I tried to no avail to send you the present message at your email address: it stubbornly bounced back, so I eventually post it to the microscopy list. I apologize to the members of this list if they should feel this is wasting bandwidth: just delete and go on to the next message ;-)
Barbara! I am afraid I did not make my question clear enough ;-) What I wanted to know was: 1) the testes you wish to fix are from what species ?
2) for what kind of microscopy are they to be processed ? I.e. T.E.M., S.E.M., or light microscopy ?
3) What's the embedding medium?
4) in the case of light microscopy, do you contemplate using some special technique (besides for recognizing the different stages of the seminiferous epithelium) which would require that some specific chemical would be either required or, on the contrary, avoided?
In my personal experience with rat testes and light microscopy (this is rather very long ago: more than 30 years! :-( ), Bouin and other picric acid containing variations thereof are not optimal fixatives for preservation of the acrosomial apparatus of spermatids, which is one of the features used by most classification methods (Clermont's among others...). The best fixatives for light microscopy of paraffin embedded tissue are those containing potassium dichromate such as, for example, Helly's fluid (Zenker + formaldehyde), but they also are followed, after rinsing and dehydration, by shrinkage which is still more important than that observed after Bouin's or Bouin-Hollande's (somewhat better) fluids.
BTW, what's wrong with picric acid? It is always shipped in 'moist condition' in order to minimize hazards. I work in our lab since 1953, with Bouin, Bouin-Allen, Bouin-Hollande, Dubosq-Brazil etc., every day, and we never experienced any problem. We store the picric acid for our needs in the form of a saturated aqueous solution which keeps for as long as you will. This stock solution is then used for preparing all picric acid containing fixatives.
There is no point in trying to eliminate the picric acid from the fixed pieces of tissue before sectioning. If the presence of picric acid in the slides should hamper subsequent staining (a rare occurrence, btw), then the easiest and expeditious way to get rid of it is to pretend your tissue was fixed with a sublimate containing fluid: at the rehydration step, your slides should undergo treatment by alcoholic iodine (or lugol solution) and sodium thiosulfate in the usual way and 'voila': no more picric acid on the slides within 2 minutes! Under no circumstances should the pieces of tissue fixed with a picric acid containing fixative undergo rinsing in water! This is sheer heresy ;-) because it induces a lot of swelling in the tissue before shrinking even more during dehydration (I never found out which histologist ever advocated such method, but I know quite a number of renowned ones who firmly condemned it, and with good reason). If you insist on rinsing the pieces of testes after fixation, then merely increasing the number of steps through 90x alcohol should do the trick; you may even add a thin layer (1-2 mm thick) of lithium carbonate on the bottom of the alcohol containing flasks: this helps dissolving part of the picric acid away.
Possibly the method which should yield the best pictures has been documented in a book entitled 'Histological and histopathological Evaluation of the Testis', by Russell,Ettlin, Sinha Hikim and Clegg: Cache River Press 1990, (15777 Bolesta, Box 129, Clearwater, Fl 34620): ISBN 09627422-0-1.
At the end of the book, there is an appendix about the materials and methods which you might find useful (although perfusion fixation of rat testes is among the most frustrating experiences I ever had, even with a lot of heparin ). This would also require embedding in epoxy resin and making large semi-thin sections, which I don't know whether you are prepared to do ;). Glutaraldehyde certainly seems to me the best choice, but then paraffin embedding would be ruled out (too hard and too brittle after paraffin embedding). HTH, and good luck. Let me know about the outcome (good, I hope) John (correction for above glitch about the alcohol/ethanol: should read alcohol 90 percent (in volume); I am still stuck with kermit and an awfull editor :( ).
*********************************************************** * Jean C. Desclin (John), Associate Prof. of Histology * * Laboratory of Histology - Faculty of Medicine * * Brussels Free University (U.L.B.) * * e-mail: desclinj-at-ulb.ac.be (internet) * * snail mail: route de Lennik 808 * * B - 1070 Brussels Belgium * ***********************************************************
this just is a test because it seems I am not able to reach the list. Please ignore and delete. I apologize for the inconvenience. desclinj-at-ulb.ac.be
The 1 MeV AEI EM-7 at the University of Wisconsin, Madison has been used for the study of biological specimens for the past 23 years. In the early 1980's, almost every subsystem of this instrument (magnetic shielding and ambient field, lens-current and HV stability, vibration isolation, LaB6 source, TV-interface, Axis-centered stereo-tilt stage, high-resolution cold-stage etc.) was upgraded with a view to performing high-resolution studies on frozen biological specimens. Images of oriented gold films at that time showed 0.14 nm spots in bright field and using an 11 mm gap (Pawley, J.B. (1984) Ultramicroscopy 13-4:387-406, describes most of these improvements in detail.)
Because it seems probable that the present management of this instrument will decide (has decided?) to decommission it in the very near future, (It was almost scrapped over last Christmas), I am posting this notice in case there are those that feel that the country has need of a second instrument with the general specifications of the Berkeley ARM or even of the very-stable Haefely 1-MEV supply that it contains.
The instrument is in operating condition, but, until recently, has received less maintenance than it should have for the past five years or so.
The questions are:
1. Do you have important projects that require higher resolution (or a large gap) than that available from 200-400kV instruments? (Keep in mind that knock-on damage may be more severe at the higher voltage.) 2. Would you be willing to travel to Madison to obtain such facilities? 3. Would you be interested in relocating the instrument to a more convenient location?
Bear in mind that, because the instrument requires a room 3 floors high and a large (100 t) vibration-isolation block and also has substantial power and cooling requirements, moving it would be a bit complex. However, it could surely be done at less than 10% of $5M cost of the new Stuttgart 1.25 MeV instrument.
It would also need stage-rods more suited for material-science applications but it is possible that the stage formerly fitted to the Cambridge HREM could be fitted without great trouble as both instruments used the same column.
The purpose of this communication is solely for the information of possible users with the aim of avoiding anyone in the future saying "If only you had told me?". I do not represent the Integrated Microscopy Resource where the instrument is installed.
Please send any responses directly to me and do so ASAP:
jpawley-at-macc.wisc.edu (James Pawley)
***************NEW ADDRESS************** Prof. James B Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr. Madison, Wisconsin, 53706. JPAWLEY-at-MACC.WISC.EDU
Dr. Keith Moulding, Materials Characterisation and Preparation Centre, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Andreas Brech Electron Microscopical Unit for Biological Sciences Department of Biology, University of Oslo. P.O.Box 1062 Blindern N-0316 Oslo 3 Norway Tel.: + 43-22 85 61 89 (work) + 43-22 43 83 23 (privat) Fax.: + 43-22 85 47 26 e-mail.: abrech-at-bio.uio.no
Please, I would like to join the list. Andreas Brech Electron Microscopical Unit for Biological Sciences Department of Biology, University of Oslo. P.O.Box 1062 Blindern N-0316 Oslo 3 Norway Tel.: + 43-22 85 61 89 (work) + 43-22 43 83 23 (privat) Fax.: + 43-22 85 47 26 e-mail.: abrech-at-bio.uio.no
Does anyone have a referrence for backscattering cross-sections of electrons in various substances as a function of energy and/or of angle? I'd really appreciate a reply either to the list, by email or if I could get a table by fax at (518) 474-8590. TIA. Yours, Bill Tivol tivol-at-tethys.ph.albany.edu
I'm looking for any recomendations for freeze-fracture units. We're putting together a grant here to obtain one and the only souce I've come up with is Bal-tec. I am looking for other possible vendors. Apparently JEOL has stopped production of their freeze- fracture unit. Specifically we're looking for units comparable to the Bal-Tec BAF 060 (not that I as of yet have anything against the BAF 060, but it is nice to look around first.)
Thank you.
Richard E. Edelmann Electron Microscopy Facility Supervisor Miami University, Oxford, Ohio
I don't think there is a unit out as yet that can match the specs and design of the new BAL-TEC unit. Cressington is another company which manufactures freeze-fracture unit. While it may not have all the features of the new BAL-TEC one, it is reported to be good for routine freeze-fracture.
************************************************************************* M.V. Parthasarathy Professor of Plant Biology Section of Plant Biology 228 Plant Science Building Cornell University, Ithaca, NY 14850 Telephone: (607) 255-1734 Fax: (607) 255-5407 E-mail: mvp2-at-cornell.edu ***********************************************************************
} I'm looking for any recomendations for freeze-fracture units. } We're putting together a grant here to obtain one and the only souce } I've come up with is Bal-tec. I am looking for other possible } vendors. Apparently JEOL has stopped production of their freeze- } fracture unit. Specifically we're looking for units comparable to } the Bal-Tec BAF 060 (not that I as of yet have anything against the } BAF 060, but it is nice to look around first.) } } Thank you. } } Richard E. Edelmann } Electron Microscopy Facility Supervisor } Miami University, Oxford, Ohio
It looks as though you have some misinformation. JEOL is making the JFD 9000 series freeze-fracture instruments. I know that John Rash has recently purchased and installed the 9010CR with the latest modifications, including a specimen stage with rapid cooling.
Both Bal-Tec and JEOL make fine instruments and are worth looking at.
John chandler-at-lamar.ColoState.EDU Fort Collins CO
Many published investigations on stress voids in aluminum/silicon alloys report using CF4/O2 plasma as the deprocessing method to remove the final PECVD oxynitride/ SiN. Does the use of this plasma chemistry generate its own voids by removing silicon precipitates which in turn leave voids possibly identified as stress voids? Precipitates that were in or near the correct orientation give the appearance (typically wedge-shaped) of a stress void. Optical inspection without deprocessing does not have adequate resolution. I'd appreciate any comments on the CF4/O2 method and a possible alternative method (other than argon backsputtering).
One of the divisions of our lab teases apart sural nerve tissue that have been kept in a 1:1 mixture of epon:propylene oxide. The samples are very easy to tease when the mixture is very fresh. The problem is that samples became back-logged and were not able to be teased right away. A quick decision had to be made, and the samples were frozen at -70C until they could be teased.
The nerve fibers need to be infiltrated to be teased but not polymerized or else they become too fragile. A problem is becoming evident that after a long period of storage, the nerve samples are being fully infiltrated and polymerized despite being frozen (which we thought would almost stop the polymerization process because it is largely dependant on heat). Realizing that hind-sight is 20/20, would it have been better to leave the accelerator out of the epoxy mixture? What affect would this have on the quality of the resin? I understand accelerator to act as a catalyst being a part of the polymerization reaction but not really consumed. Any opinions to offer?
I have been asked to make a brief (15 min) presentation at Scanning 95 in Monterey, on March 30, 1995. The topic is 'Changing Role of the Microscopy Lab in the University'. I know how my lab has changed, but would like to get some idea from a broader sample of microscopists about changes in our role etc.
My rough observations at this time include some of the following. More labs seem to be consolidating, so we keep trying to catch up with too many techniques, especially as staffs are reduced.
There are lots of other places for researchers to spend their funding than before, we have lost lots of users in biology to molecular techniques. Some of the new faculty see microscopy as 'too hard'.
There are fewer and fewer students who want to be 'microscopists'. Most seem to want to come in on Monday, drop off something, and return on Friday with a picture or two for their thesis.
The EM lab on our campus has evolved into an imaging and microscopy lab. Because of our interest in images, we have always had the best darkrooms, now we are trying to lead the way into digital imaging. This means more computers and branching out from traditional 'microscopy'.
I could probably think of other changes, but that wouldn't leave anything for you to add. Don't be shy, reply directly to me and I will try to incorporate your comments in my presentation in a constructive way. Let me know if you want specific credit for an idea, or if you prefer to have your input blended for anonymity.
If you can't make it to Monterey, I'll send a summary if you ask.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95060 emlab-at-ucsco.ucsc.edu (408) 459-2477
Reply to: RE} Re: Optical Microscopy of Tissues Dear John, Sorry about the delay in responding - I missed your message in the deluge I received while visiting labs for a few days. Check out the following papers on infrared DIC-videomicroscopy.
H.-U. Dodt, W. Zieglgansberger (1990) Brain Research 537:333-356; (1994) Trends in Neurosciences 17 (11): 453-458 (and references therein).
They looked with transmitted light at depths of up to 100 um in 300 um thick sections. Used a 63x 1.4 NA oil immersion objective and DIC optics to visualize nerve terminals. Note that the Nikon 60x 1.2 NA WATER immersion objective is now available and has advantages in deep sections (see for example Mel Brenner's application note in the November 1994 Journal of NIH Research).
Infrared penetrates well because water and tissues doesn't absorb it as much as visible. For video work a CCD (or cooled CCD) is used because they are sensitive to IR (unlike most tube cameras and your eye). DIC is the method of choice because of optical sectioning. A general review or IR cameras is: Silverman et al (March 1992) Scientific American 78-83 and 112-113. Any high quality video CCD camera will work (as long as it does not have an IR blocking filter!).
Sincerely,
Dr. George McNamara Universal Imaging Corporation --------------------------------------
Hello,
I am currently writing a grant proposal, and I have been knocking my head against the wall looking for some sort of data on the optical transmission properties of nervous tissue (the brain). Does anyone have a good reference, or, from a practical standpoint, how thick do brain tissue slices have to be to obtain good optical transmission (esp. relative to other fatty tissue slices like those from the earlobe, whose transmission characteristics I can find since people have been doing pulse oximetry.).
Thanks in advance,
John Conroy University of Utah Dept. Bioengineering
--------- Dear John,
It has been years since I worked in this area, but I suggest you might look at the literature for the optical transmission properties of retina. As a nervous tissue, its optical properties should be quite similar to the CNS as long as the measurements are not made in the vicinity of the fovea. And, of course, due the importance of retinal transparency for vision, its optical properties must be well known. For a place to start I'd have a look at the "Handbook of sensory physiology" Springer-Verlag.
The objective lens and the upper and lower column parts of our 20 year old side entry JEOL 100 C microscope need replacement. New parts, however, are rather expensive in view of the age of the instrument so we're looking for anyone who has some second hand spare parts for sale. Sincerely, Nick Schryvers tel: 32-3-2180247 fax: 32-3-2180257 EMAT, University of Antwerp Groenenborgerlaan 171 B-2020 ANTWERP Belgium
I first became aware of the recomendation to replace the epoxy resin= acelerator DMP-30 with the acelerator BDMA when Audrey Glauert published= her article "Accelerators for epoxy resins" in the RMS Proceedings (Sept, 1= 987).
The reason offered to encourage the switch from DMP-30 to BDMA was the fact= that BDMA is less viscous, therefore diffuses into the tissue sample= better. This gives a more even polymerisation and better sectioning. BDMA= also has a longer shelf life.
I followed Glauert's recomendation, replacing DMP-30 with BDMA in our resin= formula's. However, Audrey Glauert did not mention in the article that= we should double the quantity of BDMA in the formulation when switching= from DMP-30 to BDMA.
I replaced the DMP-30 with BDMA, using the BDMA at the same concentration as= I had used the DMP-30.
Generally, it worked fine. Occasionally I had 'strange embedding' problems,= ie samples not properly polymerised.
It then came to my attention that I should be using BDMA at twice the DMP-30= concentration. I did not pay much attention to this at the time as= generally everything had been working fine and I was to busy to experiment= with the change.
After recent 'flow' on the List server the topic again surfaced i.e. I= should be using BDMA at twice the DMP-30 concentration. This I have now tr= ied.
My problem is that if I do an overnight infiltration in resin with the= increased concentration of BDMA the resin is so viscous next morning it= won't even come out of the vial when tipped upside down. To change to= fresh resin is almost impossible, in fact it is easier to change the= samples to fresh resin in a new processing tube. Unfortunately even then= it is not totally successful as a large 'blob' of tacky resin stays with= the sample.
I did a little experiment as outlined below;
1. Made up resin with normal DMP-30 concentration. 2. Made up resin with BDMA at normal DMP-30 concentration. 3. Made up resin with BDMA at twice DMP-30 concentration.
Results
Viscosity (by eye) Colour To start with DMP-30 most viscous slightly orange BDMA (-at- DMP-30 conc) least viscous straw coloured BDMA (-at- 2 x DMP-30 conc)similar to BDMA -at- DMP 30 conc straw coloured
After 5 hours DMP-30 least viscous darker straw coloured BDMA (-at- DMP-30 conc) similar to DMP-30 straw coloured BDMA (-at- 2 x DMP-30 conc)quite viscous straw coloured
Overnight DMP-30 least viscous straw coloured BSMA (-at- DMP-30 conc) slightly more viscous than DMP-30 straw coloured BDMA (-at- 2 x DMP-30 conc)very very viscous straw coloured
After the overnight step both the DMP-30 mix and the BDMA mix at the DMP-30= concentration were quite acceptable, ie no difficulty replacing the old= resin with fresh resin.
The BDMA mix at the twice DMP-30 concentration ws very very tacky.
After polymerisation both the DMP-30 and the BDMA -at- the DMP-30= concentration are a light straw colour (normal expectation). The BDMA -at-= twice the DMP-30 concentration was slightly darker, although still an= acceptable straw colour.
By modifying my processing schedule such that the samples stay in 3 parts= plastic and 1 part propylene oxide overnight I can successfully process a= sample however, I am a little unhappy with the short time the sample is in= 'resin only' the following day, ie out of 3;1 and into fresh plastic first= thing in the morning, embedded and in the oven before I go home. With a= lot of samples to embed at times trying to give the samples as long as= possible in the 'resin only' makes the end of the day a bit tight for time= .
The problem becoming particularly accute when using our Lynx Automatic= Tissue processor. If using BDMA -at- twice the DMP-30 concentration I cannot= leave the resin in the processer overnight as it is to tacky the next day.
My Question ??? (at last some may say)
1. How are others out there, who are using BDMA at twice the DMP-30= concentration getting around the problem of the resin going very tacky= overnight, during infiltration?
What sort of infiltration times do you use?
2. Are any of you using BDMA at twice the DMP-30 concentration in an= automatic tissue processor?
3. Is any one else having problems with the resin if made up with BDMA at= twice the DMP-30 concentration?
Thank you in anticipation.
Allan Mitchell South Campus Electron Microscope Unit Department of Anatomy and Structural Biology Otago Medical School
Dennis Shubitowski {dennis-at-odin.morph.med.umich.edu}
Dennis,
If you omit the accelerator, it would probably work well. We use Epon-Araldite (Polyscience kit), and we routinely store the mixed resin (Epon + Araldite + DDSA), without catalyst (DMP-30), in a 4 degree C refrigerator. It will remain good for at least 6 months. When we are ready to embed, we measure out whatever volume we want, add DMP-30 to make 2%, mix, and it is fine. If you stored your nerve samples in 1:1 Epon (no accelerator):PO, and later decided to embed them, you could then embed them by standard procedures with catalyzed resin. If you have any questions about this, give me a call (763-1287), or drop over (2 blocks away).
Kent
A. Kent Christensen Department of Anatomy and Cell Biology University of Michigan Medical School Ann Arbor, MI 48109-0616 {akc-at-umich.edu}
----------------------------------------
On Thu, 2 Mar 1995, Dennis Shubitowski wrote:
} One of the divisions of our lab teases apart sural nerve tissue that have } been kept in a 1:1 mixture of epon:propylene oxide. The samples are very } easy to tease when the mixture is very fresh. The problem is that samples } became back-logged and were not able to be teased right away. A quick } decision had to be made, and the samples were frozen at -70C until they } could be teased. } } The nerve fibers need to be infiltrated to be teased but not polymerized } or else they become too fragile. A problem is becoming evident that after a } long period of storage, the nerve samples are being fully infiltrated } and polymerized despite being frozen (which we thought would almost stop } the polymerization process because it is largely dependant on heat). } Realizing that hind-sight is 20/20, would it have been better to leave the } accelerator out of the epoxy mixture? What affect would this have on the } quality of the resin? I understand accelerator to act as a catalyst being } a part of the polymerization reaction but not really consumed. Any } opinions to offer? } } Thanks, } } Dennis } } } } }
MICROSCOPY-at-AAEM.AMC.ANL.GOV Cc: Dennis Shubitowski {dennis-at-odin.morph.med.umich.edu} Message-id: {01HNOZPUD02Q93M845-at-gnv.ifas.ufl.edu} MIME-version: 1.0 X-Mailer: Windows Eudora Version 1.4.4 Content-type: text/plain; charset="us-ascii" Content-transfer-encoding: 7BIT
In reference to Dennis Subitowksi's question about expoxt accelerator, I can say that we routinely exclude the accelerator during infiltration with Epon, Epon/Araldite mixture and Epon substitutes and add it only to the second pure resin infiltration step. This allows long infiltrations in earleir steps with difficult specimens. When we have compared adding it to all steps, the results seem to be the same. Hope that helps
*****ORIGNINAL MESSAGE BELOW******** } One of the divisions of our lab teases apart sural nerve tissue that have } been kept in a 1:1 mixture of epon:propylene oxide. The samples are very } easy to tease when the mixture is very fresh. The problem is that samples } became back-logged and were not able to be teased right away. A quick } decision had to be made, and the samples were frozen at -70C until they } could be teased. } } The nerve fibers need to be infiltrated to be teased but not polymerized } or else they become too fragile. A problem is becoming evident that after a } long period of storage, the nerve samples are being fully infiltrated } and polymerized despite being frozen (which we thought would almost stop } the polymerization process because it is largely dependant on heat). } Realizing that hind-sight is 20/20, would it have been better to leave the } accelerator out of the epoxy mixture? What affect would this have on the } quality of the resin? I understand accelerator to act as a catalyst being } a part of the polymerization reaction but not really consumed. Any } opinions to offer? } } Thanks, } } Dennis } } } } } -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR EMCL 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu Gainesville, FL 32611
Due to renovations within the department, and the purchase of a confocal microscope, we have 2 transmission EMs in excess of our needs that we would like to move. Both microscopes are in good working condition, one has been under a service contract until this past year (though lightly used). The scopes:
1. Hitachi H-600 STEM (the one under service contract until recently)
2. Zeiss EM-9S TEM
We are not quite at a "No reasonable offer refused!" price level, but we would like to see these scopes get some more use and we're eager to make use of the space they are now occupying. So....if you are at all interested, contact me and we can discuss the particulars and terms of sale.
