMBC. I would like to build a good } library of reference books for their use and was wondering if anybody had } good suggestions as to what books would be a necessity and what books would } be nice to have. Also, does anybody know of a good security system for } the microscope so non-trained personnel won't have access to using the } scope. It would be nice if the system could be incorporated into the } software of the confocal system. Any suggestions would be greatly } appreciated. } } Thanks, } } Phil Rutledge, Director } Center for Electron Microscopy } University of Maryland Baltimore County } 5401 Wilkens Ave. } Catonsville, MD 21228 } Phil I have just started reading The Image Processing Handbook, second ed. by John C. Russ, CRC Press. It is a great text for digital imaging concepts and techniques. I would think it would be of value in a con focal library.
This is to let you know you about the Electron Microscopy Yellow Pages
http://cimewww.epfl.ch/emyp/
These EM Yellow Pages are intended to provide extensive and comprehensive links to electron microscopy resources inside the World Wide Web: laboratories, societies, educational resources, conferences, news and publications, software, equipment & consulting, data & databases, ...
If you know of useful Web resources which are not to be found in these pages, let me know, and I will update the list.
I am also looking for ways to improve these Yellow Pages. So if you have comments or suggestions, please email me.
Sincerely yours,
Pierre-Henri Jouneau Centre Interdepartemental de Microscopie Electronique Ecole Polytechnique Federale de Lausanne, Switzerland http://cimewww.epfl.ch
We are trying enzyme cytochemistry on spinal cord however all our attempts to slice the lightly fixed cord on vibratomes, tissue choppers etc result in spinal cord "soup". Cutting the thin slices of cord seems almost impossible. One option we thought of exploring is to lightly fix fix the cord, cryoprotect then freeze, cryosubstitute into Lowicryl and then attempt enzyme cyotchemistry on the Lowicryl semithin sections. Has anyone tried enzyme cytochemistry on Lowicryl or on other acrylic sections?
Allan Mitchell
Please reply to: richard.easingwood-at-stonebow.otago.ac.nz
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
Here is the Program for the NCSMMA Annual Meeting Sept 29 - Oct 1 at lovely Wrightsville Beach NC
For more info on Registration etc, please call Ms. Betty Gooch, Symposium Coordinator at: (919) 684-3534
DUKE UNIVERSITY MEDICAL CENTER and the NORTH CAROLINA SOCIETY FOR MICROSCOPY AND MICROBEAM ANALYSIS
present the
FOURTEENTH annual SYMPOSIUM ON ADVANCES IN MICROSCOPY
RMicroscopy Outreach: Conveying Its Science, Art and TechnologyS Holiday Inn, Wrightsville Beach, North Carolina September 29-October 1, 1995
PROGRAM Friday, 29 September 1995 4:00 - 6:00 pm REGISTRATION and refreshments - Holiday Inn at Wrightsville Beach 6:30 - 8:30 pm Barbeque - poolside at the Holiday Inn, Wrightsville Beach Courtesy of JEOL (USA) Inc. Beverages courtesy of AMRAY Inc. (Complimentary with Registration; Adult and Children Guests $10/$5)
Saturday, 30 September 1995 8:30 - 11:30 am Special Workshops/Tutorials on: Specimen Preparation for Materials Sciences - Ron Anderson Cryotechniques - Stacie Kirsch 8:00 - 10:00 am Coffee, juice and donuts - Holiday Inn 10:00 - 11:30 am Poster Session, ExhibitorsU Displays - Holiday Inn 11:30 - 12:00 Noon NCSMMA Annual Business Meeting 12:00 - l:00 pm Lunch - Casual buffet
1:00 - 1:05 pm P Welcome P 1:05 - 1:35 pm "Science and Public Policy in North Carolina Today" Quentin Lindsey 1:45 - 2:15 pm "Hands on Science Using Microscopy: the MSA MICRO Project" Michael Isaacson 2:30 - 2:45 pm P Coffee Break P 2:45 - 3:15 pm "A Delivery System for Project MICRO" Dick Ward 3:30 - 4:00 pm "Working Programs: Successes and Pitfalls" Karen Shafer 4:15 - 4:30 pm COFFEE BREAK - Informal discussion 4:30 - 5:00 pm "Navigating the Internet and Digital Imaging" Nestor Zaluzec 5:00 - 6:15 pm Workshop: TelePresence Microscopy and the Internet Nestor Zaluzec 5:00 - 6:30 pm Poster Session, ExhibitorsU Displays 7:00 pm Evening Buffet - Al Fresco at the Holiday Inn, Wrightsville Beach Supported in part by Oxford Instruments and Zeiss (Electron Optics Division)
Sunday, 1 October 1995 7:30 - 8:30 am Coffee, juice, and donuts - Meeting Room, Holiday Inn 8:30 - 9:00 am Student Awards and Presentations 9:00 - 9:30 am "An Early Visit to Inner Space--a Personal Voyage" John Watson 9:45 - 10:15 am "Imaging Muscle: The Technology, the Art, the Science" Ann Goldstein 10:30 - 11:00 am COFFEE BREAK 11:00 - 11:30 am "A Computer Network Laboratory for Microscopy Education" Gary Chandler 11:45 - 12:15 pm "Washington Outlook: Is There a Contract Out on Science?" Pat Calarco 1:00 - 2:-00 pm Lunch
** SORRY ABOUT THE ODD LETTERS APPEARING ..... THE TEXT EDITOR DOESN'T LIKE ITALICS!! MAIL SEND
Dear Fellow Microscopists, Many thanks to all who replied to me recent plea for help regarding sticking methacrylate sections to glass slides. I received lots of advice which I am still in the process of evaluating. However, the best 'solution' so far appears to be Mayere's egg albumin - which is consistently giving a section-retention rate greater than 90%. I think it just goes to prove that there is nothing new in microscopy... Best wishes to everyone! Nigel Chaffey: IACR-Long Ashton Research Station, Dept Agric. Sci., Univ. Bristol, Long Ashton, Bristol BS18 9AF, UK (email - nigel.chaffey-at-bbsrc.ac.uk)
} } We are trying enzyme cytochemistry on spinal cord however all our attempts } to slice the lightly fixed cord on vibratomes, tissue choppers etc result } in spinal cord "soup". Cutting the thin slices of cord seems almost } impossible. } One option we thought of exploring is to lightly fix fix the cord, } cryoprotect then freeze, cryosubstitute into Lowicryl and then attempt } enzyme cyotchemistry on the Lowicryl semithin sections. } Has anyone tried enzyme cytochemistry on Lowicryl or on other acrylic sections? } } Allan Mitchell
As an alternative, try cutting cryosections and running the cytochemistry on the sections, then embedding. Alternatively, put the whole slice on a grid and cover with a drop of spurr resin. Cure the resin and view the "thick" section at the highest accelerating voltage your microscope allows. We have viewed up to 1 um at 120 KeV with our Philips 400 and up to 5 um with our CM-300 at 300 KeV.
Best
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
Message-Id: {199509221704.TAA14535-at-pons.uio.no} X-Sender: finnmog-at-pons.uio.no X-Mailer: Windows Eudora Version 1.4.3 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Dear list,
We have looked mostly in vain for fluorescent standards, specifically "non-bleaching", solid, inorganic, specimens suitable for routine checks on excitation level and checks on spatial uniformity of the optical/digital imaging system.
Reports and handbooks describe the use of uranyl-glass and inorganic crystals, but the only commercial product we have found contains a 1-2 mm circular fluorescing area of unknown composition, mounted on a microscope slide. Price: USD 1400 incl VAT. Expensive?
A local contact makes Yttrium-Oxy-Sulfide crystals with discrete emission/excitation bands for use in microspectrofluorimetry. However, spectral specificity may not be required for checks on excitation level and the powder's granularity may preclude its use in testing pixel-to-pixel spatial uniformity.
Names, fax/phone-numbers and e-mail adresses of suppliers of such standards would be most helpful, perhaps to others as well.
Thanks in advance -
Finn-Mogens Haug
Department of anatomy E-mail: f.m.haug-at-basalmed.uio.no Institute of basic medical sciences Phone : +47 22 85 12 67 University of Oslo Fax : +47 22 85 12 78
} } Fellow microscopists, } I need to digitise some tem negatives with a view to carrying out 3D } reconstruction. If anyone is currently using a digitising device } capable of high resolution I would like to hear about it.
Dear Chris, We are using a Perkin-Elmer microdensitometer and/or a CCD array for these kinds of measurements.
} minimum spot size
5 micron square aperture on the P-E, ~40 micron on CCD.
} speed of scanning
Up to 40 mm/sec on P-E, but slower speeds are useful for diffraction, where there is lots of ~0 OD with a few intense spots. In this case, the slower speed gives better quantitation. The CCD can scan a micrograph in a few seconds.
} optical density range
0-4 OD on the P-E, 0-~2 on the CCD
} grey levels
~1000 on the P-E (more may be available on a newer instrument), fewer on the CCD, but I don't know how many.
