Dear Ken, You're right about not being able to legislate common sense. In the case of liquid nitrogen, I have seen LN2 slopping out of styrofoam cups onto peoples' clothes, into instruments, all over papers, etc. For all of the examples everyone can cite of *unnecessary* regulations, I am still amazed to see kitchen microwave ovens in labs being used to heat toxic chemicals, including fixatives and heavy metals, the fumes from which then "ventilate" into the open lab when the door is opened. In some cases, the oven is then used to heat coffee and/or lunch. Steven Slap 75767,640-at-compuserve.com
} Gloves, goggles, masks (and shoes) are actualy more dangerous when handling } liquid N2 than sandles and no protection.
True with the exception that the shutoff valve handle on a big LN2 tank will get cold enough to be dangerous, and does not exhibit the leidenfrost effect. Using a glove or other insulation when turning off the LN2 after filling the dewar makes good sense. } } More seriously, bureaucrats, administrators and the inexperienced should } talk to somebody who has real knowledge. } A great general rule. Reading the manual also helps, and every lab should have a safety manual on hand. Yours, Bill Tivol
I plan to return to some HRP-based histochemistry and thought I would stir up some controversy by asking what you all feel the best precipitating substrate for immunocytochemistry with peroxidase labels. I have always used DAB without metals. What is the disadvantage of using metal intensification? Does anybody have experience with the "stable" solution of DAB (e.g., Pierce Chemicals Metal Enhanced DAB Substrate Kit)? Are they worth the convenience? TIA.
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)
To: Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com} *** Reply to note of 04/01/96 07:19
We do outside analytical work over the complete spectrum of materials, chem lab and product assurance areas. We charge different rates for each instrument and basically different operations. Sample prep, data analysis, and report writing get charged, as well as actual SEM time, for example.
We figure it as "beginning to end." Think about how much time you'd have saved if you didn't have that job at all. Everything must be covered, or someone else is subsidizing it.
I have to tell you about this: after reading the mail on this topic I used the far door to enter our microscope room. I rarely use this door. It has a sign (which I've never noticed before; it must have been there for the last half-year) saying "NOTICE Safety glasses required in this area"
Wil's recent comment on the safety hazards of distilled water brought to mind some peculiar safety regulations here in MD. In reference to liquid N (which can be dangerous stuff), we were first required to wear gloves while handling the stuff. Then came goggles and soon after that, full face shields. The funny thing is, the most dangerous aspect of our handling of LN2 is that students often wear sandals in the summer and are very likely to get stung by droplets. There are no safety measures for feet protection! But now that I have mentioned it, some occupational safety officer listening in will recommend new safety procedures requiring protective booties! In the end, we can't legislate common sense, nor can we abdicate responsibility to those above.
(The opinions above are mine and of anyone who agrees with them.)
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets The Johns Hopkins University Baltimore, Maryland 21218 (410) 516-8342 (voice) (410) 516-7933 (fax) klivi-at-jhu.edu (e-mail)
We had momentary problems with the files on a Mac. We first had to locate them, our Eudora didn't tell us which folder it dropped it in. It ended up in SYSTEM\EUDORA FOLDER\ATTACHMENTS, which we should have guessed.
We could not simply double-click on it to open it. Apparently it did not have the extra information to tell the desktop what kind of file it was. Therefore, we had to first open up Word, then open the document the old-fashioned way through the FILE/OPEN menu. I have found that this is often required and usually works for documents that come through different channels.
Being a Windows man myself, I am used to opening up Word and leaving it running throughout the day so it is a simple matter to switch to Word and open a new document through the pull-down menus. Sometimes the normal associations get lost or naming conventions are not followed. But the pull-dopwns almost always work.
At 09:24 PM 3/31/96 +0000, you wrote: } Arthur, } Thanks for your time and effort--I had a problem opening the Mac file. } Would you please fax me the file? My FAX number is 814-863-1357. } } Rosemary Walsh } EM Facility } The Biotechnology Institute } 519 Wartik Lab } The Pennsylvania State University } University Park, PA 16802 ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
Microscopy List {Microscopy-at-Sparc5.Microscopy.Com}
To Jan van der Meijden
thank you for your mail,
I just checked what you recommend. The displacement is about 20 mm, so within your specifications. Despite it is within Philips specification, this does not allow to work easily from bright field-dark field and diffraction modes, because when working at about 100 000 times (usual magnification rate in order to make BF DF diff images), the movement comes to be 20*20= 400 mm, which is over my own specifications (i.e. the light goes out of the screen therefore I have to get back to 20 000 magnification in order to see where it is...).
The only reasonable way I have found is to press the intensity limit softkey when being near the cross over in order to prevent from going too low in intensity of the C2 lens. This works, but I will welcome with great pleasure another method to get rid of the problem.
On the other hand, during the alignment procedure, due to this shift problem, we do not know exactly how to center correcty the light. I am used to press at each step the normalization button (i.e. photo button), but am amazed because after the whole process (canon alignment) usually the light does NOT get back in the center of the screen. Well maybe it is within specifications, I haven't checked this out.
Thank you for every new input, I am sure that many Philips microscope users are eager to know more about the subject. For this reason I an sending a copy to the list as well.
Sincerely,
Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Luis Sole i Sabaris E-08028 BARCELONA
Tel +34 3 402 16 95 Fax +34 3 402 13 98
On Thu, 28 Mar 1996, Jan van der Meijden wrote:
} Dear Sir } } Via a detour, your request to know the C2 movement when changing C2 } has arrived on my desk. } The procedure we use in the factory is as follows: } Magnification 5800x } Focus C2 and centre the beam. } Fully over focus the beam and focus again. Centre the beam again } Fully underfocus the beam and focus again. Measure the distance from } the centre. This distance should be less than 5cm. } According to your mail this distance on your scope is about 7cm at } 20.000x and thus far within specification. } The problem is caused by the high remanence in the C2 lens and the C2 } lens not perfectly mechanically aligned in the optical axis. } To overcome the problem use one spotsize higher or lower in order to } do a light normalisation. } } Regards Jan van der Meijden } } } } } Yours sincerely } } } } Jan van der Meijden }
Dear Microscopists, Does anyone out there know of a textbook or any other reference material which covers integumental ultrastructure - cuticle, epithelium and associated structures and also oogenesis - in marine invertebrates. We have a marine science student who is studying subclass Ascothoracida who is having problems getting the ultrastructure info she needs.
Thanks in advance.
Richard
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
Hi Brian, I think that most CO2 sensors work by measuring the thermal conductivity or indirectly the heat capacity of the gas mix, this makes them awkward to use in a situation where relative humidity and temperature may fluctuate (since both of these will affect the temperature of the sensor). They also might (I don't know) need a large volume of gas to pass over them to register a change. I would suggest that you might look into three alternative strategies
i) the simplest is to use a cannister of premixed gas, presuming that you want to have a constant pCO2.
ii)Monitor the pH of the medium colorimetrically, assuming the colour change of the medium is mainly due to dissolved CO2, which may not be the case.
iii) Measure the infra-red absorbance of the gas mix, I believe that this method is used in some incubators, but I'm not sure what wavelength to use.
(i) is the easiest and cheapest assuming you don't want to vary the pCO2, if you did you could adapt it by using two bottles, one of CO2 and one of air, these methods might cause humidity regulation problems. (ii) would have to be verified, and if it worked you'd have to build your own probe (I'd have a go if you like), but you would have the benefit of regulating dissolved CO2 not just gaseous. Depending on the absorption spectrum of CO2, it might be hard to find suitable detectors and emitters for (iii).
I hope this helps
Ray
At 2:13 pm 29/3/96, Brian Burgess wrote: } Hi Everyone } We are designing an incubator to fit on a microscope to set the } temperature at 37C and control humidity and maintain a level of } CO2 at 5%. I have had a real hard time locating a CO2 } control/sensing unit. I did find one marketed by Forma } Scientific but it is lunky and quite large. I rather need a } smaller unit. As mentioned before I need to control temperature } and humidity levels. Does anyone know of a good system or a } manufacturer of such a system?
} ---------------------------------------------- } Brian Burgess } Chemical Engineering } University of Florida } e-mail: burgbr-at-che.ufl.edu
Ray Hicks ________________________________________________________________________ |University of Cambridge |Tel 01223 330149 | |Department of Medicine |Fax 01223 336846 | |Level 5, Addenbrookes Hospital |e-mail rh208-at-cus.cam.ac.uk | |Hills Road Cambridge |Web Page/ facsmac.med.cam.ac.uk | |CB2 |ftp server 131.111.80.78 | |UK | | |_________________________________|_____________________________________|
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Title: Biological Science Technician (Plants) Lab: USDA-ARS, Salinas, CA
The USDA-ARS is seeking a biological science technician (plants) (GS-404-7, 8, or 9) for the Crop Improvement and Protection Research Unit in Salinas, CA. The incumbent will share responsibilities in electron microscopy of plant virus infections for ultrastructural characteristics. The incumbent will also assist in research involving molecular, serological, and biological studies of several plant viruses infecting sugarbeet and vegetables. Candidate must have a knowledge of electron microscopy, plant virology, and knowledge of microbiological techniques. Must be a U.S. citizen. Bachelors degree is desirable. Salary is commensurate with experience ($24,610-39,140 per annum). For information regarding research program contact Gail C. Wisler or James E. Duffus (408)755-2835. For information regarding application procedures/forms contact Tom Nelson (408)755-2810. Applications must be postmarked by May 6, 1996. The USDA is an equal opportunity employer.
Bob- Energy Beam Sciences offers a range of stainless steel LN2 lab containers, from 5L up to 220L. The 10L container has a loss rate of .8 liters per day. Please contact me directly by e-mail or telephone (800-992-9037) for part numbers, prices, etc. Steven E. Slap, Vice-President 75767,640-at-compuserve.com
I understand your frustration with all the regulations that just keep coming for hazardous material in the work place, but I have to point out the flip side of this issue. I know of a lab that recently got into trouble because for years the regulators were leaving it up to the head of the lab to make sure precautions were taken. This lab was solely made up of students and the professor/lab head. The professor was leaving everything up to the students to do and learn, giving little if any dirrection to them. These students came from accounting and psycology and english-major backgrounds. They had no experience working in an environment full of hazardous substances, such as a histology lab. They were working with flourescent microscopy, without using the proper filters and hazardous substances without the proper protection, ie gloves, fume hoods. All waste was going down the drain! This is an EXTREME example, I know. I just wanted to show you how bad it can get if some sort of overseeing isn't in place.
I've worked in histology labs for 16 years and I do get tired of having the rules changing about once a year, ie- we have a built-in book self near a fire door. It's been there 3-4 years. Last month the inspectors came through and told us to take it down because it's too close to the door. They've inspected that bookcase every year and not until now was it a problem! So I do know how you feel.
Karen Pawlowski
On Fri, 29 Mar 1996, Kenneth JT Livi wrote:
} Dear Microscopists, } } Wil's recent comment on the safety hazards of distilled water brought to } mind some peculiar safety regulations here in MD. In reference to liquid N } (which can be dangerous stuff), we were first required to wear gloves while } handling the stuff. Then came goggles and soon after that, full face } shields. The funny thing is, the most dangerous aspect of our handling of } LN2 is that students often wear sandals in the summer and are very likely } to get stung by droplets. There are no safety measures for feet } protection! But now that I have mentioned it, some occupational safety } officer listening in will recommend new safety procedures requiring } protective booties! } In the end, we can't legislate common sense, nor can we abdicate } responsibility to those above. } } (The opinions above are mine and of anyone who agrees with them.) } } Kenneth JT Livi } Department of Earth and Planetary Sciences } 34th and Charles Streets } The Johns Hopkins University } Baltimore, Maryland 21218 } (410) 516-8342 (voice) } (410) 516-7933 (fax) } klivi-at-jhu.edu (e-mail) } } }
I have a general question about user fees, in particular how various people bill for hours. We use a "gentle" system with a computer interfaced to the microscope to track real hours, rather than a harsh system where one bills from the minute that the user walks in the door. As a consequence, we have comparatively low hours/week, and I have recently been criticized for this. I would be very interested in comments both from users and administrators about this - should TEM facilities bill like lawyers or not?
} Date: Sun, 31 Mar 1996 11:02:41 +0000 } From: Robert.R.Wise-at-Sparc5.Microscopy.Com } Subject: modems } To: Microscopy-at-Sparc5.Microscopy.Com } X-Sender: wise-at-vaxa.cis.uwosh.edu } MIME-version: 1.0 } Content-type: text/plain; charset="us-ascii" } Content-transfer-encoding: 7BIT } } Although not strictly a microscopy question, the members of this } list seem to know a lot about computers as well. So here goes... } } Does anyone have any recommendations on the proper modem for } connecting a Macintosh (Centris 610 with EtherNet card) at home to a local } phone line? US Robotics has a 28.8 kbps model for about $200. What does } the 28.8 stand for? Is this just a plug in or do I have to get something } like an adapter (such as an Asante' FriendlyNet Media Adapter)? Please } help the computer illiterate. } } Bob Wise
Bob, The speed of a modem is measured by Bit per Second (bps). The 28.8kbps modem will double the transmission speed than the 14.4 kbps one. The 28.8k modem uses error correction and data compression protocols called v.42bis, MNP2-5 for high speed data transmission. The installation is simple: just plug the modem cable (suppose this is a Mac modem) in the serial port marked as MODEM on your Mac, and connect phone line. You need to set up or configurate your communication software before start. EtherNet card is used for network connection, not for modem. If you have any question, please contact me. Good luck. Ya Chen
Ya Chen
Integrated Microscopy Resource (IMR)-- III M M RRRRRR
an NIH Biomedical Research Resource I M M M M R R University of Wisconsin, Madison, WI I M M M RRRRRR 1675 Observatory Drive #167 I M M R R Madison, WI 53706 I M M R R TEL : 608-263-8481 I M M R R
FAX : 608-265-4076 III M M R R
Email:YChen-at-macc.wisc.edu IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html 2nd Symposium on Integrated Microscopy: Sept. 20-22, 1996
Sender: kayton-at-ohsu.edu (Robert Kayton) Message-Version: 2 } To: Microscopy-at-Sparc5.Microscopy.Com
} This information is at least 4 years old and may not be correct. } About 4 years ago (maybe a little longer) there was a lengthy review } article in either New England Journal of Medicine or JAMA regarding } the slow human neurological disorders. At that time, there were at } least 5 recognized diseases that were classified as "Alzheimer" or } "Alzheimer-like". One of these was suspected to be of prion origin, } although by now it may be classified as something else. } } One thing to consider if you are involved in studies involving human } (or any mammalian) brain tissue is that in years past, prion diseases } were commonly misdiagnosed as Alzheimers, and as humans (even } pathologists) are not perfect, this still may be the case.
AD and prion diseases share the presence of amyloid precursor proteins (beta-PP, APrP) but they are distinct diseases, both starting at the synapse (beta-PP and PrP are proteins of the neuromuscular junction and CNS synapse). In some of the prion diseases prion amyloid plaques are seen together with paired helical filaments (NFT) making them "Alzheimer-like" as Budy writes.
********************************************************* 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 *********************************************************
Sender: kayton-at-ohsu.edu (Robert Kayton) Message-Version: 2 } To: Microscopy-at-Sparc5.Microscopy.Com
} As far as I know Alzheimers disease is not infectous---- Where did you get your } information about Alzheimers disease?
This information is at least 4 years old and may not be correct. About 4 years ago (maybe a little longer) there was a lengthy review article in either New England Journal of Medicine or JAMA regarding the slow human neurological disorders. At that time, there were at least 5 recognized diseases that were classified as "Alzheimer" or "Alzheimer-like". One of these was suspected to be of prion origin, although by now it may be classified as something else.
One thing to consider if you are involved in studies involving human (or any mammalian) brain tissue is that in years past, prion diseases were commonly misdiagnosed as Alzheimers, and as humans (even pathologists) are not perfect, this still may be the case.
W. L. Steffens, Ph.D Dept. of Veterinary Pathology College of Veterinary Medicine University of Georgia Athens, GA 30602 STEFFENS.B-at-CALC.VET.UGA.EDU Voice: (706) 542-5536 FAX: (706) 542=5828
Sender: kayton-at-ohsu.edu (Robert Kayton) Message-Version: 2 } To: Microscopy-at-Sparc5.Microscopy.Com
} Has anyone heard of the slow viruses that can survive gluteraldehyde fixation?
The infectious agents that are able to survive fixation are the prions. These agents contain protein only...no nucleic acid. Among these are Creutzfeld-Jakob, scrapie, kuru, Gersten-Straussler-Schenker, BSE (mad cow disease) and apparently some forms of Alzheimers.
It is known that formaldehyde does not inactivate them, not surprising since formaldehyde is one of the least cross-linking of all fixatives, so far as proteins are concerned. My understanding is that glut. does inactivate them, but don't take my word for it.
} On a similar note, there was an MSA-sponsored speaker a couple of years ago who } was encouraging EM labs to make money by offering virus identification sevices } to medical centers. The preparation protocol included advice to accept unfixed } (ie. infectious), unidentified material and prepare negative stained samples on } the bench in the laboratory.
We are a CLIA and State certified human clinical laboratory, licensed for negative-stain virus identification. This has always been part of our function as a veterinary EM lab, and for about 5 years we have offered it as a service to the community. We do make a considerable amount of money from it. We accept samples only from certain reference laboratories and practitioners and these are only stool samples, mostly (} 95%) from infantile diarrheas. Our submission form has an entry for any unusual precautions that must be taken (ie HIV, hepatitis, etc). Our understanding with our clients is that we don't process such samples. W. L. Steffens, Ph.D
Dept. of Veterinary Pathology College of Veterinary Medicine University of Georgia Athens, GA 30602 STEFFENS.B-at-CALC.VET.UGA.EDU Voice: (706) 542-5536 FAX: (706) 542=5828
A few months ago there was a thread on the server dealing primarily with flatbed scanners and the best technology for acquiring digital images. I may have missed out on the part of the conversation dealing with printing digital images, but perhaps one or some of you might summarize any thoughts on what technologies were considered useful.
I run an EM lab in a Pathology department of a medical school. Most of my EM is diagnostic work on kidney specimens and tumors. We printed out 4,500 images for each of the two past years. I would like to go digital, but finding a printer that will handle this load cheaply and quickly, yet with the required high quality is difficult. We plan to produce the usual 3 1/4" X 4" negatives in our scope as we always have. Then we would scan the negatives into a computer (IBM clone), storing them temporarily on a large hard disk, and ultimately archiving them on a writable CD system. The prints need not be durable as once they are studied and the case closed, they would be trashed. The negatives would be archived as would be the image files on the CD's.
The problem is that people are suggesting dye sub for quality and laser for quick & dirty. I need QUICK & QUALITY......and cheap. Printing on a dye sublimation printer would be too costly. There are some enhancement boards that can be added to laser printers, and I have a friend who has printed some of my scanned negatives on his Photoscan system, Philips's image recording system. They come close to answering the problem, but I am wondering what else there is out there. I believe that Philips's system may be LaserPix or PhotoJet Plus from XLI Corp.
I realize that I may be asking you to repeat a conversation already carried out, but if you could pass on the consensus opinions, I would appreciate it.
Joiner Cartwright, Jr., Ph.D.
Director, Electron Microscopy tel.: (713)798-4658 Department of Pathology, Rm.286-A FAX: (713)798-3945 Baylor College of Medicine Internet: joiner-at-bcm.tmc.edu One Baylor Plaza Compuserve: 71555,1206 Houston, Texas 77030 U.S.A.
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
I know that my own safety committee gets under my own skin once in a while.
But in over twenty six years of business, let me relate by far the most serious injury that occurred in our firm. Considering that we work around high voltage, use hazardous chemicals including osmium tetroxide, deal with sometimes hazardous samples, what was our most serious injury? Would you believe it involved Polaroid film?
One of the microscopists (who always claimed eye protection was not needed in an SEM lab) was opening up a case of Type 52 film. The flaps on the case, as it turned out were not creased, as they should have been, therefore the tension present in the fold, when the sealing tape on the top was slit with a razor, literally flew open, cutting the corneal surface of one of the startled technician's eyes. Fortunately sight was not lost in the eye, but there was a scar formed, and there will be the need for lifetime care under a good ophthalmologist and who knows what other impairments might show up with age.
Now in retrospect, nearly twenty years later, I still feel guilty about not enforcing to an even greater degree, perhaps with the threat of being dismissed, our own "rule" that safety glasses should be worn in the laboratory. If an accident is going to happen, generally speaking it will happen when it is least expected.
Now my point only is that some of these safety regulations, superfluous as some of them might sound, often times do have some basis of logic and rationale, my Polaroid film case being, I think, a good example.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
Sender: kayton-at-ohsu.edu (Robert Kayton) Message-Version: 2 } To: MICROSCOPY-at-Sparc5.Microscopy.Com
Sorry, I couldn't resist sharing this... It was sent to me 'as is' with no author. I'm certainly not clever enough to write this.
Ron
I am the Very Model of a Modern Teenage Cyberpunk*
I am the very model of a modern teenage Cyberpunk I rent my own apartment and it's full of electronic junk I own a VAX, a 486, I've even got a PDP I've finished Myst and Doom but I am stumped by Wing Commander III
I'm very well aquainted too with matters pornographical I have a list of image sites, both overseas and national So if you want to see a picture of that Anna Nichole Smith I'll fire up my terminal and fetch for you a naughty GIF
I'm totally an anarchist, the government I'd like to wreck, Though if they were to get blown up, who'd give to me my welfare cheque? In short if you need answers that concern your electronic junk, I am the very model of a modern teenage Cyberpunk
I know the ancient myths about RTM, Pengo and Mitnick I 'hack' into computers and I then perform a credit check I scare all my non-hacker friends with tales of cracker theivery and even though I'm spouting crap they'll listen and believe in me
I've learned to spot a troll and I've seen flames about the way I spell, I've traced badly forged cancels and seen napalm poured on AOL I've laughed at all the newbies and their flailing cries of "You all Suck!" I've been flamed by Carasso, with an anvil I have then been struck
I've hung around in alt.tasteless and seen war waged on rec.pets.cats I've spent my time in talk.bizarre and used those stupid Relay Chats In short, if you need answers that concern your electronic junk, I am the very model of a modern teenage Cyberpunk
Well postings like "MAKE.MONEY.FAST", I am now somewhat wary at, I have been "Global Killfiled" by the Joel Furr Commissariat, When rosebud posts a lengthy rant 'bout Microsoft she swears is true, I know that she is just another short lived kook without a clue
When I have learnt what progress has been made upon the Internet, When I know something more than just a smattering of netiquette, In short when I can have a world-wide soapbox on which I can stand I've got no time for other things, like beer and trips to Disneyland
My life outside the Internet is very very sad you see I cannot get my spots to fade, my social life's a tragedy, But still if you need answers that concern your electronic junk, I am the very model of a modern teenage Cyberpunk.
} Dear Microscopists, } Does anyone out there know of a textbook or any other reference material } which covers integumental ultrastructure - cuticle, epithelium and } associated structures and also oogenesis - in marine invertebrates. We have } a marine science student who is studying subclass Ascothoracida who is } having problems getting the ultrastructure info she needs.
You might browse the "Microscopic Anatomy of Invertebrates" series (Frederick W. Harrison, Burton J. Bogitsh, Eds.), Wiley-Liss, New York, 1991. It=B4s a highly comprehensive series of 15 volumes, covering in-depth ultrastructure from Protozoa up to Hemichordata, Chaetognatha, and else.
-Dietmar-
+++ Dietmar Reiter {dietmar.reiter-at-uibk.ac.at} ++++++++++++++++ +++ Dept. of Zoology and Limnology, University of Innsbruck ++++ +++ Technikerstrasse 25, A - 6020 Innsbruck, Austria +++++++++ +++ ph: (+43)-512-507-6170 (-6161), fax: ..-507-2930 (-2957) +++
Message-ID: {199604021254.HAA27062-at-IndyNet.indy.net} To: "Joiner Cartwright, Jr., Ph.D." {joiner-at-bcm.tmc.edu} , Microscopy list {microscopy-at-Sparc5.Microscopy.Com}
DOT regulations DO NOT require that an MSDS accompany every shipment. DOT does have strict requirements for packaging and labeling (exterior) that are specific to the contents. DOT also requires that shippers of hazardous material have a 24 hour telephone number (their own, or a service) that can answer questions about the material. Both DOT and OSHA require training of personel that handle hazardous materials.
Michael Lamvik asked about moving data from a Kevex 8000 to a PC. Kevex used to sell a package to do this that cost about $1000. This bought you a cable that connected to one of your printer ports on the 8000 and your PC, a diskette with Kermit, software for the 8000, and a very thick manual.
I used it many times to move images from our 8000 to a Macintosh, and it worked fine although it's not fast.
I don't know if Kevex sells this anymore since the 8000 is pretty old. A gentleman named Robert Schaller was very helpful in getting us set up with it, but this was at least 5 years ago and he may or may not still be with Kevex.
Jim Stets Air Products and Chemicals, Inc. Allentown, PA stetsjr-at-ttown.apci.com My opinions are my own, not my employer's.
We have been using the EnVision kit sold by DAKO with great success. This kit does come with a "stable" DAB solution which works very well. Under some conditions when we need to increase the intensity we use the "Quick DAB Enhancer Solution" sold by Innovex Biosciences which works very well and will not change the color of the DAB like nickle chloride.
The usual disclaimer applies to both of the companies mentioned above ie: I'm simply a satisfied customer and recieve no renueration for naming their products
Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis University of North Carolina CB#7400, Rosenau Hall Chapel Hill, NC 27599-7400 Ph. 919-966-6343 Fax 919-966-6123
If you have a digital imaging system it is possible to reduce the number of sources of trouble. Does the distortion show up on the digitally recorded image?
Joe Geller Geller Microanalytical Lab 426e Boston St. Topsfield, MA 01983-1212 508 887-7000
On Mon, 1 Apr 1996, Neuberger, Damian wrote:
} } Hello All: } } We have a JEOL JSM-6300F microscope that exhibits a not-so-slight Y-image } distortion. I hope that I can explain it clearly and that someone out there } may have suggestion(s) as to the source of the problem that we have not been } able to get resolved by field service personnel. } } Materials: (1) SEM; 2) NIST SRM 484f SEM magnification standard; 3) 2000 } copper mesh grid (Pella Cat #631C) that has been calibrated as a secondary } standard with the NIST standard; 4) Polaroid Type 53 film, 4x5". } } Conditions: Image of the mesh grid recorded at X2000, 5 keV, 25 mm WD, } aperture #4 (30 micrometers), probe current setting #8 (1x10-11), camera } aperture setting 5.6, scan rate 80 (sec/scan?). Micrograph is oriented so } that the 5" dimension is along the X-axis. Scan direction top to bottom } (Y-axis). Scan rotation is OFF, tilt correction is OFF, specimen tilt is 0, } SEI mode. } } Problem: The micrograph as well as the viewing monitors exhibit a } distortion in the Y?-axis. That is, the vertical spacing between grid bars } on one end of the X-axis of the photo is about 1.5 to 2 mm greater (out of } 74mm) at the right end than on the left end. This distortion is easily seen } in the image of the grid. (Wouldn't a photo in this email make things a } whole lot easier!) } } Solutions: That's where you knowledgeable microscopists come in. Any ideas } will be gratefully accepted and tested! } } Thanks so much, everyone. } } Damian Neuberger } neuberd-at-baxter.com } } }
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Check out DOUBLE RES IV from Laser Printer Accessories Corp., a board you install into your current laserjet II or III to get 600 dpi quality. We have one and it really does make a BIG difference. 1-800-225-4098.
Walter F. Bobrowski Subcellular Pathology Parke-Davis Pharmaceutical Research Ann Arbor, MI 48105
} Fellow microscopists - } } A few months ago there was a thread on the server dealing primarily with } flatbed scanners and the best technology for acquiring digital images. I may } have missed out on the part of the conversation dealing with printing } digital images, but perhaps one or some of you might summarize any thoughts } on what technologies were considered useful. } } I run an EM lab in a Pathology department of a medical school. Most of my EM } is diagnostic work on kidney specimens and tumors. We printed out 4,500 } images for each of the two past years. I would like to go digital, but } finding a printer that will handle this load cheaply and quickly, yet with } the required high quality is difficult. We plan to produce the usual 3 1/4" } X 4" negatives in our scope as we always have. Then we would scan the } negatives into a computer (IBM clone), storing them temporarily on a large } hard disk, and ultimately archiving them on a writable CD system. The prints } need not be durable as once they are studied and the case closed, they would } be trashed. The negatives would be archived as would be the image files on } the CD's. } } The problem is that people are suggesting dye sub for quality and laser for } quick & dirty. I need QUICK & QUALITY......and cheap. Printing on a dye } sublimation printer would be too costly. There are some enhancement boards } that can be added to laser printers, and I have a friend who has printed } some of my scanned negatives on his Photoscan system, Philips's image } recording system. They come close to answering the problem, but I am } wondering what else there is out there. I believe that Philips's system may } be LaserPix or PhotoJet Plus from XLI Corp. } } I realize that I may be asking you to repeat a conversation already carried } out, but if you could pass on the consensus opinions, I would appreciate it. } } } Joiner Cartwright, Jr., Ph.D. } } Director, Electron Microscopy tel.: (713)798-4658 } Department of Pathology, Rm.286-A FAX: (713)798-3945 } Baylor College of Medicine Internet: joiner-at-bcm.tmc.edu } One Baylor Plaza Compuserve: 71555,1206 } Houston, Texas 77030 U.S.A. }
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At 08:11 AM 4/2/96 -0400, you wrote:
} Check out DOUBLE RES IV from Laser Printer Accessories Corp., a board you } install into your current laserjet II or III to get 600 dpi quality. We have } one and it really does make a BIG difference. 1-800-225-4098. } } Walter F. Bobrowski } Subcellular Pathology } Parke-Davis Pharmaceutical Research } Ann Arbor, MI 48105 } } TEL: 313-996-7814 } FAX: 313-996-5001 } E-Mail: BOBROWW-at-AA.WL.COM
**************************** Walter -
Is the 600 dpi output of this system better than the 600 dpi output of the LaserJet 4 that we already have? There are boards that hot-rod the LaserJet 4 up to "2,400 equivalent dpi". One wonders if they will show "equivalent" deposits in my kidney specimens. Thank you for your reply.
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Scott - Here is a reply that may interest you. Joiner
At 09:35 AM 4/2/96 -0500, you wrote: } In response to your recent message posted on the newsgroup concerning } digital printers, there are several options. The first going to laser } printers, and the second being dye sub printers. The cost of laser is } obviously cheap, but not high enough quality perhaps for publication or } reports where a photo is required. Dye sub printers ex: sony makes two } digital dye sub printers. The first being model UP-D1500CN, it ouputs a } picture size of approx. 3 5/8" x 4 3/4" which is near photographic quality. } This printer sells for approx. $ 1,850.00 US and the print cost is approx. } .86 cents. for color images. The other printer from sony is the UPD-8800 } with Interface card is approx. $7,300 US and the print cost for 8.5" x 11" } is approx. $ 2.20 These printers will ouput a print in approx. 60 seconds. } I do have sample prints if you require to see some. } } Sony makes a digital thermal printer that outputs approx. a polaroid size } image on thermal paper in about 3.5 seconds, cost per print is approx. 9 cents. } } Epson makes some low cost printers, with a decent low cost image, the only } problem with laser or ink jet with these images is that it takes quite a } while to print. } } I don't know if I was helpful or not, but if I can be of any assistance, } please feel free to contact me at your convenience. } } } Best Regards, } } Philip Slakmon } Scott Scientific } } } _______________________________________________ } SCOTT SCIENTIFIC } P.O. Box 66552 Station Cavendish, } Montreal, Quebec, H4W 3J6, Canada } } Telephone: 514-485-2309 Fax: 514-485-9931 } Voice Mail: 514-888-6509 } } WWW Site: http://www.ScottScientific.com } } E-Mail: slakmon-at-scottscientific.com } info-at-scottscientific.com } sales-at-scottscientific.com } admin-at-scottscientific.com } _________________________________________________ } } ********************************** Philip -
Thank you for your reply. Dyesub, as you see, may be capable of turning out images of good enough quality to see, for example, the subtle ultrastructural changes in kidney disease, but not at a price that we can afford. And laser, although fast and cheap, cannot produce the image quality necessary to show these subtle changes. One or more of the laser enhancement boards MAY be the answer, but I am not yet convinced.
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At 01:51 PM 4/2/96 EST, you wrote:
} I run a similar facility in the Pathology Department of a Veterinary } College, but we also serve the entire College as well as some other } parts of the university. } With our SEM, we are entirely digital now...
***************** Dr. Steffens -
Thank you for your reply. The savings are one of the main reasons that I want to go digital. That includes savings in time as well as money. I am considering the LaserPix. However there are some other technologies out there that I want to look at as well.
Kevex still offers the 8000 to PC communication package using Kermit. There is also a high speed data transfer option available using a SCSI interface card. This is especially useful for transferring large image files, etc. More information is available from Kevex Customer Service at +1 (415) 591-3600.
- Jeff Allbright, Tokyo
Michael Lamvik asked about moving data from a Kevex 8000 to a PC. Kevex used to sell a package to do this that cost about $1000. This bought you a cable that connected to one of your printer ports on the 8000 and your PC, a diskette with Kermit, software for the 8000, and a very thick manual.
I used it many times to move images from our 8000 to a Macintosh, and it worked fine although it's not fast.
I don't know if Kevex sells this anymore since the 8000 is pretty old. A gentleman named Robert Schaller was very helpful in getting us set up with it, but this was at least 5 years ago and he may or may not still be with Kevex.
Jim Stets Air Products and Chemicals, Inc. Allentown, PA stetsjr-at-ttown.apci.com My opinions are my own, not my employer's.
Kevex still offers the 8000 to PC communication package using Kermit. There is also a high speed data transfer option available using a SCSI interface card. This is especially useful for transferring large image files, etc. More information is available from Kevex Customer Service at +1 (415) 591-3600.
- Jeff Allbright, Tokyo
Michael Lamvik asked about moving data from a Kevex 8000 to a PC. Kevex used to sell a package to do this that cost about $1000. This bought you a cable that connected to one of your printer ports on the 8000 and your PC, a diskette with Kermit, software for the 8000, and a very thick manual.
I used it many times to move images from our 8000 to a Macintosh, and it worked fine although it's not fast.
I don't know if Kevex sells this anymore since the 8000 is pretty old. A gentleman named Robert Schaller was very helpful in getting us set up with it, but this was at least 5 years ago and he may or may not still be with Kevex.
Jim Stets Air Products and Chemicals, Inc. Allentown, PA stetsjr-at-ttown.apci.com My opinions are my own, not my employer's.