TIA,
Phil Rutledge voice: (410) 455-3582 Email: prutle1-at-gl.umbc.edu Fax: (410) 455-3875
Dennis Shubitowski {dennis-at-odin.morph.med.umich.edu}
We had a similar experience some years ago. Not with nerve, but with other material. I can offer the following humble comments.
1. As you learned, resin does polymerize in even at freezing temperatures. We have not tried -70 C and you did not specify how far below freezing you stored your specimens. We did not test exactly why, but reasoned that since the reaction is highly exothermic and since both resin and tissue are good insulators, it is probable that local areas are heated enough to allow polymerization. I look forward to any comments on the errors in our reasoning, since this was only a guess to explain the results. 2. Leaving out the accelerator does tremendously slow down the reaction, but polymerization still occurs in absence of accelerator, and even in the cold, so don't store for grossly extended times. To polymerize infiltrate tissue with fresh resin containing accelerator (we use 2 changes, 4 hours each) after thawing tissue. This works well, although some areas may have obvious interfaces between resins of different hardness.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
On Thu, 2 Mar 1995, Dennis Shubitowski wrote:
} One of the divisions of our lab teases apart sural nerve tissue that have } been kept in a 1:1 mixture of epon:propylene oxide. The samples are very } easy to tease when the mixture is very fresh. The problem is that samples } became back-logged and were not able to be teased right away. A quick } decision had to be made, and the samples were frozen at -70C until they } could be teased. } } The nerve fibers need to be infiltrated to be teased but not polymerized } or else they become too fragile. A problem is becoming evident that after a } long period of storage, the nerve samples are being fully infiltrated } and polymerized despite being frozen (which we thought would almost stop } the polymerization process because it is largely dependant on heat). } Realizing that hind-sight is 20/20, would it have been better to leave the } accelerator out of the epoxy mixture? What affect would this have on the } quality of the resin? I understand accelerator to act as a catalyst being } a part of the polymerization reaction but not really consumed. Any } opinions to offer? } } Thanks, } } Dennis } } } } }
Richard- Why not try infiltrating your samples overnight in 100% resin with the lower concentration of BDMA, then swithching to a "short" infiltration with fresh resin made with 2X concentration BDMA. then embedding in the higher concentration of BDMA in fresh resin? just a hunch. -Mike
I performed a similar set of experiments: BDMA vs DMP-30. Although it infiltrated specimens adequately and although they sectioned fine; dealing with changes of 100% plastic that were sooooo viscous was more than I could tolerate. In spite of it's initial higher viscosity and shorter shelf life, I switched back to using DMP-30 in our PolyBed 812 resin. Doing changes of 100% plastic is no longer a headache and something to be dreaded. If someone has a solution to the overnight BDMA-plastic blob problem, I'd like to hear as well.
Cheers...........JohnA
___________________________________________________ | | | John G. Aghajanian, Ph.D. | | Worcester Foundation for Experimental Biology | | 222 Maple Avenue | | Shrewsbury, MA 01545-2737 | | | | Tel: 508 842-8921 ext. 147, 161 | | Fax: 598 842-9632 | | JOHNA-at-sci.wfeb.edu | | | |_________________________________________________|
This message came to me and I have no good answers. Any comments will be appreciated and I will forwrad to the questioners = =Greg, = =Tom Mareci suggested that you might be able to give us some technical =advice on EM's to help us solve a temporary problem we're having. = =We're wanting to move our 4.7T magnet to a location in the ARB basement =120' from Dr. Craig Tisher's EM facility, which has a Zeiss EM 10 SEM =and a Topcon/International Scientific Instruments DS-130C TEM. The SEM =specs {= 3 mG AC and {= 5 mG DC. The TEM specs {=6 mG AC. Dr. Tisher =is reluctant to have us install our magnet, which with shileding is =calculated to produce a DC magnetic field of about 17 mG in the EM room. =Tom and I are having a hard time understanding the DC spec, in light of =the earth's magnetic field being 100x greater than the 5 mG spec. =We are also finding it difficult to get good scientifically sound =answers from the EM manufacturers (at least that we NMR people can =understand!). Could you enlighten us? = =Our main questions are: Will our DC field of 17 mG really pose a problem =for the EM's? If so, how can it be remedied? How much will it cost? =How are these systems currently shielded/compensated for the earth's =magnetic field? (My guess is that at most, a quick realignment of a =passive shield will be all that's required, if there's an observable =effect.) = =Secondarily, since before and after image quality may be the best bottom- =line test of the effect (if any) of our magnet, what standard samples can =you recommend for these Q.C. checks? = =Thanks for your advice and comments. = =-Richard Briggs = =tel: 395-0680 ext 54279 (55-4279) =FAX: 395-0279 =pager: Shands # 3497 =e-mail: rbriggs-at-ufnmr.health.ufl.edu = =P.S.: You may not remember, but I met you at Tom's Christmas party. = ****************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-846-0251 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * ******************************************************************
Hello All, I have a user that is attempting double-label fluroescence for protein localization in sheep ovaries. The ovaries seem to auto-fluoresce very strongly in the FITC channel with some signal in the Rhodamine as well. They have talked to other researchers who have not seen this problem. The tissue is fixed in Carnoy's and paraffin embedded. Has anyone seen or heard of this particular tissue lighting up like a Christmas Tree? TIA,
C. Michael Stanley, Ph.D. Coordinator, Associate Director Molecular Cytology Core Facility Molecular Biology Program 2 Tucker Hall University of Missouri-Columbia Columbia, MO. 65211 (314) 882-4895 fax= 314-882-0123
In response to my invitation for a free subscription to Materials Microscopy (posted on Feb. 24), we have been receiving a large number of requests and inquiries on a daily basis via e-mail and FAX. We will try to process all your subscription requests prior to the circulation of our next issue. Due to the high volume of mail we have received, I cannot respond to your inquiries individually. However, I have tried to summarize, itemize, and respond to your most frequently asked questions as follows: a. Materials Microscopy is published and circulated at no cost among the materials microscopy community members by Promotech Associates, Inc. b. Our publication is mainly concerned with providing articles and material of interest to working materials microscopist. We would appreciate your contribution which meets this requirement. Further information in this regard can be obtained by contacting our technical editor, Dr. Patricia Labun (a materials microscopist). c. I will send a copy of our "rate sheet" to those of you who expressed interest in placing ads or promotional material. Please feel free to contact me if you require any additional info. Thank you all for your interest in Materials Microscopy.
Rene E. Nicholas Circulation/Sales Manager
Materials Microscopy P.O. Box 2014 Scottsdale, AZ 85252
TEL (602) 947-7603 FAX (602) 947-7615 MatlsMicrs-at-aol.com
I'm wondering if the possible advantages of BDMA (lower viscosity, longer shelf life) are worth all this. I'm still using a 1989 Polysciences Epon-Araldite kit (with DMP-30) that still gives excellent results with standard procedures.
A. Kent Christensen Department of Anatomy and Cell Biology University of Michigan Medical School {akc-at-umich.edu}
Hi Barbara! I tried to no avail to send you the present message at your email address: it stubbornly bounced back, so I eventually post it to the microscopy list. I apologize to the members of this list if they should feel this is wasting bandwidth: just delete and go on to the next message ;-)
Barbara! I am afraid I did not make my question clear enough ;-) What I wanted to know was: 1) the testes you wish to fix are from what species ?
2) for what kind of microscopy are they to be processed ? I.e. T.E.M., S.E.M., or light microscopy ?
3) What's the embedding medium?
4) in the case of light microscopy, do you contemplate using some special technique (besides for recognizing the different stages of the seminiferous epithelium) which would require that some specific chemical would be either required or, on the contrary, avoided?
In my personal experience with rat testes and light microscopy (this is rather very long ago: more than 30 years! :-( ), Bouin and other picric acid containing variations thereof are not optimal fixatives for preservation of the acrosomial apparatus of spermatids, which is one of the features used by most classification methods (Clermont's among others...). The best fixatives for light microscopy of paraffin embedded tissue are those containing potassium dichromate such as, for example, Helly's fluid (Zenker + formaldehyde), but they also are followed, after rinsing and dehydration, by shrinkage which is still more important than that observed after Bouin's or Bouin-Hollande's (somewhat better) fluids.
BTW, what's wrong with picric acid? It is always shipped in 'moist condition' in order to minimize hazards. I work in our lab since 1953, with Bouin, Bouin-Allen, Bouin-Hollande, Dubosq-Brazil etc., every day, and we never experienced any problem. We store the picric acid for our needs in the form of a saturated aqueous solution which keeps for as long as you will. This stock solution is then used for preparing all picric acid containing fixatives.
There is no point in trying to eliminate the picric acid from the fixed pieces of tissue before sectioning. If the presence of picric acid in the slides should hamper subsequent staining (a rare occurrence, btw), then the easiest and expeditious way to get rid of it is to pretend your tissue was fixed with a sublimate containing fluid: at the rehydration step, your slides should undergo treatment by alcoholic iodine (or lugol solution) and sodium thiosulfate in the usual way and 'voila': no more picric acid on the slides within 2 minutes! Under no circumstances should the pieces of tissue fixed with a picric acid containing fixative undergo rinsing in water! This is sheer heresy ;-) because it induces a lot of swelling in the tissue before shrinking even more during dehydration (I never found out which histologist ever advocated such method, but I know quite a number of renowned ones who firmly condemned it, and with good reason). If you insist on rinsing the pieces of testes after fixation, then merely increasing the number of steps through 90pc x alcohol should do the trick; you may even add a thin layer (1-2 mm thick) of lithium carbonate on the bottom of the alcohol containing flasks: this helps dissolving part of the picric acid away.
Possibly the method which should yield the best pictures has been documented in a book entitled 'Histological and histopathological Evaluation of the Testis', by Russell,Ettlin, Sinha Hikim and Clegg: Cache River Press 1990, (15777 Bolesta, Box 129, Clearwater, Fl 34620): ISBN 09627422-0-1.
At the end of the book, there is an appendix about the materials and methods which you might find useful (although perfusion fixation of rat testes is among the most frustrating experiences I ever had, even with a lot of heparin ). This would also require embedding in epoxy resin and making large semi-thin sections, which I don't know whether you are prepared to do ;). Glutaraldehyde certainly seems to me the best choice, but then paraffin embedding would be ruled out (too hard and too brittle after paraffin embedding). HTH, and good luck. Let me know about the outcome (good, I hope) John
*********************************************************** * Jean C. Desclin (John), Associate Prof. of Histology * * Laboratory of Histology - Faculty of Medicine * * Brussels Free University (U.L.B.) * * e-mail: desclinj-at-ulb.ac.be (internet) * * snail mail: route de Lennik 808 * * B - 1070 Brussels Belgium * ***********************************************************
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Cressington makes an excellent freeze fracture machine. Suitable for routine work and high resolution shadowing at very low temperatures. In particular the reproducibility, even of W/Ta, is very good, and attainable for everyone (user friendly).
The address is: Dr Peter A Walley CRESSINGTON Scientific Instruments Ltd 24 Chalk Hill Watford Herts WD1 4BX UK Tel 0923 220499 Fax 0923 816646
Success
Marcel Paques Unilever Research Laboratory Vlaardingen The Netherlands E-mail: Marcel.Paques-at-2488PAS.URLNL.Sprint.Com
We use a Cressington CFE- 50B which has proved excellent for reproducible rotary low-angle shadowing of peptides. The system is also very flexible for both W/Ta and Pt/C operation and is easily re-configured for a variety of research applications.
Kevin Jennings SmithKline Beecham Pharmaceuticals UK Microscopy & Flow Cytometry Section The Frythe Welwyn Hertfordshire AL6 9AR United kingdom
I am in the info gathering stage of purschasing a tissue processor for EM samples. I have info on the Reichert-Lynx unit and the one made by RMC. How do you like these units? Comments on advantages and disadvantages are welcome. I also have brouchers on a unit made by Sakura Finetek USA, Inc.. The phone number I called is no longer in working order.(213-539-5441) Is this company still around? These brouchers are probably from the early 80's. Are the any other companys that made tissue processors?
Thanks in advance
Ed Calomeni Medical College of Ohio Toledo, OH
emlab-at-opus.mco.edu
PS Sorry about the post on last friday with no message in the body. Computers can be such good friends.
I recently read W. L. Steffens comments regarding the use of Leighton tubes for embedding cell cultures. Has anyone embedded the coverslips from these tubes in LRWhite resin? I've been having trouble getting consistent polymerization of monolayers on other coverslips (eg Thermanox) and am in search of a new method for obtaining "en face" sections of monolayers for immunocytochemistry. TIA for any hints on the topic.
Christine Powers Dept. of Cell Biology University of Massachusetts Medical Center Worcester, MA 01655
Ed, I can comment on the Lynx tissue processor which we have been using since it first came out, about 8 years ago. The Lynx is made in Australia, and was originally distributed in this country by Fairleigh- Dickinson Laboratories until the distributorship was snatched up by Reichert (now Leica). We paid about 5.5K for the unit and have had little trouble with it aside from a blown power supply (they have upgraded this) and a defective exhaust fan (we fixed this). We use it routinely for processing TEM (epon and L.R. White) and SEM (through dehydration). It works very well is quite consistent. My only complaint involves the expendables for it...vials, caps, baskets, etc. Originally, it was economical to use them once and dispose of them as is intended. Once Leica became the distributor, the price of the expendable essentially tripled...some things quadrupled. As a result, we clean and recycle these components for as long as we can. Additionally, the quality control of these items has suffered immensely. There are now occasional vials that are distorted and won't fit the turntable, and lids and baskets with burrs that must be filed. If you can accept this, I still think that for the money (nearly 9K by now?) its the best one out there. Good Luck.
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
We have a JEOL 100CX scanning transmission electron microscope that we no= longer need because of more recent acquisitions. It was maintained continuously= during operation here and was in perfect working condition before we turned it o= ff in =
July, 1994. =
This is a conventional scanning transmission electron microscope with =
resolutions of 3.4 =81 TEM.
It has specimen holders for heating-cooling, double tilt (tilt range is =B1= 45=A1), =
or rotation. =
We would be deligted to entertain offers for it.
Please contact Beth Trend if you are interested.
______________________________________________***************************= ******* Beth Trend trend-at-cems.umn.edu or btrend-at-maroon.tc.= 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=
In July and September of 1994, there was some disciussion of BDMA vs. DMP-30. I saved and consolidated 13 of those messages into a single document. If anyone is interested in getting a copy, send me your e-mail address and I will zip one out to you within a day or two.
--
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
Folks, I'm aware of several programs for the Mac that simulate diffraction patterns and stereographic projections, etc. How about similar programs for the PC? Thanks in advance.
Well it may not sopund exciting to microscopists until you note that it tranlates into Noran's parent company buys Kevex's parent company! March 3rd issue of Wall Street Journal. Thermo Instrument Systems to acquire Fisons PLC's scientific instrument division. Cost $320million! Anyone know whether this covers VG too?
Just thought you guys might like to know.
-- John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu, John.F.Mansfield-at-umich.edu or jfmjfm-at-umich.edu they all reach me! URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
Hi, I know this is a little of the subject, but can anyone suggest a ftp site that has terminal emulators. We need to emulate a tektronics 4205 terminal to use our new xrd software. Any suggestions would be appreciated.
I am looking for the following pieces of used equipment either in good working order, fair condition and/or in need of minor repair:
1) glass knife breaker for LM microtomy 2) embedding oven (35-80C) 3) balance (top loading or mechanical)
Contact:
Fred A. Hayes 916-752-7712 work University of California,Davis 916-752-4701 work School of Medicine Department ofMedical Pathology; EM Lab MSIA E-mail: Davis, CA 95616 fahayes-at-ucdavis.edu
1320 Dogwood Court 916-678-6280 home Dixon, CA 95620-3227
Hi, I know this is a little off the subject, but can anyone suggest a ftp site that has terminal emulators. We need to emulate a tektronics 4205 terminal to use our new xrd software. Any suggestions would be appreciated.
Microscopy Mail {Microscopy-at-aaem.amc.anl.gov} Message-id: {01HNT9F975CY93NYWI-at-gnv.ifas.ufl.edu} MIME-version: 1.0 X-Mailer: Windows Eudora Version 1.4.4 Content-type: text/plain; charset="us-ascii" Content-transfer-encoding: 7BIT
At 08:26 AM 3/3/95 +0100, NICK SCHRYVERS wrote: } REGARDING obj. lens 100C replacement } } The objective lens and the upper and lower column parts of our 20 year old } side entry JEOL 100 C microscope need replacement. New parts, however, are } rather expensive in view of the age of the instrument so we're looking for } anyone who has some second hand spare parts for sale. Sincerely, } Nick Schryvers tel: 32-3-2180247 fax: 32-3-2180257 } EMAT, University of Antwerp } Groenenborgerlaan 171 } B-2020 ANTWERP } Belgium ********************************************************** There is a JEOL 100 C that may be headed for the scrap heap. They might be willing to give you parts. Contact Dr. Henry Aldrich, P.O. Box 110700, University of Florida, Gainesville, FL 32611 USA FAX 904-392-5922 (no e-mail) or his department chairman, Edward Hoffmann, at the same address and FAX number who gets E-mail at EMH-at-GNV.IFAS.UFL.EDU
Good Luck -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR EMCL 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu Gainesville, FL 32611
please change my address to yi_huang-at-qmgate.anl.gov. I'd like to continue the subscription at the new address. Thank you. Yi Huang Argonne National Lab building 212/c221 building 212/c221
All instruments divisions of fisons are going to Thermo Instrument Systems including VG.
Just what we all needed. I sure service and parts are going to be even easier to get.
Mike
--------------------------------------------------------------------------- | Michael Dunlap | lab (916) 752-0284 | | Facility For Advanced Instrumentation | fax (510) 422-2282 | | University of California | mrdunlap-at-ucdavis.edu | | Davis CA, 95616 | | ===========================================================================
We have successfully used LR White resin for embedment of monolayers grown in Leighton tubes. The problems encountered were the same ones you encounter when using LR White in open molds or silicone rubber molds, or just about anything other than gelatin capsules. It's most convenient to embed coverslips in flat, open molds such as the silicone rubber type. We had such problems with this (ie excluding oxygen, penetration of the molds by the resin, etc) that we began embedding the coverslips standing upright in gelatin capsules. The long narrow coverslips of the Leighton tubes are just accommodated by a 00 gelatin capsule. This neccessitates considerably more hand trimming to get to the monolayer profile, but embedding problems are eliminated. I might also add, that once the block face is prepared for cutting, the exposed coverslip may be peeled away from the embedded cells, greatly facilitating cutting. If you do this, I would also recommend mounting the sections on a support filmed grid, as the cells will be on the edge of the section. Good Luck!
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
X-Mailer: WinNET Mail, v2.30 Message-ID: {463-at-oimag.win-uk.net} Reply-To: Software department {software-at-oimag.win-uk.net} To: microscopy-at-aaem.amc.anl.gov
The following advertisement has recently been placed in the press and may be of interest to those who read this Newsgroup who would like to work in the UK:
SOFTWARE DEVELOPMENT FOR MICROANALYSIS The Software team at Oxford Instruments Microanalysis Group develop high quality instrumentation products to satisfy customers in an international marketplace. Currently we have a number of vacancies for individuals who would enjoy the challenge of solving complex problems to generate leading edge products. If you are competent in Visual Basic, C++ or C, have a basic understanding of electronics and computer interfacing and feel you could make a strong contribution in any or all of the following areas please contact us:
Practical applications experience using an SEM or microprobe. Knowledge of x-ray microanalysis and familiarity with operation of WD spectrometers. Experience in project management and identifying the "voice of the customer". Practical use of applied mathematics, numerical methods, chemometrics. Experience with real time control and hardware automation.
Successful applicants will receive an attractive benefits package and salary commensurate with capability.