} cost
A reconditioned P-E may be available for ~20,000, a CCD-type scanner can be put together for less. If you want to try either of our systems or consult with any of the group who does 3D reconstruction for a living, write to Dr. Joachim Frank, Wadsworth Center, Empire State Plaza, PO Box 509, Albany NY 12201-0509. He or one of the crew can take it from there. Good luck. Yours, Bill Tivol
I have been requested by an e-mail-deprived colleague to pass on the following:
From Bill Monroe, Electron Microscope Center, Mississippi State University
We have a request to process ice cream samples in order to visualize the spores of Bacillus licheniformis which are dispersed in the samples. The first fixation protocol (glutaraldehyde & osmium tetroxide, centrifuging at all steps) resulted in only being able to see the processed milk fat and no visible spores. Can anyone suggest a method of removing the fat and leaving the spores intact during processing?
Bill- I remember seeing a poster at Scanning '95 in the Oxford Instruments booth, they were advertising their system and had some FDA researchers images/spectra of ice-cream, sorry I don't have any further details. Contact Pat Campos (415) 578-0202 for more info. -Mike
On Fri, 22 Sep 1995, Richard F. Kuklinski wrote:
} I have been requested by an e-mail-deprived colleague to pass on } the following: } } From Bill Monroe, Electron Microscope Center, } Mississippi State University } } We have a request to process ice cream samples in order to } visualize the spores of Bacillus licheniformis which are dispersed } in the samples. The first fixation protocol (glutaraldehyde & } osmium tetroxide, centrifuging at all steps) resulted in only } being able to see the processed milk fat and no visible spores. } Can anyone suggest a method of removing the fat and leaving the } spores intact during processing? }
Nothing new in microscopy????? How about confocal, various scanning probe microscopies, acoustic, x-ray - a very short list of the microscopies developed or greatly advanced in the last decade or so. I think that there is a lot new in microscopy these days.
On Fri, 22 Sep 1995, CHAFFEYN wrote:
} Dear Fellow Microscopists, } Many thanks to all who replied to me recent plea for help regarding } sticking methacrylate sections to glass slides. I received lots of advice } which I am still in the process of evaluating. However, the best 'solution' so } far appears to be Mayere's egg albumin - which is consistently giving a } section-retention rate greater than 90%. I think it just goes to prove that } there is nothing new in microscopy... } Best wishes to everyone! } Nigel Chaffey: IACR-Long Ashton Research Station, Dept Agric. Sci., } Univ. Bristol, Long Ashton, Bristol BS18 9AF, UK (email - } nigel.chaffey-at-bbsrc.ac.uk) }
I hope I haven't been wasting my time on something that is already solved, but the previous message suggests that this is not easily available.
I have been working on measuring the illumination for quantitative fluorescence microscopy. I obtained a slab of uranyl glass some time ago, but it was too thick to be useful. (I got it from a supplier that I found from a message on the confocal mailing list.).
I read a little about toxcicity of Uranium and decided that normal care is adequate in dealing with the stuff. Uranyl glass is apparently about 1% Uranimum, and maximum inhalation is around 1 mg per day. The chemical poison effects are more important than the radioactivity.
I broke a piece of it up and ground it in a porcelain morter to produce a fine white powder. I managed to get a 10 micron thick layer of closely packed pieces mounted in acrylic media, and I programmed the XY table to move this sample in random rotary motion. This give a fairly good measurement of the illumination, and apparently the noise in this measurement is comparable to the noise from other sources.
This sample (which I got this thin by putting it in a 1 ton press while the toluene was evaporating) has been degrading: the acrylic is flowing and voids are forming. I just got some acrylic monomer and if necessary I will make a new sample of ground up glass mounted in acrylic which should be dimensionally stable after curing. However, I plan to make a uniform thin sample by melting it.
I obtained a "test kiln" for $150 from a local ceramics supply store. It has a thermocouple and goes up to 2000F (1100C). I found that a small sample of Uranyl melts and forms a sphere below 1100C. Unfortunately the slides I have been using melt around 100C lower than the Uranyl glass.
I have ordered a quartz slide and coverslip and plan to prepare a sample a micron or two thick by placing a small amount (say 5mg) of powder on the slide and melting it with a coverslip.
The CRC handbook says quartz strain point is 1070C, anealing point approx 1140C, softening point approx 1665C, so there should be no difficulty melting the uranyl glass without affecting the quartz. I plan to let it cool slowly.
Uranyl glass came from: Newport Industrial Glass, Inc. 2044-C Placentia Ave Costa Mesa, California USA 92627 Contact Bill Larson Phone (714)642-9980 Fax (714)642-4832
Quartz slide is on order from Ted Pella, Inc. tedpel-at-snowcrest.net, tedpel-at-aol.com POBox 492477 Redding, CA 96049-2477 800-237-3526 (USA) 800-637-3526(Calif) 800-243-7765 (Canada)
I am with a biotech company in St. Louis. Our products are automated microbiology and immunology systems for larger hospitals and labs. We provide reporting of the identity and susceptibility to antimicrobials of bacteria from patient samples and also the identity of viruses for certain infectious diseases. We are an international company.
We have an ad hoc team working on a home page for the WWW. Rather then using it for advertising purposes we want to have it as a resource center for microbiologists and immunologists with links to other information sites as well as monthly educational articles. For example we might have articles on organisms that gained some attention in the media such as E. coli 0157 or the ebola virus.
We are trying to locate an existing source for micrographs of both common and unusual bacteria and viruses. We would use small images of these on our home page. We would prefer to have them in a format where we could have them colorized. The idea is more of one to capture people's attention rather then expecting that the casual reader would use them for scientific purposes.
If anyone can provide assistance we could acknowledge their contribution on our home page and cover nominal reproduction and mailing expenses.
Richard- Michael Rock's post to you concerned an Oxford cryopreparation system. Such a system allows you to directly view frozen hydrated specimens. There are three manufacturers of thses systems, Oxford being one. Energy Beam Sciences is the exclusive U.S. distributor for VG Microtech (Polaron) who make another. Please fax me directly for details. Steven Slap
I am with a biotech company in St. Louis. Our products are automated microbiology and immunology systems for larger hospitals and labs. We provide reporting of the identity and susceptibility to antimicrobials of bacteria from patient samples and also the identity of viruses for certain infectious diseases. We are an international company.
We have an ad hoc team working on a home page for the WWW. Rather then using it for advertising purposes we want to have it as a resource center for microbiologists and immunologists with links to other information sites as well as monthly educational articles. For example we might have articles on organisms that gained some attention in the media such as E. coli 0157 or the ebola virus.
We are trying to locate an existing source for micrographs of both common and unusual bacteria and viruses. We would use small images of these on our home page. We would prefer to have them in a format where we could have them colorized. The idea is more of one to capture people's attention rather then expecting that the casual reader would use them for scientific purposes.
If anyone can provide assistance we could acknowledge their contribution on our home page and cover nominal reproduction and mailing expenses.
Prof. Ed Haskins (UW Botany) and I are trying to find some optical microscopy stains that will distinguish between microscopic bits and blobs of adhesives (Scotch tape -TM noted), other adhesives, paraffin (and hopefully other polymers) and cellulose.
Sudam IV shows a glimmer of promise but the red-brown color is close to the cellulose fibers.
Hello everyone I'm a biologist (botany) and have been using diamond knives (45=B0) for both semithin and ultrathin sections for many, many years. I have done some sectioning of embedded zeolite crystals, with the 45=B0 biological knife, with some success. I've noticed that people in material science are more and more interested in sectioning and some companies have diamond knives for inorganic specimens, either 35=B0, 45=B0, or 55=B0, and that they are less expensive t= han diamond knives for biological applications. Does somebody have some experience of these diamond knives for material science? What angle do you recommend? We are going to buy a new set of diamond knives for a new centre for EM and I should very much appreciate some (a lot) advice about the material science part. The samples will vary from polymers over biological specimens to organic and inorganic crystals, i e it will cover the whole range of hardness.
Somebody recently posted a note briefly describing a technique that involved embedding whole cells in Spurrs epoxy then viewing in the TEM. A drop of epoxy was placed on cells on a cover slip, cured, then viewed unsectioned at high kV (120-300 kV). Of course, I erased the original message. Could the author please send me the protocol again?
Thanks
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
Monochrome real time digital RS-422 (10 or 8 bits) and analog RS-170 10 bit CCD video cameras are presented in a informative web site, containing a question and answer section on how to go about choosing the proper camera for the application, what to look for in a camera, system configuration issues, and a complete RS-422 frame grabber selection guide listed by bus. Complete plug and play systems can be provided for PCI,Nubus,S-bus & SGI digital. -------------------------------------------------------------------------- (((((((( The URL is: http://www.edt.com/dvc/dvc.html )))))))))) --------------------------------------------------------------------------
Subject: Time: 9:28 AM OFFICE MEMO Sale: Bio prep equipment Date: 9/26/95
FOR SALE:
Life Cell CF 100 with Edwards mechanical pump, SS backing line, LN trap, LN supply line w/solenoid valve, nitrogen gas regulator and all accessories. Vintage1989, used perhaps 3 or 4 times.
Denton 502A, w manual valving and oil diffusion pump, two pairs of electrodes, LN trap w/funnel. Vintage 1988- THIS UNIT HAS NEVER BEEN USED!!! Sparkling clean. Pumps have been run only.