} I run an EM lab in a Pathology department of a medical school. Most of my EM } is diagnostic work on kidney specimens and tumors. We printed out 4,500 } images for each of the two past years. I would like to go digital, but } finding a printer that will handle this load cheaply and quickly, yet with } the required high quality is difficult. We plan to produce the usual 3 1/4" } X 4" negatives in our scope as we always have. Then we would scan the } negatives into a computer (IBM clone), storing them temporarily on a large } hard disk, and ultimately archiving them on a writable CD system. The prints } need not be durable as once they are studied and the case closed, they would } be trashed. The negatives would be archived as would be the image files on } the CD's. } } The problem is that people are suggesting dye sub for quality and laser for } quick & dirty. I need QUICK & QUALITY......and cheap. Printing on a dye } sublimation printer would be too costly. There are some enhancement boards } that can be added to laser printers, and I have a friend who has printed } some of my scanned negatives on his Photoscan system, Philips's image } recording system. They come close to answering the problem, but I am } wondering what else there is out there. I believe that Philips's system may } be LaserPix or PhotoJet Plus from XLI Corp. } I run a similar facility in the Pathology Department of a Veterinary College, but we also serve the entire College as well as some other parts of the university. With our SEM, we are entirely digital now...I don't even support film on it. With the TEM, we have a side-mounted digital camera system (JEOL TV-CAM) that is used for image acquisition, but we still use film for publication purposes. Most of the TEM images are for study purposes only, and for this we dump the image (using the system's software) to a Laserjet 4 printer through a Laserpix board. After a bit of processing, ie BCG adjustment, unsharp mask sharpening, etc., you get a "work print" that is usually acceptable. For higher quality, we also have a Codonics dye sublimation printer, on which you can put 4 images per page. For TEM, its certainly better quality than the Laserjet, but is usually not publication quality unless you do some intensive processing. SEM and light micrographs on it however are certainly publication quality. Since most images we take will not be published, we get along fine with the Laserjet prints for TEM. We do use a high quality glossy paper in it which drives up the cost of the prints to about .12 each. That still beats the 2.00+ of the Codonics. Since going mostly digital, our TEM film images have dropped from about 2000 per year to about 200 per year. The savings in tech labor, direct costs, and chemical wastes are quite substantial.
W. L. Steffens, Ph.D Dept. of Veterinary Pathology College of Veterinary Medicine University of Georgia Athens, GA 30602 STEFFENS.B-at-CALC.VET.UGA.EDU Voice: (706) 542-5536 FAX: (706) 542=5828
} } Dear Microscopists, } } Does anyone out there know of a textbook or any other reference material } } which covers integumental ultrastructure - cuticle, epithelium and } } associated structures and also oogenesis - in marine invertebrates. We hav= e } } a marine science student who is studying subclass Ascothoracida who is } } having problems getting the ultrastructure info she needs. } } You might browse the "Microscopic Anatomy of Invertebrates" series } (Frederick W. Harrison, Burton J. Bogitsh, Eds.), Wiley-Liss, New York, } 1991. It=B4s a highly comprehensive series of 15 volumes, covering in-depth } ultrastructure from Protozoa up to Hemichordata, Chaetognatha, and else. } } -Dietmar-
As well as this ref, there is:
Neville, A.C. 1975. _The Biology of the Arthropod Cuticle_. Vol, 4/5 in the series "Zoophysiology and Ecology". Springer-Verlag.
Nothing on Ascothoracida in specific, but a good general ref, and literature review up to about 1974. Phil
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
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I recently posted a query concerning printers suitable for producing high quality monochrome images from micrographs scanned into a computer....a printer that would allow me to "go entirely digital" and get me out of the darkroom, at least for all imaging except the most demanding applications. I have received a number of replies. Boiled down, the answers include:
1.) For quick & dirty, but usable work prints, a laser printer producing 600+ dpi;
2.) For high quality, a dye sublimation printer; and
3.) An intermediate would be a laser printer with one of the after market enhancement boards that runs the lasers resolution up to 1,200 dpi or "2,400 dpi equivalent", whatever that means;
4.) A fourth suggestion two or three people made was the Harris PhotoPro 2000 Gray Scale Digital Printer. This is a $10,000.00 silver salt laser printer that claims only 256 dpi, but each dot has 256 gray levels available. It was stated that the cost/print was less than $1.00.
Do any of you have any experience with this latter machine, and would you be willing to share your thoughts? Does the increased contrast resolution make up for the POSSIBLY low spatial resolution? Do I really want to spend ten big ones on a black & white printer?
Joiner Cartwright, Jr., Ph.D.
Director, Electron Microscopy tel.: (713)798-4658 Department of Pathology, Rm.286-A FAX: (713)798-3945 Baylor College of Medicine Internet: joiner-at-bcm.tmc.edu One Baylor Plaza Compuserve: 71555,1206 Houston, Texas 77030 U.S.A.
In response to last years questions raised on the performance of digital printers I tested several printers with a 1Kx1K image. Printer performances were compared at the level of whole page images and 10x enlargements of the printed images (LaserJet, Lazar Print, Dye sub,). Results are available at
In conclusion, you have to ask how many lines per inch are printed and at how many gray levels (not always the same as dots per inch), plain paper or special paper, permanence of the print, printing time, printing costs. I print 300 lpi at 256 gray levels (4,800 dpi) on plain paper in 20-45 sec for 1-10 MB of images per page at 3 cents per page with a HP LaserJet and enhancer board in archival quality.
Best regards Klaus
*****************************************
} At 08:11 AM 4/2/96 -0400, you wrote: } } } Check out DOUBLE RES IV from Laser Printer Accessories Corp., a board you } } install into your current laserjet II or III to get 600 dpi quality. We have } } one and it really does make a BIG difference. 1-800-225-4098. } } } } Walter F. Bobrowski } } Subcellular Pathology } } Parke-Davis Pharmaceutical Research } } Ann Arbor, MI 48105 } } } } TEL: 313-996-7814 } } FAX: 313-996-5001 } } E-Mail: BOBROWW-at-AA.WL.COM } } **************************** } Walter - } } Is the 600 dpi output of this system better than the 600 dpi output of the } LaserJet 4 that we already have? There are boards that hot-rod the LaserJet } 4 up to "2,400 equivalent dpi". One wonders if they will show "equivalent" } deposits in my kidney specimens. Thank you for your reply. } } } * * Joiner Cartwright, Jr. * *
Could someone tell me if there is any company delivering mechanical point counter systems. I need it for a polarizing microscope for geological samples. Olli Taikina-aho University of Oulu Institute of Electron Optics Box 400 SF-90571 Oulu ota-at-cc.oulu.fi tel. +358-81-5533142 fax. +358-81-5533149
Three-Dimensional Microscopy: Imaging Acquisition and Processing IV
Conference Chairs: Carol J. Cogswell, Univ. of Sydney (Australia); Jose-Angel Conchello, Washington Univ.; Tony Wilson, Univ. of Oxford (UK)
Program Committee: Alan Bearden, Univ. of California/Berkeley; G. J. Brakenhoff, Univ. of Amsterdam (Netherlands); Kjell Carlsson, Royal Institute of Technology (Sweden); ; Seth Goldstein, National Institutes of Health; Gordon S. Kino, Stanford Univ.; Andres Kriete, Univ. Giessen (FRG); Frederick Lanni, Carnegie Mellon Univ.
This conference will explore the rapidly developing field of three-dimensional microscopy. Consideration will be given to the characteristics of the overall system design, as well as to the topics of image formation, image recording, deconvolution in 2,3 or more dimensions, and digital methods of producing and displaying the resulting 3D reconstruction. Recent innovations in 3D microscopy are having a large impact in the biological and medical fields, as well as in materials science and the semiconductor industry. Many microscopes are now fully integrated systems, including computer hardware and software. It is hoped that the broad range of relevant topics being presented at this symposium will serve to encourage interaction among instrumentation engineers, computer image analysts, and researchers in the various fields of application.
Papers are invited in the following and related areas: - confocal microscopy - 3D image formation - instrumentation for 3D microscopy - time-resolved image acquisition systems - image processing and analysis - 3D image reconstruction - deconvolution in 2/3 or more dimensions - super-resolution - spatiotemporal reconstruction of living cells and tissues - applications of 3D microscopy in materials science - profilometry - image visualization techniques for 3D microscopy systems, including volume rendering, animation, stereoscopic and holographic displays
This conference is just one of 27 topics to be held at the BiOS '97 symposium, which is part of the larger event called Photonics West '97: 8-14 February 1997 San Jose Convention Center San Jose, California USA
TO OBTAIN ALL CALLS FOR PAPERS ELECTRONICALLY The calls for papers for all conferences in the BiOS '97 symposium (as well as all other symposia that are part of Photonics West) will be available 1 May on the SPIE Web site http://www.spie.org/web/meetings/calls/submissions.html; by anonymous FTP ftp://spie.org/meetings/calls/PW97_readme.txt; or by e-mail file retrieval send a message to info-spie-request-at-spie.org with the following in the message body: send [meetings.calls]PW97_readme.txt
For a printed BiOS '97 call for papers or other information: E-mail: PW97-at-spie.org Fax: 360/647-1445 Phone: 360/676-3290
IMPORTANT DEADLINES Paper Abstracts Due from Authors: 15 July 1996 (post-meeting proceedings) Advance Programs due from Chairs: 12 August 1996 (post-meeting proceedings) Camera-Ready Abstracts due from Authors: 9 December 1996 Manuscripts Due from Authors: 13 January 1997 (post-meeting proceedings)
GUIDELINES FOR SUBMITTING AN ABSTRACT
Send a 250 word abstract of your paper, by 15 July 1996, in ONE of the following ways:
* SPIE WEB - Complete the convenient form found at SPIE Web site: http://www.spie.org/web/meetings/calls/submissions.html
* or E-MAIL each abstract separately to: abstracts-at-spie.org in ASCII text (not encoded) format. To ensure receipt and proper processing of your abstract, write on SUBJECT line: PW '97 and Conference Chair.
Example: SUBJECT: PW97 (John Smith)
* or MAIL three copies of your abstract to: BiOS '97 SPIE, P.O. Box 10, Bellingham, WA 98227-0010 USA Shipping address: 1000 20th St., Bellingham, WA 98225 USA Telephone 360/676-3290
* or FAX one copy to SPIE at 360/647-1445 (send each abstract separately).
Be sure each abstract includes the following:
1. CONFERENCE CHAIR and CONFERENCE TITLE (submit to ONLY ONE conference) to which the abstract is submitted
2. AUTHOR LISTING (List principal author first) for each author: full name and affiliation, mailing address, phone/fax numbers, email
3. ABSTRACT/PAPER TITLE
4. ABSTRACT TEXT: 250 words
5. KEYWORDS: maximum of 5 keywords
6. BRIEF BIOGRAPHY of the principal author: 50-100 words
Please contact SPIE if you have any questions or require further information.
Marilyn E. Gorsuch, Technical Programs Manager SPIE PO Box 10 1000-20th Street Bellingham, WA 98225 Ph: 360/676-3290 Fax: 360/647-1445 e-mail: marilyn-at-spie.org
Like many of you , I have worked in EM laboratories with water (fire activiated) sprinklers overhead. In some cases I have successfully lobbied to have them removed, as I felt there was a greater danger of electrical shock or massive instrument damage if they tripped in error (beware the pipefitter). However, there is always the possibility that your EM could be the source of a fire that causes great damage and injury. How do people feel about this?
Olli Taikina-aho wrote: } } Hello friends } } Could someone tell me if there is any company delivering mechanical point } counter systems. I need it for a polarizing microscope for geological samples. } Olli Taikina-aho } University of Oulu } Institute of Electron Optics } Box 400 } SF-90571 Oulu } ota-at-cc.oulu.fi } tel. +358-81-5533142 } fax. +358-81-5533149Yes. Prior Scientific, Inc of Rockland, MA sells an electro-mechanical pointcounter. It has a number of nice features and is fairly simple. Give them a call at 617-878-8442 (FAX 617-878-8736).
John M. Libert OPELCO OPtical ELements COrporation Sterling, VA. 703-471-0080 Ext. 217
I apologize for posting this to the entire community, but I no longer have the majordomo address to unsubscribe nor the proper procedure. Please e-mail me directly to reduce bandwidth clutter.
Several people have mentioned Halon to me, but I was under the impression that Halon has fallen out of favor due to potential toxicity and suffocation.
} Like many of you , I have worked in EM laboratories with water (fire } activiated) sprinklers overhead. In some cases I have successfully lobbied } to have them removed, as I felt there was a greater danger of electrical } shock or massive instrument damage if they tripped in error (beware the } pipefitter). However, there is always the possibility that your EM could } be the source of a fire that causes great damage and injury. How do people } feel about this? } } David Rothbard } } -- } Institute of Paper Science and Technology } Our lab is equipped with a Halon fire suppression system. I can't verify that it works, as we have never needed it :)
We are using LR white for the first time in anticipation of doing some immunocytochemistry and also because I have several people who are working with bacteria and fungi. We have been fixing the bacteria in 2 + 2, trying different concentrations of buffer because the bacteria grows in a 25% sodium chloride. Then we mix the bacteria with agar, fix again, osmicate, and dehydrate to 70%. Then we mix the LR White with 70 % for infiltration, then straight LR White finally flat embed in LR White and polymerize at 60 degreesC. We wrap the embedding molds in Saran wrap (not an off-brand) before polymerization. The blocks in blue JB4 molds hardened, but have a surface stickiness. The blocks in the clear molds disappeared. The black blocks of osmicated agar are there but the epoxy is gone. Does anyone have any thoughts on this? I really like running up nice blocks of pancreas and kidney and even cardiac muscle in epoxy and taking pretty pictures in the microscope but unfortunately no-one here is doing this nice rewarding basic work.
Applications are invited for a postdoctoral fellowship on a 3-year project investigating computer modelling of macromolecular structures in plant cells. Part of this work requires preparation of TEM and immunofluorescence micrographs for comparison with computer generated images of plant cytoskeleton and plasmodesmata. We are also investigating various aspects of the interactions between these cell components and their dynamic behaviour in cells. Some confocal fluorescence microscopy may be required.
Applicants should have experience in TEM and immunofluorescence of plant material. Experience in freeze-substitution techniques and immunofluorescence of the plant cytoskeleton would be an advantage.
The position is available from 1 July, and the successful candidate will be expected to take up the appointment by 1 September 1996 at the latest. Salary will commence at $(Australian)37,345. The appointment will be for 12 months in the first instance, renewable for a further 12 months (longer if new grant application is successful). Some remuneration for removal expenses will be available.
Applications should arrive by 4 May 1996, with CV naming 2 referees.
For further information, contact
Rosemary White __ / Department of Ecology _/ \__/ \ and Evolutionary Biology / \ Monash University / Australia \ Clayton, Victoria 3168 \ ____ / phone 61-3-9905 5670 \_/ \_*_/ fax 61-3-9905 5613 __ email rgwhite-at-vaxc.cc.monash.edu.au \/
I was very impressed with the ChemExpo www site that was recommended here recently. I wonder whether there is something similar from antibody producers that would enable to locate a source of a particular antibody easily. My search for this was unsuccessful. Thanks for your help.
Sarka Lhotak EM Facility, McMaster University Hamilton, Ontario, Canada lhotaks-at-fhs.mcmaster.ca
Like many of you , I have worked in EM laboratories with water (fire activiated) sprinklers overhead. In some cases I have successfully lobbied to have them removed, as I felt there was a greater danger of electrical shock or massive instrument damage if they tripped in error (beware the pipefitter). However, there is always the possibility that your EM could be the source of a fire that causes great damage and injury. How do people feel about this?
A few years ago, we had a very bad rainstorm in Southern California. The roof of our R&D building (which houses the SEM) leaked, and the ceiling tile got so soaked that it fell in on the SEM console. When I came in the next morning,
there was a steady stream of water pouring into the keyboard and down into the electronics of the SEM. Obviously, the SEM had tripped off. I unplugged it and called the vendor (Cambridge, at the time), since we were under a service contract. End result - we allowed it to dry for 3 days and fired it up. It
worked fine, and has been operational ever since; no problems have been attributed to this incident. In summary, I believe that the risks associated with fire far outweigh the risks associated with water.
I haven't found anything on the net but an excellent (and not to expensive ) refferance is "Linscotts Directory of Immunological and Biological Reagents" the send out constant updates and is available in written and PC compatible form. The phone # for additional information is 707-544-9555.
regards, bob
Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis University of North Carolina CB#7400, Rosenau Hall Chapel Hill, NC 27599-7400 Ph. 919-966-6343 Fax 919-966-6123
I maintain an archive of most of the biological EM related material that comes across this list. Since you have net acess I will assume you can get acess to the www. Go to the web page listed at the bottom of this message and click of the "Tips & Tricks Wizard" In there is a file called "Embedding with LR White" which may be of use to you. If for some reason you do not have acess to the web or cannot read the file, let me know and I will e-mail it to you .
At 12:04 PM 4/3/96 -0600, you wrote: } We are using LR white for the first time in anticipation of doing some } immunocytochemistry and also because I have several people who are } working with bacteria and fungi. } We have been fixing the bacteria in 2 + 2, trying different } concentrations of buffer because the bacteria grows in a 25% sodium } chloride. Then we mix the bacteria with agar, fix again, osmicate, and } dehydrate to 70%. Then we mix the LR White with 70 % for infiltration, then } straight LR White finally flat embed in LR White and polymerize at 60 } degreesC. } We wrap the embedding molds in Saran wrap (not an off-brand) before } polymerization. } The blocks in blue JB4 molds hardened, but have a surface stickiness. } The blocks in the clear molds disappeared. The black blocks of osmicated } agar are there but the epoxy is gone. } Does anyone have any thoughts on this? } I really like running up nice blocks of pancreas and kidney and even } cardiac muscle in epoxy and taking pretty pictures in the microscope but } unfortunately no-one here is doing this nice rewarding basic work. } }
Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 904-392-1295 ICBR EM Core Lab fax 904-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/
Following my request for more details, Ms Kaye Patterson {kayep-at-deakin.edu.au} has sent me more information about a book I think can be very interesting for many people. For its possible general interest, I send this message to the list. Greetings George ---------------------------------------------------------- George, ... A company in America called Ward's Biology has a very large catalogue which they send to us here in Australia every year. Their address is P.O. Box 92912 Rochester, NY 14692-9012. For International Customers: Fax: 716-334-6174 Phone 716-359-2502. They have a large selection of Biological books, manuals, guides, etc. which you would probably find useful, as well as models, equipment, charts, microscope slides etc. etc. etc. They have the book you are asking about available for US$65.00, Catalogue No. 32 W 0902. The full details of the book are: Morholt, Evelyn and Brandwein, Paul F. (1986) A Sourcebook for the Biological Sciences. 3rd ed. The publisher of my 2nd ed. copy is Harcourt, Brace & World, Inc. New York, but it was published in 1966 (and also has Alexander Joseph as a third author), so I don't know what the publishing details for the 3rd ed. are. The Ward's catalogue says of the book "No biology teacher should be without this comprehensive sourcebook of techniques, procedures, demonstrations, projects, experiments and support material. Also includes sections on maintaining organisms, safety and chemical preparations. Hardcover, 813 pages, with illustrations." I can only comment about the book as a biology technician and not as a teacher. I find it useful as the book details activities and experiments which our Uni students do, along with the practical aspects which I need to get the classes ready. eg. for classes looking at Protozoans, invertebrates, etc., the book describes the best way to get material from the field, how to culture the various groups, how to prepare material for microscopic examination, anesthetization, staining and fixation techniques. The book covers many areas of Biology - anatomy, cells & tissues, photosynthesis, ingestion & digestion, transpiration, circulation, respiration, behaviour & coordination, genetics, growth & differentiation, evolution, and ecology. There is a section on special techniques - Maintaining animals useful in the classroom, Growing plants useful in the classroom and Stockroom and facilities for biology. This is the section I use the most. Hope you find all this useful. Kaye
Halon is great for stopping fires but yes, the same mechanism that extinguishes the fire will suffocate people. Sprinklers are generally OK but I prefer a standard A-B-C rated fire extinguisher in the scope room. We have had "umbrellas" mounted over the scopes for the last 20 years- aluminum tube framework covered by a heavy-duty waterproof nylon fabric. We reside in an old building and very occasionally, plumbing accidents result in water leaking throught the ceiling above the scope. Regards,
Doug Davis Staff Research Associate Electron Microscope Facility University of California Berkeley, CA 94720 (510) 642-2085, fax 643-6207 dbd1-at-uclink4.berkeley.edu
I would venture that analog instruments might be able to weather the onslaught of water better than newer instruments with lots of sensitive electronics. The best approach would be to contact computing centers and see what measures are taken to protect their instrumentation.
Secondly, some sort of risk assessment needs to be considered here. What are the odds that floods will be generated needlessly versus having a facility or instrumentation destroyed by fire? Most instances of fires affecting EM units that I have heard about were caused by fires originating other than in the EM room. Perhaps sprinklers could be used in locations other than the EM while the EM rooms could be protected by more electronically-friendly agents (Halon, carbon dioxide, etc).
Just my thoughts .....
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
I would venture that analog instruments might be able to weather the onslaught of water better than newer instruments with lots of sensitive electronics. The best approach would be to contact computing centers and see what measures are taken to protect their instrumentation.
Secondly, some sort of risk assessment needs to be considered here. What are the odds that floods will be generated needlessly versus having a facility or instrumentation destroyed by fire? Most instances of fires affecting EM units that I have heard about were caused by fires originating other than in the EM room. Perhaps sprinklers could be used in locations other than the EM while the EM rooms could be protected by more electronically-friendly agents (Halon, carbon dioxide, etc).
Just my thoughts .....
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
} David; } } A few years ago, we had a very bad rainstorm in Southern California. The roof } of our R&D building (which houses the SEM) leaked, and the ceiling tile got so } soaked that it fell in on the SEM console. When I came in the next morning, } there was a steady stream of water pouring into the keyboard and down into the } electronics of the SEM. Obviously, the SEM had tripped off. I unplugged it } and } called the vendor (Cambridge, at the time), since we were under a service } contract. End result - we allowed it to dry for 3 days and fired it up. It } worked fine, and has been operational ever since; no problems have been } attributed to this incident. In summary, I believe that the risks associated } with fire far outweigh the risks associated with water.
Yes, but rainwater is clean. I have been told (third- or fourth-hand story by now) that it is not the water in sprinkler systems as much as the rust, junk and (perhaps) rust inhibitors in the pipes that cause the major damage.
} Like many of you , I have worked in EM laboratories with water (fire } activiated) sprinklers overhead. In some cases I have successfully lobbied } to have them removed, as I felt there was a greater danger of electrical } shock or massive instrument damage if they tripped in error (beware the } pipefitter). However, there is always the possibility that your EM could } be the source of a fire that causes great damage and injury. How do people } feel about this? } Dear David, We have had one fire in an EM room--started when the LN delivery system ran dry and a solenoid valve shorted out. There are definitely other potential fire dangers, such as vacuum pumps. We do not have sprinklers. I have not heard of erronious tripping of sprinkeler sys- tems being a particular problem. All-in-all, I lean toward having a fire supression system, but there are obvious problems with a water- based one, even if it trips due to a real fire. Is there a better system for areas where large voltages and/or currents are present? Perhaps some industries have solved this problem. Yours, Bill Tivol
With regard to the thread about fire extinquishers in EM suites, we were told during our last safety inspection that we would need to get rid of our halon system the next time it needed a charge. This was mostly because the insurance companies would no longer cover such things. They require springler systems. They would rather pay the for lost equipment than the hazards of halon. Of course our safety officer had no answer to my question of whether the school would pay the 5,000 deductible or whether I would have to pay this.
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-bgsm.edu * **************************************************************
Greetings, I've read with interest the comments of others questioning liquid nitrogen safety regulations. In spite of many years of pratical experience with the stuff and being a trained safety officer, I took some time to examine the subject. I disagree with those who have problems with the regulations. The regs seem to be aimed at preventing continuous contact and the resultant injuries. In the CRC Handbook of Laboratory Safety, 3rd edition, pages 315-317, lists all the basic precautions (face shield, impervious clothing, etc.). For those who disagree, gloves are optional.....except, as Tivol pointed out, when in contact with cold metal parts. The CRC actually suggests using some sort of potholder to protect hands from the metal. The CRC also points out that efforts should be made to prevent trapping the cryogen. When trapped against the skin, LN2 will cause injury since the contact is no longer momentary and becomes continuous contact. Cuffless pants worn over the shoes/boots are reccommended, i.e.spilled LN2 that gets trapped inside a shoe with the foot may cause injury. For those who suggest sandals as appropriate should think about the possibility that with a major spill, some LN2 could be trapped beneath the foot or under the strap of a sandal, possibly causing injury. Appropriate shoes can protect the foot for short periods. If the footwear is frozen, generally it can be removed before injury occurs. If gloves are used, the fit should be loose so that they may be shed quickly...ala a hockey player's actions when he has been treated less than courteously by an opposing player. The problem I have with faculty and staff is the use of latex gloves while using LN2. The Leidenfrost effect can definitely cause injury if the latex freezes against the skin. For those who would not wear appropriate gloves, have you considered the possibility of LN2 being trapped under a watch band, or under a ring? In a major splash, could LN2 become trapped at the beltline, between the pants and shirt? The whole idea is to make sure that most any cryogen does not get trapped against the skin. If you can handle LN2 safely without protection, or have done it for years without incident, more power to you. I hope your luck holds out. Interestingly enough, Air Liquide, a national supplier of cryogens, also lists similar precautions in the use of liquid nitrogen and other cryogens. Texas Tech has (arguably) a large number of beautiful women, but they are not the ones coming to get liquid nitrogen....and of the individuals who do get LN2 from our lab, I would rather them come clothed.
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
The sprinkler next to your scope may not be the problem. We are on the second floor, but when the sprinklers went off on the sixth floor, water eventually worked its way down to us. Luckily this happened in the daytime and we had enough time to cover done the scopes and computers. No one knew how to turn off the water so there was 6 inches on the 6th floor and ceilings collapsed on the 5th floor exactly as they are designed to do during a fire. There was no fire. -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: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 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-
David, We had a great central laboratory in the basement of the Graduate School with a fire sprinkler above each of three electron microscopes. The Dean worried that the sprinklers would ruin the microscopes, so they got insurance on them and required that I keep large plastic tarps to cover them in case the sprinklers went off. They never did, howwever... One Saturday, we had a huge rainstorm and the Dean's secretary called me saying their seemed to be a "little problem in the lab." I rushed over there to find that the drains at the top of the stairs had filled with leaves and likewise the drain at the bottom and huge amounts of water, like a waterfall, was cascading down the steps and filling up the lab. The building power had gone off and the emergency lighting was on in the hallways. I called for the maintenance crew, but they were priortized to go save "expensive" student computers in another building. So there I was with my broom, frantically cleaning drains and pushing water, feeling like a character in Walt Disney's Fantasia. Because of this long and intense fight, the only thing we lost was the carpeting, papers and books on the lowest shelf. Saved the floor tile. The minimal rust on the feet of the EM's wasn't even noticeable. The lab was fine. Sprinklers a non-issue. The real devastation was done by the administration, later, when a new Dean said "What are electron microscopes good for?" eyeing the space and salary line he could use for a new Executive Secretary. But that's another story. Kind regards and good wishes to all, Beverly
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David, We had a great central laboratory in the basement of the Graduate School with a fire sprinkler above each of three electron microscopes. The Dean worried that the sprinklers would ruin the microscopes, so they got insurance on them and required that I keep large plastic tarps to cover them in case the sprinklers went off. They never did, howwever... One Saturday, we had a huge rainstorm and the Dean's secretary called me saying their seemed to be a "little problem in the lab." I rushed over there to find that the drains at the top of the stairs had filled with leaves and likewise the drain at the bottom and huge amounts of water, like a waterfall, was cascading down the steps and filling up the lab. The building power had gone off and the emergency lighting was on in the hallways. I called for the maintenance crew, but they were priortized to go save "expensive" student computers in another building. So there I was with my broom, frantically cleaning drains and pushing water, feeling like a character in Walt Disney's Fantasia. Because of this long and intense fight, the only thing we lost was the carpeting, papers and books on the lowest shelf. Saved the floor tile. The minimal rust on the feet of the EM's wasn't even noticeable. The lab was fine. Sprinklers a non-issue. The real devastation was done by the administration, later, when a new Dean said "What are electron microscopes good for?" eyeing the space and salary line he could use for a new Executive Secretary. But that's another story. Kind regards and good wishes to all, Beverly
In message {3163F416-at-hq_smtp} "Craig, Bob" writes: } } Good Morning All, } Bob Citron's response to David Rothbard reminded me on an incident we had } with an AMRAY 1000 many years ago (20?). A cooling water line on the } diffusion pump failed and filled the Plexiglass housing of the high voltage } supply with treated cooling water. Upon discovery me heart entered my } throat and visions of disaster entered my mind. As in Bob's case, the } service engineer calmly drained the water, rinsed the power supply with } deionized water and dried it with a hair dryer. Other component also have } been wet, which he dried with Kim Wipes. When he fired it up I expected } sparks to fly, etc. No way!! The microscope worked like nothing had } happened. Today the scope is 23 or 24 years old and still running! These } experiences would indicate that older analog instruments were pretty hardy. } How a modern digital instrument would react to such treatment is a good } question. } Bob Craig } OSRAM SYLVANIA INC. } Beverly, MA 01915 } craig-at-rd.sylvania.com
First of all, I'm very glad for Bob & Bob that water on their SEM's did not cause any serious problems that simple drying out did not cure.
But I must relay my experience with a wet SEM lest folks get the impression that these instuments are waterproof (no one would ever REALLY think that would they?).
In past years, we have had water accidents in labs over our heads, in some cases two floors above, and water has leaked down through cracks in their concrete floors, through our ceiling, and onto our instruments (kind of like magnetic atraction). In one case enough water got into the SEM so that a circuit board that runs the Magnification readout and a few others were damaged a bit. Even after replacing some transistors, the boards still blew out when powered up again. My service engineer had to clean dirty residue left behind by the water off the boards - many boards - with ethanol and a toothbrush to get rid of ground paths causing electrical shorts. Naturally, such events are not covered by service contracts, so we sent the bill upstairs to the offending lab. Some one up there left water aspirator running all weekend, and naturally thats a good time for paper towels to fall off the wall into the sink to plug up the drain.
We then built plexiglass "deflectors" over our SEM and TEM, big sheets suspended from the ceilings, to deflect any water coming down to the sides onto the floor. Since installation in about 1985 they have saved us twice. We feel we got off easy, as the potential was there for really serious big time damage.
The overhead panels also prevent some dust from falling onto the instruments. And they make a great conversation piece when tours come through!
If there is any potential danger in your labs from overhead water pipes or water from rooms above, I'd recommend that folks put up some kind of water sheding panels. Could save a big head ache if the water ever comes down.
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"MICROSCOPY & MICROANALYSIS 96" in Minneapolis, Minnesota, Aug.11-15, 1996
Tue, 2 Apr 1996 13:20:10 +1100 richard.easingwood-at-stonebow.otago.ac.nz (Richard Easingwood) wrote. } Dear Microscopists, } Does anyone out there know of a textbook or any other reference material } which covers integumental ultrastructure - cuticle, epithelium and } associated structures and also oogenesis - in marine invertebrates. We have } a marine science student who is studying subclass Ascothoracida who is } having problems getting the ultrastructure info she needs. } } Thanks in advance.
Richard
Do you know " Comparative Animal Cytology & Histology " written by U.Welsch and V.Storch (1976), Sidgwick & Jackson Limited, London ? This fundamental textbook translated from Germany contains "Chapter 3. Integument" and "Chapter 11. Reproductive organs and cells". Descriptions, of course, cover cuticle's ultrastructures of animals of various phyla. I think it's an available textbook for students though had been issued two decades ago.
S. Yamane
---------------------------------------------------- Shin-ichi Yamane [ a0c101u-at-cc.miyazaki-u.ac.jp ] Division of Marine Bioproduction Faculty of Agriculture, Miyazaki University, Gakuen Kibanadai Nishi 1-1, Miyazaki 889-21, Japan Phone : 0985-58-2811 ext.3356 ----------------------------------------------------
In message {199604041938.NAA17558-at-puccini.crl.umn.edu} "Gib Ahlstrand" writes:
} If there is any potential danger in your labs from overhead water pipes or } water } from rooms above, I'd recommend that folks put up some kind of water sheding } panels. Could save a big head ache if the water ever comes down. }
Sounds like a good idea ... but do you put them above or below the sprinkler heads? :)
_______________________________________________________________________ Beth Trend trend-at-cems.umn.edu http://charfac.cie.umn.edu Coordinator, Characterization Facility University of Minnesota Center for Interfacial Engineering 100 Union St SE Room 187 phone 612-624-1365 fax 612-626-7530 Minneapolis, MN 55455
In my current place of employment there are no sprinklers in the lab. I do agree with David that they are a mixed blessing. If they do open up, there is the possibility of damage to epquiment but in the real case of a fire, loss of epquiment is minor compared to a loss of a life. In my preveiously employment, the maintance folks upgraded the sprinkler system to a water sprinkler and removed the Halon system. Their reasoning was that a person will get wet, but not dead as with the Halon system. If a person is worried about electrocution with the water system, all electricty should be able to be shut off from a remote electrical substation.
Best to all, Ed Calomeni Medical College of Ohio Toledo, OH emlab-at-opus.mco.edu
} } Does anyone know of software that can calculate surface area or } surface roughness parameters from a stereo pair obtained from an SEM? } Dear Dan, If you want the true surface area--not just the projected area-- STERECON will allow you to reconstruct the volume, approximate the surface with polygonal tiles and add these up to get an area. I don't know whether it calculates roughness parameters, but these might be easy to determine from the true and projected areas. I assume you are not concerned with the fractional-dimensional properties which make the true area and the roughness dependent on the length scale. Yours, Bill Tivol
Our building (1992) was designed with a computer room and sem room next to each other but independantly run and served by a single sprinkler system where the pipe is filled with air until a fire is detected by a heat sensor (or we open a valve in a red box like a fire alarm) then a pump fills the pipe with water but the sprinkler still has to get hot enough to melt to start water flowing (each room independant)
Email me direct if you need me to find out details from the planners
Alan S. Pooley ,PhD Bivalve shell SEM & shape analysis SEM/Morphometrics lab Marine & Coastal Sciences Rutgers University 908 932 8959 ext 225 Pooley-at-ahab.rutgers.edu
Microscopy of Surfaces and Interfaces Midwest Microscopy and Microanalysis Society Workshop UWM Microscopy Open House
April 26, 1996
Please register by April 15, 1996 More information and online registration at http://www.uwm.edu/Dept/LSS/meeting.html
PROGRAM Golda Meir Library Conference Center University of Wisconsin - Milwaukee
Opening Ceremonies 8:45 - 9:00
Morning Session
9:00 - 9:30 J. Mansfield University of Michigan - Ann Arbor "Materials Science and Biological Applications of Environmental Scanning Electron Microscopy"
9:30-10:00 B. Tonner University of Wisconsin - Milwaukee "X-ray Microscopy for Chemical Analysis of Surfaces"
10:10 - 10:15 Coffee Break and Mounting of Posters Sponsored by College of Letters and Sciences
10:15 - 10:45 T. Kelly University of Wisconsin - Madison "Three-Dimensional Atom Probe Microscopy"
10:45-11:15 J. Nogami University of Wisconsin - Milwaukee STM: "Studies of Metal Growth on Silicon Surfaces"
11:15-11:45 L. D. Marks Northwestern University - Evanston Surface HREM: "Old techniques on Old Surfaces - Surprising New Results"
Open House Lunch
11:45 - 1:00 Lunch sponsored by Nissei Sangyo America and Hitachi Scientific Instruments. Poster Session
1:00 - 1:45 Poster viewing and poster competition Poster prizes are sponsored by the MMMS.