Please send a copy of your CV, and a telephone contact number to
Peter Statham , Oxford Instruments Microanalysis Group Halifax Road, High Wycombe, Bucks HP12 3SE England Telephone (01494) 442255 Fax (01494) 461033 email: software-at-oimag.win-uk.net
----------------------------------------------------------------- Please reply to this e-mail with the name of the person you wish to recieve it in the subject (i.e. FAO John Smith), as this is a shared e-mail address.
The following is a copy of the job posting that recently appeared on the University of Michigan Gopher Server UM GopherBlue.
The full URL is: gopher://gopher.itd.umich.edu:8888/00/acadaff/hris/gopher/Job%20Postings/Pro fessional%5CAdministrative/-at-39-at-RES%20ASSOC%20II%20ENGR%20%28Materials%20Scie nce%20%26%20Engineering%29
Job Title: RES ASSOC II ENGR Grade: 09 Min/Max $ 25,500/ 64,500 Posting Number: T-95-0826-LM Materials Science & Engineering Open Date: 03/06/1995 Close Date: 03/10/1995 Jobclass: 12871 Hours: 40.00
DUTIES: Operate and train users in the use of the lab instruments: a JEOL 200FX Analytical Electron Microscope, a JOEL 4000EX High Resolution electron Microscope, a Philips EM420 Transmission Electron Microscope, an ElectroScan E3 Environmental Scanning Electron Microscope, a Digital Instruments Scanning Force Microscope, a PHI 5400 X-ray Photoelectron Spectrometer and a PHI 600 Scanning Auger Microprobe; the successful candidate will not necessarily need to be proficient in the use of all instruments specified, but familiarity with at least four is essential; other duties include: aiding users with reduction and analysis of data recorded on lab instruments; aiding users with sample preparation equipment and preparing samples; help other lab staff maintain lab equipment; maintain lab darkroom and supplies; take part in collaborative projects with other members of the University; develop personal research projects as time permits.
DESIRED QUALIFICATIONS: Knowledge of Analytical Electron Microscopy, Scanning Transmission Electron Microscopy and High Resolution Electron Microscopy; TEM expertise should include detailed understanding of diffraction contract, defect imaging, selected area diffraction, high resolution imaging, high resolution image simulation and analysis; experience with computer programming in FORTRAN, C or Pascal.
MINIMUM QUALIFICATIONS: Master's degree or equivalent in a scientific related field; demonstrated prior related work experience; familiarity with computer systems such as Apple Macintoshes, PCs and/or UNIX workstations; working knowledge of general transmission electron microscopy and scanning electron microscopy; experience with standard data analysis for analytical techniques such as X-ray energy dispersive spectroscopy (XEDS), electron energy loss spectroscopy (EELS) and micro diffraction; good interpersonal skills.
Present Applications For This Position To The Following Office:
Ann Arbor Campus Employment Services Office Room G250 Wolverine Tower 3003 South State Street Ann Arbor, Michigan 48l09
(313) 764-6580
8 AM to 5 PM, Monday - Friday
John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu, John.F.Mansfield-at-umich.edu or jfmjfm-at-umich.edu they all reach me! URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
Hello Barbara! I am afraid that, under the conditions you describe for fixation, significant shrinkage should be unavoidable whatever fixative would be used. I already mentioned that picric acid 'per se' in the fixing fluid does not ensure adequate preservation of acrosomial structure which is required for easy classification of spermatids (and thus of stages of the seminiferous epithelium.) What should be avoided in any case is the presence of acids usually part of most recipes for fixatives, such as acetic acid and the like. If you use '10 percent formalin', you may experience some problems because you in fact never know the actual composition of such solution: it polymerizes on the shelf and becomes also quite acidic! You should prepare a solution of phosphate buffered (pH 7.2) 4 percent formaldehyde from paraformaldehyde powder on the same day (or the previous one) that you are sacrificing the animals (you should find recipes for making such solution in any practical handbook for microscopical technique ;-)).
Fixation through immersion of the testes in the fixative may be almost acceptable for mice testes because they are small; this won't be the case for other species, even for the rat. You may try to make small (with caution!) incisions in the albuginea with a very sharp razor blade when the testes have stayed for at least an hour in the fixative, in order to improve penetration of the formaldehyde. Another trick to improve diffusion and fixation is to add from 0.5 to 1.0 percent (in volume) of DMSO (dimethylsulfoxide) to the fixing fluid (best is to combine both...)
BUT,LAST and PROBABLY _MOST_ IMPORTANT: you should realize that formaldehyde fixation, i.e. crosslinking, proceeds exceedingly slowly! Therefore, tissues should stay in the fixative at least two weeks (one year or more won't harm!) before being further processed (two weeks probably is a conservative estimate). Assessment of pathological tissues fixed for only a few hours in NBF may frequently be required for practical reasons, but don't expect such methods to result in nice preservation of testis tissue, especially if you need estimating quality of spermatogenesis!
You also may try to enhance fixation - and shorten the duration required for fixation - by performing it in the microwave oven: several cycles of no more than one minute at a time (perhaps 10 times, tissue temperatures exceeding 50 degrees celsius should be avoided), if you are indeed in a hurry and can't wait several weeks for fixation to proceed at its usual pace. If you try this, don't forget to place a water load (a becher container with about 750-1000 ml of water) besides your fixative containing flask in the oven.
That's all what I can think of. HTH Good luck John
*********************************************************** * Jean C. Desclin (John), Associate Prof. of Histology * * Laboratory of Histology - Faculty of Medicine * * Brussels Free University (U.L.B.) * * e-mail: desclinj-at-ulb.ac.be (internet) * * snail mail: route de Lennik 808 * * B - 1070 Brussels Belgium * ***********************************************************
I was unable to reach Nick Schryvers at his E-mail address therefore my reply is below. Perhaps others will be interested:
Nick writes: } } } The objective lens and the upper and lower column parts of our 20 year old } } } side entry JEOL 100 C microscope need replacement. New parts, however, are } } } rather expensive in view of the age of the instrument so we're looking for } } } anyone who has some second hand spare parts for sale. Sincerely, } } } Nick Schryvers tel: 32-3-2180247 fax: 32-3-2180257 } } } EMAT, University of Antwerp } } } Groenenborgerlaan 171 } } } B-2020 ANTWERP } } } Belgium } } **********************************************************
} } My reply:
There is a JEOL 100 C that may be headed for the scrap heap. They } } might be willing to give you parts. Contact Dr. Henry Aldrich, P.O. Box } } 110700, University of Florida, Gainesville, FL 32611 USA FAX 904-392-5922 } } (no e-mail) or his department chairman, Edward Hoffmann, at the same } } address and FAX number who gets E-mail at EMH-at-GNV.IFAS.UFL.EDU } } } } } } Good Luck } } -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- } } Greg Erdos Phone: 904-392-1295 } } Scientific Director, ICBR EMCL } } 218 Carr Hall Fax 904-846-0251 } } University of Florida E-mail: gwerdos-at-gnv,ifas.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- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR EMCL 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu Gainesville, FL 32611
I concur with Dr. Desclin's suggestions- however, I would add that pH and osmolarity of your solutions is easy to do, and can be helpful. Perfusion fixation of the testis is also a possibility. Those are my humble additions-
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
On Tue, 7 Mar 1995, Desclin Jean wrote:
} } } Hello Barbara! } I am afraid that, under the conditions you describe for fixation, } significant shrinkage should be unavoidable whatever fixative } would be used. } I already mentioned that picric acid 'per se' in the fixing fluid } does not ensure adequate preservation of acrosomial structure which } is required for easy classification of spermatids (and thus of } stages of the seminiferous epithelium.) } What should be avoided in any case is the presence of acids usually } part of most recipes for fixatives, such as acetic acid and the like. } If you use '10 percent formalin', you may experience some problems } because you in fact never know the actual composition of such } solution: it polymerizes on the shelf and becomes also quite acidic! } You should prepare a solution of phosphate buffered (pH 7.2) 4 percent } formaldehyde from paraformaldehyde powder on the same day (or the } previous one) that you are sacrificing the animals (you should find } recipes for making such solution in any practical handbook for } microscopical technique ;-)). } } Fixation through immersion of the testes in the fixative may be } almost acceptable for mice testes because they are small; this won't } be the case for other species, even for the rat. You may try to make } small (with caution!) incisions in the albuginea with a very sharp } razor blade when the testes have stayed for at least an hour in the } fixative, in order to improve penetration of the formaldehyde. } Another trick to improve diffusion and fixation is to add from 0.5 to } 1.0 percent (in volume) of DMSO (dimethylsulfoxide) to the fixing } fluid (best is to combine both...) } } BUT,LAST and PROBABLY _MOST_ IMPORTANT: you should realize that } formaldehyde fixation, i.e. crosslinking, proceeds exceedingly } slowly! Therefore, tissues should stay in the fixative at least } two weeks (one year or more won't harm!) before being further } processed (two weeks probably is a conservative estimate). Assessment } of pathological tissues fixed for only a few hours in NBF may } frequently be required for practical reasons, but don't expect such } methods to result in nice preservation of testis tissue, especially } if you need estimating quality of spermatogenesis! } } You also may try to enhance fixation - and shorten the duration } required for fixation - by performing it in the microwave oven: several } cycles of no more than one minute at a time (perhaps 10 times, tissue } temperatures exceeding 50 degrees celsius should be avoided), if } you are indeed in a hurry and can't wait several weeks for } fixation to proceed at its usual pace. If you try this, don't forget } to place a water load (a becher container with about 750-1000 ml } of water) besides your fixative containing flask in the oven. } } That's all what I can think of. HTH } Good luck } John } } } *********************************************************** } * Jean C. Desclin (John), Associate Prof. of Histology * } * Laboratory of Histology - Faculty of Medicine * } * Brussels Free University (U.L.B.) * } * e-mail: desclinj-at-ulb.ac.be (internet) * } * snail mail: route de Lennik 808 * } * B - 1070 Brussels Belgium * } *********************************************************** }
} One of the divisions of our lab teases apart sural nerve tissue that have } been kept in a 1:1 mixture of epon:propylene oxide. The samples are very } easy to tease when the mixture is very fresh. The problem is that samples } became back-logged and were not able to be teased right away. A quick } decision had to be made, and the samples were frozen at -70C until they } could be teased. } } The nerve fibers need to be infiltrated to be teased but not polymerized } or else they become too fragile. A problem is becoming evident that after a } long period of storage, the nerve samples are being fully infiltrated } and polymerized despite being frozen (which we thought would almost stop } the polymerization process because it is largely dependant on heat). } Realizing that hind-sight is 20/20, would it have been better to leave the } accelerator out of the epoxy mixture? What affect would this have on the } quality of the resin? I understand accelerator to act as a catalyst being } a part of the polymerization reaction but not really consumed. Any } opinions to offer? } } Thanks, } } Dennis } Why do you store the fibers in resin? Isn't teasing them very messy? I teased sural nerve fibers when working for a neuropathologist. I fixed them and stored in buffer. Also I sometimes used collagenase, which made the teasing easier. } } } }
Message-Id: {MAILQUEUE-101.950307092030.384-at-vanlab.paprican.ca} To: "Griffin, Robin" {rgriffin-at-eng.uab.edu} , microscopy-at-aaem.amc.anl.gov
Hello, I have information on an "Imagist" image analysis system from Princeton Gamma Tech (PGT) which seems to be finding quite a bit of popularity. I have come across it several times in speaking to users during my hunt for an EDX system. Imagist runs on a sparc station. PGT in New Jersey, USA , can be reached at (609) 924-7310.
I have not used this system I have only seen it in use at the university here. Goodluck, Laurie
On 1 Mar, 1995 Robin Griffin wrote: } } I'm interested in finding out what the most commonly used image analysis } systems for light microscopy, materials applications. I am aware of } home-built, Beuler, Leco, and Kontron/Ibas. What else is out there and what } is used most in both University and industrial settings? } } Thanks for your help. } }
______________________________________________________________________ Laurie Frederick, A.SC.T. PAPRICAN Corrosion Control Group 3800 Wesbrook Mall The Pulp and Paper Research Vancouver, B.C. Institute of Canada Canada V6S 2L9
We are considering purchasing a scanner for TEM negatives.
1. Is this a good idea?
2. Is there a consensus on which one to buy?
Any and all input appreciated. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR EMCL 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu Gainesville, FL 32611
A regular full time position is open in The Jackson Laboratory Biological Imaging Department - Histology Laboratory. The Jackson Laboratory is a non-profit independent laboratory founded in 1929 on the premise that the causes of cancer and other diseases could be discovered through Mammalian genetic research. The Laboratory specializes in mammalian genetics using inbred laboratory mice as model systems to study health problems such as cancer diabetes, anemia, heart disease and aging. Located on a large island in the gulf of Maine and surrounded by Acadia National Park, The Jackson Laboratory is currently undergoing a major expansion of its scientific staff and its research facilities.
Applicants with a Bachelors degree and two years related laboratory experience working with Murine specimens preferred. The position includes routine histological techniques ie paraffin embedding, single and serial sectioning and cryotomy in conjunction with Immunohistochemistry techniques, staining using heavy metals as well as other special stains.
The successful candidate must be a self starter, pay attention to detail and be able to work independently with little supervision. This individual will be responsible for providing services in support of numerous diverse research projects, must interact well with multiple users and work productively in a team environment. The position includes opportunities for advancement.
Salary range is mid to high $20,000 plus benefits and is negotiable depending on level of experience.
Interested applicants should send CV to:
Joanne Bradt Employment Specialist The Jackson Laboratory 600 Main Street Bar Harbor Maine 04609 (207) 288-3371 ext. 1281 (207) 288-3371 ext. 1082 FAX jcb-at-aretha.jax.org
Dear Greg, There are at least two types of scanner to consider depending on your application. For scanning images of the usual type with many gray levels, a CCD array will give excellent results and is very fast; however, for quantita- ting ED patterns, nothing beats a scanner with a small illuminating spot. The problem with a CCD array for ED is that the highest intensities are in areas with very low transmission surrounded by areas with high transmission, so in- ternal reflections in the CCD lens, etc. will give erronious values for the intensities. Any of the high-resolution CCD arrays will work for images, since the changes in contrast are not so abrupt. Perkin-Elmer and Optronics are two names for small-illuminating-spot scanners, and doubtless there are others. We had an Eikonix (linear CCD array) at one time, but were not happy with it. Good luck. Yours, Bill Tivol
Here's a little information that I just received from polaroid that should be of interest to the microscopy community. I'd be interested in hearing any comments anyone (particularly any Polaroid reps.) might care to share with the BBS.
Recently we have been noticing that the type 55 film we are purchasing was arriving with a relatively short period before expiration. We were questioning if our supplier might have trying to unload older film on us. So I just called up Polaroid an asked them what their lead time from manufacture to expiration date was, and I was informed that for type 55 and type 665 (B&W P/N film) the expiration lead time is only 9 months.
If you allow say 2-5 months from manufacture to shipping to distributors to shipping to users (which seems very reasonable for such commercial retail) this leaves only 4-7 months before expiration! This short of an expiration date does not lend itself at all to buying in larger quantities in order to obtain any sort of a price break.
I do not know how much extension can be obtained by storage at say 4 C (May be someone out there does know). But we seem to start having problems with film only a couple of months after the expiration date even after cold storage for a few months.
You might want to consider this short time to expiration before you go and stock up on polaroid film.
Richard E. Edelmann Electron Microscopy Facility Supervisor Miami University, Oxford, OH 45056 Ph: 513-529-5712 E-mail: edelmare-at-muohio.edu
Although the journal Microscopy Research and Technique has a large editorial board, we cannot cover every research area that involves microscopy. Therefore, I would like to invite those of you who are interested in serving as ad hoc referees for articles submitted to the journal to send, by e-mail, your name, mailing (regular mail) address, phone number, fax number, and research interests using key words such as biology, materials science, pathology, microanalysis, stereology, magnetic domain, ceramics, immunocytochemistry, kidney, etc.
John E. Johnson, Jr., Ph.D. Editor-in-Chief, Microscopy Research and Technique
We are considering purchasing a scanner for TEM negatives.
1. Is this a good idea?
2. Is there a consensus on which one to buy?
Any and all input appreciated. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR EMCL 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu Gainesville, FL 32611
Ed, We have had 3 different tissue processors in the past 6 years, 3 LKB's, one Lynx, and 3 RMC's model 4189. LKB no longer manufactures a system so I will forego 'ripping' their machine. We purchased a Lynx system about four years ago. We found the same problem that W.L. Steffens had with the QC of the expendibles. We lost tissue due to improper sealing of the specimen chambers. Also the side 'openings' are relatively large and we therefore do not run tissue smaller than 0.5 mm3 in these holders. Extreme care must be used when removing tissue from the holders because tissue has a tendency to adhere to the lid of the holder making it possible to mix tissues when using the multiwell chambers. The unit itself though has been extremely reliable. About 10 months ago we purchased 3 RMC processors. These have proven not to be as reliable as the Lynx (we have had temperature and some programming problems). We therefore have had the opportunity to test RMC's service department which to date has been able to get us up and running with minimal down time. The big advantage of the RMC units are the specimen chambers which seal better and hold smaller pieces of tissue. Currently we use only the RMC units for tissue processing (approximately 3000 samples annually). The Lynx system has been modified and is used 3-4 days (and evenings) a week to immunolabel grids for our IEM program. Hope this helps.
{jon charlesworth} Electron Microscopy Facility Mayo Clinic
We are considering purchasing a scanner for TEM negatives.
1. Is this a good idea?
2. Is there a consensus on which one to buy?
Any and all input appreciated. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR EMCL 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu Gainesville, FL 32611
} noticing that the type 55 film ... with a relatively short period before } expiration } } But we seem to start } having problems with film only a couple of months after the } expiration date even after cold storage for a few months.
Please forgive the above condenstation :-}
For the last 15 years we have stored Polaroid type 55 film in a freezer without harm. This appears to negate the expiration date. We usually allow the film to thaw out completely before opening the sealed package, but sometime this process is rushed a bit and then, more often than not we have problems.
We have found that to be the situation for years, but we order it by the case and refrigerate it (not freezing). It seems to be good for at least a year past expiration and we have had some film that was several years old still work. I suspect that there could be some degradation, but for our routine SEM and light microscopy, it hasn't been a problem. Maybe Polaroid has done some testing?
***************************************************************** Michael L. Boucher Sr. Boucher-at-TCRCA.USBM.GOV Geology-Mineralogy/Chemistry Labs Ph 612-725-4614 Twin Cities Research Center Fax 612-725-4527 U.S. Bureau of Mines Center 725-4500 Department of Interior 5629 Minnehaha Avenue South Minneapolis, MN 55417-3099 U.S.A. *****************************************************************
I am going to be attempting to thin section sediment samples containing quartz and want the thin sections to be approx. 50-70nm in thickness for TEM analysis. I have to buy a diamond knife and would much appreciate input on the blade angle to choose. A representative at Polysciences told me I should go to a 55 degree knife instead of a 45, but I was wondering if I would be sacrificing much of the ability to cut sections down to 50nm. The quartz grains are approx. 200 um in diameter. Is there a particular manufacturer I should get the knife from? What is an appropriate cutting speed? Can I expect the knife to last only a VERY short time? I would appreciate any suggestions and advice. My address is: chswartz-at-MIT.edu
Does anyone on the listserver know where (prefereably in the U.S.) that I can get Carbon Putty. I'm down to the last of what I had that someone picked up at a conference in Germany a few years ago. It is basically a mixture of some kind of soft wax and carbon and is a very good specimen mounting material since there is no drying or out-gassing. Ted Pella doesn't seem to have it so maybe someone out there knows about this product and can save me a hunt.
} I do not know how much extension can be obtained by storage at say } 4 C (May be someone out there does know). But we seem to start } having problems with film only a couple of months after the } expiration date even after cold storage for a few months. } } Richard E. Edelmann
We are just finishing our last few boxes of September 1994 Type 55 film (stored in a refrigerator). It still seems to be OK.
Arthur Day, Electron Microscopy Group Ansto Advanced Materials Program Phone: 61-2-717-3457 PMB 1, Menai (Sydney), NSW, 2234 Fax: 61-2-543-7179 Australia Email: ard-at-atom.ansto.gov.au
------------------- tn8502.asc follows -------------------- Tracor 8502 users: Help with image convert to tiff
We operate a Tracor Northern (Noran) 8502 Image Analyzer and have been trying to export images to a VAX via KERMIT with no success. Has anyone done this sort of image file conversion and transfer successfully?
Background: The Tracor software can convert an image file from TN's format (.IMG) to a .TIF file (type 4, version 42). We can successfully transfer this TIFF image file, via KERMIT, to or VAX (ie. the file appears and has the appropriate size). When we try to open this TIFF image (using a PC on our LAN), we always fail regardless of the software we have employed (e.g. with Photoshop, HiJack Pro).
If you have any ideas, please contact. I have lots of additional contextual information to add to this. Thank you!