} } From Bill Monroe, Electron Microscope Center, } Mississippi State University } } We have a request to process ice cream samples in order to } visualize the spores of Bacillus licheniformis which are dispersed } in the samples. The first fixation protocol (glutaraldehyde & } osmium tetroxide, centrifuging at all steps) resulted in only } being able to see the processed milk fat and no visible spores. } Can anyone suggest a method of removing the fat and leaving the } spores intact during processing? } Dear Bill, Do you need to see the spores *in* the ice cream? If not, you could dissolve away or dilute the water-soluble part, then separate the spores either by adding organic solvent and filtering or sedimenting. I'd suggest forgetting the OsO4 until the lipid component has been removed; the OsO4 only makes the lipid darker and harder to solubilize. Spores should be hardy enough to survive some pretty harsh isolation steps. To examine the spores in situ, I'd cut frozen sections, warm to ~-40 try to infiltrate UAc (does it dissolve in an ethanol-water mixture?), recool to LN2 and see if I could see anything. Higher voltage might be a big help-- Non-disclaimer: I have a vested interest in attracting users to our 1.2 MV microscope. Good luck. Yours, Bill Tivol
Miguel- Carbon evaporation is one of the primary uses this instrument is designed for and it is a very reliable unit with good tech support from Denton. This evaporator has a value of about $13,000 in today's dollars. We are accepting offers but donations won't fly with managment -especially new, unused equipment. -Doug
-------------------------------------- (1.38.193.5/16.2) id AA14219; Tue, 26 Sep 1995 10:09:59 -0700
Is your Denton 502A system suitable for carbon evaporation ?? and how much are you asking for it ???
Subject: Time: 11:22 AM OFFICE MEMO RE} TEM of a polymer Date: 9/26/95
Huyen- Perhaps chlorosulfonic acid; it works on polyethylene. You might try the book "Polymer Microscopy" by LindaC. Sawyer snd David T. Grubb, ed. Chapman and Hall, 1987. -Doug
Does anyone know the film speed of Kodak SO-163 developed in full strength D-19 for 12 minutes using an accelerating voltage of 200 kv. So far I have been unable to find this information. Thus far, my efforts include speaking with the Tech support people at Kodak, searching the Kodak web page and reading the data provided with the film itself. Any suggestions or answers would be greatly appreciated.
THANKS,
----------------------------------------------- | Paul Chipman | | Department of Biological Science | | Purdue University | | West Lafayette, IN 47907 | | email: prc-at-bragg.bio.purdue.edu | | Phone: (317)494-5643 | | FAX: (317)496-1189 | -----------------------------------------------
To reiterate and expand on our embedding of thick cryosections:
The method we use is a modification of the Tokuyasu procedure. Lightly fixed, cryoprotected, Cryosections 1-4 um are cut and mounted onto a formvar-coated, carbon stabilized grid. The section is thawed, washed in Gey's balanced salt (this seems to preseve antigenicity of proteins better than standard buffers) and then immunostained using whatever procedures are desired. We transfer the grids to drops of solution containing the various washes and stains. After immunostaining we fix again in Glutaraldehyde, wash, osmicate, dehydrate, and then embed by blotting off excess 100% alcohol, and placing a drop of LR White or Spurr resin on the section. We blot the resin coated section between two pieces of filter paper, then place the grid on clean filter paper and heat cure the resin. Once cured the sections can be stained with lead citrate or uranyl acetate.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
We noticed recently in this place interest in preparing sticky substrates through treatment with silane. We would like to tell you about a method that we have now used routinely for about 5 years. It is based on procedures published by JP Robinson, P.Dunnill, and MD Lilly, in Biochim.Biophys.Acta 242, 659-661, 1971, and M.Buechi and T.Baechi, in J.Cell Biol. 83, 338-347, 1979. 1. The glass (or other carriers)must be absolutely clean. This can be accomplished by immersing for several hours in aqueous 20% sulfuric acid, or 20% hydrochloric acid in ethanol.I prefer washing with Bon Ami cleaning powder until the surface is hydrophylic. 2. The clean carriers are soaked for 2 h in waterfree acetone that had been stored for at least 24 h over molecular sieve). 3. Prepare a 2% solution of 3-aminopropyltriethoxysilane (Aldrich Chemical Co Milwaukee WI) in waterfree acetone. The carriers are placed in appropriate containers (glass,nalgene, polypropylene) which must be totally filled with the silane solution , and stored for 24 h at 50 degree C. If you use a 4% silution of silane, the carriers will be ready for use in 12 h. The silane treatment coats the carriers with amino groups. 4. To attach fresh cells or cell fractions to the carriers, they are rinsed in acetone and immersed for one hour in 1% glutaraldehyde. The carriers are then placed into dist. water at 4 degree C. They can be kept for about 5 days. Their surface is now coated with aldehyde groups, which bond covalently with aminogroups on the surface of cells or cell fractions. If cells or cell fractions must be fixed before attachment( in glutaraldehyde fixative) the 1% glutaraldehyde is omitted. Exposed aldehyde groups on the fixed biological materials will bond to the amino groups on the carriers. We have used sticky carriers prepared by this procedure to attach cells previously fixed in suspension to glass carriers for imaging by field emission SEM ( Malecki and Ris 1990, Scanning 13 ,82 ; Malecki and Ris 1992, Scanning 14, 76; Malecki,1991, Scanning Microscopy,Suppl.5, S 53). Silane treated coverslips were used to attach isolated amphibian oocyte nuclear envelopes for imaging of nuclear pore complexes by FESEM ( Ris, EMSA Bull.21, 54, 1991. ) Such coverslips were also used to retain sections of epon embedded biological materials, after extraction of the epon for FESEM imaging ( Ris and Malecki, 1993. J.Struct.Biol. 111,148). H.Ris and M.Malecki, Integrated Microscopy Resource, Univ. of Wisconsin, Madison WI. hris-at-facstaff.wisc.edu - Malecki-at-macc.wisc.edu
There were several responses to my query before the week-end, on "permanent standards for fluorescence microscopy".
The question was posted to three lists; comments were received there and directly. From the answers, fluorescing plastic blocks or sections, chambers containing fluorescent fluid, and uranyl glass, are practical standards for which some sources were given. Most of this may be common knowledge, so I will only send summaries directly on request.
One respondent asked if there were reasons for wanting to use an inorganic standard, except ease of storage. I had thought that inorganic standards, as uranyl glass, would be more stable under excitation, particularly when testing lamp stability over longer time. Apart from this, there is perhaps no advantage in re-usng a standard **object** rather than **formulation**. (?)
Thanks for all the advice, Finn-Mogens.
Finn-Mogens Haug
Department of anatomy E-mail: f.m.haug-at-basalmed.uio.no Institute of basic medical sciences Phone : +47 22 85 12 67 University of Oslo Fax : +47 22 85 12 78
Paul Chipman wrote: } } Does anyone know the film speed of Kodak SO-163 developed in full } strength D-19 for 12 minutes using an accelerating voltage of 200 kv. So far } I have been unable to find this information. Thus far, my efforts include } speaking with the Tech support people at Kodak, searching the Kodak web page } and reading the data provided with the film itself. Any suggestions or } answers would be greatly appreciated. } Dear Paul, Our experience with SO163 indicates that one gains a factor of 3 to 4 going from the usual developing conditions (1:2 D-19 for 4 min) to push con- ditions (D-19 full str. for 12 min). Although the film speed will vary with accelerating voltage, the ratio should not, so if you can find out the film speed under usual developing conditions for 200 kV electrons, multiply by about 3.5 to get what you want. Good luck. Yours, Bill Tivol
Does any one out there have a source for Polaroid portable film processing tanks? They are white, plastic, round, with a tightly fitting lid and have a rack on the inside that holds about twelve 4"x5" negatives.
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
Message-Id: {199509281119.MAA29334-at-pons.uio.no} X-Sender: finnmog-at-pons.uio.no X-Mailer: Windows Eudora Version 1.4.3 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
When registering to this list recently, I probably made some mistake. The problem is that after submitting a message to the list, I receive the error messages that arise when members of list are unreachable. Thus, after submitting two messages I received altogether 35 error messages. An example is enclosed at the end here.
A few days ago, I reported this to Postmaster-at-aaem.amc.anl.gov, hoping that she would be a human being, able to point out what is wrong, which she is probably not since there has been no reply in point.
Perhaps one of the list members knows what is wrong - or whom to contact about the problem.