Afternoon Session
1:45-2:15 J. M. Gibson University of Illinois - Urbana TEM: "All you ever wanted to know about building your own expensive and complicated microscope, and then waiting five years until it finally works"
2:15 - 2:45 S. Babcock University of Wisconsin - Madison TEM/HREM: "Electron Microscopy Studies of Grain Boundaries in Bicrystals"
2:45 - 3:15 V. Dravid Northwestern University - Evanston "Analytical Electron Microscopy and Holography of Interfaces: Making a Mountain out of a Mole Hill"
3:15 - 3:30 Coffee Break and Judging of Posters Sponsored by Laboratory for Surface Studies
3:30 - 4:00 N. Browning University of Illinois at Chicago "High Angle Annular Dark Field in a Dedicated STEM"
4:00 - 4:30 K.L. Merkle Argonne National Laboratory "Relaxation Modes in Metal and Oxide Grain Boundaries"
4:30- 4:45 Poster Awards Ceremony and Closing Address M. Gajdardziska-Josifovska, Workshop Organizer
Open House Tour
REGISTRATION FEES: Registration is free for the members of the Midwest Microscopy and Microanalysis Society. Members of the MicroBeam Analysis Society are members. The workshop registration fees for non-members are same as the membership fees and can be paid at the conference site (by check or cash): Regular Member ($10) Student Member ($5)
POSTERS All lectures are by invitation only. Poster contributions by students, researchers and microscope manufacturers are welcome. There are no size restrictions for the posters, however they should be mounted on a sturdy backing suitable for display on easels. The poster must be relevant for the broad topic of microscopy of surfaces and interfaces.
POSTER COMPETITION
Graduate students are particularly encouraged to present their work and they are eligible to enter a poster competition. One grand prize and two first prizes will be given by the Midwest Microscopy and Microanalysis Society. The grand prize must be used by the winner to attend a microscopy related session at a major national/international conference.
Grand Prize: $500 1st Biological Sciences:$100 1st Physical Sciences: $100
I want to find out what slide printers (for 35mm film) there are on the market with the following characteristics: - generates sharp, high res, high quality slides - has driver for Windows NT
Two apparently good slide printers are: 1. Polaroid Digital Palette HR 6000 (will have NT driver) http://www.polaroid.com/homepage.htm 2. GALLERIA made by Mirus Industries (does not have NT drive) http://www.mirus.com/
Could you please give any comments on these printers or suggest other slide printers?
Thanks, Martin
--------------- Martin K=F6hler mk-at-enk.ks.se ---------------
Kevex 8000 XEDS system 10mm sq Be detector needs new hard disk but otherwise operational.
Asking $23,000, but will consider offers. Please contact me at the address below (email, phone, snail mail).
Jfm.
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
Message-ID: {199604061622.LAA12559-at-IndyNet.indy.net} To: DANIEL C VAN HART {vanhart-at-vnet.ibm.com} , Microscopy list {microscopy-at-Sparc5.Microscopy.Com}
Since this disease has had a fair degree of expsoure on the TV recently can someone describe what it is and how it could be related to the BSE disease?
Please keep it in laymans terms if you can, I am a physicist not a physician!
Many thanks. And sorry this isn't a Microscopy question.
Jfm.
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
8:00 am - 4:00 pm REGISTRATION 12:30 pm-12:45 pm OPENING REMARKS Dennis Barr, AREMS President-Elect Jay Jerome, SEMS President
SESSION I
12:45 pm- 1:30 pm INVITED SPEAKER Margaret Ann Goldstein Baylor College of Medicine
"Imaging with microscopes and computers"
1:30 pm -2:30 pm ROUNDTABLE DISCUSSION MODERATOR: Gene Michaels, Univ of GA
"Microscopy for kids of all ages"
2:30 pm -4:30 pm POSTER SESSION
Pennington SEM evaluation of the surface of an intra-arterial sensor retrieved from critically ill pediatric patients
Johnsrude Intraerythrocytic inclusions associated with iridoviral infection in a fer-de-lance (Bothrops moojeni) snake
Vaughn The effects of exogenous calcium oxalate crystals on renal epithelial cells
Vaughn Electroreceptor organ development in larval electric eels (Electrophorus electricus)
Gambling Remote printing of large format images
Whittaker Role of electron microscopy in diagnosis of the transmission of fibropapillomas in Green turtles, Chelonia mydas. 5:00 pm- 6:00 pm AREMS BUSINESS MEETING AND RECEPTION
2:30 pm -8:00 pm VENDOR SESSION
4:00 pm -8:00 pm VENDOR SPONSORED TUTORIALS
THURSDAY
SESSION II
9:00 am - 9:45 am INVITED TALK Mike Kizer Clinch Valley College "Life Cycle Stages of H. diminuta by SEM & Freeze Cracking"
RUSKA STUDENT COMPETITION
Abstract No.
9:45 1 Effects of recombinant bovine somatotropin (rbST) on satellite cell and myofiber nuclei proliferation of beef calves. R.C. Vann.
10:00 am -10:45 am INVITED TALK Mark Teaford Johns Hopkins "Analysis of rates of tooth wear via scanning electron microscopy"
10:45-11:15 am COFFEE BREAK
SESSION III
11:15 am -12:00 pm INVITED TALK Norman Herz University of Georgia "Innovative uses of SEM in the determination of weathering of statues"
12:00 pm - 1:30 pm LUNCH ON YOUR OWN
SESSION IV
1:30 pm - 2:15 pm INVITED TALK Fred Stevie AT&T "Focused Ion Beam application in semiconductor applications"
2:15 2 Low kV analysis of uncoated and of coated artificial root caries. DG Gantt
3:00 3 Computer-assisted analysis of radial symmetry in airway epithelial cilia: a new perspective on assesment of congenital ciliary defects. JL Carson.
3:15 4 Free radical derived oxidants and endothelial cell dysfunction in a rat model of diabetic retinopathy. E Ann Ellis.
3:30 5 Fungi associated with heating, ventilating, and air conditioning (HVAC) systems in the southeastern united states. RB Simmons.
3:45 6 Negative stain electron microscopy (EM) of self-assembled gastroenteric virus capsids. CD Humphrey.
4:00 7 Ultrastructure of conida and conidium germination in the plant pathogenic fungus Alternaria cassiae. CW Mims.
4:15 pm - 4:30 pm COFFEE BREAK
4:30 pm - 5:15 pm INVITED TALK
Locke Christman FEI Co. "Focused ion beam secondary ion mass spectrometry (FIB SIMS) expands the capabilities and the applications of focused ion beam systems."
6:00 pm - 7:00 pm SOCIAL HOUR 7:00 pm- AREMS/SEMS AWARDS BANQUET FEATURING AN AFTER DINNER TALK BY: Mark Teaford Johns Hopkins "Making teeth talk"
FRIDAY
7:30 am - 9:00 am SEMS BUSINESS BREAKFAST
SESSION V
9:00 am - 9:45 am INVITED SPEAKER
Robert Price University of South Carolina, Medical School "Experimental models in the study of early heart development"
9:45 8 Microanalysis in environmental science: determination of the micro-environment of pollutants and the mechanism of remediation of contaminated soils. TB Vander Wood.
10:00-10:40 INVITED SPEAKER Michael M. Thomason, Diane Petrovich, Steve Myles Analytical Services Group, Simpsonville, SC "SEM Analysis of nonwoven fabrics used in disposable diapers and other converted products."
10:40 9 The quantification of structure within polycrystalline fiber. KE Robinson.
10:55-11:10 COFFEE BREAK
11:10 - 11:50 INVITED SPEAKER Rathna Perera " The use of electron microscopy in high performance textiles"
11:50 10 Microscopic and chemical analyses of flax and flax retting. DE Akin.
12:05-12:45 INVITED SPEAKER T.J. Stark, D.P. Griffis, P.E. Russell North Carolina State University, Raleigh, NC "H2O Enhanced focused ion beam micromachining"
12:45 CLOSING REMARKS Mark Farmer, SEMS President-Elect John Herr, AREMS President
For further information contact: 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-bgsm.edu * **************************************************************
Micro people: besides requests for copies of responses to my question, here are the responses to the question "Is there a good 1990 or later prokaryote SEM/TEM atlas?":
} A good virus morphology reference is 'Virus Morphology', CR Madeley and AM } Field, 2nd ed, published by Churchill Livingston, Longman UK, (1988 } however) ISBN 0-443-02784-6. } regards, } } Richard Easingwood } South Campus Electron Microscope Unit } School of Medical Sciences } University of Otago } PO Box 913 } Dunedin } NEW ZEALAND
} From: "Jane A. Fagerland (847) 935-0104" {FAGERLAND.JANE-at-igate.pprd.abbott.com} } Subject: Re: Atlas of prokaryotes } } Gee, maybe we should be collecting good examples and publish a book of } our own! Thanks.
I seems that you're right Jane, at least for the bacteria and archea. Maybe Madeley and Field are working on a 3rd edition? But before anyone suggests that I do it, I'm a creepy-crawlyologist. Anyone who works on prokaryotes would look at my name and wonder who I am. (Aside to John Bozzola: the authors of Brock et al. _The Biology of Micro-organisms_ are all at Southern Illinois...) Phil
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
Two weeks ago, I posted a message in response to questions about the composition of GATAN's G1 TEM Epoxy. It has been brought to my attention, thankfully, that I have mis-spoken, and should make a corrective statement. My mistake was in assuring you that GATAN's G1 Epoxy was the same as EPOXY TECHNOLOGY's 353ND product. I should have stated that it was only my personal observation that the two are similar in characteristics, and that the MSDS sheets are similar.
As GATAN has an interest in selling their G1 Epoxy, it was incorrect of me, especially as we can, in the extreme, be considered competitors, to connect their epoxy directly with EPO TECH 353ND. However, my purpose was only to direct you to a source for information on hazardous components, as requested. Please excuse my error.
MSDS information on EPO TECH 353ND can be obtained from EPOXY TECHNOLOGY by calling:(508)667-3805.
Best Regards to all, Scott D. Holt BUEHLER 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-4546
Does anyone know of a commercial source for a TEM specimen holder that has a Faraday cage? We are looking for one for a Philips EM420 and related Philips scopes. I know that Gatan, Inc has them but we were also wondering if there were any other vendors. We would also consider a used holder if one was available.
Thanks
Norm Olson ******************************************* * Norm Olson * * Department of Biological Sciences * * Lilly Hall of Life Sciences * * Purdue University * * West Lafayette, IN 47907 * * * * Phone: 317-494-5643 * * FAX: 317-496-1189 * * email: nho-at-bragg.bio.purdue.edu * * * *******************************************
My apologies to those not interested in this meeting. I am resending this announcement because in my first transmission I attached the wrong file. This submission contains pertinent information about dates, registration fees, etc. I knew I shouldn't do anything important on a Friday Afternoon.
JOINT AREMS/SEMS MEETING GREENVILLE, SC APRIL 24-26, 1996
LOCATION: Hyatt Regency, Greenville SC. 803-235-1234 Located in Downtown Greenville, SC $82/night (single or double)
OTHER HOTELS WITHIN SHORT DRIVING DISTANCE: Hampton Inn, 1-800-Hampton, 803-288-1200, $50-75 Holiday Inn, 803-297-6300, $50-75 La Quinta Inn, 1-800-531-5900, $25-50 Holiday Inn Express, 803-297-5353, $25-50
REGISTRATION FEE: $25 members/$35 non-members/$15 student (with ID) and technicians 1 Day registration: $10
THURSDAY EVENING BANQUET: $22
You may register at the meeting. For arrangements information contact: JoAn Hudson, EM Facillity, Clemson University, 803-656-2465
PROGRAM WEDNESDAY
8:00 am - 4:00 pm REGISTRATION 12:30 pm-12:45 pm OPENING REMARKS Dennis Barr, AREMS President-Elect Jay Jerome, SEMS President
SESSION I
12:45 pm- 1:30 pm INVITED SPEAKER Margaret Ann Goldstein Baylor College of Medicine
"Imaging with microscopes and computers"
1:30 pm -2:30 pm ROUNDTABLE DISCUSSION MODERATOR: Gene Michaels, Univ of GA
"Microscopy for kids of all ages"
2:30 pm -4:30 pm POSTER SESSION
Pennington SEM evaluation of the surface of an intra-arterial sensor retrieved from critically ill pediatric patients
Johnsrude Intraerythrocytic inclusions associated with iridoviral infection in a fer-de-lance (Bothrops moojeni) snake
Vaughn The effects of exogenous calcium oxalate crystals on renal epithelial cells
Vaughn Electroreceptor organ development in larval electric eels (Electrophorus electricus)
Gambling Remote printing of large format images
Whittaker Role of electron microscopy in diagnosis of the transmission of fibropapillomas in Green turtles, Chelonia mydas. 5:00 pm- 6:00 pm AREMS BUSINESS MEETING AND RECEPTION
2:30 pm -8:00 pm VENDOR SESSION
4:00 pm -8:00 pm VENDOR SPONSORED TUTORIALS
THURSDAY
SESSION II
9:00 am - 9:45 am INVITED TALK Mike Kizer Clinch Valley College "Life Cycle Stages of H. diminuta by SEM & Freeze Cracking"
RUSKA STUDENT COMPETITION
Abstract No.
9:45 1 Effects of recombinant bovine somatotropin (rbST) on satellite cell and myofiber nuclei proliferation of beef calves. R.C. Vann.
10:00 am -10:45 am INVITED TALK Mark Teaford Johns Hopkins "Analysis of rates of tooth wear via scanning electron microscopy"
10:45-11:15 am COFFEE BREAK
SESSION III
11:15 am -12:00 pm INVITED TALK Norman Herz University of Georgia "Innovative uses of SEM in the determination of weathering of statues"
12:00 pm - 1:30 pm LUNCH ON YOUR OWN
SESSION IV
1:30 pm - 2:15 pm INVITED TALK Fred Stevie AT&T "Focused Ion Beam application in semiconductor applications"
2:15 2 Low kV analysis of uncoated and of coated artificial root caries. DG Gantt
3:00 3 Computer-assisted analysis of radial symmetry in airway epithelial cilia: a new perspective on assesment of congenital ciliary defects. JL Carson.
3:15 4 Free radical derived oxidants and endothelial cell dysfunction in a rat model of diabetic retinopathy. E Ann Ellis.
3:30 5 Fungi associated with heating, ventilating, and air conditioning (HVAC) systems in the southeastern united states. RB Simmons.
3:45 6 Negative stain electron microscopy (EM) of self-assembled gastroenteric virus capsids. CD Humphrey.
4:00 7 Ultrastructure of conida and conidium germination in the plant pathogenic fungus Alternaria cassiae. CW Mims.
4:15 pm - 4:30 pm COFFEE BREAK
4:30 pm - 5:15 pm INVITED TALK
Locke Christman FEI Co. "Focused ion beam secondary ion mass spectrometry (FIB SIMS) expands the capabilities and the applications of focused ion beam systems."
6:00 pm - 7:00 pm SOCIAL HOUR 7:00 pm- AREMS/SEMS AWARDS BANQUET FEATURING AN AFTER DINNER TALK BY: Mark Teaford Johns Hopkins "Making teeth talk"
FRIDAY
7:30 am - 9:00 am SEMS BUSINESS BREAKFAST
SESSION V
9:00 am - 9:45 am INVITED SPEAKER
Robert Price University of South Carolina, Medical School "Experimental models in the study of early heart development"
9:45 8 Microanalysis in environmental science: determination of the micro-environment of pollutants and the mechanism of remediation of contaminated soils. TB Vander Wood.
10:00-10:40 INVITED SPEAKER Michael M. Thomason, Diane Petrovich, Steve Myles Analytical Services Group, Simpsonville, SC "SEM Analysis of nonwoven fabrics used in disposable diapers and other converted products."
10:40 9 The quantification of structure within polycrystalline fiber. KE Robinson.
10:55-11:10 COFFEE BREAK
11:10 - 11:50 INVITED SPEAKER Rathna Perera " The use of electron microscopy in high performance textiles"
11:50 10 Microscopic and chemical analyses of flax and flax retting. DE Akin.
12:05-12:45 INVITED SPEAKER T.J. Stark, D.P. Griffis, P.E. Russell North Carolina State University, Raleigh, NC "H2O Enhanced focused ion beam micromachining"
12:45 CLOSING REMARKS Mark Farmer, SEMS President-Elect John Herr, AREMS President
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-bgsm.edu * **************************************************************
Mr-Received: by mta RANDD; Relayed; Mon, 08 Apr 1996 12:28:42 -0600 Mr-Received: by mta MCM$RAND; Relayed; Mon, 08 Apr 1996 12:28:45 -0600 Mr-Received: by mta RANDD; Relayed; Mon, 08 Apr 1996 12:28:55 -0600 Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
Here's a simplified version of the mad cow disease story:
The connection between BSE and C-J Disease is that both are thought to be caused by prions, which are self-replicating, abnormally folded, mutated versions of a normal cellular protein. There are similar diseases in other species, the most notable being scrapie in sheep. The diseases have relatively long incubation periods (years) and result in a characteristic "spongiform" lesion in the brain. The diseases are easily diagnosed post-mortem by the presence of little holes in the brain, which can be detected with light microscopy. Prion particles themselves can be visualized with negative staining and TEM.
Great Britain has had a huge problem with BSE beginning in the late 1980's. The problem in beef was caused by feeding scrapie-infected sheep scraps to cattle, and there is good evidence that the disease jumped from sheep to cattle. The fear is that if it could make that jump, then maybe it could jump from cattle to humans. No one has thought to wonder, if that were the case, why it hasn't jumped from sheep to humans, since mutton is commonly eaten in Great Britain; but this story is not about logic - it's about sensationalism.
C-J Disease sporadically and spontaneously occurs in about 1 in a million humans. (There is another human prion disease called kuru that is transmitted by eating human brains - the solution to that health problem is pretty straightforward!) Until a few weeks ago, the British health department has been adamantly sticking to the story that there was no evidence for an increased incidence of C-J corresponding with the current epidemic of BSE in British cattle. However, they recently wavered and said they couldn't rule out a BSE connection to 10 cases of a variant form of C-J that appears to have a shorter than usual incubation period. The British press has had a heyday with that, and I noticed that even the Chicago Tribune had a headline in yesterday's paper stating that an Italian had died of mad cow disease. It's not likely that the disease would be transmitted by eating beef from infected cattle, particularly because prions reside in central nervous system tissues (brain and spinal cord), and humans don't normally eat those.
The problem in Britain right now is the proverbial tempest in a teapot. The sad part is that many farmers are losing their livelihoods because the British press was irresponsible in the way it reported the information from the health department.
Enough said, I guess, before I climb any higher on the soapbox!
Jane A. Fagerland, Ph.D. Dept. of Microscopy and Microanalysis Abbott Laboratories Abbott Park, IL 60064
} I am trying to design a special specimen holder for a high resolution TEM, } and am looking for a good alloy to use for this purpose; [snip] } one that will have sufficient strength to withstand the forces [snip], } yet is reasonably easy to machine. } We have had very good luck with aluminum, believe it or not. The machinists like it too. In the HVEM, the specimen stage is held against the translation apparatus by 1 at. applied over ~9/16" dia., and this seems not to affect the resolution, etc. Unless you need exact dimensions be- tween a fiducial point on the stage and (say) the center of the specimen, you should be able to use almost any material. If you do need the stage to have constant dimensions to an angstrom or so, how will you mount the specimen to this accuracy?
} I think I have heard that some special grades of stainless steel, and } possibly some heat-resistant alloys, are suitable for use in such } applications. Does anyone know what alloys might be candidates, or what } limiting values of magnetic properties (permeability, susceptibility, etc.) } an alloy should have? } Be very careful with stainless steels. The machining process can convert a non-magnetic form to one with magnetism. We had the shop make a pin for a device which ended up ~1.5" from the beam, and there was a magnetism problem which went away when the pin was replaced by one made of phosphor bronze--another alloy to consider if aluminum is not suitable. Yours, Bill Tivol
} can you please let me know how to obtain more information about } STERECON. Is this shareware or a commercial package? } Dear Hasso, et al., For info on STERECON, check out J. Stru. Biol. v116, pp 93-98 (1996), M. Marko & A Leith, which describes the program and its capa- bilities, or check our Web site www.wadsworth.org under SPIDER then STERECON. For further info, please ask the real expert, Mike Marko, at "marko-at-wadsworth.org". He would know the particulars of the cost of obtaining STERECON as well as any technical info. Yours, Bill Tivol
I am looking for information on high resolution cameras that would attach to a Philips CM100. I am familiar with Kodak's Megaplus 1.6 camera but was wondering what other manufacturers had to offer or if Kodak has come out with anything more recent than the Megaplus.
Thanks,
Dennis Shubitowski University of Michigan School of Dentistry dshubito-at-umich.edu
I am about to learn a technique which is very new to me. I will be embedding pulp fibres in nanoplast. I have been warned that this will tend to be very brittle for trimming. Does anyone have any experience with this, specifically with cellulose fibres? Is there a way to prevent the brittleness? Any general advice...I am green at this.
Thanks, Arlene Kingsland Pulp and Paper Research Institute of Canada
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.
#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*
This year we are happy to invite attendees to present their work during the Saturday evening poster session. There is a limit of approx. 40 posters which can be displayed, therefore they will be accepted on a first come first serve basis. A one page (8.5X11) typed abstract should accompany the registration form and fees.
Friday Evening, September 20, 1996 5:00- 6:55 Reception and Poster Set-up 7:00- 7:45 Alan Waggoner - Amersham Life Science/Carnegie Mellon University 7:50- 8:35 Andrew Dixon - Biorad Microsciences 8:40- 9:25 Fred Lanni - Carnegie-Mellon University
Saturday, September 21, 1996
8:00- 8:25 Hospitality Buffet 8:30- 9:15 David Mastronarde - University of Colorado 9:20-10:05 John Z. Kiss - Miami University of Ohio 10:10-10:40 Coffee Break 10:45-11:30 Mark Ellisman - Univ. California-San Diego 11:35-12:20 Patricia Calarco - Univ. California-San Francisco 12:25- 1:25 Lunch (on your own) 1:30- 2:15 Jeff Hardin - University of Wiscosin-Madison 2:20- 3:05 Jim G. McNally - Washington University 3:10- 3:55 Enrico Gratton - Univ. of Illinois Urbana/Champaigne
8:30- 8:55 Hospitality Buffet 9:00- 9:45 Steven M. Block - Princeton University 9:50-10:35 Conly Rieder - Wadsworth Center for Labs/Research 10:40-11:10 Coffee Break 11:15-12:00 Scott E. Fraser - California Institute of Technology 12:05-12:50 Phillip G. Haydon - Iowa State University
1:00- 2:00 IMR Tour
FEES: General Registration $ 80.00 (Includes: Opening Reception, Social and Buffet Dinner, Coffee Breaks and Materials)
Student and Local Registration $ 50.00 (Includes: Opening Reception, Social, Coffee Breaks and
Materials)
Abstract Handling Fee $ 25.00
Dinner Ticket $ 20.00
fFOR ADDITIONAL INFORMATION and PROGRAM UPDATES CONSULT OUR WEB SITE:
http://www.bocklabs.wisc.edu/imr/imr.html
TO RECEIVE A BROCHURE AND REGISTRATION FORM
WRITE: IMR Univ. Wisconsin-Madison 1675 Observatory Drive Madison, WI 53706
Following the symposium, the IMR will be conducting a 2-day workshop. We will be presenting lectures and provide "hands-on" experience for the following techniques: * Multiple-photon excitation imaging * 4D DIC imaging * Cryo-SEM * High pressure freezing * Reversible embeddment for SEM and TEM
Workshop attendence will be limited to 25 participants. A letter of application is required. Once accepted a fee of $150.00 will be due.
The Department of Molecular, Cellular and Developmental Biology at Haverford College seeks an individual to co-teach a 7 week intensive course in microscopic technique during the first quarter of 1996-1997 academic year. Part of the major core, this laboratory has an expected enrollment of between 30 and 40 students and meets in two sections over five afternoons a week. This laboratory experience should include hands-on student experience in light, fluorescence and electron microscopy with a focus upon cell and tissue structure and function. Equipment used in the course includes two Nikon Labophot fluorescent microscopes and an Hitachi H-600 EM. The ideal candidate would have a Ph.D. (although A.B.D. will be considered) and experience in electron microscopy who is interested in gaining teaching experience in a liberal arts environment.
The department is also seeking an individual to co-teach in our introductory course "The Cellular Basis of Life" in the Spring semester.
Interested candidates for either (or both) of these positions should mail or fax a cover letter and curriculum vitae as soon as possible to:
Slavica Matacic, Ph.D. or Karl Johnson, Ph.D. smatacic-at-haverford.edu kjohnson-at-haverford.edu (T) 610-896-1306 (T) 610-896-1305 Department of Molecular, Cellular and Developmental Biology Haverford College 370 Lancaster Avenue Haverford, PA 19041 (F) 610 896-4963
Additional information about the department and its program is available via the World Wide Web at http:www.haverford.edu/biology/welcome.html.
Haverford College is an affirmative action, equal opportunity employer.
Mr-Received: by mta RANDB; Relayed; Wed, 10 Apr 1996 09:38:55 -0600 Mr-Received: by mta MCM$RAND; Relayed; Wed, 10 Apr 1996 09:38:57 -0600 Mr-Received: by mta RANDC; Relayed; Wed, 10 Apr 1996 09:39:12 -0600 Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
Thanks for all the comments concerning my remarks about prion diseases. I stand corrected on my assertion that humans don't usually eat central nervous system tissue from sheep and cattle. Several folks from the UK enlightened me - sheep and calf brains are considered a delicacy in some areas of the UK and are also used as fillers in hamburger and sausage. Also, soups are made from animals parts that may still have CNS tissues attached to them.
Someone also pointed out that it was initially thought that scrapie could not jump from sheep to cattle; thus, there's precedent for inter- animal transmission, regardless of what seems likely right now.
For anyone wanting more information on BSE, there is now a BSE group. To subscribe, send an e-mail to {majordomo-at-info.aphis.usda.gov} . Leave the subject line blank, and type {subscribe bse} in the message. There is also a BSE news and information service on the Web at http://www.cabi.org/
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064
I) is there any interest in compiling a list of samples that can be used in lab classes to readily show different principles of microscope operation? One example of one of my test samples would be .4 um nucleopore filters with their nice round pores for demonstrating astigmatism and how to correct for it. Are others willing to share their ideas in this regard--I'll be happy to compile and post a summary
II) Along these same lines, does anyone have a suggestion for a test sample to use in my SEM laboratory class that clearly demonstrate the effects of changing accelerating voltage? I haven't come across an easily obtainable, reproducible sample to clearly show:
1) enhancement of surface detail at low voltage vs high votage
2) ability to view uncoated samples at low voltage without charging vs high voltage with charging
any suggestions in this regard would be welcome
steve
Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University San Diego, CA 92182-4614 phone: (619)594-4523 fax:(619)594-5676 email:sbarlow-at-sunstroke.sdsu.edu
Sorry for being a bit after the thread with this, but it would be nice to know if there is any freeware stereology programs around. I don't need anything very fancy but just something simple to use mainly for demonstrations and teaching purposes.
TIA
Stefan
Stefan Gunnarsson Microscopy & Imaging Unit, Uppsala University Norbyvagen 18A S-75236 Uppsala, Sweden tel. +46 18 182638, fax. +46 18 182683
I am looking for red/green glasses for viewing some anaglyphs that I have on slides. Ted Pella sells red/blue but I don't think they will work with my pre-existing slides.
Also, why the change from red/green to red/blue? Is this due to the "color bombardment" phenomenon that causes a strain to some viewers or are there other reasons?
Any newer published references re anaglyphs besides the 1980 article by Barber and Emerson in Scanning 3:202-206?
Many thanks.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
I am looking for red/green glasses for viewing some anaglyphs that I have on slides. Ted Pella sells red/blue but I don't think they will work with my pre-existing slides.
Also, why the change from red/green to red/blue? Is this due to the "color bombardment" phenomenon that causes a strain to some viewers or are there other reasons?
Any newer published references re anaglyphs besides the 1980 article by Barber and Emerson in Scanning 3:202-206?
Many thanks.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
(text deleted)...does anyone have a suggestion for a test sample to use in my SEM laboratory class that clearly demonstrate the effects of changing accelerating voltage?
A few years ago, Dr. David Joy wrote an article which appeared in some Hitachi literature about the advantages of low-voltage imaging. He used a common carbon-film covered TEM grid imaged at 1 or 2 kV and the same view at 20 kV. The low-voltage image clearly showed the carbon film, while at 20 kV, the dominant feature was the grid bars, and the carbon film seems to disappear.
The images were obtained at low magnification (less than X1000) so you can do this even if you don't have a FESEM.
Jim Stets Air Products and Chemicals, Inc. Allentown, PA stetsjr-at-ttown.apci.com My opinions are my own, not my employer's
Just another late word on the LN2 issue. There are crogenic gloves available which will properly protect your hands when handling liquid nitrogen or shutting off frozen valves and handling frozen hoses. Although I agree that sometimes safety issues don't necessary reflect common sense, having the gloves doesn't hurt.
By the way, it snowed yesterday, again....when will we see Spring in the Northeast?
Regards, Melanie Behrens Texaco Fuels and Lubricants Research Beacon, NY behrema -at- Texaco.com 914-838-7261
I have a biomedical engineering student that would like to analyze the spatial arrangement of collagen arrangement in cartilage. I'm a materials person and have absolutely no idea how to start, how to prepare the samples, where to look up information or if it is even possible. The papers we have show TEM analysis but we would prefer to start with SEM.
Can anyone out there point me in the right direction.
Robin Griffin Electron Optics Laboratory Manager Materials and Mechanical Engineering The University of Alabama at Birmingham
At 10:39 AM 4/10/96 -0600, you wrote: } I am looking for red/green glasses for viewing some anaglyphs that I have } on slides. Ted Pella sells red/blue but I don't think they will work with } my pre-existing slides. } } Also, why the change from red/green to red/blue? Is this due to the "color } bombardment" phenomenon that causes a strain to some viewers or are there } other reasons? } } Any newer published references re anaglyphs besides the 1980 article by } Barber and Emerson in Scanning 3:202-206? }
Hi John. This question came up recently and I archived the responses on our web page. Go to the address at the end of this message and click on the "Tips & Tricks Wizard". You should find two links that might be useful.
1." Stereo Glasses, Red/ Green" 2. "Presentation of Stereo Pairs"
If you do not have web acess or cannot read the files, please let me know and I will be happy to e-mail, fax, etc... them to you.
Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 904-392-1295 ICBR EM Core Lab fax 904-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/
Can someone remind me where the standard printer test file is located. This is the file that was being printed on different printers and then brought to the annual meetings for comparison. Thanks,
*************************************** David Garrett "DGARRETT-at-GAB.UNT.EDU" University of North Texas Dept. Biological Sciences (817)565-3964 Fax (817)565-4136 ***************************************
in regard to: 1) enhancement of surface detail at low voltage vs high voltage:
Try imaging a very ductile fracture surface (e.g. mild steel, etc.) for demonstrating high vs. low keV effects. At the higher energy, the image looks fine, but you get excessive intensity from thinner edges. At the lower energy, that excess brightness is gone and you start to see fine features in the edges as well as on (what you thought were) flatter regions. The voltage effect is quite strong for comparisons between e.g. 25 keV vs. 5 keV.
in regard to: 2) ability to view uncoated samples at low voltage without charging vs high voltage with charging:
For viewing uncoated samples, try a ceramic, even a piece of glass. You can get decent, uncharged imaging on SiO2 at around 1.5 to 2.0 keV. Above that, all the charging you could want and then some.
Bob Keller NIST Materials Reliability Division Boulder, CO
A postdoctoral position will be available at Northwestern University starting around August-October 1996. The position will involve extensive use of HREM coupled with diffraction/image simulations and crystallography on high temperature superconductors. A strong background in electron microscopy and crystallography is required, and strengths in the use of computers will be important. The ability to work with others will also be very significant, since the position will involve extensive collaborative work.
Send CV plus the names of 3 referees with (seperately) letters to: L. D. Marks Department of Materials Science Northwestern University Evanston, IL 60208, USA http://risc1.numis.nwu.edu ldm3-at-apollo.numis.nwu.edu
"Microscopy and Analysis" is published 6 times a year, in UK, European and USA editions. It is widely circulated among all types of user, both academic and commercial, in the areas of microscopy and associated analytical techniques.
For a number of years, as a service to its readers, Microscopy and Analysis has published in its UK edition a listing of laboratories selling their services. The listing provides full contact details, plus a tabulation by technique of the services offered by each laboratory, with a note on any special capabilities or expertise.
This year, we plan to extend this feature to the USA edition. Laboratories are invited to send in details of the services offered. Please send on a single page, the following information:
1. Contact name, full postal address, telephone and fax numbers, and e-mail address. 2. Indicate if your experience is in the materials or biological sciences, or both. 3. List the instrumental techniques available in your laboratory, for example TEM, light microscopy, EDX, Raman spectroscopy, etc. 4. Detail any special area of skill or expertise, for example, foreign body analysis of food, cement, asbestos fibre characterisation, etc. 5. Indicate if you would be interested in special advertising facilities linked to this feature.
The deadline for receipt of this information is 1st August, 1996. The information will be compiled, tabulated, and published in the September issue of Microscopy and Analysis. There is no charge to laboratories for this service.
Information should be faxed to +44-1372-459957.
Dr. Larry Stoter Technical Editor Microscopy and Analysis
We have been utilizing immunohistochemistry to stain for smooth muscle actin and desmin. We have noticed that actin stains all blood vessels but desmin does not. Has anyone else noticed this ? Is there a better stain to differentiate between smooth muscle and endothelial cells ?
I have followed much of the thread on negative scanners in regard to TEM negatives. It is still unclear if many biological/medical TEM labs use negative scanners, and if so which make & model scanner has proven to be suitable.
Printing the digitized image obtained from the negative scanner is the other half of the equation. What printers are in use for printing scanned negatives? I realize that viewers of digitized negatives may have different expectations than printing digitized images from other sources.
If you have experience with scanning TEM negatives (Kodak 4489) and printing, especially from PCs (as opposed to MACs) I would appreciate hearing from you.
I bought some red/blue glasses that were cheap and durable last year. I had bought some others earlier that were crap as the lenses fell off quickly. The good ones were from: Rainbow Symphony INc. 6860 Canby Ave. #120 Reseda CA 91335 800-821-5122
} At 10:39 AM 4/10/96 -0600, you wrote: } } I am looking for red/green glasses for viewing some anaglyphs that I have } } on slides. Ted Pella sells red/blue but I don't think they will work with } } my pre-existing slides. } } } } Also, why the change from red/green to red/blue? Is this due to the "color } } bombardment" phenomenon that causes a strain to some viewers or are there } } other reasons? } } } } Any newer published references re anaglyphs besides the 1980 article by } } Barber and Emerson in Scanning 3:202-206? } } }
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu
I apologize if you subscribe to both lists (confocal & microscopy) and receive this twice. I wasn't sure where to send this one.
I have two histochemistry questions:
1) How does one block endogenous biotin (the tissue in question is insect)?
2) Are there "better" mounting media for sections that have had an alkaline phosphatase reaction run on them (better than say, "permount")?