Eric Kokko Electron Microscopy and Image Analysis Agriculture and Agri-Food Canada Lethbridge Research Centre P.O. Box 3000, Lethbridge, Alberta CANADA T1J 4B1
Phone 403-327-4591 (Voice 367) FAX 403-382-3156 INTERNET kokko-at-em.agr.ca
Someone has just given me all you could ever want to know about this product. It is available from: Bal-Tec Products, Inc. 984 Southford Road P.O. Box 1221 Middlebury, CT 06762; (800) 875-3713, FAX (203) 598-3658
It is called "Leit-C-Plast; Cat. # B 801014075 Current price is $70.00 for 15g
Other properties: -high electric conductivity -long-life plasticity -high vacuum resistance -sufficient adhesive power -low sample contamination -no disturbing ED-x-ray lines -Resistance R~100 kOhm mm2/m Thanks for saving me the hunt---Jerry
"Mung II" is not carbon putty, but is very much like it: electrically conducting, a good thermal conductor, and very low vapor pressure. We use Mung in our ion mill, but in the SEMs most people prefer clips or carbon tape. You can buy mung from
Comonwealth Scientific Corp. 500 Pendleton St. Alexandria, VA 22314 703-548-0800
$160 for 5 gr.
_______________________________________________________________ Michael Rooks 607-255-2329 voice National Nanofabrication Facility 607-255-8601 fax at Cornell University rooks-at-nnf.cornell.edu Knight Laboratory Ithaca, NY 14853 USA
_______________________________________________________________ Michael Rooks 607-255-2329 voice National Nanofabrication Facility 607-255-8601 fax at Cornell University rooks-at-nnf.cornell.edu Knight Laboratory Ithaca, NY 14853 USA
You are asking about a product called "Leit-C-Plast", which is found on page 46 of the current SPI Supplies "SourceBook" as SPI #5057-AB. It is a highly conductive adhesive plastic and is designed for mounting large specimengo a very long way. I hope this information will be useful to you.
Charles A. Garber, Ph. D. PRESIDENT SPI SUPPLIES PO BOX 656 West Chester, PA 19380
"Richard E. Edelmann" {EDELMARE-at-CASMAIL.MUOHIO.EDU}
Richard Edelmann wrote " I'd be } interested in hearing any comments anyone (particularly any Polaroid } reps.) might care to share with the BBS. } } Recently we have been noticing that the type 55 film we are } purchasing was arriving with a relatively short period before } expiration. We were questioning if our supplier might have trying to } unload older film on us. So I just called up Polaroid an asked them } what their lead time from manufacture to expiration date was, and I } was informed that for type 55 and type 665 (B&W P/N film) the } expiration lead time is only 9 months. " } I have used Polaroid 665,667,55 for nearly 20 years with no problems of any continuing significance (except maybe cost !!). We have always purchased in bulk (large boxes of 50 boxes) and stored the material in a standard refrigerator, not freezing it. Usual practice would be to allow the boxes to equilibrate at room temperature before use but sometimes usage rate prevented this. The sensitivity does appear to change with temperature, but we have nver bothereded to investigate in detail. In the usual situation, we have not expereinced any problems and have both negs and prints which are old and still very good quality. Cleanliness of the holder and specially the rollers is critical to performance and they are cleaned after EVERY cartridge is exhausted and before the next one is in place. cheers jjm Professor John J. Millar Head, Department of Applied Physics Electron Microscope Unit RMIT, GPO Box 2476V, MELBOURNE AUSTRALIA 3001 +613 660 2602 fax 613 660 3837 email jjmill-at-rmit.edu.au
RE: Ultrathin sectioning material containing quartz
We have had some amount of experience in our own laboratory with the thin sectioning of these kinds of samples for TEM characterization.
There are the obvious trade offs, that is, th that a) the price is much cheaper and b) you are using the "normal" narrow angle (e.g. 45 deg.) knives and therefore you don't have to worry about compression effects.
While the ability to cut sections is somewhat related to the experience of the one doing the sectioning (and it is also an art), so long as you are patient, you should be able to do it.
If you wanted to send us one of your samples, we could produce sections for you, and essentially determine the exact conditions under which you too could easily get sections. There would be a fee of course for this, however, you would know for sure, one way or another, whether sections could or could not be made of your material,and if they could, then you would also know the exact cutting conditions.
Another important consideration would be the choice of embedding resins and we would recommend for quartz particles our SPI # 2660 SPI-PON 812 resin kit since this particular resin seems to be the most user friendly in terms of polymerizing to a hardness for which sections can be made of the embedding quartz particles.
You have also asked a question that touches on the expected longevity of the diamond knife when cutting these kinds of materials. You sort of have to think about it as an analog to the mileage expected from a pair of tires. You can buy imported tires or you can buy domestic ones, however, what really counts is how you drive your car. Be gentle, and the knife will last longer. Be abusive, and it will have a shorter life time. Obviously, cutting quartz particles is going to be "hard" on the knife. You can extend the longevity be making the particles smaller before embedding. You can get good sections faster, and therefore also less knife wear, by curing the SPI-Pon 812 resin harder right away, putting less wear and tear on the knife to get to that point.
I hope this information has been helpful to you. Let me know if you have any other questions.
Charles A. Garber SPI Supplies PO Box 656 West Chester, PA 19380 USA
The Swiss Society of Optics and Electron Microscopy, the French Society of Electron Microscopy and the Belgian Society of Microscopy have decided to hold a joint meeting in 1995. This conference will be held at the "Ecole Polytechnique Federale de Lausanne" (EPFL), Lausanne, Switzerland, from June 26 to 30, 1995. It will include two tutorial days and a three-days conference which will be mainly focused on the recent progresses in TEM and scanning probe microscopies for materials science and biological fields.
********** SCIENTIFIC PROGRAMME **********
Monday 26 and Tuesday 27 TUTORIALS, with practical training, for groups of 6 to 20 people.
- Cryo-microscopy of vitrified specimens: theory and practice (Jacques Dubochet) - Practical aspects of the methodology of image analysis in oncology (Ricardo Laurini) - TEM image simulation of crystalline materials (Pierre Stadelmann) - Electron holography (Conradin Beeli) - Analytical microscopy with a field emission TEM (Klaus Leifer) - TEM sample preparation in materials science (Daniele Laub) - Applications of large angle convergent beam electron diffraction techniques (LACBED) (Jean-Paul Morniroli)
Evening Tuesday 27 Welcome ceremony and conference opening lecture.
"Perception du monde exterieur par les systemes vivants" by Yves de Ribaupierre, University of Lausanne.
Wednesday 28, Thursday 29 and Friday 30 - Conference
- New microscopies: STM, AFM, SNOM, confocal, ultra-sound, ... (A. Engel, D. Pohl)
- Energy filtered images and electron spectroscopy (C. Colliex, D. Bazett-Jones)
- Life in extreme environment (F. Gaill)
**** BIOLOGY-PATHOLOGY SYMPOSIA ****
- Fertilization and early embryogenesis in mammals (J. E. Flechon)
- Freeze substitution, microscopy of frozen hydrated samples (J. Dubochet, S. Fakan)
- Image analysis and cancer diagnosis (R. Laurini)
- High-resolution electron microscopy of biological specimens (E. Delain, M. Muller)
- Cytoskeleton (G. Gabbiani)
- New applications of scanning probe microscopies (M. Robert-Nicoud)
**** PHYSICS-MATERIALS SYMPOSIA ****
- Electron holography (C. Beeli, D. van Dyck)
- New progress on CBED/LACBED (J. P. Morniroli, J. Steeds)
- High-Resolution microscopy of aperiodic structures (J. van Landuyt, P.Stadelmann)
- New applications of analytical microscopy: filtered images, EDS, EELS (C. Colliex, C. Humphreys)
- New applications of scanning probe microscopies (D. Pohl, D. Courjon)
**** Commercial exibition ****
A substantial commercial exhibition of scientific equipment will also be held during the conference. For information, please contact G. Peter, EPFL-CIME CH 1015 Lausanne Fax: +41 [21] 693 44 01, email: peter-at-cime.epfl.ch
********** REGISTRATION **********
The symposia will consist essentially of a small number of lectures (mainly invited, with some selected from the conference abstracts) and of poster sessions. The presentations may be given in English, or in any languages of the three national societies. However, certain parts of the presentation should be in English, at least abstracts, figure captions, transparencies... Discussion sessions will be devoted to a few hot points emerging from the poster presentations.
The registration fees for the conference, including attendance at the symposia and the abstract booklet, are 250 CHF (non-members), 200 CHF (members) and 100 CHF (students and lab technicians). The registration fees for the tutorial days will be communicated in the final circular. A maximum of 100 CHF/course is expected. A few grants have been founded by the national societies in order to encourage the participation of young scientists and technicians
********** GENERAL INFORMATION **********
The conference will be held at the Ecole Polytechnique Federale de Lausanne (EPFL), located approximately 4 km from the city centre, near (500 m) the idyllic scenery of lake Leman. Hotel rooms in different categories (from "Formule 1" to ****) have been reserved by the tourist office for the organization committee, with prices ranging from 25 (F1, 3 persons per room) to 230 CHF. A final hotel booking form will be included with the registration form.
Excursions for participants and accompanying persons are planned: e.g. a tour to the beautiful region of "la Gruyere" famous for its cheese, to a wine cellar on the coast of lake Leman !! ... These excursions will be organized in small groups of 20 to 30 persons according to their preferences.
********** ORGANIZING COMMITTEES **********
Honor committee: Walter Bollmann, Alain Gautier, Eduard Kellenberger, Bernard Vittoz
International Scientific Committee: M. Deschuyteneer (SmithKline Beecham, B), J. Dubochet (Uni. Lausanne, CH), A. Engel (Uni. Basel, CH), J. Gunter (Uni. Zurich, CH), D. Hernandez-Verdun (Inst. Jacques Monod, F), J. Lecomte-Beckers (Uni. Liege, B), J.-P. Morniroli (Uni. Lille, F), R. Portier (ENSCP, F), M. Praet (Uni. Gent, B), D. Schryvers (Uni. Antwerpen, B), P. Stadelmann (EPFL Lausanne, CH), D. Thomas (Uni. Rennes, F)
Local Scientific Committee: M. Campiche (Uni. Lausanne), J. Dubochet (Uni. Lausanne), S. Fakan (Uni. Lausanne), R. Gotthardt (EPFL Lausanne), R. Laurini (Uni. Lausanne), P. Stadelmann (EPFL Lausanne)
Local Organizing Committee: P.A. Buffat (Chairman), R. Rouquier (Secretary), C. Beeli, F. Bobard, B. Garoni, P.-H. Jouneau, D. Laub, G. Peter, B. Senior, P. Stadelmann
I am looking for information about software to read .bmp or TIFF files using a Mac. We have a Targa+ video capture board to capture images from the SEM onto a 486-66 PC. I would like to be able to import them over e-mail to a Mac Quadra 650 and paste into a Word document. Word will import the image, but there seem to be a decrease in resolution and the file size is reduced from 650K to 375K when it is converted to the Mac. I would like to here from other users who are capturing images as we are just getting the system set up. Thanks,
John Giles Honeywell Space Systems jegiles-at-space.honeywell.com (813)539-2270 phone (813)539-3630 Fax
} I do not know how much extension can be obtained by storage at say } 4 C (May be someone out there does know). But we seem to start } having problems with film only a couple of months after the } expiration date even after cold storage for a few months. } } Richard E. Edelmann
I had a problem of inside sticky #52 Polaroid film, which will expire on Sept. 1995, so that run out of our film. I found a box of old #52 film which expired on Aug. 1992 left in a drew (at room temperature) and tried to use it. To my surprise, it worked fine. For my experiences, the problem of Polaroid does not really related to the expiration date. Instead, I found a lot of problems are caused by storage. I have used a lot of good expired films and a lot of fresh films as well for 5 years.
Ya Chen Integrated Microscopy Resource Madison, WI 53706 USA Email: ychen-at-macc.wisc.edu
} We are in the process of purchasing a new printer and are looking at the } Tektronix Phaser II SDX, Kodak ColorEase PS, and Kodak 8600. Samples } provided by } Kodak and Tektronix look great. I have heard that the Mitsubishi S3600-32U } is a } great printer but cannot get any useful information, prices, or samples from } Mitsubishi or local dealers. I am curious about the Fujix 3000. Could you let } me know how or where to contact Fujix, since I have never heard of them.
You can contact Ron Saltzman, at Fujix Electronic Imaging Group at 800-736-3600 ext. 8282 (voice mail). If he's no longer with that group, you might try the general numbers, 914-789-8100 or 800755-3854, to get to a sales rep. They also advertise in some of the photo lab trade magazines.
Good luck. They are really worth a look.
John chandler-at-lamar.ColoState.EDU Fort Collins CO
Thanks to those who have helped me locate sources where I could find a material matching the description I gave for what I called "carbon putty."
Since posting the first source given to me (Bal-Tec) yesterday, I have received a few more. So, for those other subscribers who asked me to post any info. I got, here are the other sources.
Marivac LTD./Hailfax, N.S./Canada/(800)565-5821/Cat.# AS508-3/ $66.50 Canadian 15g (Approx. $50.00 American)
SGL-Carbon/Niagara Falls, N.Y./P.O.Box 667/(716)236-2859/ This company makes carbon epoxy material that sets-up for permanent attachments. It is not a soft putty after it sets but it is highly conductive and may be of some use for other purposes.
Hello All, I have a user who was told to try Zamboni Fix on their tissue. Could some kind soul tell us what it is and how to make it. I know what and Zamboni is in relation to ice rinks and I can even drive one (it's a real hoot!!) but I've never heard of the fix... TIA,
C. Michael Stanley, Ph.D. Coordinator, Associate Director Molecular Cytology Core Facility Molecular Biology Program 2 Tucker Hall University of Missouri-Columbia Columbia, MO. 65211 (314) 882-4895 fax= 314-882-0123
Ed Calomeni wrote: } I am in the info gathering stage of purschasing a tissue processor for EM } samples. } I have info on the Reichert-Lynx unit and the one made by RMC. How } do you like } these units? Comments on advantages and disadvantages are } welcome.
} Thanks in advance
} Ed Calomeni } Medical College of Ohio } Toledo, OH
We have a Lynx tissue processor which we are now very happy with. There were a few teething problems when it was first installed (the main control board and the motor control unit had to be replaced) but these problems I believe have now been sorted out at the factory and shouldn't apply to new instruments now - it was some three years ago when we bought our one. The only other problem we had was with voltage spikes from the mains. This problem was eliminated by putting an isolating transformer between the mains and the processor. We have not experienced any problems with the samples mixing upon opening up the vials (someone -sorry I forget who- mentioned this in an earlier reply). Infiltration of resin seems to be fine, something I was a little sceptical of initially, given the small holes in some of the basket types. There is a good choice of different specimen baskets anyway and I have always found one suitable for whatever specimens I happen to be processing. Hope this is helpful for making your decision. Regards, RE
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
Message-Id: {199503092054.PAA03408-at-alchemy.chem.utoronto.ca} X-Sender: pmarkiew-at-alchemy.chem.utoronto.ca (Unverified) X-Mailer: Windows Eudora Version 1.4.4 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Fellow Microscopists,
This is a general announcement which should be of interest to atomic force microscopists.
We are making the latest version of our deconvolution program MIDAS95 available for free on the Internet. The program is designed to work under Windows on Nanoscope files. It is being released as a beta version, meaning the program lacks a certain polish. The program works properly for both the Nano II and III, although testing on the latter system has been limited.
Through deconvolution, one can account for the volume occupied by the tip in the AFM images obtained. By eliminating the tip effect, the result is often a truer representation of the sample topography. Deconvolution also allows for the in situ measurement of the tip geometry. This 3D data file of the tip can be used as a check of the tip's integrity or for improvement of other AFM images. Further details are given in the README.TXT file.
The program can be accessed by the following: ftp surfturf.chem.utoronto.ca login: anonymous password: yourname-at-location Please use your E-mail address for the password. We wish to keep a record of the users of this program and notify them of any problems or updates.
The public directory has three files of interest here. One is the README.TXT file if you want to know what is in MIDAS.ZIP without having to download it. MIDAS.ZIP contains several files, including some AFM files to see how deconvolution is done and some TIFF files for viewing. Fan-xing Wei and Dr. Dan Thomas of the University of Guelph have modified our program so that it works for Burleigh Instruments. This program is available in BURL.ZIP.
If you have any difficulty, please be patient as our server allows only one user at a time and tends to lock up when doing calculations.
The best way to get bitmap and/or TIFF images into your Mac is with the NIH Image software (via ftp: anonymous-at-zippy.nimh.nih.gov directory /pub/nih- image). It can handle either format, but you will probably have to go through the IMPORT diaolog, not the OPEN dialog. Look at the documentation (also on Zippy) to guide you through. As long as there is a full valid bitmap in the image file, NIH Image can import the image and allow you to save it as a useful file. We have done a lot of image file format trouble-shooting for "standard" formats written by multiple vendors on multiple platforms around here using that capability.
A plug here: NIH Image has a nearly infinite performance to price ratio - it is free once you are on the internet. It also outperforms a whole lot of commercial stuff on any computer platform up to about the $2000 price point and the support system is unbelievably good, considering the annual maintenance non- fee. 8-).
Two hurdles are: getting the file onto your Mac (which it sounds like you are already doing) then getting a good image into Word. I do the latter by pasting the image from NIH Image into Word, then shift-shrinking the image to about 50%. The original paste goes in at 72 dpi, the shift-shrink gives the final image resolution of 144 dpi which, with a decent gray-scale printer, gives you very acceptable print quality. By the way, I __STRONGLY__ advise against using the image editor in Word. I have found it unreliable in addition to the fact that it reduces your grayscale image to 16 levels. This could explain the drop in file size (the editor converts the 8-bit image to 4-bit). A similar result would happen if you started with a 16-bit TIFF and wound up with 8-bit.
Hope this helps.
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
Message-Id: {9503092328.AA21558-at-riker.ml.wpafb.af.mil} To: WALCKSD:ML:WPAFB cc: Lloydpf:ML:WPAFB Subj: ultrathinsectioning material containing quartz In-Reply-To: Message from WALCKSD:ML:WPAFB of 3-8-95 ----------------------------------------------------------------------- Scott,
I would recommend that the knife of choice be a Diatome knife. The angle would depend on how hard the sediment is. If the sediment is very hard, then yes I too suggest a 55x knife. Using a knife with this angle still allows one to obtain sections on the order of 70nm. If the sediment is not that hard, then a 45x knife will work also. I generally use a 55x knife for all materials that I cut except for polymers, where I use a 45x knife.
I hope this helps!
Pam
---------------------- Replied Message Body ---------------------- To: LLOYDPF Subj: ultrathinsectioning material containing quartz Forwarded: Message from {chswartz-at-MIT.EDU}:ddn:wpafb of 3-8-95 ------------------------------------------------------------------
--------------------- Forwarded Message Body --------------------- To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: ultrathinsectioning material containing quartz Orig-Author: {chswartz-at-MIT.EDU}:ddn:wpafb ----------------------------------------------------------- I am going to be attempting to thin section sediment samples containing quartz and want the thin sections to be approx. 50-70nm in thickness for TEM analysis. I have to buy a diamond knife and would much appreciate input on the blade angle to choose. A representative at Polysciences told me I should go to a 55 degree knife instead of a 45, but I was wondering if I would be sacrificing much of the ability to cut sections down to 50nm. The quartz grains are approx. 200 um in diameter. Is there a particular manufacturer I should get the knife from? What is an appropriate cutting speed? Can I expect the knife to last only a VERY short time? I would appreciate any suggestions and advice. My address is: chswartz-at-MIT.edu
I'd appreciate a copy of the BDMA vs DMP-30 responses.
Regards
Mike Gregory
} In July and September of 1994, there was some disciussion of BDMA vs. DMP-30. I } saved and consolidated 13 of those messages into a single document. If anyone is } interested in getting a copy, send me your e-mail address and I will zip one out } to you within a day or two. } } -- } } 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 } }
} The best way to get bitmap and/or TIFF images into your Mac is with the NIH } Image software (via ftp: anonymous-at-zippy.nimh.nih.gov directory /pub/nih- } image). It can handle either format, but you will probably have to go through } the IMPORT diaolog, not the OPEN dialog. Look at the documentation (also on } Zippy) to guide you through. As long as there is a full valid bitmap in the } image file, NIH Image can import the image and allow you to save it as a useful } file.
As a follow-up, we use NIH Image for our image processing and also import BMP and TIFF files from PC capture systems. NIH Image will import TIFF directly using 'OPEN', and from our experience BMPs should be IMPORTed using CUSTOM, setting OFFSET to 1760 and chsing the relevant bitmap size.