} Bad address -- {RLORNB-at-ccmail.monsanto.com} } Error -- Message too old: %MULTINET-F-EHOSTUNREACH, No route to host } Bad address -- {fstewartdavis-at-ppg.com} } Error -- Message too old: Error sending MAIL command to ppg.com } Bad address -- {vit-at-scvnet.com} } Error -- Message too old: %MULTINET-F-EHOSTUNREACH, No route to host } } Start of returned message } } Message-Id: {199509221704.TAA14535-at-pons.uio.no} } X-Sender: finnmog-at-pons.uio.no } X-Mailer: Windows Eudora Version 1.4.3 } Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } Date: Fri, 22 Sep 1995 19:04:37 +0200 } To: microscopy-at-AAEM.AMC.ANL.GOV } From: f.m.haug-at-basalmed.uio.no (Finn-Mogens Haug) } Subject: Standards for fluorescence microscopy } } Dear list, } } We have looked mostly in vain for fluorescent standards, specifically } "non-bleaching", solid, inorganic, specimens suitable for routine checks } on excitation level and checks on spatial uniformity of the optical/digital } imaging system. } } Reports and handbooks describe the use of uranyl-glass and inorganic } crystals, but the only commercial product we have found contains a 1-2 mm } circular fluorescing area of unknown composition, mounted on a microscope } slide. Price: USD 1400 incl VAT. Expensive? } } A local contact makes Yttrium-Oxy-Sulfide crystals with discrete } emission/excitation } bands for use in microspectrofluorimetry. However, spectral specificity may } not be required for checks on excitation level and the powder's granularity } may preclude its use in testing pixel-to-pixel spatial uniformity. } } Names, fax/phone-numbers and e-mail adresses of suppliers of such standards } would be most helpful, perhaps to others as well. } } } Thanks in advance - } } } Finn-Mogens Haug } } Department of anatomy E-mail: f.m.haug-at-basalmed.uio.no } Institute of basic medical sciences Phone : +47 22 85 12 67 } University of Oslo Fax : +47 22 85 12 78 } } } End of returned message } } }
Finn-Mogens Haug
Department of anatomy E-mail: f.m.haug-at-basalmed.uio.no Institute of basic medical sciences Phone : +47 22 85 12 67 University of Oslo Fax : +47 22 85 12 78
"When registering to this list recently, I probably made some mistake. The problem is that after submitting a message to the list, I receive the error messages that arise when members of list are unreachable. Thus, after submitting two messages I received altogether 35 error messages."
Reply:
You are not the only one who gets these messages. Every time I submit a posting, I get many similar messages. I have taken the easiest route which is to post infrequently and answer other postings directly, not via this forum. I am sure that many others are taking this option because although questions are being posted, few of the replies are seen by all.
Additionally, it appears that I do not see all the messages posted here. I know this, not only because I pick up answers to questions I never saw but also because I never even saw two messages posted by a colleague. She warned me in advance that they were being sent and she even received replies from some subscribers, so they did get posted.
The administrators at my end are mystified by this. Perhaps our administrator could add some comments. After all, the reason we subscribe is so that we can freely exchange messages with others of similar interests. If the posting mechanisms are deterring us from adding our comments then the system may soon fall apart.
Perhaps this is all a sophisticated technological plan to filter out trivia!
Best regards, Paul Webster Yale School of Medicine
} } When registering to this list recently, I probably made some mistake. } } The problem is that after submitting a message to the list, I receive } } the error messages that arise when members of list are unreachable. } } Thus, after submitting two messages I received altogether 35 error } } messages. An example is enclosed at the end here.
most of you replied along the following lines:
} } There is absolutely nothing wrong with your setup and I believe there is } very little the site administrator could do to resolve the problem. } Actually, this is not a "problem" but the way the e-mail system works. } Since you are the originator of the message, when any of the group } subscribers moves, changes e-mail address or his/her computer is down, the } e-mail system will report the fact that your message could not be delivered } (I believe delivery is tried at least three times before the undeliverable } mail message is sent back to the originator). So. I am afraid you, as I } have in the past, have just come in contact with the "dark side" of } subscribing to an interest group. }
I have only registered with two other lists (and some news-groups) before this one and there it looks as if the liststservers filter out error messages of this kind.
} I still think the gains outweigh the losses. }
I will stay tuned. Thanks for your help.
Finn-Mogens
Finn-Mogens Haug
Department of anatomy E-mail: f.m.haug-at-basalmed.uio.no Institute of basic medical sciences Phone : +47 22 85 12 67 University of Oslo Fax : +47 22 85 12 78
Having a mail reader with mail-filtering capability really helps with the returned mail overload after posting to this and any other list. The elaborate filtering system that I have set up in my reader (Pegasus Mail) has virtually eliminated the "Undeliverable Mail", "Returned Mail", etc. as well as the equally aggrevating "Subscribe", "Unsubscribe" postings. It's even useful to filter out postings from certain individuals that you have grown weary of. (Not on this list of course!)
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia
Yes I know about the periodic Undeliverable Mail problems. I now (finally) have some new software which I am in the process of installing/debugging. If all goes well in the next few weeks the system will get better. I keep a reasonable eye on things and when I notice multiple errors from the same site (or people tell me about it) then I remove that address from the subscription list.
But as most of you know it's getting harder and harder to clone more copies of myself. The first five copies were reasonable, but now they have to grow up, although a few of them are begining to show some promise.;-)
I noticed Paul's report of mail not getting through. I need to know this type of information so if you have specifics then forward them to me.
Your Friendly Neighborhood SysOp
Nestor
P.S. I do get alot of Email (50+/day), so don't be offended if I don't reply to everything. I do need to do other things occassionally (like sleep....).
There is a rather detailed discussion of the operation of the vacuum system of the JEOL 9010 Freeze-Fracture apparatus in Section 9.1, pp. 373 - 381, of my book "Vacuum Methods in Electron Microscopy" (available from Ashgate Publishers, Old Post Road, Brookfield, VT 05036-9704, Ph: 800-535-9544, Fx: 802-276-3837). Perhaps this would be helpful to you, since, apart from specimen preparation, manipulation of the vacuum system is a major part of the operation of the instrument. Good luck, W. C. Bigelow (bigelow-at-umich.edu)
Sorry for jumping in, but I did want to make a comment on this topic. Despite the extra traffic due to undeliverable mail warnings(Yes, I get them too), I would like to encourage members of the list to respond to the list as opposed to the individule, as often as possible. Many of the questions asked, pertain to topics, I and others are interested in, and our questions haven't quite gelled yet, so the answer is informative to many of us. Further more, some of the information offered, provides different ways of accomplishing tasks that get tucked away in the "I have to try that someday" file (the Hans Ris "sticky slide" response is a good example). In a lot of the replies, even if it was the same as I might have offered is a different perspective that helps me to understand various processes better. Sometimes, in a reply, there is just a word or two that provides a solution to a problem. Lastly, some of the questions that I think I have the answer for, the alternatives offered here were solutions, I never would have thought of. From my selfish point-of view, I hope many will answer questions on the listserver for all to view, even though there will be those 30 or so undeliverable notes tomorrow to muddle through. I also thought I had made a mistake when I got a number of undeliverable mail warnings from my first reply, but talking to the system administrator calmed me down and as in another response, he was surprised there was so few for a listserver this size. Thanks to all who take the time to respond to our questions. later dlb
David Bentley Imaging Facility, Div. of Biotechnology, ARL The University of Arizona Tucson, Az 85721
Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" To: MICROSCOPY-at-AAEM.AMC.ANL.GOV
Greetings, I'm having an energetic discussion with a photographer on the proper way to indicate the true magnification on a micrograph. One denotes the print magnification by the negative magnification only. The other denotes a print magnification as the negative magnification times the enlargement factor. The former believes that no increased magnification exists unless there is a concommittant increase in resolution and that it is misleading to do otherwise. The latter believes that magnification can be increased (without necessarily increasing resolution) by enlargement and that total resultant magnification should be used to describe the micrograph; accurate measurements on the print being impossible otherwise. Anybody care to wade into the discussion?
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
To state the obvious - the two are different. Which on is expressed depends on what you want to know. Most of the time when I present photomicrographs the relevant issue is "how much smaller is the actual thing than what is shown in the picture." This is obviously a magnification issue on the image that the viewer is actually seeing.
The problem does become more fun when projecting a slide. Does someone in the audience really have a good idea of how large the projected image is? Doesn't it depend on where in the audience the person sits?
I suppose the best general solution is to include a good scale in the original preparation. I don't consider a 1 micrometer bar a good scale for most folks.
Peter D. Barnett - Forensic Science Associates - Richmond CA pbarnett-at-crl.com VOICE: 510-222-8883 FAX: 510-222-8887
Why bother with magnification at all. I always simply place a "micron" (or nanometer etc..) marker on the image. In this way the image is always calibrated regardless of what anyone else does to the print after you give it to them.
} I'm having an energetic discussion with a photographer on the } proper way to indicate the true magnification on a micrograph. One denotes } the print magnification by the negative magnification only. The other } denotes a print magnification as the negative magnification times the } enlargement factor. The former believes that no increased magnification } exists unless there is a concommittant increase in resolution and that it } is misleading to do otherwise. The latter believes that magnification can } be increased (without necessarily increasing resolution) by enlargement and } that total resultant magnification should be used to describe the } micrograph; accurate measurements on the print being impossible otherwise. } Anybody care to wade into the discussion? } }
Of course the print magnification can and should be used. I know that there is no way to increase the information contents of an image by magnifying it, but it is of no help to me when I see a print to know that the negative magnification was this or that. A scale bar included in the print is I think the best solution. Then it doesn't matter if the size of the image is changed in the publishing process, and anyone who so wishes can easily determine the resolving power of the microscope or whatever is used for the imaging of the specimen.