Thanks for your help. 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) : :...................................................................: http://www.pharm.arizona.edu/exp_path.html
I have read on the microscopy list, the suggestion of you paper on Nanoplast. Please, if possible could you send me a reprint of this article (Weyda F., 1990: Notes on the use of Nanoplast FB 101 for transmission electron microscopy. J.Electron Microsc.Tech., 16: 356-357), since I have been trying to embed magnetotactic bacteria in Nanoplast with little success. Despite this, it is very difficult to obtain a copy of this article, because of the absence of subscription of J. Electron Microsc. Tech in my country. Many thanks in advance,
Yours very truly, Ulysses Lins
} Ulysses Lins } Setor de Microscopia Eletronica } Instituto de Microbiologia Prof. Paulo de Goes } UFRJ - CCS - Cid. Universitaria } 21949-900 - Rio de Janeiro - RJ } Brazil E-mail: imvuly-at-microbio.ufrj.br you
} I have followed much of the thread on negative scanners in regard to TEM } negatives. It is still unclear if many biological/medical TEM labs use } negative scanners, and if so which make & model scanner has proven to be } suitable. } Printing the digitized image obtained from the negative scanner is the other } half of the equation. What printers are in use for printing scanned negatives? ******************************************************************************* 1. Scanning:
We use with great success for TEM negatives an AGFA Arcus II flatbed scanner with the new AGFA FotoTune 2.0.8 Software. The negatives are digitized to 14-bit with linear LUTs and 300 dpi for full frame images (overview), and at optical 1,200 dpi for detail areas, each resulting in a file size of 1Kx1Kx16-bit TIFF. The negatives should be exposed a step more than used for printing to assure covered high lights. The gray tone quality is extraordinarily good when compared to 8-bit scanning or cheap 8-bit scanners, i.e., less than 1% IR (intensity range) noise, no scan lines even at highest contrast resolution of contrasts smaller than three intensity steps (from 16,000 steps per pixel), linear LUT (same contrast distributions in highlights and shadows with positive or reversed negative acquisition mode). We routinely zoom into the data 3-5 times with a "clean" bicubic zoom in order to display details. The image quality is certainly -although I cannot prove it at this time- better than can be established in the photolab with silver halide printing.
2. Printing.
I tested several printers with a 1Kx1K image. Printer performances were compared at the level of whole page images and 10x enlargements of the printed images (LaserJet, Lazar Print, Dye sub,). Results are available at
In short, you have to ask how many lines per inch are printed and at how many gray levels (not always the same as dots per inch), plain paper or special paper, permanence of the print, printing time, printing costs. I print 300 lpi at 256 gray levels (4,800 dpi) on plain paper in 20-45 sec for 1-10 MB of images per page at 3 cents per page with a HP LaserJet and enhancer board in archival quality. I use the Lazar Print board. My source was Smart Analytical Products (301) 598-8881. You will find everything at my page. I am testing some more printers in the same manner (Epson, and a 1200 dpi Laser printer, but the results are expected not to be very different from the already tested printers), A plain LaserJet will be a good start for printing images.
Conclusion: Since over two years I have not used my state of the art photolab since digital acquisition, flatbed scanning and digital LaserJet printing fulfill all my needs for making image reports, slide templets (photographed with 120 roll film on a copy stand), and publication prints. However, my image communication is vastly improved through text imbedded image printing in reports.
Does anyone have any experience using proteinase K to unmask antigens on LRW sections? My use is for a cytoplasmic tail membrane label using endothelial cell monolayers. What would you use to quench it? Thanks for any suggestions! Shelley Landon Kaurin
Hello All: A fellow employee of mine in a different department had some interesting questions, some of which I can answer, some of which I have no experience with which to answer. I was hoping that some of you may have some knowledge you could share on this topic. These questions all relate to cloud point measurements of oils using polarized light microscopy, with a video camera and a temperature controlled stage. A drop of oil is placed on a slide with a cover slip and heated above the cloud point, then cooled 0.1 deg C/minute. The cloud point is determined as the point at which the first wax crystal forms.
Her questions were: 1) Is it more preferrable to use a polarizing scope with a full circle 360 degree rotating stage or will an X-Y scope with a polarizer and polarizing filter work just as well?
2) What kind of magnification would be needed to see the crystals form? Would 40X be sufficient? Would a zoom binocular be preferrable to fixed lenses?
3) Would there be any advantage to using a color camera over a black and white one? The oil is black, and contrast is the most necessary element to detect the crystals, not a color change.
4) What type of thermal stage is required? Mettler Toledo has a $12000 system, but is such a system overkill for this purpose? Physitemp has a TS-4 thermal stage with a 90 degree bend metal bracket with cooling fluid circulating to it. The slide fits on the flat part of the bracket. This costs about $3500. Will this stage be capable of ramping the temperature 0.1 degree C/minute? Are there other companies which sell thermal stages?
I was also wondering if anyone knows of any references on this test method. Thanks in advance for your help.
Regards, Melanie Behrens Beacon, NY behrema -at- Texaco.com 914-838-7261
I am seeking the assistance of anyone who can point me to any specific references (OSHA, NIOSH) on the proper storage of chemicals. Although our institution has made a significant investment in the purchase of approved chemical storage cabinets for acids, corrosives, and flammables, some of our technical staff are reluctant to move a small quantity of chemicals from a small storage room to a large, well ventilated preparatory room. Their concern and claim is that these storage cabinets must be vented to the outside in some manner. The chemicals of greatest concern include some concentrated acid and base solutions (HCl, NaOH) and organic solvents such as acetone and petroleum ether, in four to six liter quantities each.
I have searched the OSHA web site and have located information as to the storage of flammables. Cabinet dimensions (metal or wood) relative to the storage of specific quantities of flammable agents are noted, however, there is no mention of the need for external cabinet venting. I have been unable to find any reference as to the specific storage of acids or alkaline solutions.
In most of the labs I have encountered (including my own EM lab), chemicals are stored in the approved cabinets *without* separate external ventilation. I would appreciate any specific references (web sites, publications, etc.) that you could provide on this matter since I have a dismantled TEM that has been awaiting installation into the small storage room that these chemicals occupy, for over six months!
Please respond directly to me and I will post a summary to the group. Thanks in advance!
Stephen Beck Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829
With reference to the BSE and CJD letters. While there is of course a very faint chance that any of us can catch the any of the rarest of diseases in the same way that we can be struck by lightning, the chances of getting CJ from eating beef are marginally less that winning the big prize on the UK National Lottery. I am quite content to eat muscle beef and I buy a lottery ticket each week. I think you Americans have long had the right idea in the you never eat meat that has come from the "engine room". Leaving aside the bio-medical and econo-agricultural aspects of BSE there is of course a delicious UK political and EURO political aspect to this sorry story
Patrick Echlin Cambridge UKOn Wed, 10 Apr 1996, Jane A. Fagerland (847) 935-0104 wrote:
} Thanks for all the comments concerning my remarks about prion diseases. } I stand corrected on my assertion that humans don't usually eat central } nervous system tissue from sheep and cattle. Several folks from the UK } enlightened me - sheep and calf brains are considered a delicacy in some } areas of the UK and are also used as fillers in hamburger and sausage. } Also, soups are made from animals parts that may still have CNS tissues } attached to them. } } Someone also pointed out that it was initially thought that scrapie } could not jump from sheep to cattle; thus, there's precedent for inter- } animal transmission, regardless of what seems likely right now. } } For anyone wanting more information on BSE, there is now a BSE group. } To subscribe, send an e-mail to {majordomo-at-info.aphis.usda.gov} . Leave } the subject line blank, and type {subscribe bse} in the message. There } is also a BSE news and information service on the Web at } http://www.cabi.org/ } } } Jane A. Fagerland, Ph.D. } Dept. Microscopy and Microanalysis } Abbott Laboratories } Abbott Park IL 60064 } } }
} I apologize if you subscribe to both lists (confocal & microscopy) and } receive this twice. I wasn't sure where to send this one. } } I have two histochemistry questions: } } 1) How does one block endogenous biotin (the tissue in question is insect)? * You can try successive 20 min incubations of sections in 0.1% avidin and 0.01% biotin*
} 2) Are there "better" mounting media for sections that have had an alkaline } phosphatase reaction run on them (better than say, "permount")? * Yes, coverslip with aqueous-based medium after counterstaining* } Thanks for your help. } Doug
Good luck! Sverker
********************************************************* 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 USE TO USE A POSITIVE/NEGATIVE 35MM SLIDE FLIM TO MAKE POSITIVES OF SEM NEGATIVES - DOES ANYONE IF THIS TYPE OF FLIM STILL EXISTS? IF SO PLEASE GIVE ME ITS NUMBER AND WHO MAKES IT.
THANK YOU
Dr. David G. Gantt Phone: 1-912-681-5964 Dept. of Biology Fax: 1-912-681-0845 Landrum Box 8042 e-mail: david_gantt-at-gsvms2.cc.gasou.edu Georgia Southern University Statesboro, Georgia 30460-8042
Although I have worked with this antibody in Prostate without success, Factor VIII(von Willebrand factor), may be something to try. We feel that we didn't have success because of the lack of vascularity in the prostate to be able to observe any kind of staining patter. I have two references that talk about using this antibody. The first reference using a fluorescent Ab and the second uses it in cell culture, but either should be of some help to you. 1. Potzsch G et al Laboratory Investigation 63(6):841-852 1990
2. McGuire PG and Orkin RW Laboratory Investigation 57(1):94-105 1987.
Good luck!
Mary Laboratory Technologist Medical College of Wisconsin Department of Urology molter-at-post.its.mcw.edu
On Fri, 12 Apr 1996, BARBARA HARTMAN wrote:
} Date: Fri, 12 Apr 1996 08:52:46 -0400 } From: BARBARA HARTMAN {BARBARA.HARTMAN-at-spcorp.com} } To: histonet-at-pathology.swmed.edu, microscopy-at-aaem.amc.anl.gov } Subject: Immunohistochemistry of Desmin and Actin } } We have been utilizing immunohistochemistry to stain for smooth muscle } actin and desmin. We have noticed that actin stains all blood vessels } but desmin does not. Has anyone else noticed this ? Is there a } better stain to differentiate between smooth muscle and endothelial } cells ? }
the e-mail address of Dr. Sam Bowser of the Wadsworth Center in Albany N.Y. If someone knows it, I 'd appreciate if you could mail it to me. Sam, if your out there, please e-mail me at genexs-at-mhv.net or genexs-at-aol.com. Thankx. Cya, Gene Santagada
I have used JB-4 for regular histology of the mouse skull base and had good luck with it for toluidine blue stain only. I use 15-20 ml breakaway molds, 0.7 gm catalyst to 100 ml of soln. A, and 1.0 ml of solution B added to the embedding mixture of 100ml soln. A/0.7 gm catalyst. I put the molds into icewater to start the embedding procedure. I then overfill the molds, place the specimens into the soln., cover the surface with parafilm-making sure to force all the air bubbles out from under the parafilm, seal the edges of the parafilm to the molds with a hot spatula, and place the tray of ice water/molds into a refrigerator overnite. If heat is a problem for your or your friend's immunohistochemisty, the operation stated above must be done quickly, as the polymerization of JB-4 is exothermic. The next day, I remove the bocks from the refrig. and let them stand overnite at room tempurature before I remove the molds. You might need to let the blocks stand for a few more days before you attempt to cut them also. I cut mine on a ralf knife made from a glass microslide ( the cheaper the better) and I cut them on a standard rotary microtome with a razorblade holder to hold the ralf knives. As I stated above, I have not been able to get good staining with any stain other than toluidine blue. If anyone out there has suggestions on how to get any other stain to work with this material I'd like to know.
Karen Pawlowski On Fri, 12 Apr 1996, Michel Deschuyteneer wrote:
} Fellow microscopists, } } A colleague needs to embed nasal cavities from mice for immunocytochemistry. } His antigens are fragile and do not whistand the standard embedding } procedure with paraffin. Cryosectioning works but would preferably be } avoided because of the bone tissue and decalcification has also proven } detrimental to the antigenicity of his preparations. } } I suggested to try glycol metacrylate but I have no hands on experience with } this material. In addition, there is a variety of commercial kits available, } e.g. Polysciences' Immunobed or JB4, but the differences are not exactly } obvious to me. } Can anyone recommend a particular preparation of this medium? What are the } pros and cons? Is there some good alternative? } } Any info and/or references would be welcomed. } Thanks a bunch in advance. } Regards, } } MICHEL } } } **************************************************** } Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be } Scientist Electron Microscopy Laboratory } } SmithKline Beecham Biologicals } Rue de l'Institut, 89 B1330 Rixensart, BELGIUM } Tel: +32-2-656 9290 Fax: +32-2-656 8164 } **************************************************** } Disclaimer: the opinions expressed in this } communication are my own and do not necessarily } reflect those of SmithKline Beecham. } **************************************************** } }
I have a reliable method for hematoxylin and eosin staining of GMA sections that works well for bone and soft tissue sections. I'd be happy to mail or fax a copy of it to anyone who's interested.
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064
In my opinion the best film for making positive slides from negatives is Kodak Technical Pan Film 2415 (Cat No 129 7563 for 135-36 rollls and 129 9916 for 150 ft rolls)developed in Kodak undiluted D-19 for 4 Minutes at 20 o C. You can also vary the developer and conditions in order to adjust the contrast range--see Kodak Publication No. P-255. If you want a positive slide from a positive image try Kodak Precision Line film LPD4 (150ft rolls Cat No 157 8327) for a reasonable one step b & w transparency.
Both Kodak and Ilford make a continuous tone direct positive B&W 35 mm film. I have forgotten the specific product code, something like 6330 for the Kodak film. It's apparently what they used to use for movies.
cheers,
Rosemary White __ / Department of Ecology _/ \__/ \ and Evolutionary Biology / \ Monash University / Australia \ Clayton, Victoria 3168 \ ____ / phone 61-3-9905 5670 \_/ \_*_/ fax 61-3-9905 5613 __ email rgwhite-at-vaxc.cc.monash.edu.au \/
I'm trying to help someone localize polysaccharides/glycoproteins in/on a marine alga, first by using ConA or WGA, preferably labelled with colloidal gold. Having never used ConA or WGA, I don't have a clue as to where to begin. Can I use any glutaraldehyde? Left to my own devices I will start with 0.25%. Paraformaldehyde? Like 1 to 3%? We will be harvesting and fixing on Wednesday, April 17. After they're in blocks, I can worry about the next steps. Blocking agents? Secondary antibodies? If anyone has an opinion on how I should proceed, I would love to hear from them!
Mahalo from out here in the middle of the Pacific...
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/ * *****************************************
I have had great success using Kodak Technical Pan film 2415 (develop at maximum contrast, 4 minutes in D-19) directly through negatives sitting on a light table, using 1/2 stop bracketing.
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-trenton.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 Trenton State College, NJ 08650-4700 fax: (609) 771-2674
On Mon, 15 Apr 1996, Robert Schmitz, Biology wrote:
} } } I USE TO USE A POSITIVE/NEGATIVE 35MM SLIDE FLIM TO MAKE POSITIVES OF SEM } } NEGATIVES - DOES ANYONE IF THIS TYPE OF FLIM STILL EXISTS? IF SO PLEASE } } GIVE ME ITS NUMBER AND WHO MAKES IT. } } } } THANK YOU } } } } Dr. David G. Gantt } } } Kodak #5302 Positive Release 35mm Film } rschmitz-at-uwspmail.uwsp.edu } or } rschmitz-at-macsrv1.uwsp.edu } (note its macsrv"one" not "el") } Robert (Bob) J. Schmitz } Department of Biology, } University of Wisc. Stevens Point. } Stevens Point, Wisconsin 54481 } ph 715-346-2420 } David: I used to use Tech Pan rated at ASA 125 and got beautiful slides. Used a Nikon on automatic mode with an aperture of about f:6 on a 90mm lens. Use a good light box.
Developed in D-19, 1:1 for 3min, fixed, washed and dried. If you use Orbit Bath in your fix your fix time is cut to 2 min and wash 5 min. Orbit Bath is good for printing also. Same times for prints. For TEM slides did the same. In the past I used type 4489 EM film for my slide. Tech Pan works rather well for making B&W slides from Negs.
Shelley Landon Kaurin writes: " Does anyone have any experience using proteinase K to unmask antigens on LRW sections? My use is for a cytoplasmic tail membrane label using endothelial cell monolayers. What would you use to quench it? Thanks for any suggestions! "
I presume that the antibody currently does not label the antigen in the LR White. As you correctly guess, this is probably because the antibodies cannot gain access to the antigen. However, I an not sure that treating the sections with proteinase K will help expose the antigenic sites for you.
Firstly, the proteinase K is a protease and will not digest resins.
Secondly, if it did digest resin, how would you make sure that the proteinase K did not digest your antigen during the incubation.
May I suggest that a better approach may be to permiabilize the cells prior to embedding in resin, and then treating the sections with antibody, omitting any resin digestion. The permiabilization will wash out much of the cytoplasm but leave membrane proteins in place. Cytoplasmic tails are then accessible to antibodies (unless they are really short and close to the membrane).
The permiabilization can be done either before or after fixation and can be as simple as incubating living cells in hypotonic buffer, or incubating fixed cells in detergent.
Need more? Contact me.
Paul Webster, Center for Cell Imaging Yale University School of Medicine
} } A colleague needs to embed nasal cavities from mice for immunocytochemistry. } } His antigens are fragile and do not whistand the standard embedding } } procedure with paraffin. Cryosectioning works but would preferably be } } avoided because of the bone tissue and decalcification has also proven } } detrimental to the antigenicity of his preparations. } } } } I suggested to try glycol metacrylate but I have no hands on experience with } } this material. In addition, there is a variety of commercial kits available, } } e.g. Polysciences' Immunobed or JB4, but the differences are not exactly } } obvious to me.
} } Regards, } } } } MICHEL
I haven't tried JB4 or glycol methacrylate for immuno work, but have used butyl methyl methacrylate - it works well for plant tissue - for localising microtubules, actin and callose at least. You need to add 5-10 mM DTT to the fixative and to the embedding resin, seems to preserve the antigenicity. The reference is Baskin et al. 1992 Planta 187: 405-413 (though the procedure we use is modified from this), in which earlier work is cited also. Best results are with UV polymerisation in the cold, though some people get away with room temperature polymerisation.
good luck,
Rosemary White __ / Department of Ecology _/ \__/ \ and Evolutionary Biology / \ Monash University / Australia \ Clayton, Victoria 3168 \ ____ / phone 61-3-9905 5670 \_/ \_*_/ fax 61-3-9905 5613 __ email rgwhite-at-vaxc.cc.monash.edu.au \/
} Hello Everybody: } } I've received several comments recently about the fate of silver-enhanced } gold particles (in EM) when specimens are treated with osmium tetroxide. } The literature indicates that this process can cause a slight reduction in } the size of the particles, probably due to re-oxidation of the deposited } silver. However, the effect seems to be very variable: in some cases many } of the particles disappear, or the color fades from the sections. Has } anyone had problems with this, and has anyone found a good way to prevent } it? } } Thanks, } } Rick Powell
Have just come back from holiday and it seems noone has answered this yet. I do a lot of Ag-Au for EM and initially had just this problem. After normal embedding of the Ag enhanced tissue, there would sometimes be beautiful Ag particles, sometimes remains of Ag with holes where the particles had been, sometimes nothing at all. The problem turned out to be the uranyl acetate treatment. It appears (I am no chemist) that UAc dissolves out the Ag in some way. Not using UAc cured the problem, but obviously contrast was then hopeless. I now gold tone tissue - the Au-Ag-Au complex is completely stable in osmium and UAc. There is an increase in the contrast of the tissue, which may cause some people problems, especially if high resolution is needed, but the method really works.
Gold toning: (after method of R. Arai et al 1992, Brain Res Bull 28:343-345) after Ag enhancing, wash in water 0.05% gold chloride 10 mins/4deg water wash 0.05% oxalic acid 2 min water wash 1% sodium thiosulphate (freshly made) 1 hour water wash and embed normally
Diana van Driel Dept Ophthalmology Sydney University AUSTRALIA 2006
} Date sent: Mon, 15 Apr 1996 09:33:09 -1000 (HST) } From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } To: Microscopy Newsgroup {Microscopy-at-Sparc5.Microscopy.Com} } Subject: Need suggestions for TEM immuno glycoproteins
} Aloha, microscopists, } } I'm trying to help someone localize polysaccharides/glycoproteins in/on a } marine alga, first by using ConA or WGA, preferably labelled with } colloidal gold. Having never used ConA or WGA, I don't have a clue as } to where to begin. Can I use any glutaraldehyde? Left to my own devices } I will start with 0.25%. Paraformaldehyde? Like 1 to 3%? We will be } harvesting and fixing on Wednesday, April 17. After they're in blocks, } I can worry about the next steps. Blocking agents? Secondary } antibodies? If anyone has an opinion on how I should proceed, I would } love to hear from them! } } Mahalo from out here in the middle of the Pacific... } } 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/ * } ***************************************** } ---------------------------------------------------------------------
Hi,
Aldehyde fixation, even with gluteraldehyde, should not prevent lectin binding. We've had good success with a wide range of lectins on formaldehyde and glut. fixed human material, both LR white and araldite embedded (we prefer LR white). To visualise bound lectin, we used biotinylated lectin followed by gold labelled antibiotin. A detailed method is given in:
Localisation of alpha(2,3) and alpha(2,6) linked terminal sialic acid groups in human trabecular meshwork. SA Chapman, RE Bonshek, RW Stodart, KR Mackenzie, D McLeod. British Journal of Ophthalmology, 1994; 78:632-637. }
We modified a method described by Slot and Geuze:
JW Slot and HJ Geuze. In: Polak JM and Varndell IM, eds. Immunolabelling for electron microscopy. Amsterdam: Elsevier Scientific. 1984: 129-142.
We've also visualised lectin binding (including ConA and WGA) in resin embedded tisue at the LM level:
Glycoconjugates of the human trabecular meshwork: a lectin histochemical study. SA Chapman, RE Bonshek, RW Stoddart, CJP Jones, KR Mackenzie, E O'Donoghue, D McLeod. Histochemical Journal, 1995; 27:869-881.
This gives specificities, controls, etc, for a wide range of lectins.
Good luck!
Richard.
Richard Bonshek Lecturer/Honorary Consultant in Ophthalmic Pathology Department of Pathological Sciences University of Manchester Oxford Road Manchester M13 9PT UK
Hi, The question has been asked of me, what stain (prior to embeddment) to use to be able to see a cluster of cells embedded in LR white. Since post sectioning colloidal gold labeling is to be performed, the cells have not been osmicated. They are translucent in the polymerized block. From my understanding, the targeted gold binding site is a glycoprotein. Does anyone have any suggestions as to what stain to use to be able to visualize the cells, yet not interfer with labeling?
Please can someone help me? I'm trying to measure the grain size of TiO2 particles by SEM using a deposition of this particles from a 1% Renex 300 solution. Unhappiness I fail to separate the particles that are forming agglomerates in this solution. Can someone give me some hint?
Ubirajara Pereira Rodrigues-Filho Instituto de Quimica - UNICAMP Campinas, SP, Brazil e-mail: ubira-at-iqm.unicamp.br
Subject: Time: 7:25 AM OFFICE MEMO HowToPolish Mo Date: 4/16/96
There is a graduate student in our department who has a single crystal rod of Molybdenum. He wants to cut off thin slices, polish them so they are of known orientation, flat, and free of surface cold-work, so he can use them for some surface nucleation studies.
Is there anyone out there than could recommend a method for doing this that might be accomplished in a reasonable time and with the equipment normally found in metallography laboratories? If so, it would help him out a great deal. Information on good, easily-controlled etchants and polishing reagents for Mo might also be helpful.
Message-Id: {199604161438.QAA07474-at-pinus.slu.se} X-Sender: mekwall-at-pinus.slu.se X-Mailer: Windows Eudora Light Version 1.5.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" microscopy-at-Sparc5.Microscopy.Com
At 11.51 1996-04-15 -0600, Jane A. Fagerland (847) 935-0104 wrote: } I have a reliable method for hematoxylin and eosin staining of GMA } sections that works well for bone and soft tissue sections. I'd be } happy to mail or fax a copy of it to anyone who's interested. } } Jane A. Fagerland, Ph.D. } Dept. Microscopy and Microanalysis } Abbott Laboratories } Abbott Park IL 60064 } Dear Dr. Fagerland I would be very glad if yuo will be so kind and send me a mail about the staing method for GMA sections.
Thanks Marianne Ekwall Swedish University of Agricultural Sciences Faculty of Veterinary Medicine Dept of Anatomy and Histology} } }
I need staining ER and/OR Golgi (ER best) on cultured cells with a histochemical marker that would make the lumen electron dense. Some years ago, I accomplished this (without wanting it) with a potassium chromate stain, but can not remember the exact reference. If handy on somebody desk please send a note directly to me. Gracias.
********************************************************************* *Cesar D. Fermin, Ph.D \|T|/ Fax (504) 587-7389 * *Tulane Medical School /|M|\ Voice Mail (504) 584-2618 * *Pathology/SL79 \|C|/ Secretary (504) 584-2436 * *New Orleans La 70112-2699 /|*|\ Lab/Techn. (504) 584-2521 * * {Fermin-at-tmc.tulane.edu} ----} Prof. Pathology & Otolaryngology * *http://www1.omi.tulane.edu/departments/pathology/fermin/cdftop.html* *********************************************************************
We have a client who is fixing liver, brain, kidney and aorta with either formalin or freshly depolymerized paraformaldehyde and embedding in paraffin using our automated processor. Everything cuts fine except the liver. Fixation time has run from perfusion only, perfusion + 4 hr, perfusion + 24 hr, 4 or 24 hr immersion only. He describes the liver as "turning to dust" as he sections it. All the other tissues (fixed and processed at the same time) are cutting fine. Any liver histologists out there with insight into this problem. Thanks.
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)
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 opportuinities, 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 wildly 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 instructors 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 Location and Dates South Bay Technology - San Clemente, CA Dates: Friday & Saturday - June 14-15, 1996 Friday & Saturday - October 18-19, 1996
Previous Participants (partial list) INTEL, AMD, Motorola, LSI Logic, Conner Peripherals, Univ of Maryland, Univ of New Mexico, UNAM (Mexico), LG Electronics (Korea), Battelle, MEMC, MVA Inc., Univ of Michigan, U.S. Bureau of Mines, IBM, Naval Research Lab, Purdue Univ, Univ of Alabama, Univ of Arizona, Univ of Colorado, Univ of Wisconsin.
Class Size Due to the intensive hands-on aspects of this course, class size will be strictly limited to 10 participants.
Registration Fee: $795 (includes lunches and Friday night Dinner) $695 if registration fee paid by April 30, 1996 for June Workshop and by August 31, 1996 for October Workshop
Registration Deadline: 30 days prior to workshop
For additional Information: Monica Pflaster South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 TEL: 800-728-2233 FAX: 714-492-1499 e-mail: sbt-at-msa.microscopy.com
Registration Form
To register for the workshop, please fill out this form and send it, with registration fee to:
South Bay Technology, Inc. Workshop on Tripod Polishing 1120 Via Callejon San Clemente, CA 92673
Payment must be made in the form of a check, money order, Visa or MasterCard. Checks must be drawn on a U.S. Bank and made payable to South Bay Technology, Inc. Credit card orders by FAX may be sent to South Bay Technology at 714-492-1499.
Hello! Our EM lab has a DAGE-MTI SIT 68 video camera attached to the TEM and I am looking for a way to get those images into a Macintosh 8500 (Power PC) to analyze/process with the software IMAGE. The problem is that the SIT 68 has an oddball line scan (2:1 interlace; 875/60) so I guess I'm looking for a PCI board for the Mac that will grab and average the images. Even though it's an AV Mac, it won't take this signal as is. Does anyone have any suggestions, short of buying a new camera? Or having this one set to standard? We'd like to keep the high resolution, if possible.
Also, once I finally have images on the Mac, I'd like to print them and quit squandering time, money , environment etc in the darkroom. I've been looking into 8 X 10 video printers but am wondering if anyone has had any luck with some of the higher resolution laser printers that have come out? I only need black and white.
Like most folks, we're pretty broke so I'm trying to do this on a shoestring. Thanks for any help you can give me!
I've been receiving mail from your site, and I'd like to remove my name from your list. I didn't sign up myself, but someone else may have. I'm not even sure what your address is. If you want to talk to me about this, call me at 612-683-8732. Thanks.
Does anyone know of a good INDEPENDENT KEVEX service engineer that services the OH area? I am interested in having mapping installed on a KEVEX 8000 unit. If anyone has any suggestions please eMail me at robert414-at-aol.com. Thanks.
} Hi, } The question has been asked of me, what stain (prior to embeddment) to } use to be able to see a cluster of cells embedded in LR white. Since post } sectioning colloidal gold labeling is to be performed, the cells have not been } osmicated. They are translucent in the polymerized block. From my } understanding, the targeted gold binding site is a glycoprotein. } Does anyone have any suggestions as to what stain to use to be able to } visualize the cells, yet not interfer with labeling? } } Randy Nessler } rnessler-at-emiris.iaf.uiowa.edu
Try 0.1% Neutral Red. May need to go stronger. In water, or buffer. Phil
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
**** (Don't let the bastards wear you down) ***********************
The following message was passed on to me. I have no information as to the veracity of this report or as to the ferocity of the virus, but take note. Maybe others have more information.
This is serious stuff; thought you might like to review note I received:
"VIRUS ALERT" "VIRUS ALERT"
1. This virus alert pertains to you if you have access to on-line services such as AOL, CompuServe or any other service that allows you to download files.
2. A new TROJAN HORSE virus has emerged on the internet with the name PKZIP300.ZIP, so named as to give the impression that this file is a new version of the PKZIP software used to ZIP(compress) files.
3. DO NOT DOWNLOAD THIS FILE UNDER ANY CIRCUMSTANCES!
4. If you install or expand this file, the virus will wipe your hard disk clean and will affect modems at 14.4 baud rate and higher. This is an extremely destructive virus and there is not yet a way to cleaning up this destructive virus.
REPEAT: DO NOT DOWNLOAD ANY FILE WITH THE NAME "PKZIP300 reqardless of the extension.
5. This message was confirmed by MicroSoft security.
by tmcpop.tmc.tulane.edu (8.6.12/8.6.12) with SMTP id LAA20505 for {Microscopy-at- MSA.Microscopy.Com} ; Tue, 16 Apr 1996 11:01:00 -0500 Received: from tmcpop.tmc.tulane.edu (tmcpop.tmc.tulane.edu [129.81.174.79]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id LAA07688 for {Microscopy-at-spa rc5.microscopy.com} ; Tue, 16 Apr 1996 11:01:17 -0500 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id LA A07691 for dist-Microscopy; Tue, 16 Apr 1996 11:01:19 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by ormail.intel.com (8.7.4/8.7.3) with SMTP id SAA25166; Tue, 16 Apr 1996 18:39:24 -0700 (PDT) Received: from ormail.intel.com by relay.hf.intel.com with smtp (Smail3.1.28.1 #2) id m0u9MDN-000qDLC; Tue, 16 Apr 96 18:39 PDT
We have a Zeiss CEM 902 transmission EM. The LCD needs replacement, and Zeiss is charging about US$4000. This seemed outrageously expensive. Does anyone have experience like this? Is this reasonable pricing? Any suggestions will be welcomed. Please respond directly to mbyhsun-at-ccvax.sinica.edu.tw. Thanks very much.
Mr-Received: by mta RANDB; Relayed; Tue, 16 Apr 1996 16:56:01 -0600 Mr-Received: by mta MCM$RAND; Relayed; Tue, 16 Apr 1996 16:56:12 -0600 Mr-Received: by mta RANDC; Relayed; Tue, 16 Apr 1996 16:56:24 -0600 Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
Since I received so many requests for this procedure, I decided it would be faster to just send it via e-mail. If you have specific questions, e-mail or telephone me directly.
REAGENTS: Shandon Instant Hematoxylin: available from Shandon/Lipshaw (1-800-547-7429), catalog number 9990107. Prepare according to package instructions.
Eosin/Phloxine B: 1% aqueous eosin 50.0 ml 1% aqueous phloxine B 5.0 ml 95% ethanol 390.0 ml glacial acetic acid 2.5 ml
Scott's Solution: tap water 1 liter magnesium sulfate (anhydrous) 10 g sodium bicarbonate 2 g
PROCEDURE:
1. Filter hematoxylin solution through Whatman No. 1 filter paper. 2. Stain sections in hematoxylin - 45 minutes 3. Rinse in running tap water - 5 minutes 4. Scott's solution - 2 minutes 5. Running tap water - 5 minutes 6. 70% ethanol - 10 dips 7. Eosin/phloxine B - 20 minutes 8. 80% ethanol - 10 quick dips 9. absolute ethanol - 2 or 3 changes, 5 dips each 10. xylene - 2 changes, 5 dips each 11. mount in Permount type medium and coverslip
I've used this for bone, lung, lymph node, liver and kidney. You can vary the times in the stain solutions, depending on your preferences. Intensity of eosin can be controlled by changing the time in 80% ethanol.
Good luck!
Jane A. Fagerland, Ph.D. Dept. of Microscopy and Microanalysis D45M/AP31 Abbott Park IL 60064
Could someone suggest a reliable (permanent) water - based mountant. I wish to make wholemounts of muscle-nerve preparations which have been stored in glycerin. Any help would be appreciated.
Brett
---------------------------------------------------------------------------- Brett W. Cockman Technologist in Charge School of Dentistry University of Western Australia Voice: (619-2205834) Fax: (619-2213829) e-mail; bcockman-at-uniwa.uwa.edu.au
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
Ubirajara Iereira Rodrigues-Filho wrote the following:
Please can someone help me? I'm trying to measure the grain size of TiO2 particles by SEM using a deposition of this particles from a 1% Renex 300 solution. Unhappiness I fail to separate the particles that are forming agglomerates in this solution. Can someone give me some hint?
I am assuming you are talking about particle size of the TiO2 particles and that the problem is in getting them to disperse. Let me describe the method we have been using for more than twenty five years. It is of unknown origin but it was taught to me by a very creative and innovative microscopist many years ago who spent his career working at the DuPont Experimental Station (Robert P. Schatz).
The "secret" is the magic mixture of 60% camphor/40% naphthalene which when melted together, the two organic compounds form a eutectic which itself melts at just a few degrees above room temperature.
The procedure is to make some of the liquid of the right composition, e.g. 60%/40%, and then add on the order of 0.1-.0.5 wt% of the colloid to be dispersed, in this case, TiO2. Stubborn behavior on the part of the particles can be dealt with by way of a few minutes in an ultrasonic shaker.
With any simple eye dropper, a drop of the liquid (containing the particle dispersion) is put on the surface to be used as the substrate, and since it is at room temperature, the liquid is almost instantaneously frozen, resulting in a thin film coating on the surface. If an SEM mount, then "surface" is the surface of the SEM mount. If TEM studies are contemplated, a carbon coated glass slide would be the surface. In either case, the end result is a substrate surface covered with a thin frozen film of the eutectic system.
The interesting characteristic of the eutectic is that at room temperature, in a vacuum evaporator using ordinary rotary pump vacuum levels, the solid layer will sublime away (typically an over night procedure), leaving the colloidal particles randomly scattered around on the substrate surface. From this point, the sample can be prepared either for SEM or TEM, depending on particle size. It has been our own experience that for really careful studies on TiO2 particles, TEM will give much better images from which measurements could be taken.
With regard to the camphor and naphthalene, nothing special is needed, just ordinary off-the-shelf material of that type that is found in most chemistry laboratory storerooms.