I hope this is of use
Doug
+------------------------------------+ | Dr Douglas Arrell | | Mechanical Performance and Joining | | Institute for Advanced Materials | | 1755 ZG Petten | | Netherlands | | {ARRELL-at-JRC.NL} | | Tel. (+31) 2246 5287 | | Fax (+31) 2246 1917 | +------------------------------------+
A short message for users of (or people interested in) 'S-SIMPLY' (SHRLI-SIMulation and diSPLaY of TEM & HRTEM images, for PCs) : the freeware version, available on FTP.UNIV-LYON1.FR (/pub/dos/HRTEM) has been significantly updated, and new features are available (graphical displays, a "Make Interface" tool,...). With best regards,
______________________________________________
Thierry EPICIER GEMPPM-502, INSA de Lyon, 69621 VILLEURBANNE, France tel : (33) 72 43 84 94 (83 85) FAX : (33) 72 43 85 28 Email : EPICIER-at-CISMSUN.UNIV-LYON1.FR
Two days ago I posted a Cat.# and price for Leit-C-Plast from Bal-Tec that was given to me incorrectly. The correct Cat.# is B801014077 and the cost is $40.00 for 15g. This is now the best price I've found.
I recently purchased a Pictography 3000 printer after a extensive comparison of printer systems available on the market. The printer is attached to the network (Token ring, novell) via a pc, and is transparent configured: all pc's in the network can reach the printer. The printer is used for production of hard-copies of image files generated by various techniques: CSLM, VEM, cryo-SEM, FESEM (+EDS Noran SUN), TEM (Gatan SSC, Mac), SPM, IA (SEMPER).
We are quit satisfied.
E-mail: Marcel.Paques-at-2488PAS.URLNL.Sprint.Com Unilever research Laboratory Vlaardingen The Netherlands
Zamboni fix is an EM version of the classical LM Bouin's fix. Zamboni's original abstract didn't contain any details about how to make it up. The details are in: Stefanini et al., 1967, Nature 216:173.
A. Kent Christensen Department of Anatomy and Cell Biology University of Michigan Medical School {akc-at-umich.edu}
------------------------------------------
On Thu, 9 Mar 1995, Michael Stanley wrote:
} Hello All, } I have a user who was told to try Zamboni Fix on their tissue. } Could some kind soul tell us what it is and how to make it. I know what } and Zamboni is in relation to ice rinks and I can even drive one (it's a } real hoot!!) but I've never heard of the fix... } TIA, } } C. Michael Stanley, Ph.D. } Coordinator, Associate Director } Molecular Cytology Core Facility } Molecular Biology Program } 2 Tucker Hall } University of Missouri-Columbia } Columbia, MO. 65211 } (314) 882-4895 } fax= 314-882-0123 } } }
I have recently been shown a research report in BioTechniques, Vol 13, No 3. (1992) by Reed, J.A., Manahan, L.J., Park, C-S, and Brigati, D.JL describing an immunocytochemical method based upon capillary action. This was using the "MicroProbe" marketed by Fisher Scientific, Pittsburgh, PA.
My question is, has anyone used this system? If so what are its advantages and/or disadvantages . Also could you please give me a contact address, FAX number or E-Mail for Fisher Scientific.
Message-Id: {9503101815.AA2900-at-pho018.sb.com} To: images1 {images1-at-biosci.mbp.missouri.edu} , microscopy {microscopy-at-aaem.amc.anl.gov}
In response to this question:
Hello All, I have a user who was told to try Zamboni Fix on their tissue. Could some kind soul tell us what it is and how to make it. I know what and Zamboni is in relation to ice rinks and I can even drive one (it's a real hoot!!) but I've never heard of the fix... TIA,
C. Michael Stanley, Ph.D. Coordinator, Associate Director Molecular Cytology Core Facility Molecular Biology Program 2 Tucker Hall University of Missouri-Columbia Columbia, MO. 65211 (314) 882-4895 fax= 314-882-0123
I think that the fixative you're referring to is cited in this paper: Zamboni, L. and DeMartino, C., 1967. Buffered picric acid-formaldehyde: A new, rapid fixative for electron microscopy.
We came across Zamboni's fix when we were perfusion-fixing rat testes a couple of years ago. However, it didn't preserve Leydig cell structure as well as some of the other fixatives we tried.
Hope this info helps. (I want to hear more about driving a Zamboni!)
Bev Maleeff SmithKline Beecham Pharmaceuticals Toxicology-US, UE 0462 709 Swedeland Road King of Prussia, PA 19406 610/270-7987 610/270-7202 fax e-mail: maleeffbe-at-sb.com
The 30th annual meeting of the SouthEastern Microscopy Society (SEMS) will be held on May 17-19, 1995, at the Omni Hotel in Atlanta, GA. Scheduled invited speakers and their titles are: 1) Nestor Zaluzec, Argonne National Laboratory, Integrating Computers, Microscopy, and Microanalysis; 2) Phil Russell, North Carolina State University, Instrumentation and Application of Scanned Probe Microscopy; 3) Larry Peterson, GBI, Forensic Microscopy; 4) Terence Mitchell, Los Alamos National Laboratories, Experience with Field Emission on an SEM, STEM, and TEM; and 5) Jay Jerome, Bowman Gray School of Medicine of Wake Forest University, Exploring Ultrastructure with Quantitative 3-D Intermediate Voltage Electron Microscopy. Also on the program are pre-meeting workshops and tours, contributed papers and posters, the RUSKA student competition, and commercial exhibits. Social events are the Wednesday night Exhibitor's Mixer and the Awards Banquet, which will be held Thursday night at the Fernbank Museum of Natural History
For further information, contact Janet H. Woodward, SEMS '95 Program Chair, at (912)-945- 3152, FAX (912)-945-3155.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
My goodness I got the message several times. Also the one I sent came back to me several times. I guess the Government wants to make sure evereything gets through.
Too bad about the cuts. Frank Skelton told me about Van Closing also I guess the Farm got hit.
PCI is selling well particularly in the USA and some to Japan.
I have full internet access now. PPP with every application which is Freeware and Shareware. All windows based and relatively easy to use.
Our site in the office is working under a private account, but we haven't got the domain name yet. It will be NSCTOR. Anyway I will let you know when it comes through.
David Cockayne asked: } } WITH REGARD TO THE QUESTION ABOUT TEACHING OPTICS, CAN ANYONE ADVISE ME OF WHAT SOFTWARE PACKAGES ARE AVAILABLE FOR TEACHING ELECTRON OPTICS?
At the same time a similar question was posted on the Microscopy list (by someone different).
I would be delighted if anyone can update the following list of teaching software (in the area of microscopy) known to me:
The Institute of Materials (UK) distribute:
The Transmission Electron Microscope by Goodhew The Scanning Electron Microscope by Humphreys (John) Electron Diffraction by Goodhew The Stereographic Projection by Humphreys Analysis in the Electron Microscope by Goodhew, Humphreys and Cliff X-ray Photoelectron Spectroscopy by Watts X-ray Auger and Laser emission by Goodhew
These are all IBM PC DOS versions, available from Institute of Materials, 1 Carlton House Terrace, London, SW1Y 5DB, fax +44(0)171 839 2078 The MATTER project is currently working on greatly-improved Windows and Mac versions of most of these topics: Completion of the first two topics (TEM and SEM) is scheduled for the end of 1995. All the above are aimed firmly at teaching, not at research applications.
There is also an SEM simulation available which originated in Australia but which is supposed to be downloadable from Nestor Zaluzec's Microscopy Bulletin Board. However we have consistently failed to transfer it. Has anyone in the UK succeeded?
I also saw recently a very full simulation of a light microscope. This is aimed at biologists and was written (and is distributed) by Maurice Smith at: mol-at-molcol.demon.co.uk
If you can add to this list please reply to both lists [ microscopy and materials-l ]
Thanks
Peter
PS 120+ members of the materials list now!
------------------------------------------------------------------ ----- Professor Peter J Goodhew, Department of Materials Science & Engineering University of Liverpool LIVERPOOL Fax (44) (0)151 794 4675 L69 3BX, UK Tel (44) (0)151 794 4665 (secretary Debra) ------------------------------------------------------------------ ----- inter alia: Director of the MATTER project for educational software ------------------------------------------------------------------ -----
STUDENT COMPETITION TO BE HELD AT THE UNIVERSITY OF WISCONSIN WHITEWATER, WISCONSIN Ambrose Health Center, Room 162 Friday, March 24, 1995 PROGRAM: 1:00 PM: Dr. Ursula K. Charaf, Senior Research Scientist, Johnson Wax: BLOW IT UP! Applied Industrial Microscopy 1:45 PM: Student Presentations 3:00 PM: Dr. Robert Cardell, University of Cincinnatti Medical Center, MSA/LAS Sponsored Presidential Speaker: Modern Microscopical Approaches to Biomedical Problems 4:00 PM: Reception and Award Presentations
If you are planning to come, telephone or E-mail registrations are required by March 22. If you are planning to present your research, please send your abstract to: Dr. Lance Urven University of Wisconsin- Whitewater 800 West Main Whitewater, WI 53190 urvenl-at-uwwvax.uww.edu PHONE 414-472-5132 Reception provided by Oxford Instruments Inc., Microanalysis Group.:
Don- 1) the thin film coatings on the polycarbonate are very tightly held proprietary secrets. try some EDS or WDS. 2) try SEM to determine where these "pits" reside. 3) the "trick" is to plunge the CD into liquid nitrogen (30 sec), then give it a shot with a hammer. it shears at the interface
we are investigating the CD's by AFM and SEM here in our lab, keep in touch about further results?
-Mike
On Sun, 12 Mar 1995, Chernoff wrote:
} I am seeking advice regarding specimen preparation. I wish to } examine the physical pits created when a CD-Recordable disk is } written. I would like to understand: } - the physical arrangement of thin film coatings on the } polycarbonate substrate, i.e. what materials are stacked up, and } what their typical thicknesses are. } - what layer contains the physical pits. } - how to expose that layer for examination by SEM or AFM. } } Thanks for your help. } } DON CHERNOFF 317-251-1364 } ADVANCED SURFACE MICROSCOPY FAX: 317-254-8690 } 6009 KNYGHTON RD. E-MAIL: echernof-at-ucs.indiana.edu } INDIANAPOLIS IN 46220 Toll free: 800-374-8557 } } }
Where could I findout what was the fair market value for a JEOL 1200ex II 5 years old. Basic TEM no Scanning EDX or stuff like that. Who has the blue book??
I'VE BEEN ASKED TO ADDRESS A GROUP OF STUDENTS (LATE HIGH SCHOOL AND EARLY COLLEGE) A LOCAL COLLEGE SCIENCE CAREER DAY. I AM PLANNING ON USING A VARIETY OF SEM MICROGRAPHS TO ILLUSTRATE THE APPLICATIONS OF THE SCANNING ELECTRON MICROSCOPE TO MANY SCIENTIFIC AREAS. IF ANYONE HAS ANY PC-FORMAT IMAGES THAT THE WOULD BE WILLLING TO E-MAIL TO ME, I WOULD BE VERY GRATEFUL.
There really isn't anything equivalent to a "blue" book, since there are just not enough instruments of this type getting sold in order to get any kind of meaningful sales statistics. However, there are basically three different types of prices discussed:
a) Selling price of a used instrument from the OEM
b) Selling price from a third party service company
c) Offering price from a former owner
If you want to find out (a), the best person would be Robert Santorelli at JEOL in Peabody, MA Ph; (800) 343-6766.
For (b), you should contact Mr. Clark Houghton, Secondary Images, Winchester, OH FAX: (513) 927-5557.
For (c), you should contact again, either Mr. Houghton (above) or Mr. James Nicolino, X-ray Optics, Florida, Ph: (904) 646-3069 FAX: (904) 565-1897. Mr. Nicolino services wave length dispersive microprobe systems but has a good understanding of what different instruments fetch when sold by an owner, free of any guarantee or warranty, and such sales are generally on an "as is, where is" basis.
Hope this information will be useful to you.
Charles A. Garber SPI Supplies West Chester, PA 19380 USA
To: goodhew-at-liverpool.ac.uk Order #4457898 From: GVKM07A Date: 03/13/95 11:46 PM
Prof. Peter Goodhew
Hello! I think that the soft ware you are thinking of is called the emTutorial system, and it is indeed produced in Australia. There are actually two different system programs, emTUTOR and semTUTOR, and they have been designed to be used as a self-instruction tool, an instruction aid for class labs and for lecture demonstrations.
Also I think the commercial producers of these really outstanding software products would be quite unhappy if it should turn out that their proprietary software was distributed without any benefit to them. I could be wrong about that, but if I was a betting person, that would be my opinion.
The product can be ordered from SPI Supplies as SPI #09000-AB and has a regular price of $345. By coincidence, we have been running a "special" and until April 15, the price is $250.
There is another new product from the same Australian firm called PARFOCAL, which is a graphics file conversion program for confocal microscopy and image analysis. The regular price is $325 but this product is not current on any kind of special sale.
If you provide a FAX number, we can FAX you additional information.
Charles A. Garber SPI SUPPLIES (USA) PO Box 656 West Chester, PA 19380 USA
Ph: (800) 242-4774 Toll free in USA only (610) 436-5400 Regular phone number (800) 526-6562 Toll free in Canada only - rings in USA
FAX: (610) 436-5755 1-800-55-7780 Toll free FAX from Ireland only 0800-44-0873 Toll free FAX from New Zealand 0800-89-3066 Toll free FAX from UK
I have received the following information from the organizers and wanted to publicize it to anyone interested in attending or participating:
First International Conference on Electron Microscopy and Advances in REsearch in Different Fields of Science
September 2-4, 1995 Ismailia-ETAPT
Sponsored by Electron Microscopy Center Suez Canal University Ismailia EGYPT
Special Topics of the Conference: 1] Role of EM in diagnostic virology 2] Role of EM in diagnosis of tumors cytology and urinary stones 3] Role of EM in ultrastructure pathology of the lung (non neoplastic conditions) 4] X-ray microanalysis: Applications particularly metallurgical, mineralogical, and biological 5] Scanning EM of plants, animal, insects, and mineral material 6] Study of biological macromolecules from their characteristic electron diffraction patterns 7] Skin pathology by EM 8] Morphological identification of antigens by EM 9] Different low temperature methods for biological EM 10] Safety measures and maintenance needed for EM
There will be an equipment exhibition in conjunction with this meeting.
Registration for foreigners will be US $150 inclusive of full board during the time of the meeting.
For further information, contact the organizer:
Prof. Dr. Khalifa Ibrahim Khalifa Electron Microscope Center Suez Canal University Ismailia EGYPT
X400-Received: by mta merit in /PRMD=internet/ADMD=telemail/C=us/; Relayed; Tue, 14 Mar 1995 03:40:10 -0500 X400-Received: by /ADMD=TELEMAIL/C=US/; Relayed; Tue, 14 Mar 1995 03:38:06 -0500 X400-Received: by /PRMD=SMXFL2/ADMD=TELEMAIL/C=US/; Relayed; Tue, 14 Mar 1995 03:41:49 -0500 X400-Received: by /PRMD=LANGATE/ADMD=TELEMAIL/C=GB/; Relayed; Tue, 14 Mar 1995 03:41:00 -0500
To: GVKM07A From: Peter Goodhew Date: 03/13/95 06:01 AM
David Cockayne asked: } } WITH REGARD TO THE QUESTION ABOUT TEACHING OPTICS, CAN ANYONE ADVISE ME OF WHAT SOFTWARE PACKAGES ARE AVAILABLE FOR TEACHING ELECTRON OPTICS?
At the same time a similar question was posted on the Microscopy list (by someone different).
I would be delighted if anyone can update the following list of teaching software (in the area of microscopy) known to me:
The Institute of Materials (UK) distribute:
The Transmission Electron Microscope by Goodhew The Scanning Electron Microscope by Humphreys (John) Electron Diffraction by Goodhew The Stereographic Projection by Humphreys Analysis in the Electron Microscope by Goodhew, Humphreys and Cliff X-ray Photoelectron Spectroscopy by Watts X-ray Auger and Laser emission by Goodhew
These are all IBM PC DOS versions, available from Institute of Materials, 1 Carlton House Terrace, London, SW1Y 5DB, fax +44(0)171 839 2078 The MATTER project is currently working on greatly-improved Windows and Mac versions of most of these topics: Completion of the first two topics (TEM and SEM) is scheduled for the end of 1995. All the above are aimed firmly at teaching, not at research applications.
There is also an SEM simulation available which originated in Australia but which is supposed to be downloadable from Nestor Zaluzec's Microscopy Bulletin Board. However we have consistently failed to transfer it. Has anyone in the UK succeeded?
I also saw recently a very full simulation of a light microscope. This is aimed at biologists and was written (and is distributed) by Maurice Smith at: mol-at-molcol.demon.co.uk
If you can add to this list please reply to both lists [ microscopy and materials-l ]
Thanks
Peter
PS 120+ members of the materials list now!
------------------------------------------------------------ ------ ----- Professor Peter J Goodhew, Department of Materials Science & Engineering University of Liverpool LIVERPOOL Fax (44) (0)151 794 4675 L69 3BX, UK Tel (44) (0)151 794 4665 (secretary Debra) ------------------------------------------------------------ ------ ----- inter alia: Director of the MATTER project for educational software ------------------------------------------------------------ ------ -----
-------- Original message header follows -------- From goodhew-at-liverpool.ac.uk Mon Mar 13 06:01:17 1995 [PIM 3.2-342.56] Received: from AAEM.AMC.ANL.GOV (aaem.amc.anl.gov [146.139.72.3]) by inetgate.prodigy.com (8.6.10/8.6.9) with SMTP id GAA64180 for {GVKM07A-at-mail.prodigy.com} ; Mon, 13 Mar 1995 06:01:17 -0500
To: MICROSCO:CENTRAL cc: DRStadden:R_D:Armstrong Subj: AFM Learning Curve ------------------------------------------------------------------
One of our researchers is looking into buying an AFM, primarily for studying latex coatings. As a microscopist (PLM,SEM,EDX) in the analytical group, I'm interested in the broader applications we might find for our entire product line.
These are my questions:
1.) What is the "typical" learning curve for the AFM for non- microscopists vs. microscopists?
2.) How useful is it for one to have access to other types of microscopies to correlate with AFM results?
3.) Does the usual rule of "fewer operators, less downtime" apply to any greater or lesser degree with the AFM?
I look forward to any thoughts and experiences you can share.
Dave Stadden DRStadde+aCENTRAL%ARMSTRONG-at-MCIMAIL.COM Phone 717-396-5109 FAX 717-396-5865
I am the author of "Virtual SEM", an earlier version of which called "SUPERSEM" is available through FTP from the EMMPDL library.
"Virtual SEM 1.2" is an update of this earlier package and is much enhanced. I'm a bit short of time but the recent discussion (Prof. Goodhew, and Chuck Garber - regards to both) I think is in part mistaking my software for that available commercially through SPI and others.
The latest version will be passed to Nestor and John Mansfield in about a fortnight at the Scanning meeting and I hope they will see fit to update the old version. It is public domain and comments are most appreciated. To summarise it is a MAC product utilising real images controlled through a "SEM console" simulation. It includes tutorial information and two self assessing tests, the results from which are automatically stored on file. We have been using it for 18 months and feedback has been good.
The drawback is that it is now around 35 meg in size and I expect it to reach 40 meg soon. It only needs 4 meg of ram to run on, for example, a MAC IIsi or better. I developed most of it on a powerbook 180C.
The size is a problem for FTP and as noted in earlier versions I can supply it on a multisession CD. To do this I am charging US$75 to cover writing and postal costs only. I will upgrade to later versions and hope to be able to at postal cost, if I have to hire a bod to do it then would have to cover the time but cost would still be minimal. We also have a virtual EDS in development and plan to fully integrate the two.
I will try and answer any queries. Those that have contacted me earlier, I have not lost your requests, am working my way thru as fast as I can - please be patient.
Brendon Brendon J. Griffin Centre for Microscopy and Microanalysis The University of Western Australia Nedlands, WA, AUSTRALIA 6907 ph 61-9-380-2739 fax 61-9-380-1087
I am an Associate Professor at The Rockefeller University interested in obtaining access to the microscopy listserver. A colleague told me that this would be a good medium to advertise postdoctoral and technical positions that may be available in my laboratory.
Please let me know if I need to submit any more information. My address is: William A. Muller, MD, PhD Box 176 The Rockefeller University 1230 York Avenue New York, NY 10021 Phone (212) 327-8104 Fax (212) 327-8875
I have heard rumors that there will be an international microbeam analysis meeting in Australia early in 1996. Does anyone know if this is so; and if it is, do you have any information about when and where it will be?
I'm staining spinal cord and medulla's neurons from rats' fetuses, 10 days old, with the following primaries antibodies: Rabbit anti Glycine 1:2500, 1:4000; Rabbit anti GABA 1:100, 1:200; Rabbit anti GAD 1:300, 1:500; and anti-synaptophysin 1:150, 1:300. I'm using FITC 1:100 or Rhodamine 1:100 as my secondary antibodies. I keep getting crossover between FITC and Rhodamine despite the different dilutions of the primary antibodies I have used. Does anyone has any suggestion? Thanks in advance,
Ciprian Almonte Research assistant Medical College of Pennsylvania Dept. of Anatomy and Neurobiology 3200 Henry Ave. Philadelphia, PA 19129 Voice: (215) 842-4081
We've been trying to measure "integrated density" of some fluorescent images and found significant differences in the numbers calculated in v. 1.53 and 1.57. We have 1.53 on a quadra 950 and 1.57 on an 8100. Working with the same image, 1.53 returns numbers approximately 2,000,000 and 1.57 gives us numbers about 400. The mean intensity, Std dev, background, number of pixels...are identical in both versions. LUTs are also the same in both versions. These are TIFF images acquired on a DOS machine. I'm not sure if it's related to the integrated density difference, but in 1.57 we can "open" the TIFF files, while in 1.53 we have to "import" the TIFF file.