} Sorry for jumping in, but I did want to make a comment on this } topic. Despite the extra traffic due to undeliverable mail warnings(Yes, I } get them too), I would like to encourage members of the list to respond to } the list as opposed to the individule, as often as possible. Many of the } questions asked, pertain to topics, I and others are interested in, and our } questions haven't quite gelled yet, so the answer is informative to many of } us. Further more, some of the information offered, provides different ways } of accomplishing tasks that get tucked away in the "I have to try that } someday" file (the Hans Ris "sticky slide" response is a good example). In } a lot of the replies, even if it was the same as I might have offered is a } different perspective that helps me to understand various processes better. } Sometimes, in a reply, there is just a word or two that provides a solution } to a problem. Lastly, some of the questions that I think I have the answer } for, the alternatives offered here were solutions, I never would have } thought of. } From my selfish point-of view, I hope many will answer questions on } the listserver for all to view, even though there will be those 30 or so } undeliverable notes tomorrow to muddle through. I also thought I had made a } mistake when I got a number of undeliverable mail warnings from my first } reply, but talking to the system administrator calmed me down and as in } another response, he was surprised there was so few for a listserver this size. } Thanks to all who take the time to respond to our questions. } later dlb }
} } Additionally, it appears that I do not see all the messages posted here. I know } this, not only because I pick up answers to questions I never saw but also } because I never even saw two messages posted by a colleague. She warned me in } advance that they were being sent and she even received replies from some } subscribers, so they did get posted. } } The administrators at my end are mystified by this. Perhaps our administrator } could add some comments. After all, the reason we subscribe is so that we can } freely exchange messages with others of similar interests. If the posting } mechanisms are deterring us from adding our comments then the system may soon } fall apart.
I may have one answer to this point. I used a few months ago a soft called POPMAIL, and actually there WERE messages that never went through. Then once I could notice that all of these undetected mail had one commun feature (by looking directly in the inbox.mbx directory): There was always one blank line in the header: for any cryptic reason the soft was unable to recognise them as mails. However the mails files did exist, and by removing this blank line (using the EDIT program of MS DOS for example) I could get them back to life. I have now switched to PINE and believe things are far easier. Which mail system are you using?
I'm looking for information regarding the American Society for Cell Biology. Could someone please tell me who to contact to get registration and accomodation details?
Thanks, Felicity EM Unit, QUT. Brisbane, Australia
Advertisment forwarded to the microscopy mail reflector on behalf of a colleague. Please address all enquiries to the numbers and addresses listed below - not to me thanks, Mark Aindow
Postdoctoral Position Available at The University of Birmingham, IRC in Materials for High Performance Applications, Birmingham, UK
Synthesis of Oxide Ceramic Precursor Sols for Structural and Functional Ceramics
A Research Fellow (RA1A grade) is required to work on the hydrothermal synthesis and emulsion synthesis of ultrafine nanometre size oxide ceramic precursor sols for structural and functional ceramics with the desired chemical stoichiometry, phase composition and homogeneity. The compositions will be chosen as being representative of single-, dual-, and multi-cation oxide ceramics, and will need to be characterized physically, structurally and chemically using advanced analytical techniques such as TEM/EDX, XRD, PEELS etc.
In the first instance this is a two-year EPSRC funded core project in the IRC in Materials for High Performance Applications, The University of Birmingham. There is the possibility of a project extension. Thus, the initial appointment will be for 2 years from December 1995 to December 1997. Starting salary will be in the range of 14,317 to 17,446 pounds sterling per annum depending on relevant ability and expertise.
Preliminary enquiries and requests for further particulars should be directed to Dr. C.B. Ponton by telephone on + 44 -121-414 5226; FAX on +44-121-414-3441, or preferably by Email to C.B.Ponton-at-bham.ac.uk
The post would suit ideally a person who could meet all the following essential criteria; and at least some, if not all of the desirable criteria.
Essential :
A good working knowledge of transmission and scanning electron beam techniques, particularly TEM and/or STEM chemical analysis (EDX) and structure determination techniques is essential.
The ability and desire to extend undergraduate and/or graduate knowledge of aqueous and/or organo-aqueous reaction chemistry to the hydrothermal and emulsion synthesis of ceramic precursor sols, and to develop hydrothermal, i.e. high temperature and pressure process engineering skills.
Proven ability to carry out scientific research and to communicate the results of the research by scientific journal publications and conference presentations.
Ability to work in a team and interact with workers in other scientific/engineering disciplines.
Computer literate as regards computer-based word-processing, data logging and analysis.
Desirable :
Experience of applying electron beam chemical analysis techniques such as TEM and/or STEM EDX and (P)EELS with ELNES applied to nanometre size ceramic particulates.
Familiarity with advanced chemical and spectroscopic characterisation techniques including quantitative powder XRD and one or more of MAS MRI, FTIR etc.
Familiarity with inorganic and organometallic reaction chemistry including reaction thermodynamics and kinetics as well as ceramic powder processing science and technology including powder pressing, sintering and mechanical property testing.
Does anybody have any crystallographic information, or know of a source, of lead lanthanum titanate pyrochlore phase. This phase is often refered to in the literature, however aquiring precise structural data has proved difficult. Thank, *************************************** ************ Lee Smith ************ ******* School of Materials ******* ******* University of Leeds ******* ******* Leeds, LS2 9JT, UK ******* ***************************************
I always consider the print magnification to be the neg. mag times the enlargement factor. The resolution should only be calculated from a neg. therefore resolution is mostly mote when looking at a print.(except when publishing).
Best wishes, Ed Calomeni Medical College of Ohio Toledo, OH emlab-at-opus.mco.edu
Message-ID: {MAILQUEUE-101.950929144237.384-at-physics.uwc.ac.za} To: microscopy-at-aaem.amc.anl.gov
Hi
Just my two cents worth of knowledge:
Magnification on the print is the total magnification, ie negative magnification times enlargement factor. It might be that it is an empty magnification, ie no further detail becomes visible due to poor resolution on the micrograph. However the negative own grain structure is usually far smaller than detail visible (resolved?) on the micrograph, ie if the micrograph is done well, you should be able to get additional detail at enlargerment factors up to 10 times. In either case you do not change anything on the original resolution of the micrograph, only the visibility thereof becomes better (that's why you look at a micrograph through a manifying lens to see the detail on the micrograph not visible to the naked eye).
Hope it clears up some confusion, Dirk
} Greetings, } I'm having an energetic discussion with a photographer on the } proper way to indicate the true magnification on a micrograph. One denotes } the print magnification by the negative magnification only. The other } denotes a print magnification as the negative magnification times the } enlargement factor. The former believes that no increased magnification } exists unless there is a concommittant increase in resolution and that it } is misleading to do otherwise. The latter believes that magnification can } be increased (without necessarily increasing resolution) by enlargement and } that total resultant magnification should be used to describe the } micrograph; accurate measurements on the print being impossible otherwise. } Anybody care to wade into the discussion? } } } } Charles J. Butterick (Chuck) }
Prof Dirk Knoesen U U W W CCCCCC Department of Physics U U W W C University of the Western Cape U U W W W C Private Bag X17, Bellville 7535 U U W W W W C South Africa UUUUUU W W CCCCCC Tel:+21 959 2266. Fax:+21 959 3474. Internet: dirk-at-physics.uwc.ac.za
--Boundary (ID EukuE+CAP3e/3Urph/Qb5A) Content-type: TEXT/PLAIN
Working in a metallographic lab and and electron optic lab the best way to label images with an indicator for subsequent measurement is to use a micron marker (calibrated of course). Regardless of mag. direct measurements can then be taken with better accuracy. Enlarge the image and you enlarge the measuring marker equally as well.
That being said mag. and res. are obviously two different "critters". By defintion magnification increases an object's size so that we can observe it but resolution allows us to separate fine detail in that object. Who cares how large we make an object if we can't resolve any features! Image size will be increased by enlargement but USEFUL mag. is not, nor is resolution increased! Take a 4x5 inch 500x print of a certain microstructure and measure a feature, divide by the mag to get "true" size. Now enlarge the print by 4 times, remeasure the feature and divide by the same mag. What happens? Your numbers don't match! True if you include the enlargment factor, the numbers will match but you have not increased the mag. You have actually used "empty magnification" to make the object larger.
I think the differences lies in the interpretation of magnifiction. Microscopists interpret mag as an increased image size that supplies useful information. I suppose photographers refer to magnification as enlarged images.
The lesson learned, there is useful mag and empty mag and don't use magnification as the be all and end all for measurement purposes!
} Greetings, } I'm having an energetic discussion with a photographer on the } proper way to indicate the true magnification on a micrograph. One denotes } the print magnification by the negative magnification only. The other } denotes a print magnification as the negative magnification times the } enlargement factor. The former believes that no increased magnification } exists unless there is a concommittant increase in resolution and that it } is misleading to do otherwise. The latter believes that magnification can } be increased (without necessarily increasing resolution) by enlargement and } that total resultant magnification should be used to describe the } micrograph; accurate measurements on the print being impossible otherwise. } Anybody care to wade into the discussion? }
Magnification is magnification and resolution is resolution and never the twain shall meet. The negative magnification can be just as empty magnification as the print magnification. IT IS IMPERATIVE TO GIVE THE MAGNIFICATION OF THE "IMAGE" THE VIEWER IS LOOKING AT. As you pointed out this is the only way for readers to make measurements from the "image" they have in front of them. I don't submit manuscripts to journals which have the nasty habit of changing the size of micrographs and including in the caption something like "reduced from the original magnification of X". This is a meaningless phrase. It gives me no information except the warning not to trust the image. Even if the image is pointlessly magnified above the effective resolution, you still need the magnification information to judge the science.