We have found this technique to be useful in the dispersion of a wide range of inorganic particles or any other particle that would not be effected by the camphor/naphthalene mixture.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
On Fri, 12 Apr 1996, Michel Deschuyteneer wrote:
} Fellow microscopists, } } A colleague needs to embed nasal cavities from mice for immunocytochemistry. } His antigens are fragile and do not whistand the standard embedding } procedure with paraffin. Cryosectioning works but would preferably be } avoided because of the bone tissue and decalcification has also proven } detrimental to the antigenicity of his preparations. } } I suggested to try glycol metacrylate but I have no hands on experience with } this material. In addition, there is a variety of commercial kits available, } e.g. Polysciences' Immunobed or JB4, but the differences are not exactly } obvious to me.
} Can anyone recommend a particular preparation of this medium? What are the } pros and cons? Is there some good alternative?
These kinds of questions are addressed in the following two publications:
Stirling, John W., Histochemical Journal 24, 190-206 (1992)
Gerrits, Peter O., Eppinger, Bernhard, van Good, Harry, and Horobin, Richard W., Cells & Materials, 1, No.3, 189-198) 1991
There can be some major differences in different GMA or HMPA based kits coming from different sources. The first of the two above papers gives specific mention about "low acid" GMA. The exact acid level can depend on the purification process being used, the quality of the packaging, and of course age of the product and the conditions of storage. The acid level can make a big difference in the quality of the final results. For LM, the acid component shows up as much higher levels of background staining.
Standing in the SPI Supplies exhibit booth at trade shows might not be the most perfect of scientific methods to draw conclusions, but the impression I get is that because GMA has a viscosity like water, when it comes to "bone tissue", it infiltrates quite nicely. Some have told me "better than anything else". Another advantage of GMA is that absolutely no alcohol dehydration is needed, since the monomer acts as its own dehydrator. This is the presumed explanation for there being a generally greater retention of antigenicity than with other commonly used resins. The biggest perceived negatives associated with the use of GMA seem to be the longer than normal learning curve frequently reported in terms of learning how to work with it the first time. Toxicity is often times cited as a negative, however, relative to most of the other acrylates used in microscopy, it surely is not any worse, and some have told me it does not effect them as much. Cost is also a factor since the GMA and HPMA kits are certainly more expensive than some alternative resin systems.
Disclaimer: SPI Supplies is the original commercial supplier of low acid GMA and HPMA kits for both light and electron microscopy and we would love to see more people enjoying the benefits of low acid GMA and low acid HPMA embedding.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
Message-Id: {199604171315.OAA14894-at-mailspool.liv.ac.uk} Comments: Authenticated sender is {t.g_reed-at-fs2.mt.umist.ac.uk} GVKM07A-at-PRODIGY.COM (DR CHARLES A GARBER)
Referring to Chuck Garber's ingenious method for dispersing particulates for microscopy, what are the safety/health issues (if any) of pumping the camphor/naphthalene mixture into the atmosphere?
Message-Id: {2.2.16.19960417153721.21ef4c84-at-email.uncc.edu} X-Sender: sfzane-at-email.uncc.edu X-Mailer: Windows Eudora Pro Version 2.2 (16) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Hi Everyone, There are a couple of questions which I would like to ask those of you who are associated with small EM labs in a university setting. Those of you who are kind enough to respond, would you share also the number of people served by your facility and the number of technicians employed?
1. Do your labs have a director?
2. If so, is that director a 9 month employee or a 12 month employee?
3. Is the director a faculty member or a staff employee?
4. What are the directors responsibilities?
5. If your labs do not have a director, is there a technician who assumes a managerial position?
6. What are the duties of the technician/manager?
I will be most appreciative for all responses.
Sandra Zane Sandra F. Zane, EM Tech. sfzane-at-email.uncc.edu Dept. of Biology, UNCC Ph.(704)547-4051 9201 University City Blvd. Fax (704)547-3128 Charlotte, NC 28223
I am in the process of replacing a sputter coater used for our SEM samples. I am seriously considering three units: 1) Desk II with carbon yarn evaporation accessory from Denton; 2) EFFACoater and EFFA Mark II Carbon Coater from Ernest F. Fullam Inc; 3) SPI Module Sputter/Carbon Coating System from SPI Supplies.
I would like to hear from anyone who has used one or more of these systems. What did you like best, hate worst, overall performance, reliability.... If you've used more than one, how would you compare them?
I'm considering including a thickness monitor. If you have one, do you find it useful or a waste of money?
If you would rather not have you response included in a summary, please so indicate.
My thanks to all who take the time to respond. ------------------------------------------------------------------------------ Bede Willenbring H.B. Fuller Company
There is a quick alternative to H & E staining of GMA sections. This works on sections that are 0.5 to 2 microns thick. The stain is called Lee's methylene blue-Basic fuchsin Stain. This stain takes about 1 minute and gives some real nice staining similar to H & E. If you want anymore info about this, contact me at psic-at-uclink4.berkeley.edu
Camphor and naphthalene are both materials that are commonly used as moth balls, and as such have been left laying around all over the place in closets, drawers and storage chests in many homes throught the world for many years. Probably the actual level of danger from them is minimal, but if you bring the matter up, OSHA can probably find a lot of restrictions to apply (as in the case for shipping distilled water into a laboratory).
We had a dean once who used to tell us, "If you don't want to hear the answer you know I'll have to give, then don't ask the question!"
I've gotten liver to section by facing off the block to expose the tissue and then soaking the blocks in ice water for about an hour. If you want to do serial sectioning, you may need to soak the block again as you cut deeper into the block.
Hope this helps you.
Jean Ross Central Microscopy Research Facility Univ. of Iowa 85 EMRB Iowa City IA 52242 (319)335-8142 Web site: http://www.uiowa.edu/~cemrf
On Tue, 16 Apr 1996, Tom Phillips wrote:
} We have a client who is fixing liver, brain, kidney and aorta with either } formalin or freshly depolymerized paraformaldehyde and embedding in } paraffin using our automated processor. Everything cuts fine except the } liver. Fixation time has run from perfusion only, perfusion + 4 hr, } perfusion + 24 hr, 4 or 24 hr immersion only. He describes the liver as } "turning to dust" as he sections it. All the other tissues (fixed and } processed at the same time) are cutting fine. Any liver histologists out } there with insight into this problem. Thanks. } } 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 is serious stuff; thought you might like to review note I received:
"VIRUS ALERT" "VIRUS ALERT"
1. This virus alert pertains to you if you have access to on-line services such as AOL, CompuServe or any other service that allows you to download files.
2. A new TROJAN HORSE virus has emerged on the internet with the name PKZIP300.ZIP, so named as to give the impression that this file is a new version of the PKZIP software used to ZIP(compress) files.
3. DO NOT DOWNLOAD THIS FILE UNDER ANY CIRCUMSTANCES!
4. If you install or expand this file, the virus will wipe your hard disk clean and will affect modems at 14.4 baud rate and higher. This is an extremely destructive virus and there is not yet a way to cleaning up this destructive virus.
REPEAT: DO NOT DOWNLOAD ANY FILE WITH THE NAME "PKZIP300 reqardless of the extension.
5. This message was confirmed by MicroSoft security.
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
Tony Garratt-Reed asked the following question:
Referring to Chuck Garber's ingenious method for dispersing particulates for microscopy, what are the safety/health issues (if any) of pumping the camphor/naphthalene mixture into the atmosphere?
Glad you raised the issue, it was one I have not thought about for a long time. Of course as Wil Bigelow pointed out, part of what we are talking about is just moth balls. And since such a small quantity is actually being sublimed, the total volume is just one drop from an eye- dropper, whenever I have myself walked into the lab area when this was being done, I have never even smelled the naphthalene. But to be on the safe side, I guess good practice would suggest that pumps be vented to the outside, right?
Of course the whole set up when preparing the eutectic should be done under a vented hood as should any work involving organic chemicals.
But since nothing seems to be present at levels high enough to smell, perhaps incorrectly, we have not worried about it further.
With regard to my "teacher" of the technique, Bob Schatz, who is no longer with us, he used to say that "the best things in life are those that come free, and the best techniques in the EM lab are those that don't require expensive materials". How right he was.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
At 11.17 1996-04-17 -0700, dbd1-at-uclink4.berkeley.edu wrote: } To All Who Are Interested, } } There is a quick alternative to H & E staining of GMA sections. This works } on sections that are 0.5 to 2 microns thick. The stain is called Lee's } methylene blue-Basic fuchsin Stain. This stain takes about 1 minute and } gives some real nice staining similar to H & E. If you want anymore info } about this, contact me at psic-at-uclink4.berkeley.edu } } Hallo } Iwould like to have more info about the staining. } Marianne Ekwall
Comments: Authenticated sender is {t.g_reed-at-fs2.mt.umist.ac.uk}
Dear Mark
There are two issues regarding light element analysis, namely, does the X-ray reach the detector, and is the system capable of recording it.
Regarding the first issue, there is more to the arrival of x-rays at the crystal than just the "thin window". What is the window made of? A polymer window can be quite good at transmitting B x-rays, because the boron energy is below that of the carbon absorption edge, provided the window is not too thick. However, on older detectors there is, I am sure, a thick layer of ice, which may be microns thick. This will do a fine job of absorbing boron x-rays (and any other low-energy ones, too). Modern detectors (from some manufacturers, at least) are quite able to withstand a cycle to room temperature and a pump-out, and at least one manufacturer fits a small heater to the crystal to allow it to be warmed, to sublime the ice. If you have a TN5500, though, your detector may be fairly old.
Regarding the recording of the x-rays, one major problem is system noise, which also degrades the spectral resolution. All manufacturers have made steady progress at reducing the noise with the passing of the years. However, noise can also be seriously compromised by installation peculiarities, or inadvertent ground loops, etc. This could be especially the case on an older installation where, perhaps, a succession of people have been responsible for it since the original installation, so that the special arrangements made to reduce the noise originally have been forgotten. Some systems allow you to view the "zero" peak which enables you to judge directly the system noise. I don't know if the 5500 does this - many systems cut the spectrum off a few channels above zero.
Yet another problem arises because of the very low x-ray yield from light elements. If you analysing for boron in the presence of heavier elements, than the boron peak is superimposed on the bremsstrahlung from the heavier material. I did a "back-of-the-envelope" calculation once that said that in LaB6 the boron concentration is about 4 times the minimum detactable concentration!. This would not be such a problem if you are analysing boron metal, but could still be an issue for boric acid.
In the SEM, of course, you will only get a tiny boron signal, even though the counts in the rest of the spectrum may be high, because of the very large absorption of the boron x-rays in the sample. You can minimise the effect by operating the SEM at a very low voltage ( you only need 600eV electrons to excite the boron), but of course you will lose source brightness and electron intensity (or, conversely, spatial resolution) by doing this.
MARK DARUS (216) 266-2895 wrote: } } } Hello, } My EDX is a TN-5500 and I'm trying to determine if it is sensitive, } enough to detect Boron. I have some standards, and in a moment I'm going to } set up some boric acid and look into it. Just placing it in the chamber and } getting the counts up, plus switching it to thin window doesn't give me } overwhelming success from an SPI metal standard that I have. } I'm sure there is much more to it than that. Is there any advice that you may } have to offer. My beam current settings are 1, 5, 10, 20 & 30 KeV. } } Mark Darus } } Darus-at-cle.dnet.ge.com
A word of caution! Boric acid has high vapor pressure and will sublime under SEM vacuum conditions contaminating your column. I would suggest some other compound to test.
--
Naresh Shah
University of Kentucky Consortium for Fossil Fuel Liquefaction Science (CFFLS) and Department of Chemical and Materials Engineering (CME) 533 South Limestone Street, Room 111 Lexington, KY 40506-0043 Phone: (606)257-5119, Office: (606)257-4027 FAX: (606)257-7215 e-mail: naresh-at-pop.uky.edu
Greetings people: Can anyone e-mail me, or post, their experiences and opinions of microwave processing for TEM. I work in a diagnostic pathology lab. We primarily want to speed-up embedding time; but your comments concerning any stage of processing will be appreciated.
Does anyone know if there is a dual filter available for viewing FITC and TEXAS RED at the same time? We have a Leitz Orthoplan 2. A student is interested in purchasing this filter (perhaps used ?), if the cost is reasonable. Thanks .....also thanks for all the help with the Au target for sputter coating. Linda lfox1-at-wpo.it.luc.edu
We have been using antigen retrieval with desmin immunostain on mice tissue and are getting some nice staining but not quite what we expected. Our literature search has not turned up anything specific involving the antigen retrieval and desmin , does anyone know of any references or had any experience with this scenario ?
Your mail was returned as undeliverable. My company manufactures replacement computer control systems for the JAMP-30 Auger system, but not parking stages. That is available from JEOL.
You may contact Geller MicroAnalytical Laboratory at sales-at-gellermicro.com.
Your mail was returned as undeliverable. My company manufactures replacement computer control systems for the JAMP-30 Auger system, but not parking stages. That is available from JEOL.
You may contact Geller MicroAnalytical Laboratory at sales-at-gellermicro.com.
Mr-Received: by mta RANDB; Relayed; Thu, 18 Apr 1996 08:08:36 -0600 Mr-Received: by mta MCM$RAND; Relayed; Thu, 18 Apr 1996 08:09:11 -0600 Mr-Received: by mta RANDB; Relayed; Thu, 18 Apr 1996 08:09:38 -0600 Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
Bede,
We have used a Desk II for several years and it has performed extremely well. The only maintainence we have done is replace worn out targets and chnage the pump oil once a year.
Joe Neilly Abbott Laboratories Dept. of Microscopy and Microanalysis North Chicago, IL 60064
I was just reading a "How to" chapter about problems of this sort. The text is A manual for Histologictechnicians, Third edition, by Ann Preece, Little, Brown and Co. Pubs., 1972. They state the reasons for this crumbling could be inadequate fixation or incomplete dehydration. Sometimes the tissue in the block is just too hard. The suggest trying to soften the tissue first by placing it in water with a little detergent added (1/2 tsp. per 100 ml H2O) and let it soak for up to 3 hours. If this doesn't work, you may have to deparafinize the block and refix and dehydrate and reembedd the block. Depending on how the tissue was originally fixed, i.e. cardiac perfusion, the liver just may have not gotten completely exposed to the fixative. I haven't tried these tricks myself yet so if you have any luck, let me know. Good luck.
Karen Pawlowski
On Tue, 16 Apr 1996, Tom Phillips wrote:
} We have a client who is fixing liver, brain, kidney and aorta with either } formalin or freshly depolymerized paraformaldehyde and embedding in } paraffin using our automated processor. Everything cuts fine except the } liver. Fixation time has run from perfusion only, perfusion + 4 hr, } perfusion + 24 hr, 4 or 24 hr immersion only. He describes the liver as } "turning to dust" as he sections it. All the other tissues (fixed and } processed at the same time) are cutting fine. Any liver histologists out } there with insight into this problem. Thanks. } } 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) } } }
Hello All, I have recently strted trying to do some quantitative backscattered imaging of lead zirconium titanate (PZT). I'm getting rather puzzled by the results and wondered if anybody had any ideas...
I naively took some papers at face value which state that backscattered intensity is directly proportional to the mean atomic number of the material. However, I am getting about twice the backscattered signal from PZT that I expect. The composition has been confirmed by microprobe analysis, so I guess that complex compounds and/or oxides behave differently from single elements and binary metals (?) I also see very strong differences in contrast between a nanocrystalline phase (pyrochlore) and a large grained phase (perovskite), although both are _supposed_ to be of the same composition. Am I actually seeing composition differences or does the grain size have a large effect on backscattered intensity? Both effects are present when using a TEM (accelerating voltage 20-100kV) and a FEGSEM (accelerating voltage 0.5-25kV).
Any ideas, references or advice would be very welcome!
Many thanks in advance,
Richard Beanland, GMMTL Caswell, Towcester, Northants NN12 8EQ UK.
Dear Gene: There are several published reviews reporting how microwave-accelerated processing methods are incorporated into routine histopathologic practice. A recommended review is: 1. Leong, A. S.-Y. Microwave fixation and rapid processing in a large throughput histopathology laboratory. Pathol 23: 271-273, 1991.
Reviews of microwave Fixation for TEM and LM are reported in: 1. Login, G. R., and A. M. Dvorak. Methods of microwave fixation for microscopy. A review of research and clinical applications: 1970-1992. Prog Histochem Cytochem 27/4: 1-127, 1994. 2. Kok, L. P., and M. E. Boon. Microwaves for microscopy. J Microsc 158: 291-322, 1990. 3. Leong, A. S.-Y. Microwave technology for morphological studies. Cell Vision 1: 278-288, 1994.
A review of microwave-accelerated embedding is in: 1. Giammara, B. Microwave embedment for light and electron microscopy using Epoxy resins, LR White, and other polymers. Scanning 15: 82-87, 1993.
Two books describing how to get started using microwave techniques: 1. Kok, L. P., and M. E. Boon. Microwave Cookbook for Microscopists. Leyden: Coulomb Press, 1992. 2. Login, G. R., and A. M. Dvorak. The Microwave Toolbook. A Practical Guide for Microscopists. Boston: Beth Israel Hospital, 1994.
Gary Login (my e-mail address is at the end of this message)
In message {960418095422_472715458-at-emout13.mail.aol.com} writes: } Greetings people: } Can anyone e-mail me, or post, their experiences and opinions of microwave } processing for TEM. I work in a diagnostic pathology lab. We primarily want } to speed-up embedding time; but your comments concerning any stage of } processing will be appreciated. } } Gene Santagada } genexs-at-mhv.net } genexs-at-aol.com } } Kalvin Electron Microscope Lab. } Lenox Hill Hospital }
Gary R. Login, D.M.D., D.M.Sc. Assistant Professor of Oral Pathology Beth Israel Hospital Department of Pathology 330 Brookline Avenue Boston, Massachusetts, 02215
Subject: Time: 7:21 AM OFFICE MEMO Safety Regs & water Date: 4/18/96
Following my comments on the safety regulations and the use of champhor and naphthalene, several people have asked if OSHA does indeed require an MSDS for water, and apply the usual chemical safety regulations to it. I assure you that they actually and in fact do. Here is a message on the subject I received recently from one of my ex-students: - - - - - - - - - - - - - - - "At Lockheed, the chem lab had a high resolution chromotography system requiring triple distilled water. The bottles had to be shipped with an MSDS, they were labeled "Hazardous", and they had to be inventoried as a "Chemical". Likewise, an NBS EDS standard of stainless steel had to be handled as a "Hazardous Material" because it contained chromium!" - - - - - - - - - - - - - - Further support can be found in a column written by Mike Ryoko, a syndicated columnist whose writings appear in many newspapers throughout the country, a couple of days ago. In this column he referrs to a similar situation that was called to his attention in the Chicago area. He of course makes a number of witty comments about the situation.
There are, of course, many other examples of situations where safety regulations have been enforced to a ridiculous end. I heard of one laboratory where a bottle of sand, which was unfortunately had the chemical formula SiO2 written on it, was declared a hazardous substance (presumeably because it contained the element silicon).
We used to have a foundry in our department which used large quantities of ordinary sand for various purposes. Again some of this material was in bags labeled "Silica Sand", and when workers who were cleaning out the foundry area spotted this they called in a full safety crew complete with gas masks, sealed suits, and all related equipment, to dispose of the material. This was done in the summer, and undoubtedly some of the workers spent their weekends picnicking at the beach without giving the hazard of doing so a second thought.
In a high school not far from here a student accidentally knocked over a graduated cylinder, spilling about 100 ml of ethyl alcohol. The entire school was closed down for the remainder of the day while the local fire department was called in to clean up the spill. It was later learned that several teachers celebrated the unexpected vacation by stopping at a local bar and consuming more alcohol than was spilled at school.
These are only a few examples that I know of. There undoubtedly many more floating around, and it would perhaps be interesting to hear about them.
I don't mean to imply that I am against appropriate safety regulations, and I do recognize that the imposition of such regulations have been very beneficial in protecting people in a great number of situations. However, in some instances, largely because of the ignorance and perversity of those who formulate and enforce these regulations, the system has been pushed to ridiculous extremes.
In closing, I should correct myself. I said that camphor was commonly used in moth balls. I think this is incorrect. The two common ingredients for moth balls are, I believe, naphthalene and para-dichlorobenzene. I know that naphthalene is still used in this way, because I recently bought some naphthalene moth balls to evict a possum from a hole it had dug under my front porch. However, the para-dichlorobenzene may not be so commonly used in this way any more because it is a chlorinate phenyl compound. Camphor, on the other hand, has been commonly used in the stuff you spread on your lips to help heal chapping, and for years was used as a non-prescription medicinal in such things a camphorated oil, nazal decongestants, cough lozengers, etc.
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
Mark Darus wrote:
My EDX is a TN-5500 and I'm trying to determine if it is sensitive, enough to detect Boron. I have some standards, and in a moment I'm going to set up some boric acid and look into it. Just placing it in the chamber and getting the counts up, plus switching it to thin window doesn't give me overwhelming success from an SPI metal standard that I have. I'm sure there is much more to it than that. Is there any advice that you may have to offer. My beam current settings are 1, 5, 10, 20 & 30 KeV. ==================
There is just no better "standard" than a solid lump of pure boron! It is inert and it won't be changed by the electron beam. Boric acid will be unstable and as others pointed out, it will sublime causing all kinds of other problems.
Based on our own experience fielding technical service calls, you should check to make sure no one has inadvertently applied too thick of a carbon coating. People don't admit to doing this but it does happen! This has in fact happened before, with some element of frequency, where the carbon was thick enough to absorb all exiting B x-rays. If that is indeed the case, then read the User Manual that came with the SPI standard for instructions for cleaning it up and returning it to a state where you will see the x-rays. Note: Great care must be taken or you can run the risk of damaging the electron beam lettering opposite each standard item.
Another point: You have listed "beam currents" but these are really accelerating voltages. You did not mention the current, but if you see any x-ray continuum at the B (K alpha) energies, then one can assume the detector is working. To be sure about that, check to make sure you can see the slightly more energetic C (K alpha) from the carbon standard.
More often than not, the problem is simply that the detector, for one reason or another is just not "up" to it. But you should check whether it is a transient kind of thing (e.g. ice) or something more serious.
Additionally, and no one likes to think this way, what if something horrible did happen and something did get mixed up along the way? But "it" still does have to be "something" and the x-rays from that something surely should be able to be measured and the material actually present identified.
Hope this information might be helpful to you.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
Dear Microscopists, We have two Reichert OMU2 ultramicrotomes which we would like to give away (either whole or whatever pieces people want) to anyone prepared to cover freight costs.
They are mid 1960's vintage, one is complete and one is missing a few bits, like the binoculars. They both still have their antivibration tables. I know it is long shot, but they may be useful for someone out there...otherwise to the tip they go.
Contact me if interested.
Yours faithfully,
Richard
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
This is a slightly facetious input re. safety data sheets!
In the UK there is a very useful pair of publications from BDH, a chemicla supply company (used to be known as British Drug House, I believe). These are collcted data sheets. Yes folks, there is one for water. Here are a few salient points which all users of the substance should bear in mind at all times:
1. colourless liquid - maybe that implies it is rare and difficult to see if dropped! 2. Against solubility in water: miscible in all proportions. 3. fire and explosion hazard: not applicable (thats a relief - although if it caught fire I suppose one could foolishly attempt to extinguish with more water?). 4. Health hazard: no significant hazard expected, may be irritating to the eyes. 5. Toxicity: no data. 6. Carcinogenicity: no evidence of carcinogenic properties. 7. First aid - eyes: irrigate thoroughly with water(!) 8. First aid - lungs: remove from exposure. 9. First aid - skin: wash off thoroughly with soap and water (work that one out!). 10. First aid - mouth: wash out thoroughly with water. In severe cases obtain medical attention(!) 11.Reactive hazards: violent reaction with acyl halides, alkali and alkaline earth metals .... 12. Spillage disposal: wear appropriate protective clothing - listed are gloves, goggles, face shield and protective apron. Luckily, respirators are not needed.
Have a nice day (but watch out for water raining down!).
Keith Ryan (Local Safety Advisor - among other hats worn!) (used to be a good EM guy - there's a cry from the heart!) Plymouth Marine Laboratory Citadel hill Plymouth PL1 2PB, England
Jolanta Mesjasz asked about the availability of aluminum index or finder grids.
As some of you know, I am responsible for the laboratories of Structure Probe, Inc., parent company of SPI Supplies. We would love to be able to sell finder grids and other specialty products in aluminum, but we have not been able to develop products that meet our quality requirements. The reason, unfortunately, is very simple but also very basic.
Most TEM grids are made by electrodeposition. The copper, nickel, gold and other precious metal grids which we and our competitors offer are plated. While there is a lot of technology involved in the process, it is basically very simple, and it allows control over bar width and other details to an incredible degree; letters, numbers, fiduciary marks and other identifications are easy compared to controlling bar width within a range of micrometers of the nominal dimension.
For aluminum, tungsten, molybdenum, beryllium and stainless steel grids, however, the production process uses etching. There are several problems, but they come down to problems in the resolution of the imaging process, the thickness of the grid material and the nature of the etching process. We tear our hair out trying to put out product which LOOKS like a grid, with bars, etc. Details like letters and numbers are simply beyond our capability at this point; we do not know of a process which can produce these details at a price anyone is willing to pay.
If a reader knows how to produce an aluminum (or other etched) finder grid, we (and I suspect our competitors, as well) would be most happy to talk with you about setting up production.
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 2262 e-mail: AWBlackwoo-at-aol.com WWW: http://mail.cccbi.chester.pa.us/spi/spihome.html
} I am in the process of replacing a sputter coater used for our SEM samples. } I am seriously considering three units: 1) Desk II with carbon yarn } evaporation accessory from Denton; 2) EFFACoater and EFFA Mark II Carbon } Coater from Ernest F. Fullam Inc; 3) SPI Module Sputter/Carbon Coating } System from SPI Supplies. } } I would like to hear from anyone who has used one or more of these systems. } What did you like best, hate worst, overall performance, reliability.... If } you've used more than one, how would you compare them? } } I'm considering including a thickness monitor. If you have one, do you find } it useful or a waste of money? } } If you would rather not have you response included in a summary, please so } indicate. } } My thanks to all who take the time to respond.
We purchased a Denton Desk II with the carbon yarn evaporation accessory acouple of years ago and have had no problems with it. The only things that I have had to do in terms of maintenance is give it one oil change a year, target changes, and cleaning of the glass cylinders & various removable stage parts.
Peling Melville
-------------------------------------------------------------- Peling Fong Melville Senior Scientific Assistant Interdepartmental Facilities American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 U.S.A. ****************************** E-mail: peling-at-amnh.org Work #: (212) 769-5469 FAX #: (212) 769-5495
In a past life Sandy Brady and I gave several workshops on GMA processing and staining. If I remember we gave out a very comlete handout of staining techniques. I don't know if I can easly locate a copy but there should be some copies floating aroud somewhere. Perhaps Sandy has an extra copy or two. regards bob
Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis University of North Carolina CB#7400, Rosenau Hall Chapel Hill, NC 27599-7400 Ph. 919-966-6343 Fax 919-966-6123
I have been asked to image crystals of nearly pure sodium sulfate. As it is a drying agent which contains 10 molecules of water per molecule of Na2(SO)4 I'm a little concerned about putting it in the SEM. I have put it in a drying oven for 2 days and I was planning to put the stub under vacuum for a while before coating it with gold. Are there any other precautions or preparations I should do?
I'm assuming that you are working with rat or mouse models. We do a lot of rat, mouse and fish liver work in our lab. All of our histology processing is contracted out so I had to develop SOP's for processing and sectioning. If you want I can fax these out to you. We've had no sectioning problems (over 5 years now) since these SOP's were written. regards, bob
Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis University of North Carolina CB#7400, Rosenau Hall Chapel Hill, NC 27599-7400 Ph. 919-966-6343 Fax 919-966-6123
Hi All, A department here at the University of Iowa contacted us about finding a new home for their used Zeiss 10C. It has been under service contract until just recently. Since they have decided not to renew the service contract, they are interested in finding someone who is willing to pay for the system to be dismantled, shipped, and reassembled. These are the only costs, as they will give to the instrument to said party. They have pressing needs for the space that the scope occupies, and thus are considering junking it. It would almost be a shame to recycle a fully functional microscope...... If interested, contact Kenneth Moore (kenneth-moore-at-uiowa.edu) or myself either by email or phone (319-335-8142).
I think that the fire safety issue is important enough to continue the thread. An incident happened in my building last week which reminded us of the importance of smoke detectors. An instrument which had been left running and unattended many times in the past began to melt down when on this occasion was left unattended for 15 minutes. Part of the instrument became red hot, melted a hole in the bench top, and sent thick, black, acrid smoke into the room and two adjacent ones. Directly above the instrument are located air intake and exhaust vents which pulled some smoke and presumably heat out of the room. The heat activated sprinkler (designed to be activated at 135F, we think) did not activate, presumably because heat was exhausted away. Fortunately, a student returned to discover the situation. Later we extracted the following information from the large gathering which included 7 firemen, several safety and facilities management personnel: smoke detectors are not required under current fire safety codes in this city. Our building is two years old. The nearest smoke detector was approx. 100 feet away in the lobby. I suspect that the lack of smoke detectors in laboratories is not unique to this building or this city. The safety office here on campus has at least taken this matter under consideration. We also realized that we have taken equipment reliability for granted.
The Microscopy and Microanalysis Program in the Materials Characterization Laboratory at GE Corporate Research and Development, Schenectady, NY, has an opening for a Microanalyst/Microscopist at the Professional/Lead Professional (BS/MS) level. The primary duties associated with this position involve the execution of research projects involving Electron Backscatter Pattern (EBSP) analysis of crystalline materials in the Scanning Electron Microscope (SEM). Additional duties may involve research conducted using a variety of other electron imaging and analysis equipment, including transmission electron microscopes and electron microprobes. The Microscopy and Microanalysis Program at GE CRD is one of the world's leading centers for the application of EBSP techniques to the analysis of texture and orientation. Two dedicated SEMs are used for this capability. The present position involves the operation and maintenance of that equipment, and the management of project and sample flow through the facility.
The Materials Characterization Laboratory is involved in research into the structure and composition of materials in support of development programs both at GE CRD and at GE businesses. A wide range of materials, including metals, ceramics, composites, polymers, and coatings/surface modifications, are analyzed by this group. Staff members are expected to work independently with a high level of expertise, and to become involved with a number of major project teams. Good communication skills, both written and oral, are extremely important.
A BS or MS in Materials Science or a closely-related field is required. Prior experience with electron beam instruments, particularly transmission and scanning electron microscopes, is highly desirable. Some familiarity with crystallography and computer usage/programming in the Windows environment is also desired.
Resumes and other information can be sent to:
Ernest L. Hall Manager, Microscopy and Microanalysis Program Room K1-2C27 GE Corporate Research and Development PO Box 8 Schenectady, NY 12301 Fax: 518-387-6972 E-mail: hallel-at-crd.ge.com
Does anyone know whether the PKZIP300 virus reported by Scott Walck is directed at Macintosh or IBM computers?? or can it infect both?? W. C. Bigelow (bigelow-at-umich.edu)
} Hello All, } I have recently strted trying to do some quantitative backscattered } imaging of lead zirconium titanate (PZT). I'm getting rather puzzled by the } results and wondered if anybody had any ideas... } } I naively took some papers at face value which state that backscattered } intensity is directly proportional to the mean atomic number of the material. I just taught my radiation sciences class about nuclear backscatter- ing (it was used as an analytical method on the moon, BTW), and the text-- Nuclear and Radiochemistry, Friedlander, et al., p430--states that the cross section is proportional to Z^2. Since it is a Coulomb scattering, and since e-e back scattering would have zero energy, I would expect electron back scattering to have the same Z-dependence as proton backscattering. Per- haps David Joy will comment.
} However, I am getting about twice the backscattered signal from PZT that I } expect. The composition has been confirmed by microprobe analysis, so I guess } that complex compounds and/or oxides behave differently from single elements } and binary metals (?)
This shouldn't be the case for backscattering. There might be minor differences for low-angle scattering due to differences in the valence elec- tron distribution--the low-order CBED amplitudes are sensitive to this.
} I also see very strong differences in contrast between a nanocrystalline } phase (pyrochlore) and a large grained phase (perovskite), although both are } _supposed_ to be of the same composition. Am I actually seeing composition } differences or does the grain size have a large effect on backscattered } intensity?
Does the effect vary with the orientation of the large grains?
} Both effects are present when using a TEM (accelerating voltage 20-100kV) } and a FEGSEM (accelerating voltage 0.5-25kV).
I am not surprised that you see no dependence on voltage. Yours, Bill Tivol
Symposium W: Interfacial Engineering for Optimized Properties Fall 1996 Materials Research Society Meeting December 2-6, 1996
Description: Interfaces often exert a controlling influence on the properties of materials. Consequently, there is a great desire to engineer these interfaces in order to optimize beneficial properties and eliminate those that are deleterious. This optimization is usually achieved by fabrication or processing methods to control the composition, structure, or crystallography of the interface. This four day symposium will address these topics in metals, ceramics, and composites. The discussion will primarily center on internal interfaces, including grain boundaries, interphase interfaces, and film/coating-substrate interfaces. The papers will include those which discuss problems which have been addressed by interfacial engineering and those that discuss the development of methods of interfacial engineering.
The topics to be covered, as they apply to interfaces, include the following: o Strength and toughness o Cohesion and adhesion o Corrosion and embrittlement o Fracture and fatigue o Electrical and optical properties o Creep and diffusion o Reactions and mobility o Measurement and characterization
Partial list of invited speakers : Yet-Ming Chiang (MIT); Gino Palumbo (Ontario Hydro-electric); Fennell Evans (University of Minnesota); Michael F. Henry (GE CRD): Vinayak Dravid (Northwestern); Ali Argon (MIT); Kathi Alexander (ORNL)
Abstracts must be received at MRS Headquarters no later than June 21, 1996 and must follow the standard MRS abstract model. Abstract templates and additional information on the meeting can be obtained from MRS (E-mail: fall96-at-abstracts.mrs.org; WWW: http://www.mrs.org; Phone: 412-367-3003; Fax: 412-367-4373) or from the symposium organizers.
Symposium Organizers:
Clyde L. Briant Div. of Engineering Brown University PO Box D Providence, RI 02912 Phone: (401) 863-2626 Fax: (401) 863-7677 E-mail: briant-at-engin.brown.edu
C. Barry Carter Dept. of Chem. Eng. & Mat. Sci. Univ. of Minnesota 421 Washington Ave., SE Minneapolis, MN 44544 Phone: (612) 625-8805 Fax: (612) 626-7246 E-mail: carter-at-cems.umn.edu
Ernest L. Hall GE CRD PO Box 8 Room K1-2C27 Schenectady, NY 12301 Phone: (518) 387-6677 Fax: (518) 387-6972 E-mail: hallel-at-crd.ge.com
} Mark Darus wrote: } } My EDX is a TN-5500 and I'm trying to determine if it is sensitive, } enough to detect Boron. I have some standards, and in a moment I'm } going to set up some boric acid and look into it. Just placing it in } the chamber and getting the counts up, plus switching it to thin window } doesn't give me overwhelming success from an SPI metal standard that I } have. I'm sure there is much more to it than that. Is there any advice } that you may have to offer. My beam current settings are 1, 5, 10, 20 } & 30 KeV.