Is there something obvious I'm overlooking? Any suggestions are appreciated. Thanks
I'm staining spinal cord and medulla's neurons from rats' fetuses, 10 days old, with the following primaries antibodies: Rabbit anti Glycine 1:2500, 1:4000; Rabbit anti GABA 1:100, 1:200; Rabbit anti GAD 1:300, 1:500; and anti-synaptophysin 1:150, 1:300. I'm using FITC 1:100 or Rhodamine 1:100 as my secondary antibodies. I keep getting crossover between FITC and Rhodamine despite the different dilutions of the primary antibodies I have used. Does anyone has any suggestion? Thanks in advance,
Ciprian Almonte Research assistant Medical College of Pennsylvania Dept. of Anatomy and Neurobiology 3200 Henry Ave. Philadelphia, PA 19129 Voice: (215) 842-4081
A general posting since I've had a bunch of questions all about the same thing.
Yes the EMMPDL/MASLIB/PD Shareware libraries are avaiable via anonymous FTP. The site is
WWW.AMC.ANL.GOV (= 146.139.72.10 )
Login via standard FTP protocols with Username=Anonymous Password=Your Email Address
but please note that you CANNOT access these files using a WWW Browsing program like Mosiac, Netscape or any other, even though they have FTP capabilities. You must use a standard FTP protocol client program. which accesses the conventional FTP ports.
The reason for this is that the although WWW server and the FTP server are both on the same Computer (WWW.AMC.ANL.GOV=146.139.72.10) they reside on two different disk partitions which are NOT linked. The FTP server looks in one place and the WWW server another. When you login to the WWW site, your browser (client) program has only access to the disk space assigned to the WWW server hence it (the WWW program) will NEVER see the FTP files and vice versa! The reasons for this are a combination of security and convenience on my part, but with the explosion of the WWW I've created yet another headache for myself.
So for all of you that have tried and failed this is the most common reason. If you don't understand this call me next week sometime and I'll explain, or better yet come to the Telecommunications Tutorial which I'll be giving at the August MSA meeting.
In retrospect I should have put them on different machines with different DN's and then the problem would not have happened, but the damage is done and until I get a chance to come up for air it will have to stay that way. There is yet another alternative which is to define an alias on the DNS for the FTP server. If I can figure out the right person to contact I'll try that route but be guarenteed it will not be soon.
Just a reminder..... Abstracts/Papers for the August MSA meeting are due March 15th i.e. probably the day you receive/read this message.
If anyone has information on the Microscopy Meeting in August could you please send information on contacts for the meeting.
Also, I am looking to attend regional meetings and would love to find a calendar of meetings and places with contacts. I would be attending for training and exibiting.
Thank You all!!!
P.S. all that are interested about the LaserMaster ImagePrinter 1800 DPI the "NEW" price is $ 6,995.00 and an upgrade from a 60 mhz processor to a 100 mhz. WOW, 1K off plus upgrade.
Greg Begin - LaserMaster Imaging Specialist in Scientific/Medical Imaging
Just read the posting by Ciprian Almonte concerning crossover between FITC and Rhodamine and had a couple of questions about what he is doing. What are your FITC and Rhodamine linked too--goat anti-rabbit??? Are you trying to do double labelling??? Or is it that you are getting fluorescence of the FITC at the rhodamine settings?? More information is required to help. Yours Mark Elliott, PhD UBC-Pulmonary Research Laboratory, St. Paul's Hospital Vancouver BC Canada
You need to "fine tune" your microscope filters. Do you perhaps have an older Olympus?? Call your local microscope rep and ask for the correct filters to narrow the band pass of the dichroic filters. Grace
I think this may have been discussed by this group earlier but at the time we were not worried about it so I didn't keep a record of the responses. One of our tech's asked me to post this: We are looking for a plastic medium which can replace Epon in Biological TEM. In our lab we regularly use Epon 812 for its good stainability of membranes. Recently however, we have just too many samples to infiltrate and Epon is just too viscous for multiple tissue transfers. The result is extensive numbers of labour hours. Please let us know if you have succcessfully worked with any plastic that has equal stainability of the membranes but is less viscous. Thanks Mark Elliott UBC-Pulmonary Research Laboratory St. Paul's Hospital Vancouver BC Canada
} Just read the posting by Ciprian Almonte concerning crossover between } FITC and Rhodamine and had a couple of questions about what he is } doing. What are your FITC and Rhodamine linked too--goat } anti-rabbit??? Are you trying to do double labelling??? Or is it } that you are getting fluorescence of the FITC at the rhodamine } settings?? More information is required to help. } Yours } Mark Elliott, PhD } UBC-Pulmonary Research Laboratory, } St. Paul's Hospital } Vancouver BC Canada
FITC and Rhodamine are linked to anti-rabbit, and I'm planning to do double labelling. My major problem is that I'm getting flourescence of the FITC at the rhodamine settings and viceversa. Thanks, to all of those that have replied so far. I appreciate all their suggestions. Ciprian
Ciprian Almonte Research assistant Medical College of Pennsylvania Dept. of Anatomy and Neurobiology 3200 Henry Ave. Philadelphia, PA 19129 Voice: (215) 842-4081 E-mail almonte-at-medcolpa.edu
Try using more selective excitation filters (narrower band pass). Another suggestion is to use a "Red Cut" Filter. This greatly supresses the secondary peak that is associated with FITC. Contact you local microscope supplier or Omega Optical at (802) 254-2690. Good Luck,
Hello all, We have a user wanting to look at the inside of Polycaprolactone Microcapsules in the SEM. We are not wanting to do sections for the TEM. I have tried to fracture the microcapsules in liquid N2, after embedding them in resin, but they dont fracture through the capsule, only around it. Also, I have tried to section the block face of the capsules in resin, but resin seems to infiltrate into the capsule and so no internal structures can be seen. These capsules are between 100 microns - 20 microns in diameter, and have a melting point around 65oC. Has anyone tried this before and had success? The references I have contain no details about preparation of their specimens. Any help would be appreciated.
G'day Subscribers (actually in it's now about 11 pm in Chicago)
Here's the info that was posted about the IUMAS meeting that I dug out of the Archives. You will have to check with Dave Williams of Lehigh (dbw1-at-lehigh.edu) if you want further details.
Also the WWW site is also starting to get fairly busy, lately we've been averaging ~ 250 connections/day. So if it appears slow, consider yourself forwarned.
The FTP server crashed sometime on Monday Evening. So if anyone tried to get to the libraries, it was a useless battle. As of a few minutes ago it was back up and running.
Why does this always happen just before deadlines for MSA abstracts?
It must be the Ides of March syndrome, but then again maybe it was a Leprechaun.
MICROBEAM ANALYSIS SOCIETY STUDENT SCHOLARSHIPS TO MAS-1995 (Breckenridge, CO) and IUMAS-1 (Sydney Australia)
All students (and faculty) involved in microanalysis-related research, should note a remarkable opportunity for travel to two forthcoming microanalysis conferences. The Microbeam Analysis society is offering student scholarships to the 29th Annual MAS meeting in Breckenridge CO (August 6-11, 1995). Student papers must be submitted, in standard MAS-MSA format to Dr. E. Etz, NIST, Division 837, Bldg. 222, room Q-113, Gaithersburg, MD 20899 by March 15, 1995. The best submitted papers will be awarded funds towards attending the annual meeting in Breckenridge. Any more information about the program can be obtained from Dr. Etz at etz-at-gapnet.nist.gov
What makes this scholarship offer extraordinary is that the best three papers given by student scholarship winners at the Breckenridge meeting will be awarded a minimum of $500 towards the cost of attending the 1st International Union of Microbeam Analysis Societies Conference in Sydney, Australia, February 5-9, 1996. These scholarships are only open to student members of MAS, and student application forms for MAS are available in past issues of Microbeam Analysis, the MAS journal. Student membership is a great bargain at $2.50, and doesn't require that the advisor be a MAS member - although if you aren't, I ask that you consider joining.
I will mail you more information with appropriate details about the meetings and the paper format etc., but if you have any immediate questions, please don't hesitate to contact me by email (dbw1-at-lehigh.edu).
Can anyone out there point me in the right direction for finding information on the health risks associated with epoxy resin dust produced by jewler's saws, jig saws, dremel tools, files, sand paper, etc.. Yes, I have looked at the various MSDS for the resins but they haven't provided much real world information (I at least haven't run into the problem of a tanker truck spillage of NSA).
Richard E. Edelmann Electron Microscopy Facility Supervisor 352 Biological Science Building Miami University, Oxford, OH 45056 Ph: 513-529-5712 E-mail: edelmare-at-muohio.edu
We use double stick tape to fracture the Microcapsules. On a SEM stub we mount an stip of tape. On tape that is covering the stub we add a small amount of dried microcapsules. Using an exta piece of tape cover the tape and capsules and rip off. This will fracture many of the microcapsule so that you will be able to look inside and outside many of them. Coat the stubs and view in the SEM.
Good luck
Mike
--------------------------------------------------------------------------- | Michael Dunlap | lab (916) 752-0284 | | Facility For Advanced Instrumentation | fax (510) 422-2282 | | University of California | mrdunlap-at-ucdavis.edu | | Davis CA, 95616 | | ===========================================================================
I have done many TEM preparations on cell monolayers, using tissue culture dishes and/or glass coverslips. There is a lab here that would like to do light microscopy and TEM on a single cell in a monolayer. I assume this can be done, if I had a coverslip that was etched with a marker grid. This marked grid pattern would have to show on the epoxy face, after the coverslip was separated from the embedding media. Can anyone tell me if there are glass or plastic coverslips that have some type of etched grid pattern that could be used in this way and where they could be ordered in the U.S.
I would prefer working with a plastic/tissue culture type of coverslip, as they are easier to remove before sectioning.
thanks Louisa Howard Ripple Electron Micrscope Facility Dartmouth College Hanover, N.H 03755 Louisa.Howard-at-Dartmouth.edu
I have done many TEM preparations on cell monolayers, using tissue culture dishes and/or glass coverslips. There is a lab here that would like to do light microscopy and TEM on a single cell in a monolayer. I assume this can be done, if I had a coverslip that was etched with a marker grid. This marked grid pattern would have to show on the epoxy face, after the coverslip was separated from the embedding media. Can anyone tell me if there are glass or plastic coverslips that have some type of etched grid pattern that could be used in this way and where they could be ordered in the U.S.
I would prefer working with a plastic/tissue culture type of coverslip, as they are easier to remove before sectioning.
thanks Louisa Howard Ripple Electron Micrscope Facility Dartmouth College Hanover, N.H 03755 Louisa.Howard-at-Dartmouth.edu
I have done many TEM preparations on cell monolayers, using tissue culture dishes and/or glass coverslips. There is a lab here that would like to do light microscopy and TEM on a single cell in a monolayer. I assume this can be done, if I had a coverslip that was etched with a marker grid. This marked grid pattern would have to show on the epoxy face, after the coverslip was separated from the embedding media. Can anyone tell me if there are glass or plastic coverslips that have some type of etched grid pattern that could be used in this way and where they could be ordered in the U.S.
I would prefer working with a plastic/tissue culture type of coverslip, as they are easier to remove before sectioning.
Thanks.
Louisa Howard Ripple Electron Micrscope Facility Dartmouth College Hanover, N.H 03755 Louisa.Howard-at-Dartmouth.edu
If you are going to look at cells in a light microscope, it is a poor idea to use plastic coverslips. You will not get a good image in Differential Interference Contrast or Polarized light...as the cover slip may be strained, and so forth. N.Allen
Hello Microscopists, Although I can't help Richard Edelmann with his question regarding resin dust except to say that we do use dust masks when sawing or cutting blocks; I would like to pose a question closely related to this. How many of you have your curing ovens vented to a hood? We have a VERY low volumn of resins being cured at any given time and have sought, somewhat unsuccessfully, information that would help us determine at what level it would be necessary to do this. As Richard said, the MSDS sheets don't provide much real world information. I did call one of the EM suppliers (I don't recall which one now), and was told that venting to a hood would be necessary only in an industrial situation. I would be very interested in hearing your opinions and how you handle this problem. I would also be willing to summarize any responses and post that to the list.
Thank you very much, Sandra Zane
Sandra F. Zane, EM Tech. Biol. Dept. UNCC Charlotte, NC 28223 sfzane-at-unccvm.uncc.edu Fax (704) 547-3128
We use glass gridded coverslips for tracking cells. You can get them from Bellco Glass, Inc. P.O. Box 340 Edrudo Road Vineland, New Jersey 08360 Cat. # 1916 -92525 Etched gridded coverslips 1 case approx. $30.00.
Patty Jansma Tel:602-621-6671 plj-at-manduca.neurobio.arizona.edu Arizona Research Labs Division of Neurobiology University of Arizona
On 16 Mar 1995, Louisa Howard wrote:
} I have done many TEM preparations on cell monolayers, using tissue culture } dishes and/or glass coverslips. There is a lab here that would like to do light } microscopy and TEM on a single cell in a monolayer. I assume this can be done, } if I had a coverslip that was etched with a marker grid. This marked grid } pattern would have to show on the epoxy face, after the coverslip was separated } from the embedding media. Can anyone tell me if there are glass or plastic } coverslips that have some type of etched grid pattern that could be used in } this way and where they could be ordered in the U.S. } } I would prefer working with a plastic/tissue culture type of coverslip, as } they are easier to remove before sectioning. } } Thanks. } } Louisa Howard } Ripple Electron Micrscope Facility } Dartmouth College } Hanover, N.H 03755 } Louisa.Howard-at-Dartmouth.edu
Hello Microscopists, Although I can't help Richard Edelmann with his question regarding resin dust except to say that we do use dust masks when sawing or cutting blocks; I would like to pose a question closely related to this. How many of you have your curing ovens vented to a hood? We have a VERY low volumn of resins being cured at any given time and have sought, somewhat unsuccessfully, information that would help us determine at what level it would be necessary to do this. As Richard said, the MSDS sheets don't provide much real world information. I did call one of the EM suppliers (I don't recall which one now), and was told that venting to a hood would be necessary only in an industrial situation. I would be very interested in hearing your opinions and how you handle this problem. I would also be willing to summarize any responses and post that to the list.
Thank you very much, Sandra Zane
Sandra F. Zane, EM Tech. Biol. Dept. UNCC Charlotte, NC 28223 sfzane-at-unccvm.uncc.edu Fax (704) 547-3128
Glass coverslips etched with an alphanumeric grid are available from Bellco Glass, Inc. (800-257-7043). I have successfully grown cells in culture on these coverslips for LM and SEM analysis after coating the coverslips with either poly-lysine or collagen. Identified cells are relatively easy to re-identify when going from one system to the other. I have not embedded cells grown on these coverslips, so I do not know if the grid will show up on the surface of the plastic when the coverslip is removed.
Jeff Thompson, Ph.D. Director, Electron Microscope and Image Analysis Center California State University San Bernardino, CA 92407 jthompso-at-wiley.csusb.edu
I must confess I'm a bit perplexed by Z.Q. Liu's question about x-sections of Nano-tubes. If they are truely "nanometer" in size why would you attempt to x-section them? Are they actually larger? All the EM I've seen about nanotubes did not involve x-section, but simple direct imaging.
Okay so let me ask the dumb question. How big are they really?
What is the appropriate way to digitize TEM negatives for HRTEM work at resolution of approx.0.23 nm? Please give approximate cost of the digitizing hardware when possible. Leszek Kepinski
***************************************************************** Leszek Kepinski Institute of Low Temperature and Structure Research, Polish Academy of Sciences, P.O.Box 937, 50-950 Wroclaw, Poland tel:(4871) 350 21 ext.153 fax:(4871) 44 10 29 email: kepinski-at-highscreen.int.pan.wroc.pl *****************************************************************
} I have been asked to analyse some micas for Rb and from a } preliminary examination it looks like I should use the L alpha } line - except that the Si K alpha is rather close making the } Rb sit on the slope to the right of it. } } Anyone have any advice on backgrounds etc? } } I am using TAP xtals, 25 kV,55 nA with a counting time of } 60 seconds for peak and background - is this reasonable? } } Machine is Cameca SX50.
25kV, 55 nA sounds pretty high for routine mica analyses, though it might be OK to analyse the Rb at this condition after analysing the major elements. At 25kV, you could use the Rb KA line on an LIF crystal and avoid the Si overlap. The counting times sound reasonable, though you may have to increase them if you are really interested in "trace" analysis of low levels of Rb.
I suspect that you will always obtain "false" Rb when analysing Rb LA on TAP in the presence of high Si in your sample. The Si KA peak is about -750 steps from the Rb LA line (and about +200 from the Rb LB line). This is far enough that most of the Si peak will be resolved, but there will probably be some tail effect of the Si under the Rb LA peak. You can check for this be analysing SiO2. This will give you the worst case Si interference for silicates. You could continue this process and draw up a working line (curve) of wt.% Si vs. wt.% measured Rb by measuring various silicates with differing Si contents and no Rb. You could then use this curve to correct for your analyses of Rb on your unknowns.
The SX50 software should also give you the chance to use a background "slope" instead of having to take background counts at both +bkgd and -bkgd positions. The "slope" on the older CAMECA MBX software was defined as (bkdg+ + bkgd-)/(2*bkgd+). Thus, a "flat" background would have a slope of 1.000, while a background that increases with lower sin(theta) will have a slope } 1.000. Using a background slope will not solve the tail effect of the Si under the Rb LA, but may help you in using the Rb KA line on LIF, where the bkgd- position may be out of range for the spectrometer.
Good luck!
Carl
Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2005 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA ----------------------------------------------------- (313) 936-1550 (voice) **** Next time, **** (313) 763-4690 (FAX) *** take the *** chender-at-umich.edu (e-mail) **** train! **** -----------------------------------------------------
Dear Nestor, John Cowley gave me reprints of some of his recent papers in which there are images of nanotubes which are on the order of 10 nm across (O.D.). He and his co-workers have found different structures at various places in the nanotubes, so I can see why someone might wish to section them (although it is hard to imagine how this can be done without perturbing (destroying?) the structures). John would likely send you reprints if you asked him. Yours, Bill Tivol
Dear Leszek, In order to retain the original resolution (0.23 nm), the digitization should have a pixel size corresponding to no more than half that resolution. That is, if the magnification of the negative is 100,000x, the pixel size must be ~10 micrometers. This can be done on (among other brands) a Perkin-Elmer flatbed microdensitometer (cost ~ $100,000). A much cheaper alternative is a CCD camera. By arranging to image the negative with the CCD so that the pixels are ~10 micrometers you can get the required resolution. This means that a 1000*1000 CCD array will look at a 1 cm**2 area. This can be done at a cost of ~$50,000 (maybe less--I'm no expert here). You also save scan time with the CCD; however, quantitation of intensities--especially for ED patterns etc. where there are regions of very high OD surrounded be regions of low OD-- is much worse with CCD than with a flatbed scanner. Good luck. Yours, Bill Tivol
Subject: Time: 8:16 AM OFFICE MEMO nanotube x-sect. Date: 3/17/95
Can standard TEM images normal to the axis of the nanotube determine unambiguously the cross sectional shape of tubes (i.e. whether the tubes are facetted, circular cylinders, oval-shaped, hollow or filled with amorphous material, etc?). We think there are 2 ways to get this information: direct imaging of cross-sections or electron holography (in which case you don't need cross-sections). The problem with cross-sections is the potential development of artifacts during the preparation process. We suggest strongly to use electron holography (see for example: J. Mater. Sci., 29 (1994) 5612-5614 (this is on Pd particles, but the principle is the same). Also see our paper which includes nanotubes in the Proceedings of the 1st Int'l Workshop on Electron Holography, Elsevier, in press.
Bill Tivol said in his email that "the digitization should have a pixel size corresponding to no more than half that resolution". I think that this is a typo: you need at least 2 sampling points for a given wave. Therefore if one wants to quantify 0.23nm spacings in HREM the smallest sampling is half this (Nyquist limit) and } 6 samplings is more standard.
I've repeated the original question below, in case someone's missed the thread. I suspect that Liu is trying to make cross-sections to characterize growth mechanisms. Nanotubes, like asbestos fibers, are easily examined in plan-view, since their high aspect ratio virtually guarantees that they will lie flat on a film with the long axis parallel to the film surface. Most of the pictures that I have seen claiming to be nanutubes in cross-section had no supporting evidence that they were anything but an essentially spherically symmetric "bucky onion."