The other alternative is to include a measuring standard bar in the image. This avoids all such problems.
That is my two cents worth. Obviously, this is something I feel very strongly about.
Thanks for listening-
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
Dear Charles, I, too, go along with the use of a scale bar to indicate magnification (could this agreement among the members of this list be why MSA asks for scale bars :-)). There is, however, a reason that the neg mag and print enlarge- ment factors could be useful in some circumstances. For high mags and largish grain, these numbers could tell one how much graininess is due to the film and how much is due to noise. In this case, it is the neg mag which is important, and the scale bar alone is not sufficient. Yours, Bill Tivol
Hello: I will forward to you a message that I received from Ms. Kathy King about the Annual Meeting of the American Society for Cell Biology. I hope that's what you are looking for.
Adriana Rodriguez Secao de Biologia e Melhoramento Vegetal Centro de Energia Nuclear na Agricultura Universidade de Sao Paulo, Brazil
On Fri, 29 Sep 1995, Felicity Lawrence wrote:
} G'day. } } I'm looking for information regarding the American Society for Cell Biology. } Could someone please tell me who to contact to get registration and } accomodation details? } } Thanks, } Felicity } EM Unit, QUT. Brisbane, Australia } }
This is a test of my mail system response and a short comment on the ongoing discussion. It seems perfectly obvious that if one can't see what is being blown up in the print, it is useless. I don't know anyone that makes prints of things that can't be seen! If one can see it in the print magnification is always useful as it gives one the scale, if not why make the print. I am a failure analyst and an amateur photographer, I'm somewhat baffled by the logic of this discussion.
Have a very nice day. I am enjoying the mail I receive from this listserver and I hope in the future to contribute something more useful than this comment.
Thanks for the many replies to my query regarding Polaroid processing buckets ("PN-10; Clearing Tank For 665/55 Film", for something like $39.95, this includes the tank, the rack and a pound of sodium sulfite). Several people suggested contacting Graphic Center, P.O. Box 818, Ventura, CA. 93002. Phone : 1-800-336-6096. Others said that Polaroid stopped making them several years ago and they are unavailable. I called the Graphic Center and their tape machine said they will be closed from Sept 28-Oct 8 (maybe they all took a road trip to pick up a new supply of PN-10s). I'll try them again after the 8th and let ya'll know if I am successful.
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
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The magnification vs. resolution multilog has been interesting and many have extolled the virtues of scale bars. Let me point out that scale bars are not as appropriate for those images having a significant depth demension. An example might be a photograph of you ten feet from the camera with the Washington Monument visible five miles away in the background. A scale bar would not be able to indicate both your size and the size of the monument. Scanning electron micrographs (secondary electron imaging) are such images and it is best to express the magnification for these numerically.
Joiner Cartwright, Jr., Ph.D. Director, Electron Microscopy Department of Pathology, Rm.286-A Baylor College of Medicine One Baylor Plaza Houston, Texas 77030 U.S.A. tel.: (713)798-4658 FAX: (713)798-3945 joiner-at-bcm.tmc.edu Compuserve: 71555,1206
Resolution is how much detail your system was able to put into the micrograph. The optimal resolution of a system is usually defined by the familiar equation, R=(0.61&)/(u sin -at-).
Magnification is how much the original object has been enlarged, usually negative magnification times enlargement factor. Your negative magnification, of course, is how many times larger the structures shown in your negative are compared to the original structures you were photographing (or "electrographing"). A micron bar is useful, since it shows how long a micron would be in terms of the original structure.
Further magnification of a negative can make the detail more easily visible, but it can't create new detail (in other words, it can't increase the resolution of the original negative).
A. Kent Christensen, University of Michigan, {akc-at-umich.edu}
---------------------------------------
On Thu, 28 Sep 1995, Charles J. Butterick wrote:
} Greetings, } I'm having an energetic discussion with a photographer on the } proper way to indicate the true magnification on a micrograph. One denotes } the print magnification by the negative magnification only. The other } denotes a print magnification as the negative magnification times the } enlargement factor. The former believes that no increased magnification } exists unless there is a concommittant increase in resolution and that it } is misleading to do otherwise. The latter believes that magnification can } be increased (without necessarily increasing resolution) by enlargement and } that total resultant magnification should be used to describe the } micrograph; accurate measurements on the print being impossible otherwise. } Anybody care to wade into the discussion? } } } } Charles J. Butterick (Chuck) } Electron Microscopy Center } Department of Cell Biology } and Biochemistry } Texas Tech University Health } Sciences Center } 3601 4th Street } Lubbock, Texas 79430 } } vox (806) 743-1633 } fax (806) 743-1219 } email emccjb-at-ttuhsc.edu or } chuck-at-micron1.lubb.ttuhsc.edu } } }
PINE is an exceptional mail reader for UNIX, but there are much more intuitive and more easily configurable mail readers available in Macintosh and Windows format.
I currently use the Eudora (available on both formats) from QUEST. There is a freeware version and a commercial version. Both use TCP/IP software to interface with the POP mail server and make replying, editing, building group mail aliases, and sending attached files easy! They both provide the capability for multi-user use from the same machine (currently 7 people use this copy - each with separate preferences) as well as the capability to have multiple mailboxes. The commercial version will sort (or block) mail by subject or kewords in the body of the mail message and place it in the proper mailbox and do a couple more things.
________________________________________________ Kirk A. Rogers krogers-at-materials.ecn.purdue.edu 317-494-8751 office http://materials.ecn.purdue.edu/~krogers Purdue Unuversity, School of Materials Engineering, 1289 MSEE building, W. Lafayette, IN 47907-1289
I for one prefer not to put markers in my images, as I believe they distract from the art aesthetics.
--------------------------------------
REGARDING Res/Mag hoopla To the group,
This is a test of my mail system response and a short comment on the ongoing discussion. It seems perfectly obvious that if one can't see what is being blown up in the print, it is useless. I don't know anyone that makes prints of things that can't be seen! If one can see it in the print magnification is always useful as it gives one the scale, if not why make the print. I am a failure analyst and an amateur photographer, I'm somewhat baffled by the logic
of this discussion.
Have a very nice day. I am enjoying the mail I receive from this listserver and I hope in the future to contribute something more useful than this comment.
Respectfully, Bob Lawrence
------------------ RFC822 Header Follows ------------------ Received: by qmgate_backup.anl.gov with SMTP;29 Sep 1995 10:35:37 -0500
As a tangent to the current thread on magnification and resolution, our photographer today inquired about the difference between a macrograph (e.g., a photograph made using a macro lens) and a micrograph. Anyone want to jump in on this?
Can anyone recommend a decent shutter crontroller for a Zeiss Axiovert microscope that has epi-fluor and DIC? Hopefully one that is not too expensive and manually operated. Thanks, Jim Ryan Biology Department Hobart and William Smith Coleges Geneva, NY 14456 ryan-at-hws.edu
There is often confusion about resolution and magnification. One can produce very high magnification of the image by the enlargement factor, without increasing resolution. (fuzzy, unsharp...) The resolution mainly depends on the image forming instrument (LM, TEM, SEM, AFM...) AND the specimen, the enlarger is only a tool to provide additional magnification for the human eye (it has limits also!) to view the detail of the image. There are many other factors influence the resolution of the image, many excellent books have been published on this subject.
To indicate the total or final magnification (enlargement factor X image magnification on the negative) on the print, is to apply the appropriate "micron" marker. The marker also "calibrates" all slides, hard copies and computer images.
Laszlo J. Veto Electron Microscopist Research Centre Summerland, B.C. Canada V0H 1Z0 604-494-7711 voice 604-494-0755 fax Veto-at-bcrssu.agr.ca e-Mail
Whoa, people, you must all remember something important when labeling or viewing images with magnifications bars. That bar is only useful for measurements *in the plane of the film*! This is a problem with SEM micrographs. You are looking at a 2-D representation of a 3-D object, and two structures that may seem to be connected or overlapping or near each other (due to the effects of foreshortening) may, in fact, be far apart. WHen training researchers on the SEM I make sure that they understand this and get used to tilting and rotating the stage a lot to see the true relationships between structures. I also make sure they understand that trying to measure a structure or the distance between structures that are not in the same plane (the plane of the viewing screen or the micrograph) is bogus. A few examples usually gets the idea across. Having given the warning, however, I still tell them to put the micron bar on there, because without it I'm easily lost! The editors of some journals prefer only the field magnification for SEMs, which is correct. Micron bars would be great if the audience understands their limitations.
What do you editors out there think?
Aloha, Tina
***************************************** Tina (Weatherby) Carvalho * Biological Electron Microscope Facility * University of Hawaii * (808) 956-6251 * tina-at-ahi.pbrc.hawaii.edu * http://www.pbrc.hawaii.edu/bemf/ * *****************************************
Can anyone recommend a shutter controller for a zeis microscope that has epi-fluorescence and dic? I would appreciate the names and phone #'s of companies producing reasonably priced shutters. Thanks Jim Ryan Biology Dept. Hobart & William Smith Colleges Geneva, NY 14456 ryan-at-hws.edu
The American Society of Cell Biology is meeting in Washington, DC from December 9 - 13th. Their EMail address is ascbinfo-at-ascb.faseb.org. Have a pleasant visit.