Just a few comments about the composition of the detector window material on EDS detectors. The mass absorption coefficients (mac) for Boron Ka radiation by different absorbing elements are (these values from table 14.3 of Goldstein et al):
Absorber mac H 1,723 Be 69,937 B 2,861(?) C 5,945 N 10,118 O 15,774
So B Ka is strongly absorbed by a Be window on your EDS detector, and this (as you know) is the reason that B Ka cannot be detected with this normal window material. Note that carbon is fairly transparent to B Ka, either as an EDS window material via a diamond window, or as a conductive coating layer on your (nominally non-conductive) specimen. You should be able to detect B Ka on a carbon-coated sample and/or with a diamond window just fine; too thick a carbon coat will absorb just about anything, but comparatively it is not too bad. If you have a BN window, then the absorption is a little stronger than for the diamond window (average the macs for B and N). Even slightly worse is for the case of an ice layer (think of it as a layer of oxygen).
Now if you have a too thick carbon coat, and a thick layer of ice on the detector crystal, and maybe you are using a thin Be window or a thicker BN window, then B Ka detection is going to be difficult. As mentioned before, reconditioning the EDS detector to get rid of the ice layer is mandatory before attempting to do light element work like this.
Secondly, the pulse processor has to be optimized for processing low energy x-ray pulses -- if the discriminator is set too high you are filtering out all the low energy x-ray pulses, and if the time constant is set incorrectly you may not be getting the best throughput on the detector. Refer to the manuals at this point.
Suffice to say that optimization of the detector should be done on a pure element (lump boron) before trying to look at lower levels in your samples.
It was also pointed out by Chuck Garber: } There is just no better "standard" than a solid lump of pure boron! It } is inert and it won't be changed by the electron beam. Boric acid will } be unstable and as others pointed out, it will sublime causing all } kinds of other problems.
Note however, from the mac values above, that B Ka is strongly absorbed by many materials (for example, the mac for B Ka by Silicon is 83,702), so you may see very strong absorption in a B-Si alloy (in fact, you may not observe a peak at all for lower concentrations of boron because it is absorbed entirely within the sample!). In this example, the absorption correction via the "A" part of the ZAF correction is very high for your sample, compared to your standard (assuming you used pure boron). The bottom line here is that you absolutely *must* use a standard that is as close as possible to your samples for good quantitative correction. Of course, the available boron standards are pure boron, boron nitride (which is typically polycrystalline and is not a good conductor; single crystal BN is much better but hard to find in pieces larger than a few microns), and numerous borosilicate glasses (which of course have the problem of absorption by silicon). For EDS work you probably have to use pure boron to avoid peak overlaps, but for WDS analysis things like BN can be used.
The magnitude of the absorption correction is directly related to the accelerating voltage used, so if you can do analysis at 10 KV rather than 15 KV, you are better off. You can work at lower voltages (like your 5 KV setting), but the sample surface must be really clean, and you then get into problems where aspects of the correction algorithms are not really valid for this range of accelerating voltages.
As a final note, it is relatively meaningless to look at EDS spectra of pure elements when shopping for an EDS system to do light element analysis. It is the performance on typical multi-element samples that really matters. For example, I was looking at the possibility of studying synthetic vs. natural emeralds (Be3Al2Si6O18) including inspection of the Be Ka peak by EDS. Even with a freshly reconditioned detector on *open window* mode, with the SEM probe current full up, no Be peak was observed. So Be measurement was just not meant to be on an EDS-SEM system. It turns out that even by WDS, using a Mo4C analyzing crystal (red-hot for Be and B), and the probe current full up on our microprobe (400 nA -at- 10KV), and analyzing at low accelerating voltage, and a clean high vacuum turbo-pumped system, that Be in emerald is barely detectable (count times of up to an *hour* were used!). I'll just say that boron analysis is a walk in the park compared to beryllium analysis...
Anyway, good luck!
Paul Carpenter
+----------------------------------------------------+ | Paul K. Carpenter paulc-at-gps.caltech.edu | | Division Analytical Facility | | Geological and Planetary Sciences MC 170-25 | | California Institute of Technology | | Pasadena, CA 91125 | | 818-395-6126 (X-ray Lab) 818-568-0935 (Dept. FAX) | +----------------------------------------------------+
I would like to know if any one has investigated gas permeability properties of plastic/rubber hose. I'm trying to set up a microscope perfusion system with flow from a solution with CO2 at a specific concentration. The problem is that the silicon rubber hose used for interconnection between the parts, cannot keep CO2, which very quickly escapes into surrounding air.
Has anyone heard of a gas impermeable hose (inner diameter ca 1 mm or less) or
Best regards
============================================= Mikael Gustafsson MD, PhD Dept Med. Microbiology and Dept Internal Medicine, Cardiology section University Hospital of Linkoping S 581 85 LINKOPING SWEDEN
Randi, I suggest transferring your semithin sections to a water bath at about 50 Cand pick up on acid clean slides (this is most important!). Dry on a hot plate and then place in hot oven (about 60 C +) for at least an hour. This will guarantee that your sections stay on the slide. I further suggest you remove the resin by treatment with potassium ethoxide (saturated KOH in ethanol; make up at least 24 hours before use then filter): 1. Treat with K-ethoxide for about 3 - 4 mins. 2. Wash well with 3 changes of absolute ethanol. 3. Hydrate through 95%, 70% to distilled water and then dry. 4. Stain with toluidine blue or any other stain!
If you need more details please ask.
. . Brett W. Cockman Technologist in Charge School of Dentistry University of Western Australia Voice: (619-2205834) Fax: (619-2213829) e-mail; bcockman-at-uniwa.uwa.edu.au
We just purchased a new Mohr-Pro print processor but have not been able to use it because our building engineer is concerned that we must vent the unit. The manual does not address this issue and there is no outlet or attachment for venting the chemistry fumes. We have had several processors over the years, with no exhaust vent and have had no problems. Would you please share with us whether you have your print processors connected to a vent system, how you accomplished it, if it is an OSHA requirement, if your system came with an outlet made specifically for venting chemistry fumes and any other pertinent information. Thanks for your assistance.
Donna Wagahoff SIU School of Medicine Springfield, Il. 217-782-0898 fax 217-524-3227
A good sample to test the detector capability is solid boron nitride. It not only gives you the boron K-alpha peak, but also the nitrogen K-alpha for reference.
D. Clark Turner Director, Thin Film Products Group MOXTEK, Inc. 452 West 1260 North Orem, Utah 84057 phone (801) 225-0930 email moxtek-at-moxtek.win.net
} } } } Hello, } My EDX is a TN-5500 and I'm trying to determine if it is sensitive, } enough to detect Boron. I have some standards, and in a moment I'm going to } set up some boric acid and look into it. Just placing it in the chamber and } getting the counts up, plus switching it to thin window doesn't give me } overwhelming success from an SPI metal standard that I have. } I'm sure there is much more to it than that. Is there any advice that you may } have to offer. My beam current settings are 1, 5, 10, 20 & 30 KeV. } } Mark Darus } } Darus-at-cle.dnet.ge.com }
As already suggested, cleaning the slides is important. For different plastics, soaking slides for 30 min. in 50% bleach (commercial bleach mixed 1:1 with dstilled water), or in 70% ethanol containing 1% HCl, will do the trick instead of messing about with chromic acid. Wash several times with destilled water after cleaning.
What are you coverslipping with? Some mountants cause wrinkling unless slides are very clean or the mountant is cut 1:1 with xylene.
With the above approaches, we get reliably flat sections and etch only when necessary for immunocytochemistry.
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
On Sat, 20 Apr 1996, Randi Olsen wrote:
} Hello, } I wonder if there is any help somewhere out there: } Lately we have been fighting some catfish larvaes, embedded } in Epon/Araldite. Because we need sections of the whole } length og larvaes up to 4 weeks old, we need big semithin } sections, and they dont want to strech properly. I have } small foldes (or waves) in the sections wisible at x40 that } make it impossible to get sharp photographs at this } magnification. (The foldes also trap some air between the } glass slide and the sectiokn). Is there anything we can do } to make the sections flatter? } } Thanks in advance. } } Randi Olsen } Department of Electron Microscopy } University of Troms=F6 } Norway } } }
I am trying to collect information from the various manufacturers of Backside Emission Microscopes. If anyone can provide me with names and phone numbers of appropriate manufacturers, I would greatly appreciate it.
Best regards-
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: 73531.1344-at-compuserve.com sbt-at-msa.microscopy.com
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy.
Can anyone provide me with the URL for the ASTM home page?
Thank you!
Best regards-
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: 73531.1344-at-compuserve.com sbt-at-msa.microscopy.com
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy.
Why does the engineer think the processor needs to be vented? There is very very little mixture of chemicals (ie ammonia fumes). If he/she is so concerned let them design and make a vent for it. The least that should be done is to make sure there is enough room ventalation. I myself you not worry.
Best of Luck, Ed Calomeni Medical College of Ohio Toledo, OH 43699 emlab-at-opus.mco.edu
Our safety people required us to have an emergency exhaust fan (with a manual switch) installed in the darkroom in which the processor is located. Other than occasional testing, the fan has not been operated in the ten years since it was installed.
Mike O'Keefe National Center for Electron Microscopy Berkeley, CA 94720 --------------------------------------
Why does the engineer think the processor needs to be vented? There is very very little mixture of chemicals (ie ammonia fumes). If he/she is so concerned let them design and make a vent for it. The least that should be done is to make sure there is enough room ventalation. I myself you not worry.
Best of Luck, Ed Calomeni Medical College of Ohio Toledo, OH 43699 emlab-at-opus.mco.edu
Please post the following on the Microscopy Server:
Biological SEM and X-ray Microanalysis
The 1996 Lehigh Short Course, in addition to teaching the theory and basic prractical skills associated with scanning electron microscopy and x-ray microanalysis, also provides a thorough understanding of how specimens should be prepared for imaging and analysis.
Special emphasis is placed on providing answers to the problems associated with the preparation, microscopy, and analysis of biological, organic, hydrated and bio-medical materials such as plastics, paints, polymers, elastomers, insulators, cements, resins, lubricants, pharmaceutical agents, foods and dairy products, textiles, paper, archeological remains and the whole spectrum of plant, animal and human tissues. The very nature of these samples, which are generally beam-sensitive, frequently wet, and invariably hetereogeneous, require an appreciation and understanding of a wide range of new techniques and methods. The Lehigh Short Course is uniquely placed to provide the necessary information and hands-on instrumentation to address these problems.
Specific instruction will be given in low temperature and low voltage microscopy and analysis; sample preparation and specimen coating; immuno- and histo-chemical and staining techniques to localize specific chemical ligands; non-invasive preparative methods and techniques for diffusible and soluble substances; environmental microscopy and analysis, and the strategies and tactics to employ when examining and analysing beam-sensitive samples. Special biological lectures by Dr. Patrick Echlin, University of Cambridge.
For a free brochure contact:
Sharon Coe Materials Science Department Lehigh University 5 East Packer Avenue Bethlehem, PA 18015 Phone: 610/758-5133 Fax: 610/758-4244 e-mail: slc6-at-lehigh.edu
We have an externally venting overbench extraction hood over both the print processor and an adjacent wet area. This appears to work well and certainly eliminates the smell of the warm chemicals.
All photographic processing should be carried out with some form of fume extraction, something that is often ignored. Our photographic section installed such hoods in their darkrooms 12 or 13 years ago after one of the photographers developed respiratory problems carrying out conventional hand printing..
Ian Hallett
(Donna Wagahoff writes ...) } We just purchased a new Mohr-Pro print processor but have not been } able to use it because our building engineer is concerned that we must } vent the unit. The manual does not address this issue and there is no } outlet or attachment for venting the chemistry fumes. We have had } several processors over the years, with no exhaust vent and have had } no problems. Would you please share with us whether you have your } print processors connected to a vent system, how you accomplished it, } if it is an OSHA requirement, if your system came with an outlet made } specifically for venting chemistry fumes and any other pertinent } information. Thanks for your assistance. } } Donna Wagahoff } SIU School of Medicine } Springfield, Il. } 217-782-0898 } fax 217-524-3227
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
Hello Robert, In answer to your questions: 1. I usually use a cotton bud to pick up the sections and 'roll' them onto the surface of the waterbath. 2. I havn't used spurr's resin or mixtures with other epoxy's but it is worth a try!. 3. I add KOH ot cover the base of a 1lt reagent bottle (screw top) and fill with ethanol (absolute). The solution goes brown after a day or so , then filter before use. It keeps reasonably well. Please note that it is very corrosive!!!. 4. The original reference is: Imai, Y et al (1968): Removing method of resin for histological section following halogenation. Electron Microsc. vol 17,84
The Spring meeting of the Central States Microscopy Society will take place on June 21, 1996 in Carbondale, IL at the Giant City State Park Lodge. The theme of the meeting will be "Ride the Wave" (e.g., technological wave). Innovative uses of new and established technologies (microwave, LM, SEM, TEM, scanned probes, etc) are invited. Biological and physical science presentations are welcome.
Cash prizes for best student presentations. Typically this ranges from $50-100, depending upon number of entrants and quality of work presented. Abstracts must be received 3 weeks prior to meeting to be considered in the student competition. Follow MSA abstract guidelines.
CSMS corporate sponsors are invited to participate by means of talks and demonstrations of equipment. Contact me well in advance of the meeting, however, if you require tables, space or special electrical setups. There will be no charge for CSMS corporate sponsors.
Programs, lodging recommendations, directions to meeting site will be sent several weeks in advance of the meeting.
Speakers and presentors: please contact John Bozzola for topics/timing no later than May 21, 1996.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
We have a Rapiline print processor which came with a (very small) built-in fan to extract fumes from the tanks... and blow them into the room. The installation did not include connecting this to any extract system although the technician indicated that this could be done. Unlike your experience however, we found the fumes unpleasant so we had our workshop make elbows and ducts to connect the output to the building extract sustem and we have noticed that the air is much fresher in the darkroom as a result. I don't know if the fumes were unsafe before but it is certainly more pleasant to use be in that room now and we feel that it was a worthwhile modification. I suppose whether or not is worthwhile depends a lot what photographic chemistry you are using and at what temperature (we use 27degC which is relatively cool). I wouldn't assume that because the manual doesn't mention it that it isn't worth doing, my expectation of manuals gets lower with every new bit of equipment we buy.
Best regards,
Richard
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
I need a little help. I am using the NBT/BCIP method to detect alkaline phosphatase in indirect immunomarking on plant tissue sections. I want to mount these sections permanently and I have tried DPX but the reduced tetrazolium salt just (re)crystallize and any specific staining is lost. Any suggestion about what to use instead of DPX is much appreciated.
Nanoprobes, Inc is looking for independent sales representatives to carry their immunoreagent product line. This position(s) are commission only. Please send inquires/resumes to Ms. Tempel at Nanoprobes, Inc. 25 E. Loop Rd. Ste 124, Stony Brook NY 11790-3350,Phone: 516 444 8815 or Fax 516 444 8816; E.mail: nano-at-mail.lihti.org.
Nanoprobes, Inc is looking for independent sales representatives to carry their immunoreagent product line. This position(s) are commission only. Please send inquires/resumes to Ms. Tempel at Nanoprobes, Inc. 25 E. Loop Rd. Ste 124, Stony Brook NY 11790-3350,Phone: 516 444 8815 or Fax 516 444 8816; E.mail: nano-at-mail.lihti.org.
I have good success picking up semi-thin sections with a fine wire loop that I attached to a wooden stick. With a high meniscus approach the section from below and lift up. The section can then be deposited on a water droplet (on a slide) either by inverting the loop on the water droplet or by floating the section off. Heat slide GENTLY to avoid wrinkles and dry the section.
For em: I usually transfer the section to a beam capsul filled with water, and put it in a warm oven (at 37-45 deg.C), to remove the wrinkles. I then use a plastic film-(e.g. parloidin) coated grid to pick up the section. This prevents "electrostatic jumping" of the section when it is dry.
Good luck, Ada
ADA L. OLINS THE UNIVERSITY OF TENNESSEE-OAK RIDGE and THE BIOLOGY DIVISION, ORNL P.O. BOX 2009 TEL: 423 574 1269 OAK RIDGE, TN 37831-8077 FAX: 423 574 1274
Richard Easingwood wrote something in his recent post regarding ventilation and film processors that struck a raw nerve. He wrote: "I wouldn't assume that because the manual doesn't mention it that it isn't worth doing, my expectation of manuals gets lower with every new bit of equipment we buy."
We have just received CE (European Community) certification for our laboratory microwave processor after many months and many thousands of dollars. CE compliance with electrical safety regulations has been required since Jan 1, 1996, for *all* electrical equipment exported into Europe.
Many of the CE regulations relate to what must be in the users' manual. Everything that is required for the manual must be translated into several languages, in addition to conforming to complicated rules regarding content and format. The end result is that we've cut our very thorough manual of almost 200 pages down to fewer than 20 pages at the urging of our CE certification consultant.
I would expect that the trend that Richard points out in his post will continue to worsen as more international manufacturing companies deal with the new CE regulations.
Does anyone know of a source for antibodies to rabbit myeloperoxidase?? Can't find them in Linscott's or the MSRS catalogue. Do any of the antibodies to human cross-react with rabbit??
Thanks Mark Elliott Pulmonary Research Lab, UBC Vancouver Canada
Have a customer interested in EM courses. I suggested the Lehigh course. Would like to know who to have her contact to get info on the Lehigh course. TIA Cal Montgomery
Hello, I am doing immunocytochemical experiments on mouse cochlea using frozen sections. I find that I must block endogenous peroxidase in these tissues. Most protocols I see require dehydration to 100% EtOH or MeOH when blocking with H2O2. Cochlear tissue is very delicate and there is a risk losing some of the fine structure with these additional steps. My questions are: why can't I just block with H2O2 in my buffer? is the alcohol necessary for penetration allowing better access of the H2O2 to the tissue? These are 10 micron sections. Thanks in advance. Sincerely, Gary
Our lab has been asked to help on a possible project studying mammalian brain tissue. The material will be collected in the field( a few thousand miles fom our lab) and storage times could be up to 3-4 weeks, before the material reaches the lab for processing. I have an excellent fixation method for brain tissue, but its fairly involved and the solutions must be made up fresh; not a real possibility for this field work. In the past, I have had great success, using PIPES buffer for the glutaraldehyde fixation, whenever mammalian em material had to be stored for 3-4 days at 4 degrees C. I would make up the fixative bottle of 4% GTA in 0.15M PIPES, pH 7.35 and send it out. They would add their samples and mail(at -at-4 degrees C) them to me. I've also had suceess using the GTA/PIPES fixation and then transferring samples to a buffer of 0.1M Na Cacodylate with 0.15M Sucrose, pH 7.4. and holding the sample for a week, at 4 degrees C. Has anyone had experience in keeping a sample for 4 weeks before final processing. Would it be better to store the sample: 1. in PIPES buffer, after GTA/PIPES fixation? 2. 0.1M Na Cacodylate with 0.15M Sucrose, pH 7.4. after GTA/PIPES fixation? 3. leave in GTA/PIPES fixative for the entire time at 4 Degrees C. ( I worry about the staibility of the GTA over such a long time) 4. Some other method, that works with mammalian tissue.
Any suggestions or thoughts on long term fixation/storage would be greatly appreciated. thanks Louisa Howard
SUMMER COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221)
NASSAU COMMUNITY COLLEGE
A five week course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered over the first summer session, between May 28 and June 27, 1996. The class meets from 8:00 am to Noon four days per week (usually Monday thru Thursday).
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $78 per credit.
Registration for summer courses can be conducted via phone by calling (516) 572-7131 or (516) 572-7372 or (516) 572-7425, Monday through Thursday 2:30pm to 7:00pm (This phone service ends on May 2, 1996)
More information about summer registration and course offerings is available at our web site {http://www.sunynassau.edu}
The catalog description is specified below. If you have further questions, you should e-mail me directly at the address below.
CATALOG DESCRIPTION BIO 221: Transmission Electron Microscopy 4 cr. Prerequisites: BIO 109-110 or equivalent, CHE 151-152 or equivalent. An introduction to the basic principles of transmission electron microscopy including tissue preparation, microscope (TEM) operation, black & white photography, and micrograph interpretation. The entire laboratory is devoted to the development of skills and preparative techniques involved with the operation of an actual transmission electron microscope. (3 lecture, 3 laboratory hours). Laboratory fee applies.
Stephen Beck Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829
} Northern CA Society for Microscopy (NCSM) Meeting Announcement } } Where: Roche Biochemicals (formerly Syntex), Gallery } Conference Center Bldg A-2 } 3401 Hillview Ave, Palo Alto CA } } When: Thursday, May 9, 1994, from 2:30 to 8:45 PM } } } 2:30 -3:00 Registration } Speakers } 3:00-3:45 Hank W. Bass, UC Berkeley, Molecular and Cellular } Biology, "Analysis of Meiosis using 3-D } Deconvolution Light Microscopy". 4:00-4:45 David Blake, } NASA Ames Research Center, Space Sciences Division } "Electron Microscopy of Astrophysical Ice" } 5:00 - 6:30 Social Hour and Local Society A filiate meeting } sponsored by JEOL Instruments } 6:30-7:30 Dinner. Choices: Marinated Stripped Loin with sundried } tomatoes, Lemon chicken with caper sauce, or roasted } vegetable lasagna, } each with garlic mashed potatoes, summer squash medley, } mixed green salad, dinner rolls, and dessert. } 7:45 - 8:45 Paul Carpenter, Manager Analytical and Geology, Cal } Institute of Technology, "XRay and Optical } Methods as Applied to the Study of Gemstones." } } Please help us to plan accurately. Make your reservation before Friday MAY } 3rd } Fax reservation(s) with meal preference to Laura Knoff at (510) 486-4750. } E-Mail address = KNOFF-at-LIPOVX.LBL.GOV } Phone = (510) 486-4088, if leaving a message please speak slowly and clearly. } Price: $17 for Loin, $14 for Chicken, $13 for Lasagna,$8 for student } members. } Avoid standing in line by pre-paying. Send your check, payable to NCSM, } to NCSM c/o Laura Knoff } LawrenceBerkeley National Lab } Bldg 1, Room 264 } 1 Cyclotron Rd } Berkeley, CA 94720 Sorry about the format (or lack of it).
} Date: Tue, 23 Apr 1996 12:37:02 -0800 } To: microscopy-at-Sparc5.Microscopy.Com } From: knoff-at-lipovx.lbl.gov (Laura Knoff) } Subject: No. Cal Micro. Meeting } Cc: } Bcc: } X-Attachments: } } } Northern CA Society for Microscopy (NCSM) Meeting Announcement } } } } Where: Roche Biochemicals (formerly Syntex), Gallery } } Conference Center Bldg A-2 } } 3401 Hillview Ave, Palo Alto CA } } } } When: Thursday, May 9, 1994, from 2:30 to 8:45 PM } } } } } } 2:30 -3:00 Registration } } Speakers } } 3:00-3:45 Hank W. Bass, UC Berkeley, Molecular and Cellular } } Biology, "Analysis of Meiosis using 3-D } } Deconvolution Light Microscopy". 4:00-4:45 David } } Blake, NASA Ames Research Center, Space Sciences Division } } "Electron Microscopy of Astrophysical Ice" } } 5:00 - 6:30 Social Hour and Local Society A filiate meeting } } sponsored by JEOL Instruments } } 6:30-7:30 Dinner. Choices: Marinated Stripped Loin with } } sundried tomatoes, Lemon chicken with caper sauce, or } } roasted vegetable lasagna, } } each with garlic mashed potatoes, summer squash } } medley, mixed green salad, dinner rolls, and } } dessert. } } 7:45 - 8:45 Paul Carpenter, Manager Analytical and Geology, Cal } } Institute of Technology, "XRay and Optical } } Methods as Applied to the Study of Gemstones." } } } } Please help us to plan accurately. Make your reservation before Friday MAY } } 3rd } } Fax reservation(s) with meal preference to Laura Knoff at (510) 486-4750. } } E-Mail address = KNOFF-at-LIPOVX.LBL.GOV } } Phone = (510) 486-4088, if leaving a message please speak slowly and clearly. } } Price: $17 for Loin, $14 for Chicken, $13 for Lasagna,$8 for student } } members. } } Avoid standing in line by pre-paying. Send your check, payable to NCSM, } } to NCSM c/o Laura Knoff } } LawrenceBerkeley National Lab } } Bldg 1, Room 264 } } 1 Cyclotron Rd } } Berkeley, CA 94720 } Sorry about the format (or lack of it). }
} Date: Tue, 23 Apr 1996 12:37:02 -0800 } To: microscopy-at-Sparc5.Microscopy.Com } From: knoff-at-lipovx.lbl.gov (Laura Knoff) } Subject: No. Cal Micro. Meeting } Cc: } Bcc: } X-Attachments: } } } Northern CA Society for Microscopy (NCSM) Meeting Announcement } } } } Where: Roche Biochemicals (formerly Syntex), Gallery } } Conference Center Bldg A-2 } } 3401 Hillview Ave, Palo Alto CA } } } } When: Thursday, May 9, 1994, from 2:30 to 8:45 PM } } } } } } 2:30 -3:00 Registration } } Speakers } } 3:00-3:45 Hank W. Bass, UC Berkeley, Molecular and Cellular } } Biology, "Analysis of Meiosis using 3-D } } Deconvolution Light Microscopy". 4:00-4:45 David } } Blake, NASA Ames Research Center, Space Sciences Division } } "Electron Microscopy of Astrophysical Ice" } } 5:00 - 6:30 Social Hour and Local Society A filiate meeting } } sponsored by JEOL Instruments } } 6:30-7:30 Dinner. Choices: Marinated Stripped Loin with } } sundried tomatoes, Lemon chicken with caper sauce, or } } roasted vegetable lasagna, } } each with garlic mashed potatoes, summer squash } } medley, mixed green salad, dinner rolls, and } } dessert. } } 7:45 - 8:45 Paul Carpenter, Manager Analytical and Geology, Cal } } Institute of Technology, "XRay and Optical } } Methods as Applied to the Study of Gemstones." } } } } Please help us to plan accurately. Make your reservation before Friday MAY } } 3rd } } Fax reservation(s) with meal preference to Laura Knoff at (510) 486-4750. } } E-Mail address = KNOFF-at-LIPOVX.LBL.GOV } } Phone = (510) 486-4088, if leaving a message please speak slowly and clearly. } } Price: $17 for Loin, $14 for Chicken, $13 for Lasagna,$8 for student } } members. } } Avoid standing in line by pre-paying. Send your check, payable to NCSM, } } to NCSM c/o Laura Knoff } } LawrenceBerkeley National Lab } } Bldg 1, Room 264 } } 1 Cyclotron Rd } } Berkeley, CA 94720 } Sorry about the format (or lack of it). }
Am interested in finding out the best overall fixation for Kidney (medulla and cortex). Interested in good preservation (who isnt) as well as defined membrane boundries. Specifically looking at mitochondria.
Probing & Structure has a large fully illustrated microscopy catalogue on the www. Prices are shown in Australian dollars. Most items are shipped FIS. Currency converter, numerous good links and many MSDS documents are also available. It's worth a bookmark
} } Any suggestions or thoughts on long term fixation/storage would be greatly } appreciated. } thanks } Louisa Howard
I have had no problems with long term (years) storage of retina in cacodylate buffer (no additives) after glutaraldehyde fixation. Long term (weeks) storage in glut/cacodylate is also not a problem - the tissue doesn't harden excessively - though I've never done it with brain. All at 4 deg C of course.
Diana van Driel Dept Ophthalmology Sydney University AUSTRALIA 2006
In response to Cal Montgomery and any other people interested in EM (SEM) courses they should contact Sharon Coe tat e-mail address {slc6-at-Lehigh.EDU} who will be able to give you details of the all singing, all dancing, bells and whistles LEHIGH SHORT COURSE which will covers everything you want to know and were afraid to ask about SEM and x-ray microanalysis of every conceivable type of specimen (as can be seen it does not, alas, cover typing)
Patrick Echlin Cambridge UKOn Tue, 23 Apr 1996, cal Montgomery wrote:
} Have a customer interested in EM courses. I suggested the Lehigh course. } Would like to know who to have her contact to get info on the Lehigh course. } TIA } Cal Montgomery } }
If anyone out there has got advice/recommendations/experience on the preparation and use of standards for TEM+EDX we would be most interested to hear from you.
Though our emphasis will ultimately be on biological applications, any input would be of interest at this stage.
Tony Bruton Centre for Electron Microscopy University of Natal, Pietermaritzburg Private Bag X01, Scottsville 3209 KwaZulu Natal, South Africa
For postmortem human brain tissue, which is also delicate, we use 1% H2O2 in PBS for 1 hour at 4C. This incubation is done without shaking, because the peroxide is also somewhat harsh on the tissue.
} } } } Hello, I am doing immunocytochemical experiments on mouse cochlea } } using frozen sections. I find that I must block endogenous peroxidase } } in these tissues. Most protocols I see require dehydration to 100% } } EtOH or MeOH when blocking with H2O2. Cochlear tissue is very delicate } } and there is a risk losing some of the fine structure with these } } additional steps. My questions are: why can't I just block with H2O2 } } in my buffer? is the alcohol necessary for penetration allowing better } } access of the H2O2 to the tissue? These are 10 micron sections. } } Thanks in advance. Sincerely, Gary } } } } } } zajic-at-umich.edu
Mr-Received: by mta SRVR05.MUAS; Relayed; Wed, 24 Apr 1996 09:32:29 -0400 Mr-Received: by mta SRVR05; Relayed; Wed, 24 Apr 1996 09:32:30 -0400 Mr-Received: by mta SRVR01; Relayed; Wed, 24 Apr 1996 09:34:22 -0400 Disclose-Recipients: prohibited MSA MICROSCOPY MAILING LIST {MICROSCOPY-at-Sparc5.Microscopy.Com} Message-Id: {4129320924041996/A20679/SRVR05/11A4C2601C00*-at-MHS} X-Envelope-To: Greg2NJ-at-aol.com, MICROSCOPY-at-MSA.MICROSCOPY.COM Autoforwarded: false Mime-Version: 1.0 Content-Type: TEXT/PLAIN; CHARSET=US-ASCII Content-Transfer-Encoding: 7BIT Importance: normal Priority: normal Sensitivity: Company-Confidential Ua-Content-Id: 11A4C2601C00 X400-Mts-Identifier: [;4129320924041996/A20679/SRVR05] Hop-Count: 2
Retrograde perfusion fixation via abdominal aorta. See: Griffith, LD et al. (1967). The ultrastructure of the functioning kidney. Lab. Invest. 16, 220-246. Best regards,
Walter F. Bobrowski Subcellular Pathology Parke-Davis Pharmaceutical Research Ann Arbor, MI 48105
TEL: 313-996-7814 FAX: 313-996-5001 E-Mail: BOBROWW-at-AA.WL.COM } Greetings: } } Am interested in finding out the best overall fixation for Kidney (medulla } and cortex). } Interested in good preservation (who isnt) as well as defined membrane } boundries. Specifically looking at mitochondria. }
To expand upon Louisa Howard's question of long term storage -she is asking about storage in cacodylate buffer.
The question has recently been asked here about long term storage in phosphate buffers. We have a variety of fish, rat and cow tissues that were collected (or will be collected) "just in case" EM may be needed. Some of these tissues have been around for at least 10 years and will likely be here another 10 years.
What are the current thoughts on storage for long lengths of time? Would it be better to keep the tissues in fixative as opposed to buffer? Would cacodylate buffer be preferable to phosphate (or is there something else)? Any special techniques for processing old tissues and getting useable results? How long have tissues been kept around and still been successfully worked with?
Thanks for any advice from all the experts on the list.
Rosemary Harris Electron Microscopy Laboratory Alberta Environmental Centre
Message-Id: {199604241519.KAA06313-at-watson.bcm.tmc.edu} X-Sender: joiner-at-bcm.tmc.edu (Unverified) X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 09:02 PM 4/23/96 -0400, you wrote:
} Greetings: } } Am interested in finding out the best overall fixation for Kidney (medulla } and cortex). } Interested in good preservation (who isnt) as well as defined membrane } boundries. Specifically looking at mitochondria. } } ********************** We get good results fixing renal biopsies in 3% glutaraldehyde in 0.09 M PIPES, pH 7.2 followed by 2.0% osmium in PIPES. We then enbloc stain with saturated aqueous UAc for 45 minutes and embed in Spur's resin. The tissue remains in the glutaraldehyde for varying times up to a day because these are medical specimens coming from various hospitals.
Joiner Cartwright, Jr., Ph.D.
Director, Electron Microscopy tel.: (713)798-4658 Department of Pathology, Rm.286-A FAX: (713)798-3945 Baylor College of Medicine Internet: joiner-at-bcm.tmc.edu One Baylor Plaza Compuserve: 71555,1206 Houston, Texas 77030 U.S.A.
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We have a Bal-Tec CPD030. It has been used very little. This is because we cannot get it to drain. It might take all night for the alcohol-CO2 mixture to empty. The instructions seem very straigt-forward. We fill partially with 100% ethanol (with or without a sample), cool to 8 degrees C, let in the carbon dioxide (yes, the tank is the kind with the dip tube), then press the medium out button. At first it seems to empty, then gets very slow. Has anyone had a similar problem? The valves seem to work as I hear a loud click when pressing buttons. The filters are clean. Any other ideas? thanks.
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There is a list of microscopy short courses and in-house trainers on the WWW at http://www.mwrn.com/product/train.htm The list has direct links to information on the WWW and e-mail addresses. Lehigh information is included.
Susanne Pignolet Brandom MicroWorld Resources and News http://www.mwrn.com/ spb-at-wwa.com 847-548-6522
At 04:00 PM 4/23/96 -0400, you wrote: } Have a customer interested in EM courses. I suggested the Lehigh course. } Would like to know who to have her contact to get info on the Lehigh course. } TIA } Cal Montgomery } } }
I recall that some NASA sponsored studies of long term fixation found the best results leaving tissue in the glutaraldehyde fix. This may even have been presented at an EMSA meeting in the mid 1970's but I haven't found the reference yet.
} } If anyone out there has got advice/recommendations/experience on the } preparation and use of standards for TEM+EDX we would be most } interested to hear from you. } } Though our emphasis will ultimately be on biological applications, any } input would be of interest at this stage. } Dear Tony, A few years ago, the late, lamented Chuck Fiori produced some lithium borate glass standards especially designed for average Z to be equal to that of tissue. I think someone at NIST can tell you more. Good luck. Yours, Bill Tivol
In reply to Tony Bruton's request for info on EDS/TEM standards:
There is a National Institute of Standards and Technology Standard Reference Materials (SRM) specifically designed for the X-ray analysis on the TEM, though not for biological applications, and unluckily it is not free.
Here is a brief description of the material, so that you can see if you might be interested:
The standard is called "SRM 2063 Microanalysis Thin Film" and consists of a thin film of sputtered mineral glass supported by a carbon film and a copper TEM grid. The composition is certified as listed below and was determined by several independent techniques after the glass was deposited on the grids. Thus the certified composition values take into account sample preparation. The thickness of the glass (76 nm) and density (3.1 gm/cm**3) are reported (though not certified). No preparation is necessary for use in the TEM.