In order to get a true nanotube cross section, one will first probably have to anneal the mass to remove all lesser fullerenes, including the onions. Then break the mass up and embed in a hard epoxy. Although ultramicrotomy isn't a bad idea, there is no real way to align the nanotubes and I'm not certain what effect this would have on cutting. It is probably better just to sandwich between silicon and do a standard cross-section by mechanical polishing, dimpling, and ion-milling. It is going to be hard to prevent rapid milling of the epoxy as opposed to the tubes, so it is important to mechanically thin as far as possible before ion-milling (hence, the silicon as an optical guide).
That's my 2 cents worth.
Daniel L. Callahan Department of Mechanical Engg. and Materials Science Rice University dlc-at-owlnet.rice.edu ---------- Forwarded message ----------
Regarding the recent debate on negative scanning: The Ektron scanner is {$30K, and gives a highly linear image up to 4096 x 4096 pixels. It will permit accurate density corrections to be done for best quantitative results (see Voelkl, et al., Ultramicroscopy 55 (1994) 75-89, for all you could ever want to know about scanning negatives and density corrections etc.)
L.D. Marks is correct--I meant a pixel size corresponding to twice the reso- lution, or half the size. I always seem to have trouble writing about things for which less is better :-). Yours, Bill Tivol
Microscopical Society of Canada 22nd Annual Meeting
The MSC Executive and the Local Organising Committee cordially invite you to attend and participate in the 22nd Annual Meeting of the Microscopical Society of Canada. This meeting will be held in the University Centre Building, University of Ottawa, Ottawa, Ontario, Canada, June 4-7, 1995. A varied and interesting scientific program has been planned and will consist of a combination of interdisciplinary symposia presented by speakers from around the world, separate physical and biological symposia, oral and poster presentations, workshops on TEM specimen preparation of materials and cryo- SEM specimen preparation and X-ray analysis, and commercial exhibits.
Local Organising Committee
Jim Corbett (Chairman, Secretary, University of Ottawa Liaison) John McCaffrey (Treasurer, Space Management, Social Program, Accommodation) Jeff Fraser (Commercial Exhibits, Social Program, Accommodation) Graham Carpenter (Scientific Program Chair, Materials) Larry Arsenault (Scientific Program Chair, Biology) Peter Sewell (Corporate Liaison, Commercial Exhibits) Kamal Botros (Scientific Program) Louise Weaver (Scientific Program) Paula Allan-Wojtas (Registration) Shea Miller (Registration)
DEADLINE FOR RECEIPT OF ABSTRACTS: March 31, 1995
DEADLINE FOR PRE-REGISTRATION: May 1, 1995
For further information contact:
Program: Registration:
Jim Corbett Shea Miller or Paula Allan-Wojtas Department of Physics Centre for Food and Animal Research University of Waterloo Agriculture and Agri-Food Canada Waterloo, Ontario Room 2016, K.W. Neatby Bldg. Canada, N2L 3G1 Central Experimental Farm Ottawa, Ontario, Canada, K1A 0C6 Tel: (519) 885-1211 Tel: (613) 957-4347, ext. 7908 (Shea), 7970 (Paula) Fax: (519) 746-8115 Fax: (613) 943-2353 e-mail: corbett-at-physics.watstar.uwaterloo.ca e-mail: millers-at-ncccot.agr.ca allanwojtasp-at-ncccot.agr.ca
PRELIMINARY MEETING PROGRAM
INTERDISCIPLINARY SYMPOSIUM ADVANCES IN MICROSCOPY
J. Watson (Michigan State University) Events in science and microscopy J. Hillier (Princeton University) Early developments in EM A. Eades (University of Illinois) Electron microscopes of the future O. Krivanek (Gatan Inc) Nanoscale elemental and chemical mapping M. Hansen (FEI Inc.) A comparison of LaB6 and CeB6 thermionic cathodes I.A. Rauf (Queen's University) STM, AFM, CTEM and STEM imaging of polycrystalline tin-doped indium oxide thin films H.J. Kreuzer (Dalhousie University) Lensless low-voltage electron holography
MATERIALS SCIENCES
D. Dorset (University of Buffalo) Direct structure determination by electron crystallography A. Eades (University of Illinois) RHEED: progress towards the extraction of quantitative information P. Midgley (University of Bristol) Electron diffraction for structure determination D. McCombe (McMaster University) Scanning tunnelling microscopy of semiconductor surfaces and interfaces D. Perovic (University of Toronto) Direct imaging of semiconductor dopants by EM M. Loretto (University of Birmingham) Analytical TEM of advanced materials G. Weatherly (McMaster University) Microstructures of SiC-reinforced Mg casting alloys C. Hansen (Queen's University) The application of environmental SEM to studies of concrete
BIOLOGICAL SCIENCES
W. Chiu (Baylor College of Medicine) Cryo-imaging P. Ottensmeyer (University of Toronto) Imaging and computer analysis H. Hagler (University of Texas) Calcium imaging P. Ingram (Research Triangle Inst) Elemental mapping P. Echlin (Cambridge University) Cryo-methods
WORKSHOPS
TEM specimen preparation for Materials Science Topics include: Ion-beam sputtering Cleaving Ultramicrotomy Cross sections, etc.
Cryo-SEM specimen preparation and X-ray analysis Topics include: Fracturing and cryo-planing for morphometric and elemental analysis Cryo-conductive coating Standards for frozen hydrated specimens Light element detection and quantification
Satellite Workshop (June 8, Central Experimental Farm, Ottawa) Applications of Microscopy in Agricultural Research Topics include: Preparative techniques 3-D view of the cell Microtubules Soils in agricultural use Electron microscopy in dairy research Entomology Image analysis-seed quality Image analysis of SDS-PAGE gels Spore development Immunogold labelling Food microstructure
Remember, if the camera is on the ragged edge of 8 bit performance and you want to add gain for more sensitivity, even the smallest amount of gain might put you in the 7 bit range if you we not there already. If you have 10 bit capability on your board why not use a 10 bit camera. * Also a camera specification of 60dB is not a 10 bit camera only an 8. * Also there are many types of noise in a sensor besides thermal noise. Cooling a sensor will reduce (thermal noise only) and allow you to integrate longer. I would double check the impression some make that when a 50db sensor is cooled it offers a better s/n of 10 or 12 bits vs just 7. The reduction of one type of noise (thermal) is all that cooling provides. * Other types of sensor noise besides dark current include photon, nyquest, shot, gain noise, and flicker noise etc. * The signal to noise of the sensor at room temperature is a good measure of what you can expect. ** Look at the quantum efficiency at the nm range needed. ** look at the spectral response of the sensor. ** There are different grades of sensors available relative to dead pixels and gray level variation on other manufacturers cameras. *Digital RS-422 eliminates pixel jitter for sub-pixel accuracy if it is needs versus the genlock action between the digitizer board and the camera as they try to lock to each other in the RS-170 analog mode. *Digital cameras of the higher end variety typically offer 7.5 frames/sec vs 30 (real time) or even less depending on the amount of pixels you need to process. If you wish to do real time imaging 30fps, 7.5fps will not do. Also for fluorescence the shortest duration exposure is preferred. * The camera has to have a good performance to cost ratio. At the low end there are plenty of security type cameras out there, but will they offer you the performance you require. * The big point here is that the higher the signal to noise, the quieter the signal, the more gain that can be added to the signal/ which adds noise as a result, but allows the user to get more sensitivity. Thus if you start with the quietest / highest signal to noise obtainable the better off you are when trying to get more gray levels with minimum sensitivity or more signal to noise allows more gain to be added to achieve a set sensitivity far superior to a signal to noise start point less than the DVC.
The following features listed below are offered by the DVC camera line:
General Statement: Analog video RS-170 focus *Most digitizer boards are typically 8 bits some go higher with digital RS-422 input. With Dc's 10 bits of signal on the analog RS-170 video, this offers first of all more top end room for adding more sensitivity and still maintaining 8 bits/ 256 gray levels for the digitizer board. *You will see more digitizer boards with 10 bit capability on the analog RS-170 input. What will you use as a real time 30 frame / sec camera to match that feature. With most ( real time ) cameras being in the 7 bit range, they would better qualify for security applications. There are some 8 bit cameras, but they only offer 256 gray levels with not much room to add gain for more sensitivity. The DVC camera line offers 10 bits or 1024 gray levels output for those boards along with simultaneous digital RS-422 on the digital models One of the digitizer boards with a 10 bit analog front end is Mutech. They are the only one presently that has that capability for now and others will follow soon.
* Very high signal to noise } 62dB at .5 lux at 30fps/ real time * Frame transfer 1/2" format sensor with 755 horizontal by 484 vertical pixels, or 565 TV lines horizontal by 350 TV lines vertical resolution. * The only upgradable research camera with the researchers funding limitations in mind using the same high quality sensor on all 3 models. Upgradability from the RS-170 analog unit to the dual output 8 bit digital or to the dual output 10 bit digital models with RS-170 analog. (This allows the camera to grow with the researcher) and his budget. See part 3 of 3 next.
And Digitizer Selection Guide For Benefit of Microscope Users Group
Monochrome Digital And Analog Video Camera Line By DVC Company Part 1 of 3 due to capacity limitations of e-mail ***************************************************************** With all due respect: If anyone is offended by a guide to choosing a CCD video camera with a new product release that is applicable for the users in this group ( HIT YOUR DELETE KEY NOW! ) There are some users that might appreciate this information. (This was done with good intentions) so any providers of negative feedback have been given the DELETE KEY OPTION and need not waste net time, thank you. Valid questions are appreciated and I will try to answer them relative to the volume I receive. Included is list of RS-422 digitizer boards in part 2 and 3. * Please address any inquiry directly to dvcco-at-aol.com, not the main file server. ***************************************************************** Dear Microscope Users,
I wish to share some data with you on analog and digital monochrome cameras that I thought you might be able to make use of when reviewing your present or future camera situation. Some of the below information you might be aware of, but others might not. If anyone belongs to other pertinent groups and believes this information to be of benefit to that particular group we would appreciate you letting us know of them.
Monochrome cameras offer much superior gray level and resolution performance along with higher signal to noise specs than the typical color versions. Everyone has different needs and the data below is for researchers who have decided to utilize the benefits of monochrome cameras. Adding pseudo colors to the many more gray levels attainable with a monochrome camera might be a option for you, or adding a tunable liquid crystal filter in line with the camera for serial Red, Green, Blue could also be a option if accurate correlation of the pixel is important. For monochrome applications a electronic tunable filter exists that is very selective with 5,7,10,15,20 ,35 or 50nm band pass filters from 450 to 1050 nm range tunable electronically! See 2 of 2 for info on tunable filters.
Some things to look for in a research grade monochrome camera are the following: * The highest signal to noise possible in a real time 30 frames/sec. unit. Signal to noise is the ultimate measure of a camera and no amount of manipulation, bells and whistles, or 20 different knobs to adjust, will change the bottom line which is the amount of gray levels obtainable. * No geometric distortion- CCD cameras offer basically no distortion while any tube camera gives you distortion in the } 5% range. My opinion is that to use a tube camera where a ccd unit could be substituted is just qualitative, not quantitative microscopy. * No dead pixels on your sensor and lowest pixel variation with no false fill-ins of pixels with data from the pixel next to it by use of memory circuits due to imperfections in the manufacturing process. Different types of CCD cameras have their uses. The 2 main types are frame transfer (FT) and interline transfer (IT) types for microscopy. CID types are also out there but have uses more in machine vision applications. Interline transfer sensors have large gaps between the pixels due to the construction. They offer 100% fill factor by the use of plastic micro lenses that help their sensitivity due to smaller well capacity, but still have the ( same smaller wells.) Plastic does not offer good performance down in the UV range below 400nm, and have other limitations at the near IR range. 1/3 format sensors have even less well capacity and I feel are only produced to provide better yields/profit for the manufacturers for security applications.
Frame transfer sensors which DVC uses have dynamic range of 70dB to start and have no dead pixels and no false fill-ins. The wells are very large and the dark current is {30 electrons at 25C, room temperature. The pixels are contiguous, or within angstroms of electrical separation. Larger wells offer higher signal to noise. Double correlated sample and hold circuits built right into the sensor help reduce noise further on some frame transfer sensors. Frame transfer units offer one half the vertical resolution when doing integration and shuttering due the field transfer technique used. Spectral range with glass face plate attached 400-1100nm. Customers doing (laser) work have reported response in the low 200nm range with face plate removed and up to 1300nm.
Typical bit vs signal to noise ranges: 7 bits = 44dB to technically 50dB but more like 54dB realistically 8 bits = 55dB to 61dB or 256 gray levels 10 bits = 62dB+ for 1024 gray levels. **10 bits offers 4 times the performance of an 8 bit camera. * Another words a camera specification of 50dB falls into the 7 bit range even though manufacturers and sales people might have you believe otherwise. If you have an 8 bit board why not use a 8 bit or higher camera.
Video Camera Selection Guide and DVC Product Release 3 of 3
* Sensitivities in the 1x10-4fc range at over 50% video. When other (IT) type cameras barely see an image these DVC units are offering a perfect image with extra gain to spare and no vertical line noise. Estimate performance at about 5-6 ND filter better sensitivity than 768 type interline transfer sensors. * The only 10 bit digital camera with dual outputs RS-170 & RS-422 and at real time video rates! 30 frames per second * The only camera that has no sensor board. Sensor is physically mounted to the front plate of the camera. Cooling above dew point can then be done to have a truly quantitative camera in that temperature of sensor can be lowered from room temperature or regulated at a constant temperature or our -40C unit can be added. * No need to cool the camera in most applications/ save the money. * The DVC camera line was designed from the ground up to be a digital output camera not a security camera. We use linear well regulated multiple voltage linear power supply and solid C-Mounts with a 360 degree compression ring holding the c-ring firmly in place, not just a little set screw that tends to loosen with time and strip out the threads. * 30dB of additional gain can be added for manual internal adjustable operation accessible via a port from outside the camera, or external gain and offset, or computer controlled gain! 30dB gain vs the others at 20dB because the } 62db performance can fully utilize the 30db range. Where with more than 20db on the IT types you just might end up with snow. * Many other modifications can be done and our units do not come with AGC unless you specify automatics. Automatics are for security cameras. DVC will be responsive to your requirements from the end user to the OEM. * Glass face plate removal to allow the user to easily go down to 300nm or so. Customers with laser applications have reported response in the low 200nm range obviously at low quantum efficiencies.
The DVC camera line is excellent for any high signal to noise or low light application and in real time!
DVC fits a niche between the too pricey and slow non real time digital only video cameras and the security type cameras which don't offer enough of what you need. If you like the resolution DVC can offer our flexibility in being able to upgrade to the digital models is nice to have. If any of you are interested in having a digital camera that works with your Silicon Graphics Indy /Indigo 2 or Sun system, let us know. DVC cameras are made in the USA and have a two year warranty.
* Three models of monochrome camera which are all upgradable to the DVC-10. DVC-0A - RS-170 only with DB-37 connector for remote gain and upgradable to the digital output RS-422 versions offering both outputs. DVC-8 - 8 bit RS-422 digital with RS-170 simultaneously. DVC-10 -10 bit RS-422 digital with RS-170 simultaneously.
*** Below you will find a list of RS-422 digitizer boards that are compatible to the DVC camera line, others are in the works.
****** Compatible Digitizers for RS-170 analog monochrome video are any manufacturer. RS-170 is standard, typically from a BNC connector. Does your board have capability to process RS-170 at 10 bits? The DVC analog output will offer this. Remember if running 10 bit analog RS-170 to a 8 bit board you will only see 8 bits.
Some of these boards have either 8 bit or 10 bit and higher capability. *Compatible digitizers for RS-422 digital video that are plug and play with the DVC digital camera line are listed below:
Manufacturer A-Z Platform Board Name/Model Alacron IBM Alacron Bit Flow IBM/MAC Raptor-VL Coreco IBM F-64, OC-500 Datacube IBM MV-200 Dipix IBM P-360, XPG-1000 Epix IBM 4 Meg, Model 12 EDT SUN S-bus EDT Hyperspeed IBM Hyperspeed Imaging Technology IBM 15040, AFG, MFG Imagraph IBM HI-Def Matrox IBM Magic, 1280, 640, L/C MuTech IBM 10 bit analog video in Precision Digital (PDI) IBM/MAC IMAXX-PCI, SD Univision IBM Piranha
Let DVC know if you wish, the following information: Which digitizer board you are using or would like to use, or need help on. If you have interest in the PCI bus IBM, MAC, Sun, or SGI Indy or Indigo2 computer used If DVC can help answer some of your technical concerns. If your interest is analog or digital or both If your interest is in tunable electronically selectable filters call DVC.
***************************************************************************** ***New DVC Camera Release estimated 4/95 will be the following: Same features as above except now the digital RS-422 can be programmed or reprogrammed for the VINO digital bus on the ((( SGI- Silicon Graphics Indy/Indigo 2 workstations))) Imagine being able to move from SGI to RS-422 digital compatibility with simultaneous RS-170 analog video! Info on our cameras will be on the SGI Web soon. *****************************************************************************
Feel free to e-mail, fax, or call DVC for more information. If DVC can not reach you, we can not help you, so we chose to put this info on the net in hopes of helping anyone that needs it with a cost effective research grade video camera product that we feel, has a definite focus to this microscope group. DVC has heard many of your problems and can be helpful, and possibly offer a solution. Stay in touch.
Sincerely,
Richard Klotsche DVC Company e-mail: dvcco-at-aol.com phone: (619) 444-8300 fax: (619) 444-8321
} I have heard rumors that there will be an international microbeam analysis } meeting in Australia early in 1996. Does anyone know if } this is so; and if it is, do you have any information about when and } where it will be?
The meeting is the first meeting of the International Union of Microbeam Analysis Societies (AKA IUMAS).
It will be held at the University of Sydney, Sydney, NSW Australia for Feb 5 to Feb 9 1996 (Preceeded by workshops) in conjunction with our 14th E.M. Conference and 9th LM symposium. All welcome. Warm weather, water, cool beer. David Cockayne is the chair.
Information on the WWW at http://www.bio.uts.edu.au/em.html
For low cost, high capacity, high resolution and high quality, _nothing_ beats film!
ÒUnfortunatelyÓ if you want to do digital image processing on micrographs then they must be digitised. As previous answers have indicated there is a choice between flat bed scanners and CCD cameras. ThereÕs no doubt that the flatbed scanner produces better data, but the cost is very high. (In addition to purchase price you have maintenance, and so on.). Bill TivolÕs first reply overestimated the cost of the CCD camera solution. You can get away with well under $10000 for camera, lens, stand and light box, a tenth of the price for flat bed scanners.
Optronics made a rotating drum scanner some years ago, but I think itÕs disappeared now. Does anyone know? What did/does it cost?
The Ektron scanner mentioned by Larry Allard is new to me. Does the referenced article give details? Who makes it?
Lab quality CCD cameras often have a geometric distortion between x and y directions of a few percent. This must be measured and corrected for in software. Another limitation is nonlinearity of the response with OD, if you are planning on intensity measurements. Again, it must be measured (with a stepwedge of standard ODs) and corrected for.
CCD cameras have problems with ED patterns, as Bill also pointed out. The very sharp spots with very high gradients cause trouble. I know of some people who are using CCD camera for ED digitising, maybe they can comment in the List about performance.
Another solution is to miss out film and have the CCD camera on the microscope. Horribly expensive, but it has certain advantages. Is anyone digitising ED patterns in this way, and what sort of performance are they getting? I know of one group who is doing so, with success. Are the specifications of on-line CCD cameras so much better than lab ones?
As my old dad used to say ÒYou get what you pay forÓ when confronted with The Sun newspaper at 20p and The Times, at 30p.
I would like to thank everyone that replied to my query concerning networking a Tektronix Phaser IISDX dye sublimation printer. Following is a summary of the problem and the responses.
I wished to put the printer on a Novell network so that it could be accessed from macs, PCs, and Unix boxes. I had purchased Tektronix's Internal Ethertalk card (~$650).
I was told by several listserver users and multiple people at Tektronix that this was absolutely impossible to network the printer under a Novell server using the Ethertalk card. Tektronix put me on to a company in California called Milan that makes a network interface box that should have allowed me to use the parallel port of the printer as the network port. The cost of this box was ~$850.
I was told by several listserver users that it was trivial, no problem, plug it in and it works regarding hooking the printer up to the network.
Finally, I convinced my network administrator to set up a que on one of the network servers and to activate a faceplate so that I could connect my printer to the net. The result .... Novell works fine with Tektronix's Ethertalk card. I have printed from all three platforms and they all work.
The moral of the story: Keep asking your question until you get the answer you want to hear!
p.s. For all of those highly experienced network users who told me in no uncertain terms that I was a fool for not checking with my network administrator before purchasing the printer .... Our local network here has, conservatively speaking, 5,000 users. The network administration is organized with a flowchart of personnel that would make the Ex- Soviet politburo blush. I did try to go through logical channels before making the purchase and was told, "we won't guarantee that it will work, the best we can offer is for you to buy the printer and we'll test it out when it gets here." I suppose this is probably an indication of what the future of networking will be (once everybody's LAN gets so huge).