Eudora requires a POP server. If your mail server doesn't support that protocol, like here at University of Washington, then you can't use Eudora.
Glen MacDonald Hearing Development Laboratories University of Washington Seattle, WA 98195-6515 (206)543-8360 glenmac-at-u.washington.edu
On Fri, 29 Sep 1995, kirk rogers wrote:
} Yves, } } PINE is an exceptional mail reader for UNIX, but there are much more } intuitive and more easily configurable mail readers available in Macintosh } and Windows format. } } I currently use the Eudora (available on both formats) from QUEST. There } is a freeware version and a commercial version. Both use TCP/IP software } to interface with the POP mail server and make replying, editing, building } group mail aliases, and sending attached files easy! They both provide the } capability for multi-user use from the same machine (currently 7 people use } this copy - each with separate preferences) as well as the capability to } have multiple mailboxes. The commercial version will sort (or block) mail } by subject or kewords in the body of the mail message and place it in the } proper mailbox and do a couple more things. } } More info can be found via the following: } } QUALCOMM Enterprise Software Technologies (QUEST) Sales Administration at } (800) 2-Eudora, (619) 658-1291, } } OR quest-rep-at-qualcomm.com. } } OR ftp.qualcomm.com/quest/product_literature } } OR http://www.qualcomm.com/quest } } -Kirk } } ________________________________________________ } Kirk A. Rogers } krogers-at-materials.ecn.purdue.edu } 317-494-8751 office http://materials.ecn.purdue.edu/~krogers } Purdue Unuversity, School of Materials Engineering, } 1289 MSEE building, W. Lafayette, IN 47907-1289 } } }
} Charles J. Butterick (Chuck) wrote: } I'm having an energetic discussion with a photographer on the } proper way to indicate the true magnification on a micrograph. One denotes } the print magnification by the negative magnification only. The other } denotes a print magnification as the negative magnification times the } enlargement factor. The former believes that no increased magnification } exists unless there is a concommittant increase in resolution and that it } is misleading to do otherwise. The latter believes that magnification can } be increased (without necessarily increasing resolution) by enlargement and } that total resultant magnification should be used to describe the } micrograph; accurate measurements on the print being impossible otherwise. } Anybody care to wade into the discussion?
Magnification is the ratio of the size of a feature in the image to that in the original object. This can be increased by repeated enlargement without improving resolution. (empty magnification) To make accurate measurements possible, one should include a scale marker in the image. If there is no scale marker, the total magnification should be used to describe the print.
Having participated in this list since the beginning, I have noted that there are a lot of small, but good, bits of practical information that come down the line. We see question like "How do I do....... ? How do I make......? Where do I find...? etc.
Here in our lab we often ask if anyone can recall what was said a few weeks back on solving some problem. Often we have forgotten to save the message or print it out.
We also see the same questions repeated every few months on the list as new people join and as we older dudes forget.
Therefore we are going to make a concerted effort to archive all the good ideas and advice we come across on this list. We will add our own collection to it as we go. And we want your input.
Since it is just as easy to make the archive available to the world as to keep it to ourselves we want all of you in on it.
So as an experiment (duration undetermined) we will begin editng and accumulating what we have come to call "Tips & Tricks" for biological microscopy in a file available over the internet. We hope someone in materials microscopy will choose to do likewise for their colleagues.
The file is currently in an embryonic stage but given the proper gestation should grow to be a valuable resource.
If this task is being done at another site, please tell me ASAP so we don't waste too much time duplicating the efforts of another/.
Feel free to send us material, unsolicited, that you think appropriate. Either send text via e-mail or give us a URL that can serve as a link from our page. Include literature citations whenever possible so that the right person gets the credit.
You can find us at www.biotech.ufl.edu/~emcl. Go down the page and click the Wizard.
There is not much there yet, so give us some time to get going.
..........and keep those cards and letters coming.
Regards, Greg Erdos -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
In any respectable treatise on optics (including photgraphy books) MAGNIFICATION is defined as image size divided by object size ("size" is a linear dimension as opposed to area)
therefore the print magnification is the total magnification or the negative magnification times the enlargement factor
Marcelle Gillott uwm
On Thu, 28 Sep 1995, Charles J. Butterick wrote:
} Greetings, } I'm having an energetic discussion with a photographer on the } proper way to indicate the true magnification on a micrograph. One denotes } the print magnification by the negative magnification only. The other } denotes a print magnification as the negative magnification times the } enlargement factor. The former believes that no increased magnification } exists unless there is a concommittant increase in resolution and that it } is misleading to do otherwise. The latter believes that magnification can } be increased (without necessarily increasing resolution) by enlargement and } that total resultant magnification should be used to describe the } micrograph; accurate measurements on the print being impossible otherwise. } Anybody care to wade into the discussion? } } } } Charles J. Butterick (Chuck) } Electron Microscopy Center } Department of Cell Biology } and Biochemistry } Texas Tech University Health } Sciences Center } 3601 4th Street } Lubbock, Texas 79430 } } vox (806) 743-1633 } fax (806) 743-1219 } email emccjb-at-ttuhsc.edu or } chuck-at-micron1.lubb.ttuhsc.edu } } }
Mathematically the second way is absolutely correct. In practice, useing magnification bar is more convenient and convention. =================================================================== } Greetings, } I'm having an energetic discussion with a photographer on the } proper way to indicate the true magnification on a micrograph. One denotes } the print magnification by the negative magnification only. The other } denotes a print magnification as the negative magnification times the } enlargement factor. The former believes that no increased magnification } exists unless there is a concommittant increase in resolution and that it } is misleading to do otherwise. The latter believes that magnification can } be increased (without necessarily increasing resolution) by enlargement and } that total resultant magnification should be used to describe the } micrograph; accurate measurements on the print being impossible otherwise. } Anybody care to wade into the discussion? } } } } Charles J. Butterick (Chuck) } Electron Microscopy Center } Department of Cell Biology } and Biochemistry } Texas Tech University Health } Sciences Center } 3601 4th Street } Lubbock, Texas 79430 } } vox (806) 743-1633 } fax (806) 743-1219 } email emccjb-at-ttuhsc.edu or } chuck-at-micron1.lubb.ttuhsc.edu ================================================
Xiaogang
**************************************** * Xiaogang Xie * * Department of Geology and Geophysics * * Louisiana State University * * Baton Rouge, LA 70803 * * Office (504)388-2240 * * Fax (504)388-2302 * ****************************************
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At 01:34 PM 9/29/95 -0400, you wrote: } } As a tangent to the current thread on magnification and resolution, our } photographer today inquired about the difference between a macrograph } (e.g., a photograph made using a macro lens) and a micrograph. Anyone } want to jump in on this? } } James Martin } } ************ OK, I'll give it a try. I am going to say that a camera with a macro lens....IF it is set up to record images larger than the photographed subject (ie. 1:1+).....is equivalent to a microscope with a film-back. Therefore your photographer's "macrograph" is the same as a micrograph. I think that macrograph may be a misnomer, or at the most, jargon. Many photographers use macrography to mean recording images between what a "standard" camera lens can do and what a close-up lens can do up to an image ratio of 1:1, ie. not magnifying.
Howzzat?
Joiner Cartwright, Jr., Ph.D. Director, Electron Microscopy Department of Pathology, Rm.286-A Baylor College of Medicine One Baylor Plaza Houston, Texas 77030 U.S.A. tel.: (713)798-4658 FAX: (713)798-3945 joiner-at-bcm.tmc.edu Compuserve: 71555,1206
Message-Id: {199509301540.LAA06149-at-post-ofc02.srv.cis.pitt.edu} To: "microscopy-at-AAEM.AMC.ANL.GOV" {microscopy-at-AAEM.AMC.ANL.GOV}
Felicity: Here is the information I received from Ms. Kathy King about hotel reservation and meeting program. I hope that helps Adriana Rodriguez BMV/CENA/University of Sao Paulo, Brazil
Follows is hotel information and a summary of the meeting. Let me know if you need additional information. -Kathy ================ PARTICIPATING HOTELS AND HOTEL RESERVATION INSTRUCTIONS
14. Renaissance Washington DC Hotel, 700, $108, $128, $148, Metro Center (Hotel is across the street from Convention Center) [Club rates - Single $128, Double $148, Triple $168 (continental breakfast, and other amenities included in Club rate)]
*Convention Center is located one block from the Metro Center stop at the 11th & G Streets exit.
========================
HOTEL RESERVATION INSTRUCTIONS
To make hotel reservations call: 1-800-535-3336 (US & Canada) or 1-202-842-2930 (Washington, DC & International)
DEPOSIT: A $100/per room deposit is required for all reservations. The deposit amount is payable by credit card or check.
CREDIT CARD: Your credit card will be charged immediately. Most major credit cards are accepted. Your room confirmation will be sent upon acceptance of your credit card charge.
CHECKS: An invoice for the $100/per room deposit will be mailed to you. Payment must be received within 15 days of the invoice date or your reservation will be cancelled. Do not send payment without an invoice stub. Your room confirmation will be sent upon receipt of your check.
CHANGES: Prior to November 10, all changes should be made through the ASCB housing service. After November 10, changes should be made directly with the hotel.