Table of Concentration Values for SRM 2063 Mineral Glass Element Concentration (% wt.) O 43.2 Mg 7.97 Si 25.34 Ca 11.82 Fe 11.06
The composition allows for a range of relative sensitivity values to be determined for many commonly analyzed elements and x-ray lines. Thus the standard can be a primary, traceable way of checking other in-house standards you may use. The standard is robust under many handling and beam conditions, though very high beam dose may cause a change in composition (as noted on the certificate of analysis.) We have used one standard grid for about seven years in our laboratory to monitor/compare several instruments and detectors.
For more information about obtaining the standard you may contact:
Standard Reference Materials Program National Institute of Standards and Technology Gaithersburg, MD 20899-0001 USA
I read the current set of postings about "the best fixative for...", and "the effect of long term storage on..."and see that many researchers are using PIPES adjusted to a pH above 7.2 to buffer their fixatives.
This buffer has a pKa of 6.8 so would anyone like to make a comment on the buffering capacity of this buffer above pH 7.0?
If we have to use a "Good" buffer for fixation, we choose HEPES which has a higher pKa. We get exceptional morphology in cryosections even if we only use formaldehyde. We also used it at a conc. of 300mM recently to fix some shark tissue with similar results. We never did get much from using PIPES.
As for tissue storage, there should be no problem in storing tissue for long periods in fixative, if the samples are to be examined only for morphology. The samples will slowly get harder and more brittle the longer they are in gluteraldehyde but if they are embedded in resin it will not be a problem. There might be some extraction of soluble material with time but that will only help with increasing the final contrast of the specimen.
If the samples are only to be fixed in formaldehyde then the fixative should not be removed. I have heard unconfirmed stories of 10 - 20 year old samples, stored in formaldehyde at RT, being taken off the shelf and successfully labeled with antibodies. I guess it all depends on what the antigens were.
Best regards
Paul Webster, Center for Cell Imaging Yale University School of Medicine.
The question of which solvent to use depends upon what you are trying to block. The methanol denatures the peroxidase's catalytic site such that the peroxide cannot be split or released once it has been bound. Many years ago I compared peroxidase inhibition on bone marrow needle biopsies between peroxide solutions with PBS, ethanol, methanol and acetone. PBS and ethanol gave modest reduction of endogenous peroxidase, and PBS is probably sufficient for most tissues. Methanol gave the most complete inhibition of the variety of peroxidatic activities in marrow. Acetone actually appeared to potentiate it in some leucocytes
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
Gary H. Zajic wrote:
} Hello, I am doing immunocytochemical experiments on mouse cochlea } using frozen sections. I find that I must block endogenous peroxidase } in these tissues. Most protocols I see require dehydration to 100% } EtOH or MeOH when blocking with H2O2. Cochlear tissue is very delicate } and there is a risk losing some of the fine structure with these } additional steps. My questions are: why can't I just block with H2O2 } in my buffer? is the alcohol necessary for penetration allowing better } access of the H2O2 to the tissue? These are 10 micron sections. } Thanks in advance. Sincerely, Gary } } } zajic-at-umich.edu }
I think that the inlet diaphram maybe clogged. I am now with RMC and still help people with problems with BAL-TEC / Balzers equipment. Please feel free to call me at RMC, 520-889-7900. I can talk you through your difficulties if you call.
P.S I am available to help any BAL-TEC/ Balzers users out there, just E-Mail me or call.
} Date: Wed, 24 Apr 1996 11:04:54 -0500 } From: Joyce Craig {Chicago.State.University-at-uxa.ecn.bgu.edu} } Subject: critical point dryer problems } To: Microscopy-at-aaem.amc.anl.gov } X-Sender: bafpjec-at-pop3.ecn.bgu.edu } X-Mailer: Windows Eudora Light Version 1.5.2 } Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } } We have a Bal-Tec CPD030. It has been used very little. This is because we } cannot get it to drain. It might take all night for the alcohol-CO2 mixture } to empty. The instructions seem very straigt-forward. We fill partially } with 100% ethanol (with or without a sample), cool to 8 degrees C, let in } the carbon dioxide (yes, the tank is the kind with the dip tube), then press } the medium out button. At first it seems to empty, then gets very slow. } Has anyone had a similar problem? The valves seem to work as I hear a loud } click when pressing buttons. The filters are clean. Any other ideas? thanks.
Joyce, In my opinion, it looks like the problem is that the exhaust pipe is jammed. Maybe it is too thin or dirty. If this is the case, in the beginning it will work fine. After a while, it will freeze and jam. Under normal conditions, you can hear loud sound and see CO2 gas and ice is expelled when you open the venting value. If this does not happen, it was probably jammed. Ya Chen
Ya Chen
========================================================================== Cryo/SEM Coordinator Integrated Microscopy Resource (IMR)-- III M M RRRRRR an NIH Biomedical Research Resource I M M M M R R University of Wisconsin, Madison, WI I M M M RRRRRR 1675 Observatory Drive #167 I M M R R Madison, WI 53706, USA I M M R R TEL : 608-263-8481 I M M R R FAX : 608-265-4076 III M M R R Email:YChen-at-macc.wisc.edu ========================================================================== IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html 2nd Symposium on Integrated Microscopy: Sept. 20-22, 1996
} Date: Wed, 24 Apr 1996 18:28:18 } To: Joyce Craig {Chicago.State.University-at-uxa.ecn.bgu.edu} } From: Sylvia Francis Zalzal {franciss-at-ere.umontreal.ca} } Subject: Re: critical point dryer problems } } We have in our lab and for over three years the same CPD030 and we never had this problem. The dehydrant/Co2 exchange should not cause any trouble,and the sound should not be loud when you depress the buttons. However we had prior to this Bal-tec 030 we had a cpd020 and a similar problem occured the reason was " frozen valves " Maybe when cooling the chamber do not cool under 11 degrees. Try it then call for service check the valves. } } Good luck=20 } Sylvia=20 } } } } } At 11:04 4/24/96 -0500, you wrote: } } We have a Bal-Tec CPD030. It has been used very little. This is because= we } } cannot get it to drain. It might take all night for the alcohol-CO2= mixture } } to empty. The instructions seem very straigt-forward. We fill partially } } with 100% ethanol (with or without a sample), cool to 8 degrees C, let in } } the carbon dioxide (yes, the tank is the kind with the dip tube), then= press } } the medium out button. At first it seems to empty, then gets very slow. = =20 } } Has anyone had a similar problem? The valves seem to work as I hear a= loud } } click when pressing buttons. The filters are clean. Any other ideas? thanks. } } } Sylvia Francis Zalzal Chef de Laboratoire Laboratoire de Microscopie Electronique Facult=E9 de m=E9decine dentaire Universit=E9 de Montr=E9al
} Can anyone provide me with the URL for the ASTM home page? } } Thank you! } } Best regards- } } David Henriks TEL: 800-728-2233 (toll-free in USA) } South Bay Technology, Inc. 714-492-2600 } 1120 Via Callejon FAX: 714-492-1499 } San Clemente, CA 92673 USA e-mail: 73531.1344-at-compuserve.com } sbt-at-msa.microscopy.com } } Manufacturers of Precision Sample Preparation Equipment and Supplies for } Metallography, Crystallography and Electron Microscopy. } }
Any University interested in obtaining an old, but restorable ARL-SEMQ Microprobe should contact Advanced MicroBeam at 72714.265-at-compuserve.com or phone: 330 394-1255, fax: 330 394-1834.
Advanced MicroBeam, Inc. 4217C King Graves Rd. PO Box 610 Vienna, OH 44473 USA
Jonathan Krupp gave me your eMail address. I have a TEM to sell and would like to list it on the MSA BB/mail listserver (I am a member of the MSA). Would you please let me know the format for the listserver? Also, would you please send instructions for subscribing to the list via your Web site? Thank you.
Kurt H. Albertine, Ph.D. Director, Research Microscopy Facility University of Utah Health Sciences Center Salt Lake City, UT 84132
Alternate-Recipient: prohibited Auto-Forwarded: prohibited Content-Return: allowed Disclose-Recipients: prohibited Conversion: allowed Importance: normal Priority: normal Sensitivity: Normal
We are looking to buy a new critical point drier to replace our old one that cooled adiabatically (by venting CO-2) and notice that some CPDs are now available that cool electronically by the Peltier effect. This seems like a big advantage but does anyone with experience with both types care to comment?
Gary Radice 804-289-8107 (voice) Department of Biology 804-289-8233 (FAX) University of Richmond Richmond VA 23173 USA
Valve could be freezing. Can you get to it to warm it up?
Ron
----------
We have a Bal-Tec CPD030. It has been used very little. This is because we cannot get it to drain. It might take all night for the alcohol-CO2 mixture to empty. The instructions seem very straigt-forward. We fill partially with 100% ethanol (with or without a sample), cool to 8 degrees C, let in the carbon dioxide (yes, the tank is the kind with the dip tube), then press the medium out button. At first it seems to empty, then gets very slow. Has anyone had a similar problem? The valves seem to work as I hear a loud click when pressing buttons. The filters are clean. Any other ideas? thanks.
Hi all, I wanted to post more information about the Zeiss TEM that the University of Iowa's Biology dpartment would like to give away. I spoke with Shelley Plattner this morning, as he is in charge of the instrument, and is the person to contact. He can be reached at (319)-335-1070, or email at shelley-plattner-at-uiowa.edu. He says that it is a 10A, not a 10C. Zeiss knows that they are trying to give the scope away, and estimates that it would take about 40 hours of labor -at-$150.00/hr. Zeiss will make sure that the instrument works upon relocation. Shipping would be extra, depending on the carrier and distance. Zeiss might be willing to discount the cost if the person purchases a service contract. You could discuss this with them by calling 1-800-233-3334. Lam, the Zeiss service engineer who has maintained this instrument can be reached at extension 725 at the above 800 number. He can answer question in regards to what would be needed to hook up the microscope. Thanks to those who have expressed interest, and have tolerated the run around on our end.
---Paul Webster wrote: I read the current set of postings about "the best fixative for...", and "the effect of long term storage on..."and see that many researchers are using PIPES adjusted to a pH above 7.2 to buffer their fixatives.
This buffer has a pKa of 6.8 so would anyone like to make a comment on the buffering capacity of this buffer above pH 7.0? --- end of quoted material ---
I agree that HEPES would be much better to use at pH 7-3-7.4, given its pKa of 7.4. We use it often in SEM preps. The reason we use PIPES for TEM, is that it appears to cause less extraction in mammalian and plant tissues. This is strictly anectdotal on our part, although Coetzee and van Der Merwe, J of Microscopy135(2) 147-158, 1984, showed that extraction in plant tissue was greater using HEPES than using PIPES. I'm definitely willing to try HEPES buffer on mammalian brain tissue, as other posters have mentioned using HEPES for TEM, with success. Can I use the same molarity as with PIPES? I currently use a buffer concentraion of 0.1M PIPES pH -at-7.3, with a gluatraldehyde concentration of 3-4%. Also, I agree with other posters about the toxicity of Na Cacodylate, although the HEPES and PIPES buffers, supposedly have carcinogenic problems. If we work in EM, we have to take all neceesary precautions, when working with these compounds. I think the PIPES or HEPES buffers might be a better choice for field work. thanks. Louisa Howard EM facility- Remsen 240 Dartmouth College Hanover, NH. 03755
I must admit I use HEPES buffer, pH 7.2 for my fixatives with great luck and therefore have not been tempted to change it but I think a good argument to use PIPES can be made. PIPES has a pKa of 6.8 and HEPES has one of 7.5. Thus, for a target of pH 7.2, PIPES is 0.4 pH units off and HEPES is 0.3 units. A good rule of thumb is to use a buffer whose pKa is within 0.5 units of the desired pH. HEPES may look to be marginally better than PIPES but fixation solutions tend to acidify during the fixation step so by having a buffer whose pKa is on the low side of the starting pH, one would get increased protection at reasonable buffer concentrations (i.e., the pH would have to drop 0.9 units before being } 0.5 units from PIPES pKa but by only 0.3 units before being } 0.5 units from HEPES pKa). I think both are "Good" buffers in more than one sense and both are definitely superior to cacodylate in many ways (e.g., cost, toxicity, environmental). Furthermore, I believe cacodylate has a pKa of around 6.3! My feeling has always been the continued prevalence of cacodylate buffers simply demonstrates that morphologists are some of the most old-fashioned scientists around and least willing to change. That's my two cents worth.
} I read the current set of postings about "the best fixative for...", and "the } effect of long term storage on..."and see that many researchers are using PIPES } adjusted to a pH above 7.2 to buffer their fixatives. } } This buffer has a pKa of 6.8 so would anyone like to make a comment on the } buffering capacity of this buffer above pH 7.0? } } If we have to use a "Good" buffer for fixation, we choose HEPES which has a } higher pKa. We get exceptional morphology in cryosections even if we only use } formaldehyde. We also used it at a conc. of 300mM recently to fix some shark } tissue with similar results. We never did get much from using PIPES. } } As for tissue storage, there should be no problem in storing tissue for long } periods in fixative, if the samples are to be examined only for } morphology. The } samples will slowly get harder and more brittle the longer they are in } gluteraldehyde but if they are embedded in resin it will not be a problem. } There might be some extraction of soluble material with time but that will only } help with increasing the final contrast of the specimen. } } If the samples are only to be fixed in formaldehyde then the fixative } should not } be removed. I have heard unconfirmed stories of 10 - 20 year old samples, } stored in formaldehyde at RT, being taken off the shelf and successfully } labeled } with antibodies. I guess it all depends on what the antigens were. } } Best regards } } Paul Webster, } Center for Cell Imaging } Yale University School of Medicine.
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)
since long ago I've been tempted to ask all of you, specially to those giving TEM/SEM services as the only work activity, your opinion about the type of services to offer in an institution which is fully oriented to this activity:
1) How do you consider giving services in the areas of biological materials and materials science at the same time?
2) Should one focus the work into specific subjects within a given area?
3) How do you define, in advance, the type of work you would be able to do? or you wait to be asked for a specific task before deciding to do it or not?
These may sound elemental questions to you but I've never had the chance to hear people from other systems to give their opinion on that.
Now, for those doing research and giving services...
A) How do you combine both activities?
B) How much time, or effort, do you put on each one of these activities?
C) About the services to offer, should they be only related to the subject of your research work?
I would appreciate very much any comments and I promise to make a compilation of all answers for those interested in the subject. Thanks in advance.
............................................................................ Silvia Montoro Centro Regional de Investigacion y Desarrollo Santa Fe Argentina
I am working with kangaroo heart muscle, and my aim is to find mitochondrial volume density and also mitochondrial numbers, but I'm having a problem with the uranyl acetate and lead citrate staining. I am staining with 4% uranyl acetate for 1 hour under a lamp, then lead citrate for 4 minutes, yet my sections are very pale under the transmission microscope, with very little contrast in many cases. Has anyone else had this problem? I am wondering if this is a characteristic of this kind of muscle or if it is me and my methods! If you have any suggestions please RESPOND TO
Dr. SUNIL TEWARI S.Tewari-at-unsw.edu.au Patrick Marks Electron Microscope Unit University of New South Wales Sydney 2053 Australia Email : P.Marks-at-unsw.edu.au
} To: microscopy-at-aaem.amc.anl.gov } From: p.marks-at-unsw.edu.au (patrick marks) } } Hi Netter, } } I am working with kangaroo heart muscle, and my aim is to find mitochondrial volume density and also mitochondrial numbers, but I'm having a problem with the uranyl acetate and lead citrate staining. I am staining with 4% uranyl acetate for 1 hour under a lamp, then lead citrate for 4 minutes, yet my sections are very pale under the transmission microscope, with very little contrast in many cases. Has anyone else had this problem? I am wondering if this is a characteristic of this kind of muscle or if it is me and my methods! If you have any suggestions please RESPOND TO } } } Dr. SUNIL TEWARI } S.Tewari-at-unsw.edu.au } Patrick Marks Electron Microscope Unit University of New South Wales Sydney 2053 Australia Email : P.Marks-at-unsw.edu.au
} On Wednesday April 24th, Joyce Craig writes: } } } We have a Bal-Tec CPD030. It has been used very little. This is because we } } cannot get it to drain. It might take all night for the alcohol-CO2 mixture } } to empty. The instructions seem very straigt-forward. We fill partially } } with 100% ethanol (with or without a sample), cool to 8 degrees C, let in } } the carbon dioxide (yes, the tank is the kind with the dip tube), then press } } the medium out button. At first it seems to empty, then gets very slow. } } Has anyone had a similar problem? The valves seem to work as I hear a loud } } click when pressing buttons. The filters are clean. Any other ideas? } } thanks. } } } Hi Joyce, } } We have a Balzers CPD030 and I've experienced the same problem that you } have. What I have had to do is check the inlet diaphram. 9 times out of } 10 something has clogged it. I'll take it out and holding the diaphram } firmly, I "blast" it with an air duster. That usually does the trick. } } Peling Melville } } -------------------------------------------------------------- } Peling Fong Melville } Senior Scientific Assistant } Interdepartmental Facilities } American Museum of Natural History } Central Park West at 79th Street } New York, NY 10024-5192 U.S.A. } ****************************** } E-mail: peling-at-amnh.org } Work #: (212) 769-5469 } FAX #: (212) 769-5495
-------------------------------------------------------------- Peling Fong Melville Senior Scientific Assistant Interdepartmental Facilities American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 U.S.A. ****************************** E-mail: peling-at-amnh.org Work #: (212) 769-5469 FAX #: (212) 769-5495
Sunil Tewar asked for some advice about lead citrate and low contrast - I could also use some tips! My problem is that lead citrate gives variable and unpredictable contrast in the TEM. I am using LX 112 (plant tissue),osmium, and 4% uranyl acetate for 45-60 minutes; over the years lead citrate staining times have varied from 8 min to 25 min. Right now I'm staining around 15 min - was at 18 for a while but suddenly everything was too dense.
I 've read that the pH of the final solution is very important - that it should be pH 12.0 +/ 1 0.1 When I make it up however the pH is never less than 12.4! I 've been reluctant to introduce HCl into the picture so I haven't yet tried reducing the pH, consequently don't know if this is the problem. I do know that the amount of NaOH in the rinse water is very important - I use about 5 drops of 1N in 40 mls of water - more than that bleaches out the tissue. ( Can't remember off the top of my head what that means in terms of pH). There appears to be no connection between staining problems and the batch of resin, the type of tissue or the fixation.
One interesting note: when I moved to a new building a year ago, I couldn't get any staining at all for three months! Non microscopists in the building were also having unrelated problems, ultimately traced to impurities in our central distilled water source. Once this was fixed our problems disappeared.
Has anyone else had problems getting contrast with lead citrate? If so, how did you fix it?
Thanks, Peggy .
} Hi Netter, } } I am working with kangaroo heart muscle, and my aim is to find mitochondrial } volume density and also mitochondrial numbers, but I'm having a problem with } the uranyl acetate and lead citrate staining. I am staining with 4% uranyl } acetate for 1 hour under a lamp, then lead citrate for 4 minutes, yet my } sections are very pale under the transmission microscope, with very little } contrast in many cases. Has anyone else had this problem? I am wondering if } this is a characteristic of this kind of muscle or if it is me and my } methods! If you have any suggestions please RESPOND TO } } } Dr. SUNIL TEWARI } S.Tewari-at-unsw.edu.au } Patrick Marks } Electron Microscope Unit } University of New South Wales } Sydney 2053 } Australia } Email : P.Marks-at-unsw.edu.au
Tom, I just had to let you know that you made my day. The comment about microscopists being resistant to change describes my boss to a T. He recently gave me the task of determining the best way to screen for mucins in tissue sections, and when I came back, after reading ALOT of info on mucins, with the suggestion of using lectin staining and immunohistochemistry (which we have done before for other material), he pronounced that I would have to work out a technique using ONLY histochemistry, as he felt the other methods wouldn't work. Karen P.
} I read the current set of postings about "the best fixative for...", and "the } effect of long term storage on..."and see that many researchers are using PIPES } adjusted to a pH above 7.2 to buffer their fixatives. } } This buffer has a pKa of 6.8 so would anyone like to make a comment on the } buffering capacity of this buffer above pH 7.0?
PIPES is one of a few buffers suitable for work with cytoskeletal components (e.g. microtubules). It is often used to buffer fixativesfor light & electron microscopy to preserve cytoskeletal structure.
****************************************************** Steve Rogers Dept. of Cell & Structural Biology and the Beckman Institute - Optical Visualization Facility University of Illinois -at- C/U srogers-at-delphi.beckman.uiuc.edu ******************************************************
I am helping the Montpelier (Vermont) High School maintain and use a JSM T-300 SEM as part of the science labs. The SEM is linked via its TV output to a Scion frame grabber in a Macintosh running NIH Image. The science teaching with whom I am working, Dave McGraw, is involved with the UA Center for Image processing in Teaching, so this fits in with things that are already being done in the school. All is well except we need two things. 1. If there is a used T-300 out there that is available for the cost of shipping, we could use it for spare parts. 2. We are in need of a used sputter coater, too. We have a vacuum pump and can make some repairs. If you have one available, please let me know.
Thanks in advance to any who respond.
Chuck ************* Charles P. Daghlian, Ph.D. Director, Rippel E. M. Facility Dartmouth College, HB 7605 Hanover, NH 03755 phone 603-646-1039
Polaroid Corp. in Waltham, Ma. USA is looking for a Ph.D. or M.S. in chemistry, materials science, physics or engineering with training in polymer science and experience in TEM and sample prep. SEM, LM, XPS, XRD, AFM all plusses. STRONG Communication skills and computer skills desirable. Send inquiries to:Lynne Garone E. Mail:GaroneL-at-Polaroid.com
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I would like to apologize if I affended anyone on this list with my "airing" my frustrations. It was inappropriate. Tom's comment just struck a cord. I realize setting up new protocols for any one tissue is time consuming and often the "why fix it if it works" attitude is the best. But, sometimes the old ways don't work as well as you need them to, then new methods may be necessary.
Seeking ideas for building a low cost microscope hot stage. It doesn't have to be pretty, only accurate and reliable. We're a small non-profit museum lab, so I will only ever see a Mettler stage in my dreams. I have scavenged most of the components necessary for generating and control T from a PE GC and a melting point apparatus, but am looking for a design for the stage and insulator jacket (probably water flow).
Any thoughts appreciated. Thanks.
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center
9:00 - 9:45 AM Understanding Video Signals: Cameras, Recording Formats and Printers MARTY HARALDSON, Alpha Video, Edina, MN
9:45 - 10:30 AM Coffee Break \ Vendor Displays
10:30 - 11:15 AM Capturing, Storing and Organizing Digital Image Files MARK SANDERS, Imaging Center, College of Biological Sciences University of Minnesota, St.Paul Campus
11:15 - 12:45 PM A buffet lunch will be served at the Sheraton Inn
12:45 - 1:00 PM MMS Business Meeting and Election of Officers
1:00 - 1:45 PM Microscopy Resources on the Internet Dr. STUART MCKERNAN, Center for Interfacial Engineering, Characterization Facility, University of Minnesota
1:45 - 2:15 PM Coffee Break \ Vendor Displays
2:15 - 3:00 PM Public Domain Shareware, Microscopy and the Internet Dr. DAVID BRIGHT, 1996 MSA Traveling Speaker Research Chemist, Microanalysis Group NIST, Gaithersburg, MD
3:00 - 4:30 PM Vendor Displays
********************************************************************* Please make your reservation in advance! NO LATER THAN MONDAY, MAY 20, if you plan to attend the Symposium (preferred, or pay at the door).
Contact Stuart McKernan at (612) 626-7942, stuartm-at-maroon.tc.umn.edu or Dwight Erickson at (612) 736-2830, usmmm214-at-ibmmail.com
Symposium Fee: $20.00 current regular MMS members 95/96, $10.00 student members 95/96 $30 non-member(confers regular membership), $15.00 non-member students (confers student membership).
Vendors: A number of vendors of digital and other microscopy equipment will be present at the Symposium. If your company would like to have table space for product display, please contact Symposium Vendor Liaison: Diana Kittleson at Pillsbury TPC Labs, 330 University Ave. S. E., Minneapolis, MN 55414, Phone: (612)330-1898 Fax: (612)330-8266
Stuart McKernan stuartm-at-maroon.tc.umn.edu CIE Microscopy Center, University of Minnesota Office: (612) 626-7942 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
Tom Phillips said: } I must admit I use HEPES buffer, pH 7.2 for my fixatives with great luck } and therefore have not been tempted to change it but I think a good } argument to use PIPES can be made.
Hi, I'd like to chime in on this thread. I have been using PIPES as a buffer, but only in fixations for immunolocalizations. I had used it for morphological studies, but following a conversation with MV Parthasarathy (EM Director, Cornell), I switched back to either phosphate or cacodylate. My original interest stems from the following report: Salema R, I Brandao 1973 The use of PIPES buffer in the fixation of plant cells for electron microscopy. J Submicr Cytol 5:79-96. As I work exclusively with plant material, primarily leaves, it appeared to be best. The paper presents results indicatng that PIPES is the least extractive of buffers currently in use at the time. However, the work by Coetzee and van der Merwe (Coetzee J, CF van der Merwe 1987 Some characteristics of the buffer vehicle in glutaraldehyde-based fixatives. J Microsc 146:143-155.) indicates that the least extraction was found with phosphate-buffered glutaraldehyde. Again, their work was conducted using plant material (bean leaves) and may not be directly applicable to animal tissue, which was the subject of the original question. As most people know, phosphate is not compatible with added salts, hence, cacodylate would then be the buffer of choice. I would agree that morphologists are conservative in their approach to new techniques. I was extremely sceptical of the use of microwaves in fixations, until I was able to use one of the units myself. I am now convinced that this is a very powerful technique and yields fixation images equal to or superior to conventional protocols. The time-saving aspect is a tremendous bonus! I was loaned a Pelco unit by Phillip Slakmon of Scott Scientific for a number of months and have used it for more than a dozen fixations, which included standard protocols for both morphological and immunocytochemical studies. I have polymerized Spurr's resin, Epon-Araldite, LR White, and mixtures of Spurr and Epon, all with success (good infiltration, polymerization, sectioning, staining, and stability in the beam as criteria). Note that the polymerizations were conducted under water (!) in BEEM capsules. Has to be tried to be believed, I guess. Anyway, I would hope that Partha would add his comments to this discussion on buffers, particularly regarding the suitability of PIPES for morphological work.
Dwight U. Beebe E-mail: beebed-at-ere.umontreal= .ca Institut de recherche en biologie v=E9g=E9tale Voice: 514-872-4563 Universit=E9 de Montr=E9al FAX: 514-872-9406 4101, rue Sherbrooke est Montr=E9al, Qu=E9bec H1X 2B2 Canada
We need to find a supplier of stereo viewers, the cardboard-mounted polarizing kind. I've looked in every EM supply house catalog that I have and can't find them anymore. I'd appreciate any help in finding these. Thanks.
Several months ago I posted a request for microscope manufacturers to contact me with information on costs for epi-fluorescence upgrades to polarizing light microscopes. The information will be presented at a national conference.
So far, I have information only from Olympus. Anybody else like to be represented?
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center Williamstown, MA
Greetings fellow microscopist: Our usually dependable refrigerator turned on us one day recently. It dropped to 0C and our 4% aqueous osmium tetroxide froze. Does anyone know if this is still good to use. Please drop me a line if anyone has had similar experiences. Thanks Charlie Murphy
On Fri, 26 Apr 1996 cmurphy-at-GGPL.ARSUSDA.GOV wrote:
} Greetings fellow microscopist: Our usually dependable refrigerator turned on us one day recently. It dropped to 0C and our 4% } aqueous osmium tetroxide froze. Does anyone know if this is still good to use. Please drop me a line if anyone has had similar } experiences. Thanks Charlie Murphy }
Yes -
I usually keep my osmium solutions frozen. They appear to work just fine -
We have also had this problem intermittently. It's almost always been solved by making up new lead citrate, being scrupulous about purity of distilled water, following the recipe exactly, and making sure the pH is above 11 (usually ends up higher than this anyway - but may not stay that high). There are various lead citrate recipes, once we had to switch to the original Reynolds recipe for some reason, after a while someone tried the usual recipe and it worked again.... Staining times have varied too, usually it depends on how old the lead solution is - increasing time for older solutions.
With plant and animal tissue, we use alcoholic U acetate for only 10-15 min (made fresh from 4% aqueous - add equal quantities of 4% UA, distilled water and 100% ethanol, leave for 10 min till no more bubbles), and no NaOH in rinse water - we just warm it slightly - too warm and sections lose contrast.
good luck!
Rosemary White __ / Department of Ecology _/ \__/ \ and Evolutionary Biology / \ Monash University / Australia \ Clayton, Victoria 3168 \ ____ / phone 61-3-9905 5670 \_/ \_*_/ fax 61-3-9905 5613 __ email rgwhite-at-vaxc.cc.monash.edu.au \/
} Polaroid Corp. in Waltham, Ma. USA is looking for a Ph.D. or M.S. in chemistry, materials science, physics or engineering with training in polymer science and=20 } experience in TEM and sample prep. SEM, LM, XPS, XRD, AFM all plusses. STRONG Communication skills and computer skills desirable. Send inquiries to:Lynne=20 } Garone E. Mail:GaroneL-at-Polaroid.com } =20
Dear Sir,
I would like to express my interest in your proposal. Enclosed is my=20 resume. As it indicates, I have a solid physicochemical background, both=20 from my education at the Moscow University and my subsequent professional= =20 experience including training in polymer science.
I am currently working as a visiting scientist at the Department of=20 Chemistry, University of Antwerp (U.I.A.), Belgium in close collaboration= =20 with Prof. Dr. Renaat Gijbels and Prof. Dr. Willem Jacob. At this=20 department well as earlier at the Photochemistry Department of the=20 N.N.Semenov Institute of Chemical Physics, Russian Academy of Sciences I am primarily involved in analytical electron microscopy studies of=20 silver halide photographic systems, colloids and small particles.
I am familiar with the following methods and techniques: conventional and= =20 energy-filtering TEM, SEM, STEM, EDX, EELS, image analysis, IR-,=20 UV-VIS-spectrometry, Raman-spectroscopy, electron spin resonance,=20 scanning tunneling microscopy, working with PC.
I am deeply impressed by your proposal. I am hard working, and I feel=20 strongly that my motivation and enthusiasm could be of benefit to your=20 programs.
I look forward to hearing from you.
Sincerely Yours,
Vladimir Oleshko =20 Curriculum Vitae of Vladimir P. Oleshko, Ph.D. Personal data Date of birth: September 19, 1953. Place of birth: Kobishcha, Bobrovitsky district, Chernigovsky region, Ukraine. Martial status: married with son and daughter. Fluent in Russian and in English.
Permanent address: Photochemistry Department, N.N. Semenov Institute of Chemical Physics, Russian Academy of Sciences, Novatorov Str. 7a, 117421 Moscow, Russia. =20 Tel.: 7 (095) 9363950 (office); Fax: 7 (095) 936 3950;=20 Tel.: 7 (095) 5504730 (home).=20
Current address: Micro- and Trace Analys is Centre (MiTAC), Chemistry Department, University of Antwerp (U.I.A.), Universiteitslpein 1, B-2610 Antwerpen-Wilrijk, Belgium. Tel.: +32-3-820.23.64; Fax: +32-3-820.23.76;=20 E-mail: oleshko-at-uia.ua.ac.be=20
Education=20 1974 B.S. (Chemistry) Moscow State University, Moscow, USSR. Honours in Chemistry.=20
1976 M.S. (Physical Chemistry) Moscow State University, Moscow, USSR.=20 Honours in Physical Chemistry.=20
1983 Ph.D. (Chemistry) Moscow State University, Moscow, USSR.=20
Ph.D. Thesis "Application of spin trapping to investigation of adsorption and catalytic processes".
Professional Experience 1971-1976: Student, Chemistry Department, Moscow State University.=20 Extended university courses and practical training in inorganic, analytical, organic and physical chemistry, polymer science and chemical technology and additional extended courses in high mathematics, physics and physical chemistry (catalysis and chemical kinetics, adsorption phenomena, electrochemistry, physical chemistry of solid state, quantum chemistry, and physical instrumental methods in chemistry).=20
1976-1982: Postgraduate student, N. I. Kobozev Laboratory of Catalysis and Gas Electrochemistry, Physical Chemistry Division, Chemistry Department, Moscow State University.=20 Experience in physical chemistry, catalysis, adsorption phenomena, kinetics of chemical reactions, vacuum and adsorption techniques and characterization of oxide based catalytic system s by electron spin resonance using spin probe and spin trap techniques, infrared-, and electron spect rometry in the ultraviolet and visible regions and computer simulation of donor-acceptor interactions on the oxide surfaces.=20
1982-1984: Junior Research Fellow, Research Institute of Chemical Technology, Moscow region, USSR.=20 Experience in polymer science of composite polymer systems and characterization of polymer molecular dynamics and stability by spin probe technique.=20
1984-1988: Head of research group, Senior Research Fellow, Experimental Designing Bureau "Horizon", Moscow, USSR. =20 Experience in materials science and characterization of superconductors (Nb-Ti alloys, Nb3Sn, and high-temperature superconductor Y-Ba-Cu-O systems) by scanning (SEM) and transmission electron microscopy (TEM) and energy-dispersive X-ray microanalysis (EDX).=20
1988-1993: He ad of research group, Senior Research Fellow, Photochemistry Department, N.N.Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow, Russia. =20 Experience in imaging science and chemical physics of record media (silver halide photographic systems, thin films, small particles and nanoclusters, photosensitive semiconductor dispersions and their characterization by analytical electron microscopy (AEM) methods (TEM/SEM/STEM/electron diffraction/EDX), image analysis and scanning tunneling microscopy.=20
1992.08.-11.: Visiting Scientist, Korea Research Institute of Chemical Technology, Daedeog-dangi, Taejeon, South Korea. =20 Experience in photographic science and technology of silver halide emulsions.=20
1993.08.-to present: Visiting Scientist, Micro- and Trace Analysis Centre (MiTAC), Chemistry Department, University of Antwerp (U.I.A.), Antwerpen-Wilrijk, Belgium. =20 Experience in energy-filtering transmission electron microscopy (EFTEM) and electron energy-loss spectroscopy (EELS) and scanning energy-dispersive X-ray microanalysis (STEM/EDX) of disperse many-particle systems (silver-halide-based photographic systems, metal and metal sulfide colloids, polynuclear cluster coordination compounds, nanocrystalline films) and Monte Carlo simulations of electron beam-solid interactions.=20
Number of papers: 51 scientific publications.
Invited reviews: Characterization of complex silver halide photographic systems by means of analytical electron microscopy //Microbeam Analysis. 1995. V.4. N.1, pp. 1-29 (first author).=20
Scanning Microanalysis: In Handbook of Microscopy. Eds. by S. Amelinckx et al., VCH, 39 p. 14 ill. (1996) (submitted, first author).
Presentations:=20 20 presentations at international and national scientific congresses and conferences.=20
Research interests 1. Surface phenomena, adsorption, photochemistry, photophysics and catalysi= s=20 at surfaces of solids, nanochemistry and nanophysics of molecular organized= =20 systems.
2. Analytical electron microscopy (energy-filtering TEM/ESD/EELS, STEM/SEM/EDX), image analysis and scanning tunneling microscopy of disperse matter.=20
3. Formation of ultradisperse metal and metal sulfide phases on microcrystals of silver halide emulsions at different stages of the photographic process i.e., ripening, exposure, and development.=20
Main professional accomplishments=20 1. Elucidation of mechanisms of acid-base interactions on oxide surfaces during radical catalytic reaction by spin traps and spin probes.=20
2. Development of a technique for evaluation of a surface oxide bacidity by the probing reaction of catalytic radical decomposition of 2-methyl-2-nitrosopropane.
3. Development of a technique for estimation of fine structure, core, shell and outer sizes of drops of color coupler dispersions in color photograph ic materials by combined positive-negative staining of ultrathin sections following stereological reconstruction of volume sizes.= =20
4. Elucidation of mechanisms of selective and non-selective reduction of silver halide emulsions with a series of developers (black-and-white, color, volume and surface developers) and with NaBH4. Demonstration of formation of percolation networks of silver filaments and particles in the course of development with an active volume developer.
5. Demonstration of fractal aggregation of products of chemical sensitization on tabular microcrystals of silver halide emulsions in the course of the ripening.=20
6. Development of the optimal strategy of structural and analytical characterization of complex silver halide photographic systems by the combination of AEM and image analysis techniques.=20
7. Evaluation of crystalline and electronic structure and elemental composition of advanced double structure tabular microcrystals of silver halide emulsions by the combination of cryo-EFTEM/ESD/EELS and cryo-STEM/EDX techniques. Development of a modified log-ratio EELS technique for estimation of the local crystal thickness.=20
Professional Societies=20 Russian D. I. Mendeleev Chemical Society=20 Belgian Society for Microscopy European Microanalysis Society
References=20 Prof. Dr. Renaat H. Gijbels=20 Micro- and Trace Analysis Centre (MiTAC), Co-Director Department of Chemistry, University of Antwerp (U.I.A.), Universiteitsplein, 1, B-2610 Antwerpen-Wilrijk, Belgium.=20 Tel.: +32-3-820.23.60 Fax: +32-3-820.23.76
Prof. Dr. Willem A. Jacob=20 Electron Microscopy Centre, Head Department of Medicine, University of Antwerp (U.I.A.), Universiteitsplein, 1, B-2610 Antwerpen-Wilrijk, Belgium. Tel.: +32-3-820.25.09 Fax: +32-3-820.26.03=20
Prof. Dr. Gustaaf Van Tendeloo EMAT, University of Antwerp (R.U.C.A.),=20 Groenenborgerlaan 171, B-2020 Antwerp, Belgium. Tel.: +32-3-218.02.62 Fax: +32-3-218.02.57.
Prof. Dr. Valery V. Lunin Department of Chemistry, Dean Moscow State University, Vorob=92evy Gory, 119899 Ms cow, Russia Tel.: 7 (095) 939 45 75, 7 (095) 939 35 71 Fax: 7 (095) 932 00 67.
Prof. Dr. Michael V. Alfimov Department of Photochemistry, Director N.N. Semenov Institute of Chemical Physics, Russian Academy of Sciences Novatorov Str., 7a,=20 117421 Moscow , Russia. Tel: 7(095) 936 77 53 Fax: 7(095) 936 12 55.
The following book may be of interest in regard to this discussion thread:
"Health Hazards for Photographers" by Siegfried & Wolfgang Rempel. Published by Lyonn (? may be an abbreviation) in 1993. ISBN 1558211810
I have not seen this text myself as it is too expensive in this part of the world - only $16-95 in the US though.
Perhaps Donna Wagahoff's building engineer was thinking of X-ray processors rather than ordinary photographic machines when he expressed concern over the extraction of fumes. I have no experience of X-ray processors myself, but understand that they have caused problems for a lot of users - isn't formaldehyde used as a hardener in these machines?
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department School of Medicine University of Auckland Private Bag 92019 Auckland New Zealand
} We need to find a supplier of stereo viewers, the cardboard-mounted } polarizing kind. I've looked in every EM supply house catalog that I have } and can't find them anymore. I'd appreciate any help in finding these. } Thanks. }
One possible solution:
Reel 3-d Enterprises Inc...Po Box 2368...Culver City, CA 90231-2368 Phone: (310)837-2368
I just posed a similar question for red/greed glasses. Several kind respondants gave me the name of the above company which sells all kind of 3D materials - including the polarizing glasses. I received the r/g glasses the next day. They accept charge cards or checks only (no PO's, etc). Prices are so reasonable, that I just ordered them on my own charge card.
I am just a happy patron with no financial interests in the company.
Peace -
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
Hello Charlie, I store my aqueous osmium solutions (usually 4%) in a frozen state and have had no problems to date. It is a convenient way to keep the osmium over a longer period as I only need to use it occasionally and saves the fridge interior from being inadvertently redecorated with the 'black' precipitate. Regards, . . Brett W. Cockman Technologist in Charge School of Dentistry University of Western Australia Voice: (619-2205834) Fax: (619-2213829) e-mail; bcockman-at-uniwa.uwa.edu.au
To expand upon Louisa Howard's question of long term storage -she is asking about storage in cacodylate buffer.
The question has recently been asked here about long term storage in phosphate buffers. We have a variety of fish, rat and cow tissues that were collected (or will be collected) "just in case" EM may be needed. Some of these tissues have been around for at least 10 years and will likely be here another 10 years.
What are the current thoughts on storage for long lengths of time? Would it be better to keep the tissues in fixative as opposed to buffer? Would cacodylate buffer be preferable to phosphate (or is there something else)? Any special techniques for processing old tissues and getting useable results? How long have tissues been kept around and still been successfully worked with?
Thanks for any advice from all the experts on the list.
Rosemary Harris Electron Microscopy Laboratory Alberta Environmental Centre
The last few years I have been involved in research on localized (near nanometer resolution) strain characterization using two-beam electron diffraction contrast imaging (an operational mode of transmission electron microscopy). See any book on transmission electron microscopy for the basics.
Part of this project was to develop a software package allowing for the analysis of strain fields of arbitrary geometry. This software allows one to model a microscopic strain field with various mathematical methods (including finite elements) and subsequently verify this model by comparison with experimental observations in the TEM.
At this moment we would like to know whether anybody would be interested in this software (SIMCON, i.e. SIMulating CONtrast images). If there are enough people interested we will probably arrange for a shareware software release. If you are indeed one of the interested please let me know by e-mail. To keep track of SIMCON you might check on http://www.mtm.kuleuven.ac.be/~janssens/simcon.html
Koenraad G. F. Janssens, Dr. Ir. Katholieke Universiteit Leuven (KUL) Departement Metaalkunde en Toegepaste Materiaalkunde (MTM) de Croylaan 2, B-3001 Leuven Belgium Tel: +32-16-32.1232 Fax: +32-16-32.1992 Koenraad.Janssens-at-mtm.kuleuven.ac.be
With regard to all the dicussion on fixatives, buffers,etc; maybe it's time to suggest two references:
1.Coetzee,J.and van der Merwe, C.F.,Some characteristics of the buffer vehicle in gluteraldehyde-based fixatives.Journal of Microscopy,Vol 146 Pt2, May 1987,pp143-155.
2.Coetzee,J. and van der Merwe, C.F.,The influence of processing protocol on the ultrastructure of bean leave cells.S.African Journal of Botany,1986 52(2)
Alan Hall Unit for Electron Microscopy University of Pretoria Pretoria. Tel: +27+012-420 3297 Fax: +27+012-420 3266
} } __________________________________________________________________________ _____ } } } } } HERE IS SOMETHING THAT WE ALL NEED TO TAKE ACTION ON. PASS THIS ON } } } } TO YOUR FRIENDS. } } } } } } } } } THE NEO-NAZIs are trying to start a news group. Their purpose is to } } } } get } } } } } their message of hate out to young people using the Internet. Its } } } } } originator, Mr. Burdi, is confident they will get the "yes" votes they } } } } need. } } } } } He said: "Let me be perfectly blunt and state that we have more than } } } } enough } } } } } 'net-nazis' to win this thing hands-down. But every one of you must } } } } vote } } } } } 'yes'". } } } } } } } } } } Mr. Burdi's confidence is disturbing, so please give this letter the } } } } widest } } } } } possible distribution and help us defeat this news group with a your } } } } "No" } } } } } vote. } } } } } } } } } } News groups are public discussion areas of the Internet and their } } } } formation } } } } } requires enough support from the Internet community. EACH AND } } } } EVERYONE OF } } } } US } } } } } HAS ONE VOTE when it comes to creating a new Usenet group. Please } } } } vote } } } } } against this news group. } } } } } } } } } } In order to vote, E-mail: music-vote-at-sub-rose.com. Your mail message } } } } should } } } } } contain only one statement: I vote NO on rec.music.white-power. } } } } } } } } } } Vote counting is automatic. Failure to follow these instruction may } } } } mean } } } } } that your vote does not get counted. If you do not receive an } } } } acknowledgment } } } } } of your vote within three days, contact the vote taker about the } } } } problem. } } } } } } } } } } Please distribute this message. } } Dr. Sheldon M. Schuster } Professor and Program Director } Box 110580 } University of Florida } Gainesville, FL 32611-0580 } Phone -- 352-392-8408 } Fax -- 352-392-8598 } } } ******************************************************* Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
Slide making machine: We have a Montage FR1 slide making machine that worked fine for us in older Macs with systems older than 7.5.2. My secretary told me that the company went out of business and that the phone was disconnected.
1) Does anyone knows if the compony was bought? 2) Has anyone newer drivers that possible works on newer macs?
I would appreciate any information you can provide and solution. As now, I contemplate hooking it up to an older mac.
********************************************************************* *Cesar D. Fermin, Ph.D \|T|/ Fax (504) 587-7389 * *Tulane Medical School /|M|\ Voice Mail (504) 584-2618 * *Pathology/SL79 \|C|/ Secretary (504) 584-2436 * *New Orleans La 70112-2699 /|*|\ Lab/Techn. (504) 584-2521 * * {Fermin-at-tmc.tulane.edu} ----} Prof. Pathology & Otolaryngology * *http://www1.omi.tulane.edu/departments/pathology/fermin/cdftop.html* *********************************************************************
Why is this on a microscopy newsgroup? If we are all going to start posting about our particular political hobby horses, just let me know. I got a million of 'em.
Charlie was worrying whether frozen osmium tetraoxide was still good for fixation. Michael and Brett have their osmium stored frozen. A colleaque in another lab. liked to store 4% aqueous oxmium in the freezing compartment. She had had broken ampules on several occasions. This came to light only after her retirement when her successor asked me how I stored mine. I make aqueous solution, seal 0.5ml in each ampule and leave them in the fume hood; no more osmium fume in the fridge and no black cooling compartment to worry about.
Although no longer listed in the catalog, Ted Pella still carries polarizing glasses for stereo viewing- or at least did a month ago.
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-bgsm.edu * **************************************************************
On Fri, 26 Apr 1996, John Chandler wrote:
} We need to find a supplier of stereo viewers, the cardboard-mounted } polarizing kind. I've looked in every EM supply house catalog that I have } and can't find them anymore. I'd appreciate any help in finding these. } Thanks. } } John } chandler-at-lamar.ColoState.EDU } } }
At 10:16 AM 4/25/96 EDT, you wrote: } } Our lab would like to convert our Zeiss Axiomat from film to digital. } We have no direct experience, and would appreciate any comments or } experiences that would help us research the best route to take, both } hardware and software, but particularly the former. } } I believe that this topic has been discussed recently, but unfortunately } I was not concerned until just now! If anyone has saved either their own } previous postings or those of others, I would appreciate if you would } email to me directly if you consider it repetitious to repost.
Hi Pat, I maintain an archive of most of the biological and computer related questions and responses that appear in this list. If you go to the www address at the end of this page and click on the "Tips & Tricks Wizard" you should find a discussion called "Acquiring digital images". Let me know if you do not have www access and I will be hapy to E-mail it to you.
Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 904-392-1295 ICBR EM Core Lab fax 904-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/
} } } } HERE IS SOMETHING THAT WE ALL NEED TO TAKE ACTION ON. PASS THIS ON } } } } TO YOUR FRIENDS. } } } } } } } } } THE NEO-NAZIs are trying to start a news group. Their purpose is to } } } } get } } } } } their message of hate out to young people using the Internet.........etc.
------------------------ This forum (microscopy) is for the discussion of microscopy related material. Political issues and so forth have NO place here. This is a place where scientists can discuss science.
We all have opinions concerning issues of censorship and other such controversial things concerning the Internet. There are appropriate forums for addressing your concerns......but not here.
We have a 2 yr old Edwards 2-stage direct drive pump (Model E2M-12, serial 20372) that needs to be repaired (keeps blowing fuses). Anyone know of a good repair place who might be able to handle this? Many thanks.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
} Greeting: } } } Does anyone know the OSRAM Inc. telephone# & address? We need bulb } replacement for our microscope. } Yhank you. } } Joseph Fu } NIST Rm.A117 Bldg.220 } Gaithersburg, Md. 20899 } Tel:301-975-3495 } e mail: jofu-at-enh.nist.gov } Fax: 301-869-0822
Try:
ESV Inc. (309)-347-6685
Tom
Thomas Moninger (moninger-at-emiris.iaf.uiowa.edu) Central Microscopy Research Facility 85 EMRB University of Iowa Iowa City, IA 52242 319-335-8142 FAX 319-335-8049
Message-Id: {9604291839.AA1097-at-pho903.sbphrd.com} To: microscopy {microscopy-at-Sparc5.Microscopy.Com} {Beverly_E_Maleeff-at-sbphrd.com}
Philadelphia Society for Microscopy Meeting Notice May 1996
DATE: Thursday, May 9, 1996
PLACE: Laboratory for the Research of Science and Materials (LRSM) Building, 33rd and Walnut Street, Philadelphia, PA. Parking is available behind the LRSM Building after 5:00 PM.
TIME: 5:30 Social hour, hosted by our meeting sponsor.
6:30 Dinner
7:30 PM Speaker:
Electron Microscopy in the Study of Copolymers: Blend Miscibility and Shear-Induced Defects
Dr. Karen I. Winey Department of Materials Science and Engineering University of Pennsylvania Philadelphia, PA
Polymers containing two or more types of monomer units are copolymers. The different monomer units can be distributed along the polymer chain in a variety of sequences which produce random, alternating, or block copolymers. Both the sequence distribution and the relative amounts of each monomer can produce significant changes in morphological, chemical and mechanical properties. Two examples will be given from our studies of copolymers which illustrate the value of electron microscopy. (1) We have developed a method which combines transmission electron microscopy, image analysis and mass balance to construct quantitative ternary phase diagrams. (2) Shear-induced defects, reminiscent of kink bands, have been imaged using low voltage scanning electron microscopy. The use of electron microscopy as a tool in these studies will be discussed.
DINNER:
COST: Members $12.00 Student members $6.00 Non-members $15.00
MENU: Barbequed chicken breast Barbequed beef ribs Hamburgers with all the trimmings Red bliss potato salad Pasta salad with fresh vegetables
Brownies Fresh fruit
Reservations will be taken by Ms. Pat Overend at the University of Pennsylvania, 215/898-8337. Deadline for reservations will be Friday, May 3. If you have any questions regarding the meeting please feel free to contact Rollin Lakis at 215/898-2013 or lakis-at-sol1.lrsm.upenn.edu. Cancellations must be received by Ms. Overend no later than 5:00 PM, May 3, 1996.
Please pass this information to anyone who is interested.
{bold} {fontfamily} {param} Times {/param} {bigger} {bigger} {bigger} TECHNICAL SUPPORT OPPORTUNITIES {/bigger}
The University of Michigan {/bigger} {/bigger} {/fontfamily} {/bold} {fontfamily} {param} Times {/para= m} {bigger}
{bold} {bigger}
Department of Materials Science and Engineering {/bigger} {/bold} =20
Due to recent and pending retirements, three technical support positions are open. The department seeks to hire a technical support team for the research and teaching needs of the department. The positions that are to be filled are outlined as follows:
Maintenance, service and repair of instruments and research equipment including electronics, vacuum systems, computer-based systems, mechanical testing equipment. This includes a number of scanning & transmission electron microscopes, an Auger system and X-ray photoelectron spectrometer. Assist with the maintenance and service of=20 X-ray diffraction systems, including a computer controlled Rigaku system. Installation and networking of computers, interfacing computers to research instrumentation, installation of data acquisition packages (e.g. LabVIEW) and programming data acquisition modules for such packages. Review and improve the safety program of the department.=20 Minimum requirement: B.S. in engineering or equivalent, plus three years of relevant experience.
Maintenance and service of two scanning electron microscopes (one scope is a field emission instrument) and two=20
X-ray diffraction systems, including a computer controlled Rigaku system. Operate and train students on proper use of above mentioned instruments. Operate and maintain departmental multimedia facilities (presentation computers, VCRs, TVs, TV cameras, CD-ROM drives, etc.). Assist with review and improvement of departmental safety program.=20 Minimum requirement: B.S. in science, engineering, or equivalent, plus three years of relevant experience.
{bold} {bigger} Computer Systems Specialist I/II, T-96-1176 {/bigger} {/bold}
Responsibilities include:
Assist in the planning, operation and maintenance of the hardware and software (network and stand alone) for departmental computer systems.=20 Provide technical advice to faculty, support staff and graduate students. Coordinate the implementation of problem resolutions for system hardware, software and network attachments and other operational needs; arrange for hardware repair; maintain a schedule of system back-ups and recovery procedures; load, test and implement upgrades to existing and/or new applications; provide user training on upgraded or new applications. Evaluation and acquisition of software packages.=20 Installation and networking of computers, interfacing computers to research instrumentation, installation of data acquisition packages (e.g. LabVIEW) and programming data acquisition modules for such packages. Operate and maintain departmental multimedia facilities (presentation computers, VCRs, TVs, TV cameras, CD-ROM drives, etc.).=20 Assist with review and improvement of departmental safety program.=20 Minimum requirement: B.S. in science or engineering, plus two years of relevant experience.
Interested individuals should submit 2 copies of resume to: Employment Services, The University of Michigan, G300 Wolverine Tower, 3003 S. State Street, Ann Arbor, MI 48109-1281. Reference the Job # listed next to the job title above. A non-discriminatory, affirmative action employer.
{bigger}
{/bigger} {/bigger} {/fontfamily}
John Mansfield
North Campus Electron Microbeam Analysis Laboratory
On the behalf of The International Society of Molecular Morphology
Would like to take the opportunity to announce the
FOURTH INTERNATIONAL CONFERENCE & WORKSHOP on MOLECULAR MORPHOLOGY
June 3-4, 1996 - Conference June 5 -6, 1996 - Workshop in
Montreal, Canada
CONFERENCE FEATURING Advances in Principles, Techniques and Applications in Research and Diagnosis of:
- In Situ PCR - In Situ Hybridization - Immunohistochemistry - Immunogold-Silver Staining - Immunogold Electron Microscopy - Microwave Immunohistochemistry - Atomic Force Probe Microscopy - Confocal Microscopy - Antigen Retrieval - Image Analysis
Call for Abstract Submission DEADLINE: APRIL 15, 1996
The abstract should be typed single space on white paper within 9 x 7 inch (23 x 18 cm) typed space. Total of two pages per abstract. Photographs and references may be included. Please follow the style of CELL VISION, in which the Proceedings will be published. Abstracts should be submitted in duplicate.
THREE HANDS-ON WORKSHOPS (at the laboratories of Dept. of Anatomy, University of Montreal) 1 In Situ PCR and In Situ Hybridization 2 Immunogold EM and Immunogold-Silver Staining 3 Microwave Fixation, Antigen Retrieval and Immunohistochemistry
June 2, 1996 (Sunday), 6.30-9.30 pm: An optional preparation lecture on "Molecular Biology for the Uninitiated"
ORGANIZING COMMITTEE CO-CHAIRMEN Jiang Gu, M.D., Ph.D. Deborah Research Institute Browns Mills, New Jersey, USA
Moise Bendayan, Ph.D. University of Montreal Montreal, Canada
MEMBERS Virginia Anderson, M.D. Health Science Center at Brooklyn State University of New York Brooklyn, New York, USA
Gerhard Hacker, Ph.D. Institute of Pathology General Hospital, University of Salzburg Salzburg, Austria
Lawrence DeBault, Ph.D. Oklahoma University Health Center Oklahoma City, Ok, USA
Shahla Masood, M.D. University of Florida Health Science Center Jackonsville, Florida, USA
Robert Day, Ph.D. University of Montreal Montreal, Canada
David Kersten, M.D. Ealing London, UK
---------------------------------------------------------------------------- -------- ADVANCED REGISTRATION FORM (please print and use photocopies for additional forms)
CHOICE OF WORKSHOP (circle one): 1 2 3 ---------------------------------------------------------------------------- - ADVANCED REGISTRATION FREE (for one of three): - For the two-day conference $250 (US) - For the two-day workshop $350 (US)
Make check payable to CELL VISION. A 15 % discount for members of the "International Society of Molecular Morphology" and students (with proven ID). A 15% discount will be reimbursed upon becoming a member of the society before or at the conference. Please book the hotel room directly by calling The Best Western Hotel in Montreal, Canada (800) 361-3000 or Fax (514) 861-4089. You can obtain a special discounted room rate at $79 (Canadian) per day (rate includes breakfast) by identifying yourself as a participant of the conference/workshop. Student dormitory available at University of Montreal (15-minute subway transporation to conference location, 5 minute walk to workshop location) at $35 (Canadian) per day by calling (514) 343-6531.
Send abstract, registration form, and registration fee to: CELL VISION-JOURNAL OF ANALYTICAL MORPHOLOGY, EDITORIAL OFFICE, DEBORAH RESEARCH INSTITUTE 1 Trenton Road, Browns Mills NJ 08015-1799, USA Phone: (609) 735-0477 Fax: (609) 735-0478
For further information please direct your inquiries by email to: morphology-at-scottscientific.com _______________________________________________ SCOTT SCIENTIFIC P.O. Box 66552 Station Cavendish, Montreal, Quebec, H4W 3J6, Canada
A Reminder: This forum should not to be used to post non-Microscopy/Microanalysis related announcements, requests, and/or informational items. There are sufficient number of public forums in which these issues may be freely and openly debated and discussed to your heart's desire.
Please refrain from using the Microscopy ListServers for those purposes, however, noble you may believe your motivations are....
There is no need for the ListServer community to echo this, as enough bandwidth has already been wasted, and enough mailboxes cluttered.
A colleague wishes to differentiate between living and dead cells. Does anyone have a simple and reliable method (or if not simple at least reliable).
The cells are from plant cell suspension cultures that have been subjected to various environmental stresses.
Thanks in advance
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
His mail address is: Philosophical Magazine Department of Physics and Astronomy University of Leicester University Road, Leicester LEI 7RH England
If someone knows, please tell me.
Many thanks!
Zhen Quan Liu --------------------------------------------------------------------- *Important!!! ALL TELEPHONE NUMBERS in Beijing will be changed from May 8, 1996 from 7 digitals to 8 digitals!!! The way of changing is by putting 6 infront of the initial number. Therefore my tel since then will be Tel: 6275 1427(office) 6275 3727(home) Fax: 6275 1615 The codes for China and Beijing will not be changed, they still are: China 86 Beijing 10 -------------------------------------------------------------------- ** Sorry, my E-mail software cannot show me the address of coming mails. Please tell me your E-mail address within the text. --------------------------------------------------------------------- Zhen Quan Liu (Ph.D) Tel:(86) 10 275 1427(Office) Physics Building (86) 10 275 3727(Home) EM Lab. Email: zqliu-at-pku.edu.cn Physics Building Email(home) wl-at-ibmstone.pku.edu.cn Peking University Fax (office): (86) 10 275 1615 Beijing 100871, China
I already have the information needed and is enclosed below for anyone else may need it.
The company was bought by the employees. Their new name is Montage Graphics, the address is 2730 Scott Boulevard, Santa Clara CA 95050. The phone is Sales - 800-416-4166, Email sales-at-montagegraphics.com, Technical support 408-654-0684, Tech email tech-at-montagegraphics.com
Mei Lie Wong Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 email wong-at-msg.ucsf.edu
} We have a 2 yr old Edwards 2-stage direct drive pump (Model E2M-12, serial } 20372) that needs to be repaired (keeps blowing fuses). Anyone know of a } good repair place who might be able to handle this? Many thanks. } } } ############################################################################# } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Southern Illinois University } Carbondale, IL 62901-4402 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ############################################################################# } We had a Balzers rotary pump rebuilt by Atlantic Vacuum Repair -at-201-623-1115. They did a great job, and I would recommend that you give them a call. They are at 96 Harper St., Newark, NJ. 07114. I'm one satisfied customer.
I have not tried differentiating between living and dead eucaryotic cells, but maybe it would be a good idea to contact Molecular Probes Europe BV. They are selling a number of live/dead fluorescence staining kits for both procaryotic and eucaryotic cells.
(unfortunately they are not going to pay me for posting their address) Molecular Probes Europe BV, 2333 AA Leiden, The Netherlands Phone: +31-71-233378; Fax: +31-71-233419
Stefan
* Stefan Andreatta * Institute of Zoology and Limnology * University of Innsbruck; Austria
Patrick Drexel or Tom Freda at Prior Scientific, 800-877-2234
Irv Toplin at Carl Zeiss, 800-356-1090
Jim Dutkiewctz at Meiji, 800-832-0060.
All these references feature products in The Microscope Book. The Spring Edition is on press. Would you like a copy?
Please call us if there is anything more we can do to assist.
Elinor Solit, Director of Publications, The Cambrex Group 800-440-0311
On Fri, 26 Apr 1996, James S MArtin wrote:
} Several months ago I posted a request for microscope manufacturers to } contact me with information on costs for epi-fluorescence upgrades to } polarizing light microscopes. The information will be presented at a } national conference. } } So far, I have information only from Olympus. Anybody else like to be } represented? } } James Martin } Director of Analytical Services and Research } Williamstown Art Conservation Center } Williamstown, MA }
Subject: Time: 7:36 AM OFFICE MEMO RE:VacPmpRepair Date: 4/30/96
Most manufacturers of vacuum pumps will provide repair and maintenance services for their pumps. Usually, to save time in getting you back into operation, they will make an exchange with you, immediately shipping you a used, but fully reconditioned pump, for the one you send in for repair. Having the pump serviced by the manufacturer should be about the most reliable way of ensuring that it receives proper attention. The phone number of the Edwards service unit is 716-695-6354
I passed on the inquiry regarding Peltier vs adabiatic coolong of CPDs to Tony King, Product Specialist at VG Microtech, who manufacture the Polaron range of critical point dryers which we sell. This is his reply:
RE: Re: Peltier vs adabiatic CPD
One of the main reasons for changing from adibiatic cooling was due to health and safety, 10% CO2 in air can cause permanent brain damage. Secondly the cooling of a large vessel suitable for a number of samples can be slow and difficult to control. If you use a peltier cooled device the big advantage is that by reversing the polarity the device acts as a heater with a limited range so it is safe and efficient for this very application.
The only draw back that we have seen is that in a warm area the excess heat must be removed by additional cooling. We used air previously but again this was inefficient so we went to water.
As soon as you add peltier cooling and heating you can run a 3 term controller and any one that wants to be clever can heat or cool in precise stages!.
I hope this helps
Regards,
Tony King Product specialist VG Microtech/ Polaron range
Tel: +44 (0)1825 746251 Fax: +44 (0)1825 768343
Disclaimer: The views and opinions expressed are not necessarily those of Fisons plc or VG Microtech.
Dear John, Try letting your maintance shop do it. I was informed by my old job maintance men that a direct drive pump is good for about 2-3 rebuilds. The belt driven pumps can take about 8-10 rebuilds.
Best of luck, Ed Calomeni Medical College of Ohio Toledo, OH 43699 emlab-at-opus.moc.edu
Attached is a technical meeting announcement for anyone interested
MAMAS (Mid-Atlantic Microbeam Analysis Society) Meeting at the National Institute of Standards and Technology Gaithersburg, MD on Thursday, May 9, 1996 10:30 am- 2:15 pm Lecture Room D, Administration Bldg.
10:30 Coffee and Doughnuts
10:45 Joe Geller, Geller Microanalytical Laboratory, Standards for the SEM Laboratory, Preparation for ISO-9000
12 noon Lunch
1:15 John Armstrong, NIST, Evaluating and Choosing Standards for Quantitative Microbeam Analysis
For more information, contact Ryna Marinenko (301)975-3901, FAX(301)216-1134, email:ryna.marinenko-at-nist.gov
Eric B. Steel, Leader e-mail: eric.steel-at-nist.gov Microanalysis Research Group Office: 301-975-3902 N.I.S.T. FAX: 301-216-1134 Bldg. 222/Rm A113 Gaithersburg MD 20899
I would just like to thank all ( and there were many ) scientist that took the time to send me information regarding the frozen 4% osmium. The information came so quickly that I was able to carry out an experiment with complete confidence of the outcome. By the way it was unanimous. The OSO4 is O.K. if it has been frozen. Thanks again. Sincerely Charlie Murphy
} Greeting: } } } Does anyone know the OSRAM Inc. telephone# & address? We need bulb } replacement for our microscope. } Yhank you. } } Joseph Fu } NIST Rm.A117 Bldg.220 } Gaithersburg, Md. 20899 } Tel:301-975-3495 } e mail: jofu-at-enh.nist.gov } Fax: 301-869-0822 } Joe: Have you tried Bulbman Inc. in Reno, Nevada? They have had every type of bulb I've needed. The cost is usually a fraction of what the manufacturer charges. So far they have been the least expensive of any other people I've tried. The people are pretty good handling customers and their needs. Excellant to deal with. Their number is: 1-800-648-1163 Fax: 1-800-548-6216 One time I had to replace a bulb, the bulb had no recognizable numbers to give them. I faxed them the type and model of the microscope and they were able to give me the right bulb. Hope this helps.
Joe Fu wrote: } } Greeting: } } Does anyone know the OSRAM Inc. telephone# & address? We need bulb } replacement for our microscope. } Yhank you. } } Joseph Fu } NIST Rm.A117 Bldg.220 } Gaithersburg, Md. 20899 } Tel:301-975-3495 } e mail: jofu-at-enh.nist.gov } Fax: 301-869-0822
A colleague wishes to differentiate between living and dead cells. Does anyone have a simple and reliable method (or if not simple at least reliable).
The cells are from plant cell suspension cultures that have been subjected to various environmental stresses.
Thanks in advance
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
Here is a reminder and some clarifying information.
The Microbeam Analysis Society is sponsoring their first Topical Conference immediately prior to the Microscopy and Microanalysis 96 meeting in Minneapolis this year. The Topical Conference is entitled:
{bold} "Microscopy & Microanalysis Resources on The World Wide Web"
It will be held in The Minneapolis Convention Center on
SATURDAY THE 10TH of AUGUST 1996.
Note that the ad in Microscopy Today contained a typo stating that the meeting was to be on the Sunday. The meeting has been placed on SATURDAY to avoid any conflict with the M&M Tutorials.
The starting time will be 9:00am and the conference will last all day.
The morning will be a series of presentations which will cover the following:
1. Basic introduction to the Web
2. What is available on the Web generally.
3. What is available on the Web for Microscopy & Microanalysis
4. How to create your own Web Page.
Invited Presenters include:
Greg Meeker - U.S. Geological Survey.
Marc De Graef - Carnegie Mellon University.
Darcy Clark - University of Michigan (formerly University of Queensland).
John Mansfield - University of Michigan.
The afternoon will be focussed on a hands-on workshop where there will be a minimum of 15 computer workstations connected to the Internet (via a T1 line)
for attendees to use. The morning's presenters will be available to advise attendees on all aspects of accessing and using the Web.
Attendance is FREE to MAS members
Attendance is $35 for Non Members
If you join The Microbeam Analysis Society you may attend free, membership is $25 per year.
You may register by:
1. Filling out the card that was contained in the last issue of Microscopy Today
and mailing it in to the address noted on the card.
2. You may register electronically if you are a member. Connect to:
http://www.microanalysis.org/mas/regtopconf.html
3. Non members may complete the form at:
http://www.microanalysis.org/mas/regtopconf.html, print it and mail it to the address supplied in the form.
4. Members & Nonmembers may send their:
Name
Full address
Business phone
FAX
email address
and an indication of their computer and Web expertise to John Mansfield
at the address at the end of this message.
Non-Members should enclose a check for $35 (drawn on a US bank) made out to The Microbeam Analysis Society.
I have been using Cy5 with great luck on my confocal but now need to take some B&W photos using my epi-fluorescence rig. I know something like Tri-X pan would be virtually insensitive to the far red emission of Cy5. Does anybody have practical experience with Tmax and Cy5 or is there a better choice of B&W film for this purpose. Thanks.
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)
Sender: kayton-at-ohsu.edu (Robert Kayton) Message-Version: 2 } To: Microscopy-at-Sparc5.Microscopy.Com
James,
Try the following companies and people:
Patrick Drexel or Tom Freda at Prior Scientific, 800-877-2234
Irv Toplin at Carl Zeiss, 800-356-1090
Jim Dutkiewctz at Meiji, 800-832-0060.
All these references feature products in The Microscope Book. The Spring Edition is on press. Would you like a copy?
Please call us if there is anything more we can do to assist.
Elinor Solit, Director of Publications, The Cambrex Group 800-440-0311
On Fri, 26 Apr 1996, James S MArtin wrote:
} Several months ago I posted a request for microscope manufacturers to } contact me with information on costs for epi-fluorescence upgrades to } polarizing light microscopes. The information will be presented at a } national conference. } } So far, I have information only from Olympus. Anybody else like to be } represented? } } James Martin } Director of Analytical Services and Research } Williamstown Art Conservation Center } Williamstown, MA }
Message-Id: {9605010047.AA03572-at-asuka.nal.go.jp} Comments: Authenticated sender is {davies-at-asuka.nal.go.jp}
I work at a research institute in Japan in the field of materials science. I have experience in TEM and SEM but recently have been trying to take micrographs using an Olympus SZH optical microscope. This microscope has a "zoom" lens from nominally 7.5 to 64.
My problem is that this microscope appears to exhibit something similar to astigmatism even at the lowest magnification. We also have an Olympus STM5-BD which works perfectly. With the SZH I have tried to realign the optical system but there is little improvement. Another department in my institute also has an SZH with exactly the same problem.
Does anyone know if this problem is common with the SZH microscope and how it may be corrected.
Please could you send any thoughts and suggestions to me at DAVIES-at-ASUKA.NAL.GO.JP
Any ideas would be gratefully received. Thank you.
} If } you use a peltier cooled device the big advantage is that by reversing } the polarity the device acts as a heater with a limited range so it } is safe and efficient for this very application. } } The only draw back that we have seen is that in a warm area the } excess heat must be removed by additional cooling. We used air } previously but again this was inefficient so we went to water. } } As soon as you add peltier cooling and heating you can run a 3 term } controller and any one that wants to be clever can heat or cool } in precise stages!. } Dear Tony, We have a darkroom rinsing tank which is heated/cooled by peltier devices. One caution with using these devices is that most of the commer- cial controllers are duty-cycle type, i.e., they turn from full off to full on for a fraction of the time which depends on the difference between the set point and the temperature being controlled. This kind of controller is harmful to peltiers. Our electronics shop designed a controller which puts out a voltage which is proportional to the set-point-temperature difference, so it adjusts slowly as the temperature reaches the set point and never has sudden changes in output. With this kind of controller, the peltiers have lasted many years without apparent damage. Yours, Bill Tivol
Please enter my subscription to your service. Please use the following e-mail address: szenk-at-kevex.com. Should you have any questions, please contact me.
Thanks for your continued efforts to satisfy the needs of the microscopy consortium.
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