CANCELLATIONS/REFUNDS: Cancellations made prior to November 10 should be made through the ASCB housing service and will be refunded in full by the ASCB housing service. Cancellations made after November 10 and PRIOR to 72 hours before arrival should be made through the hotel which will issue the refund. A $10 fee will be deducted from your deposit.
Reservations must be cancelled 72 hours prior to arrival or the entire $100 deposit is forfeited.
Please have the following information available prior to calling for reservations:
1. Name of convention: ASCB 2. 1st, 2nd, and 3rd choice of hotel 3. Arrival/departure dates 4. Number of rooms requested 5. Type of room (single, double, twin, etc.) 6. Number of persons in party 7. Arrival time 8. Credit card type, account number, and expiration date 9. Names of all occupants of room 10. Address to which confirmation is to be sent 11. Telephone number 12. Fax number (if you would like a confirmation faxed)
INQUIRIES: ASCB Housing Service 1212 New York Avenue, NW 6th Floor Washington, DC 20005
International participants may make reservations by faxing the information requested above to Convention Housing s 24-hour fax number at 202-289-8079. A written confirmation will be sent. Please be sure to include a fax number. ===================== AMERICAN SOCIETY FOR CELL BIOLOGY THIRTY-FIFTH ANNUAL MEETING PROGRAM SUMMARY Saturday, December 9 Wednesday, December 13, 1995 Washington Convention Center Washington, DC
SATURDAY, DECEMBER 9 11:00AM - 8:00PM Registration 2:00PM - 4:00PM Educating the Public: Ideas for Scientists as Advocates 1:00PM - 5:00PM Special Interest Subgroup Meetings 3:00AM - 5:00PM Education/Minorities Affairs Information Booth 6:00PM - 7:00PM PUBLIC POLICY ADDRESS 6:30PM - 9:00PM Posters on Display 7:00PM - 8:00PM SCIENCE KEYNOTE Eric Kandel, Columbia University, Genes, Synapses, and the Cell Biology of Long-Term Memory 8:00PM - 10:00PM Reception 8:00PM - 10:00PM Student Reception
SUNDAY, DECEMBER 10 7:00AM - 8:00PM Congressional Liaison Committee Breakfast (date and time subject to change, see official Program) 7:00AM - 6:00PM Exhibitor Showcase 7:30AM - 5:00PM Registration 7:30AM - 9:00PM Posters on Display 8:00AM - 9:30AM SYMPOSIUM I The Cell Cycle and Cancer Leland Hartwell, University of Washington (Chair), Cell cycle checkpoints Kim Nasmyth, Institute of Molecular Pathology, Vienna, Cyclin-dependent kinases and the cell cycle Carol Greider, Cold Spring Harbor Laboratory, Telomerases in cell immortalization and cancer 9:00AM - 4:00PM Exhibits Open 9:00AM - 5:00PM Education/Minorities Affairs Information Booth 9:30AM - 10:30AM Complimentary coffee and cookies in Exhibit Hall 9:45AM - 10:15AM Education Coffee Break Forum 10:30AM - 12:00PM SYMPOSIUM II Pattern Formation and Evolution Cynthia Kenyon, University of California, San Francisco (Chair), Pattern formation in C. elegans Sean Carroll, University of Wisconsin/HHMI, Evolution and development of the insect body plan Cliff Tabin, Harvard Medical School, Evolution and development of the vertebrate body plan 11:30AM - 3:00PM Minorities Poster Session and Awards 12:00PM - 3:00PM Poster Presentations 12:30PM - 3:00PM High School Program 2:00PM - 3:00PM E.E. Just Lecture 3:00PM - 3:30PM Refreshment break in Exhibit Hall 3:30PM - 5:45PM Minisymposia 1-6 6:00PM - 7:00PM E.B. Wilson Award Presentation 8:00PM - 9:30PM Exhibitor Tutorials 8:00PM - 10:30PM Film Session 8:00PM - 10:00PM NIH Peer Review
MONDAY, DECEMBER 11 7:30AM - 5:00PM Registration 7:30AM - 9:00PM Posters on Display 7:00AM - 10:00PM Exhibitor Showcase 8:00AM - 10:00PM ASCB/Carl Zeiss, Inc. Run 8:00AM - 9:30AM SYMPOSIUM III Cell Adhesion in Differentiation and Disease Richard Hynes, MIT/HHMI, Integrin and extracellular matrix function in development Mary Beckerle, University of Utah (Chair), Cell adhesion-dependent signalling Judah Folkman, Harvard Medical School, Endogenous inhibitors of blood vessel growth 9:00AM - 4:00PM Exhibits open 9:00AM - 5:00PM Education/Minorities Affairs Information Booth 9:30AM - 10:30AM Complimentary coffee and cookies in Exhibit Hall 9:45AM - 10:15AM Education Coffee Break Forum 10:30AM - 12:00PM SYMPOSIUM IV The Evolution of Eukaryotic Sex Lawrence Hurst, University of Cambridge, Selfish genetic elements and their role in evolution: the evolution of sex and some of what that entails Ursula Goodenough, Washington University (Chair), Sex in simple eukaryotes Robin Lovell-Badge, Medical Research Council, London, Sex determination in mammals: the role and evolution of the sry gene on the Y chromosome 12:00PM - 2:00PM Women in Cell Biology Luncheon 12:00PM - 3:00PM Poster Presentations 1:00PM - 2:00PM NSF/Support for Improving Education in Cell Biology 1:00PM - 2:30PM Graduate Student Symposium 3:00PM - 3:30PM Race awards at Zeiss booth 3:00PM - 3:30PM Refreshment break in Exhibit Hall 3:30PM - 5:45PM Minisymposia 7-12 5:30PM - 6:30PM ASCB Business Meeting 6:30PM - 7:30PM Women in Cell Biology Business Meeting and Awards Presentation 7:30PM - 11:00PM Social, National Museum of Natural History
TUESDAY, DECEMBER 12 7:00AM - 7:00PM Exhibitor Showcase 7:30AM - 5:00PM Registration 7:30AM - 9:00PM Posters on Display 8:00AM - 9:30AM SYMPOSIUM V How Molecular Motors Work Ron Vale, University of California, San Francisco/HHMI (Chair), Mechanical and structural studies of the kinesin motor: are two heads better than one? Ivan Rayment, University of Wisconsin, Structural basis of myosin motility Mary Porter, University of Minnesota, Mutational analysis of dynein regulation 9:00AM - 4:00PM Exhibits open 9:00AM - 5:00PM Education/Minorities Affairs Information Booth 9:30AM - 10:30AM Complimentary coffee and cookies in Exhibit Hall 9:45AM - 10:15AM Education Coffee Break Forum 10:30AM - 12:00PM SYMPOSIUM VI Neuronal Connections: Establishment and Plasticity Marc Tessier-Lavigne, University of California, San Francisco/HHMI (Chair), Guidance of developing axons to their targets Story Landis, Case Western Reserve University School of Medicine, Interactions between pre- and postsynaptic cells in the initial establishment of synaptic junctions Jeff Lichtman, Washington University School of Medicine, Cellular basis of activity- dependent synaptic rearrangements 12:00PM - 3:00PM Poster Presentations 1:30PM - 3:00PM Practice of Science Panel 3:00PM - 3:30PM Refreshment break in Exhibit Hall 3:30PM - 5:45PM Minisymposia 13-18 7:30PM - 8:30PM Fourteenth Keith R. Porter Lecture in Cell Biology 8:30PM - 10:00PM Exhibitor Tutorials
WEDNESDAY, DECEMBER 13 7:30AM - 3:30PM Registration 7:30AM - 3:00PM Posters on Display 8:00AM - 9:30AM SYMPOSIUM VII Membrane Assembly from the Nucleus to the Cell Surface Randy Schekman, University of California, Berkeley/HHMI (Chair), Protein sorting during vesicle budding Douglass Forbes, University of California, San Diego, Nuclear assembly and transport Richard Scheller, Stanford University/HHMI, Mechanism and regulation of membrane fusion 9:00AM - 3:00PM Exhibits open 9:00AM - 12:00PM Education/Minorities Affairs Information Booth 9:30AM - 10:30AM Complimentary coffee and cookies in Exhibit Hall 9:45AM - 10:15AM Education Coffee Break Forum 10:30AM - 12:00PM SYMPOSIUM VIII Understanding and Controlling Cell Biology through Synthetic Molecules Roger Tsien, University of California, San Diego/HHMI (Chair), Dissection of a nitric oxide signalling cascade using caged compounds Heidi Hamm, University of Illinois, Chicago, Use of synthetic peptides to investigate sites and mechanism of G-protein action in signal transduction Stuart Schreiber, Harvard University/HHMI, Chemical approach to understanding and controlling signal transduction 12:00PM - 3:00PM Poster Presentations and Special Poster Session 1:00PM - 1:30PM Refreshment break in Exhibit Hall 1:00PM - 2:00PM NIH/A Walk on the Wild Side 3:30PM - 5:45PM Minisymposia 19-24 6:00PM Meeting ends
On Fri, 29 Sep 1995, Felicity Lawrence wrote:
} G'day. } } I'm looking for information regarding the American Society for Cell Biology. } Could someone please tell me who to contact to get registration and } accomodation details? } } Thanks, } Felicity } EM Unit, QUT. Brisbane, Australia } }
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