Sorry for wasting the bandwidth, but last week a 3 part information guide on a particular ccd camera was posted to this forum or the confocal microscopy list server. A disk crash lost my copy of this posting as well as the address of the sender before I had a chance to review it. If the sender could repost a copy directly to me, I would appreciate it. Again, sorry for cluttering your mailbox.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
Well, I was cleaning my email folders on the mainframe and came across this postponed message. Here it is, hope it can still be useful.
Regards, Glen
There is always celloidin, its disadvantages are a lengthy embedding process. It produces a lot of shrinkage in animal tissue, the plant cell wall should resist gross shrinkage, but there may be vacuolization.
For material that has been stained prior to embedding you might try embedding in an epoxy like Pelco's EMBed. You can cut up to 50 microns by either a carbide tungsten knive or using a standard steel knife. These are real knives, not those disposable blades. You also need a solidly built microtome. The carbide knife only needs a slow cutting stroke. The steel knife needs an extremely slow stroke or you can watch the metal forming the knife's edge be ripped away before your very eyes. A small tacking iron, like those used for dry mounting photographs, can be clamped on the end of the knife with a C-clamp. Heat the knife to about 50 deg. C and the plastic will cut more easily.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
On Tue, 8 Nov 1994, Mike Folsom wrote:
} Folks - } } A quick question - } } I'm trying to embed plant material is some type of matrix that will } allow me to make fairly thick sections, say 25 - 50 um. } } I've used paraffin and the tissue starts to fracture when section } thickness gets greater than 20 - 25 um. Frankly besides embedding } the tissue in plastic and then grinding it down I'm not sure } what else to do. } } I'd appreciate any suggestions - } } Michael } } _______________________________________________________________________________ } M.W.Folsom/Biology/UNM/Albuquerque,NM~87131/505.277.4277/mwfolsom-at-hydra.unm.edu } }
Message-Id: {199503210352.AA02291-at-shrike.adl.soils.csiro.au} X-Sender: mcc332-at-shrike.adl.soils.csiro.au X-Mailer: Windows Eudora Version 2.0.3 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Jay Jerome {jjerome-at-isnet.is.wfu.edu}
} Date: Sun, 19 Mar 1995 03:52:28 -0500 } From: DVCCO-at-aol.com } To: Microscopy-at-aaem.amc.anl.gov } Subject: Video Camera Selection Guide1of3 } } Subject: Guide to Choosing A CCD Video Camera With New DVC Product Release } And Digitizer Selection Guide For Benefit of Microscope Users Group } } Monochrome Digital And Analog Video Camera Line By DVC Company } Part 1 of 3 due to capacity limitations of e-mail } ***************************************************************** } With all due respect: If anyone is offended by a guide to choosing a } CCD video camera with a new product release that is } applicable for the users in this group ( HIT YOUR DELETE KEY NOW! ) } There are some users that might appreciate this information. } (This was done with good intentions) so any providers of negative feedback } have been given the DELETE KEY OPTION and need not waste net time, thank you. } Valid questions are appreciated and I will try to answer them relative to } the volume I receive. Included is list of RS-422 digitizer boards in part 2 } and 3. } * Please address any inquiry directly to dvcco-at-aol.com, not the main file } server. } ***************************************************************** } Dear Microscope Users, } } I wish to share some data with you on analog and digital monochrome cameras } that I thought you might be able to make use of when reviewing your present } or future camera situation. Some of the below information you might be aware } of, but others might not. If anyone belongs to other pertinent groups and } believes this information to be of benefit to that particular group we would } appreciate you letting us know of them. } } Monochrome cameras offer much superior gray level and resolution performance } along with higher signal to noise specs than the typical color versions. } Everyone has different needs and the data below is for researchers who have } decided to utilize the benefits of monochrome cameras. Adding pseudo colors } to the many more gray levels attainable with a monochrome camera might be a } option for you, or adding a tunable liquid crystal filter in line with the } camera for serial Red, Green, Blue could also be a option if accurate } correlation of the pixel is important. For monochrome applications a } electronic tunable filter exists that is very selective with 5,7,10,15,20 } ,35 or 50nm band pass filters from 450 to 1050 nm range tunable } electronically! } See 2 of 2 for info on tunable filters. } } Some things to look for in a research grade monochrome camera are the } following: } * The highest signal to noise possible in a real time 30 frames/sec. unit. } Signal to noise is the ultimate measure of a camera and no amount of } manipulation, bells and whistles, or 20 different knobs to adjust, will } change the bottom line which is the amount of gray levels obtainable. } * No geometric distortion- CCD cameras offer basically no distortion while } any tube camera gives you distortion in the } 5% range. My opinion is } that to use a tube camera where a ccd unit could be substituted is just } qualitative, not quantitative microscopy. } * No dead pixels on your sensor and lowest pixel variation with no false } fill-ins of pixels with data from the pixel next to it by use of memory } circuits due to imperfections in the manufacturing process. } Different types of CCD cameras have their uses. } The 2 main types are frame transfer (FT) and interline transfer (IT) } types for microscopy. CID types are also out there but have uses more in } machine vision applications. } Interline transfer sensors have large gaps between the pixels due to the } construction. They offer 100% fill factor by the use of plastic } micro lenses that help their sensitivity due to smaller well capacity, } but still have the ( same smaller wells.) Plastic does not offer good } performance down in the UV range below 400nm, and have other limitations } at the near IR range. 1/3 format sensors have even less well capacity } and I feel are only produced to provide better yields/profit for the } manufacturers for security applications. } } Frame transfer sensors which DVC uses have dynamic range of 70dB to start } and have no dead pixels and no false fill-ins. The wells are very } large and the dark current is {30 electrons at 25C, room temperature. } The pixels are contiguous, or within angstroms of electrical separation. } Larger wells offer higher signal to noise. Double correlated sample and } hold circuits built right into the sensor help reduce noise further on } some frame transfer sensors. Frame transfer units offer one half the } vertical } resolution when doing integration and shuttering due the field transfer } technique used. } Spectral range with glass face plate attached 400-1100nm. Customers } doing (laser) work have reported response in the low 200nm range with } face plate removed and up to 1300nm. } } Typical bit vs signal to noise ranges: } 7 bits = 44dB to technically 50dB but more like 54dB realistically } 8 bits = 55dB to 61dB or 256 gray levels } 10 bits = 62dB+ for 1024 gray levels. **10 bits offers 4 times the } performance of an 8 bit camera. } * Another words a camera specification of 50dB falls into the 7 bit range } even though manufacturers and sales people might have you believe } otherwise. If you have an 8 bit board why not use a 8 bit or higher } camera. } } } --------------------------------------------------------------------------- Stuart G. McClure, | Post small : P.B.#2, Glen Osmond, CSIRO Division of Soils, | Sth Australia, AUSTRALIA, 5064. Adelaide Laboratories, | Post large : Waite Rd, Urrbrae, SA, Australia | Sth Australia, AUSTRALIA, 5064.
Phone: (08) 303-8484 International use +61-8- instead of (08) Fax: (08) 303-8550 Email: Stuart.McClure-at-adl.soils.csiro.au ---------------------------------------------------------------------------
Message-Id: {199503210352.AA02295-at-shrike.adl.soils.csiro.au} X-Sender: mcc332-at-shrike.adl.soils.csiro.au X-Mailer: Windows Eudora Version 2.0.3 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Jay Jerome {jjerome-at-isnet.is.wfu.edu}
} Date: Sun, 19 Mar 1995 03:54:20 -0500 } From: DVCCO-at-aol.com } To: Microscopy-at-aaem.amc.anl.gov } Subject: Video Camera Selection Guide2of3 } } DVC Video Camera Guide And Release 2 of 2 } } Remember, if the camera is on the ragged edge of 8 bit performance and } you want to add gain for more sensitivity, even the smallest amount of } gain might put you in the 7 bit range if you we not there already. } If you have 10 bit capability on your board why not use a 10 bit camera. } * Also a camera specification of 60dB is not a 10 bit camera only an 8. } * Also there are many types of noise in a sensor besides thermal noise. } Cooling a sensor will reduce (thermal noise only) and allow you to } integrate longer. I would double check the impression some make that } when a 50db sensor is cooled it offers a better s/n of 10 or 12 bits vs } just 7. } The reduction of one type of noise (thermal) is all that cooling } provides. } * Other types of sensor noise besides dark current include photon, nyquest, } shot, gain noise, and flicker noise etc. } * The signal to noise of the sensor at room temperature is a good measure } of what you can expect. } ** Look at the quantum efficiency at the nm range needed. } ** look at the spectral response of the sensor. } ** There are different grades of sensors available relative to dead } pixels and gray level variation on other manufacturers cameras. } *Digital RS-422 eliminates pixel jitter for sub-pixel accuracy if it is } needs versus the genlock action between the digitizer board and the } camera as they try to lock to each other in the RS-170 analog mode. } *Digital cameras of the higher end variety typically offer 7.5 frames/sec } vs 30 (real time) or even less depending on the amount of pixels you need } to process. If you wish to do real time imaging 30fps, 7.5fps will not } do. Also for fluorescence the shortest duration exposure is preferred. } * The camera has to have a good performance to cost ratio. At the low end } there are plenty of security type cameras out there, but will they offer } you the performance you require. } * The big point here is that the higher the signal to noise, the quieter } the signal, the more gain that can be added to the signal/ which adds noise } as a result, but allows the user to get more sensitivity. } Thus if you start with the quietest / highest signal to noise obtainable } the better off you are when trying to get more gray levels with minimum } sensitivity or more signal to noise allows more gain to be added to achieve } a set sensitivity far superior to a signal to noise start point less than } the DVC. } } The following features listed below are offered by the DVC camera line: } } General Statement: Analog video RS-170 focus } *Most digitizer boards are typically 8 bits some go higher with digital } RS-422 } input. With Dc's 10 bits of signal on the analog RS-170 video, this offers } first of all more top end room for adding more sensitivity and still } maintaining } 8 bits/ 256 gray levels for the digitizer board. } *You will see more digitizer boards with 10 bit capability on the analog } RS-170 } input. What will you use as a real time 30 frame / sec camera to match that } feature. With most ( real time ) cameras being in the 7 bit range, they } would } better qualify for security applications. There are some 8 bit cameras, but } they only offer 256 gray levels with not much room to add gain for more } sensitivity. The DVC camera line offers 10 bits or 1024 gray levels output } for those boards along with simultaneous digital RS-422 on the digital models } One of the digitizer boards with a 10 bit analog front end is Mutech. They } are the only one presently that has that capability for now and others will } follow soon. } } * Very high signal to noise } 62dB at .5 lux at 30fps/ real time } * Frame transfer 1/2" format sensor with 755 horizontal by 484 vertical } pixels, or 565 TV lines horizontal by 350 TV lines vertical resolution. } * The only upgradable research camera with the researchers funding } limitations in mind using the same high quality sensor on all 3 models. } Upgradability from the RS-170 analog unit to the dual output 8 bit } digital or to the dual output 10 bit digital models with RS-170 analog. } (This allows the camera to grow with the researcher) and his budget. } See part 3 of 3 next. } } --------------------------------------------------------------------------- Stuart G. McClure, | Post small : P.B.#2, Glen Osmond, CSIRO Division of Soils, | Sth Australia, AUSTRALIA, 5064. Adelaide Laboratories, | Post large : Waite Rd, Urrbrae, SA, Australia | Sth Australia, AUSTRALIA, 5064.
Phone: (08) 303-8484 International use +61-8- instead of (08) Fax: (08) 303-8550 Email: Stuart.McClure-at-adl.soils.csiro.au ---------------------------------------------------------------------------
Message-Id: {199503210352.AA02299-at-shrike.adl.soils.csiro.au} X-Sender: mcc332-at-shrike.adl.soils.csiro.au X-Mailer: Windows Eudora Version 2.0.3 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Jay Jerome {jjerome-at-isnet.is.wfu.edu}
} Date: Sun, 19 Mar 1995 03:54:55 -0500 } From: DVCCO-at-aol.com } To: Microscopy-at-aaem.amc.anl.gov } Subject: Video Camera Selection Guide3of3 } } Video Camera Selection Guide and DVC Product Release 3 of 3 } } * Sensitivities in the 1x10-4fc range at over 50% video. When other (IT) } type cameras barely see an image these DVC units are offering a perfect } image with extra gain to spare and no vertical line noise. Estimate } performance at about 5-6 ND filter better sensitivity than 768 type } interline transfer sensors. } * The only 10 bit digital camera with dual outputs RS-170 & RS-422 and at } real time video rates! 30 frames per second } * The only camera that has no sensor board. Sensor is physically mounted } to the front plate of the camera. Cooling above dew point can then be } done to have a truly quantitative camera in that temperature of sensor } can be lowered from room temperature or regulated at a constant } temperature or our -40C unit can be added. } * No need to cool the camera in most applications/ save the money. } * The DVC camera line was designed from the ground up to be a digital } output camera not a security camera. We use linear well regulated } multiple voltage linear power supply and solid C-Mounts with a 360 } degree compression ring holding the c-ring firmly in place, not just a } little set screw that tends to loosen with time and strip out the } threads. } * 30dB of additional gain can be added for manual internal adjustable } operation accessible via a port from outside the camera, or external gain } and offset, or computer controlled gain! 30dB gain vs the others at 20dB } because the } 62db performance can fully utilize the 30db range. Where } with more than 20db on the IT types you just might end up with snow. } * Many other modifications can be done and our units do not come with AGC } unless you specify automatics. Automatics are for security cameras. } DVC will be responsive to your requirements from the end user to the OEM. } * Glass face plate removal to allow the user to easily go down to 300nm or } so. Customers with laser applications have reported response in the low } 200nm range obviously at low quantum efficiencies. } } The DVC camera line is excellent for any high signal to noise or low light } application and in real time! } } DVC fits a niche between the too pricey and slow non real time digital only } video cameras and the security type cameras which don't offer enough of what } you need. If you like the resolution DVC can offer our flexibility in being } able to upgrade to the digital models is nice to have. } If any of you are interested in having a digital camera that works with your } Silicon Graphics Indy /Indigo 2 or Sun system, let us know. } DVC cameras are made in the USA and have a two year warranty. } } * Three models of monochrome camera which are all upgradable to the DVC-10. } DVC-0A - RS-170 only with DB-37 connector for remote gain and upgradable } to the digital output RS-422 versions offering both } outputs. } DVC-8 - 8 bit RS-422 digital with RS-170 simultaneously. } DVC-10 -10 bit RS-422 digital with RS-170 simultaneously. } } *** Below you will find a list of RS-422 digitizer boards that are compatible } to the DVC camera line, others are in the works. } } ****** Compatible Digitizers for RS-170 analog monochrome video are any } manufacturer. RS-170 is standard, typically from a BNC connector. } Does your board have capability to process RS-170 at 10 bits? } The DVC analog output will offer this. Remember if running 10 bit analog } RS-170 to a 8 bit board you will only see 8 bits. } } Some of these boards have either 8 bit or 10 bit and higher capability. } *Compatible digitizers for RS-422 digital video that are } plug and play with the DVC digital camera line are listed below: } } Manufacturer A-Z Platform Board Name/Model } Alacron IBM Alacron } Bit Flow IBM/MAC Raptor-VL } Coreco IBM F-64, OC-500 } Datacube IBM MV-200 } Dipix IBM P-360, XPG-1000 } Epix IBM 4 Meg, Model 12 } EDT SUN S-bus EDT } Hyperspeed IBM Hyperspeed } Imaging Technology IBM 15040, AFG, MFG } Imagraph IBM HI-Def } Matrox IBM Magic, 1280, 640, L/C } MuTech IBM 10 bit analog video in } Precision Digital (PDI) IBM/MAC IMAXX-PCI, SD } Univision IBM Piranha } } Let DVC know if you wish, the following information: } Which digitizer board you are using or would like to use, or need help on. } If you have interest in the PCI bus } IBM, MAC, Sun, or SGI Indy or Indigo2 computer used } If DVC can help answer some of your technical concerns. } If your interest is analog or digital or both } If your interest is in tunable electronically selectable filters call DVC. } } ***************************************************************************** } ***New DVC Camera Release estimated 4/95 will be the following: } Same features as above except now the digital RS-422 can be programmed or } reprogrammed for the VINO digital bus on the } ((( SGI- Silicon Graphics Indy/Indigo 2 workstations))) } Imagine being able to move from SGI to RS-422 digital compatibility } with simultaneous RS-170 analog video! } Info on our cameras will be on the SGI Web soon. } ***************************************************************************** } } Feel free to e-mail, fax, or call DVC for more information. } If DVC can not reach you, we can not help you, so we chose to put this info } on } the net in hopes of helping anyone that needs it with a cost effective } research } grade video camera product that we feel, has a definite focus to this } microscope group. DVC has heard many of your problems and can be helpful, } and } possibly offer a solution. Stay in touch. } } Sincerely, } } Richard Klotsche } DVC Company } e-mail: dvcco-at-aol.com } phone: (619) 444-8300 } fax: (619) 444-8321 } } } } } } } --------------------------------------------------------------------------- Stuart G. McClure, | Post small : P.B.#2, Glen Osmond, CSIRO Division of Soils, | Sth Australia, AUSTRALIA, 5064. Adelaide Laboratories, | Post large : Waite Rd, Urrbrae, SA, Australia | Sth Australia, AUSTRALIA, 5064.
Phone: (08) 303-8484 International use +61-8- instead of (08) Fax: (08) 303-8550 Email: Stuart.McClure-at-adl.soils.csiro.au ---------------------------------------------------------------------------
With reference to embedding plant material for thick(ish) sections. We routinely use LR White (London Resin White acrylic resin for plant material and can certainly cut sections up to 15-20um. The nice thing about LR white is that is like the British, tolerant and kind to specimens and you need only dehydrate to 80% EtOH and still get godd embedding. You can use all sorts of light microscopy stains on sections cut from such material.
Greetings from a beautiful spring morning in Cambridge where all the daffodils are in full bloom along the Backs.
Patrick Echlin
Multi-Imaging Centre School of Biological Sciences
On Mon, 20 Mar 1995, Glen Macdonald wrote:
} Well, I was cleaning my email folders on the mainframe and came across } this postponed message. Here it is, hope it can still be useful. } } Regards, } Glen } } } There is always celloidin, its disadvantages are a lengthy embedding } process. It produces a lot of shrinkage in animal tissue, the plant } cell wall should resist gross shrinkage, but there may be vacuolization. } } For material that has been stained prior to embedding you might try } embedding in an epoxy like Pelco's EMBed. You can cut up to 50 microns } by either a carbide tungsten knive or using a standard steel knife. } These are real knives, not those disposable blades. You also need a } solidly built microtome. } The carbide knife only needs a slow cutting stroke. The steel knife } needs an extremely slow stroke or you can watch the metal forming the } knife's edge be ripped away before your very eyes. A small tacking iron, } like those used for dry mounting photographs, can be clamped on the end of } the knife with a C-clamp. Heat the knife to about 50 deg. C and the } plastic will cut more easily. } } Glen MacDonald } Hearing Development Laboratories RL-30 } University of Washington } Seattle, WA 98195 } (206)543-8360 } glenmac-at-u.washington.edu } } } On Tue, 8 Nov 1994, Mike Folsom wrote: } } } Folks - } } } } A quick question - } } } } I'm trying to embed plant material is some type of matrix that will } } allow me to make fairly thick sections, say 25 - 50 um. } } } } I've used paraffin and the tissue starts to fracture when section } } thickness gets greater than 20 - 25 um. Frankly besides embedding } } the tissue in plastic and then grinding it down I'm not sure } } what else to do. } } } } I'd appreciate any suggestions - } } } } Michael } } } } _______________________________________________________________________________ } } M.W.Folsom/Biology/UNM/Albuquerque,NM~87131/505.277.4277/mwfolsom-at-hydra.unm.edu } } } } } }
I'm a recent subscriber and am wondering if perhaps there is another listserver that might be closer to my fields of interest, i.e. micro-manipulation of cellular and sub-cellular particles either via mechanica l or optical trapping techniques. If anyone knows of a group that is more involved in these areas, I would appreciate if you could reply to me directly.
WINTHROP UNIVERSITY Electronic Mail Message Date: 21-Mar-1995 12:50pm EST From: Kenneth Gregg GREGGK Dept: Biology Tel No: 803-323-2111
TO: Remote Addressee ( _smtp%"microscopy-at-aaem.amc.anl.gov" )
WINTHROP UNIVERSITY Electronic Mail Message Date: 21-Mar-1995 11:31am EST From: Kenneth Gregg GREGGK Dept: Biology Tel No: 803-323-2111
TO: Remote Addressee ( smtp%"microscopy-at-aaem.amc.anl.gov" )
Greetings, My guess is that many members of the list teach a histology course at the University level. I would like some comments on a method of teaching the laboratory section of histology which we are considering. We find that many students are rapidly bored when looking at tissue/organ sections in the light microscope. They also seem to lack the ability to know when they have acquired an acceptable level of expertise. As an antidote to this we are kicking around the following idea -- and we would like to know if any members of the list have tried similar approaches, and with what success?
We are thinking about having individual students prepare computer-assisted-instruction lessons on topics such as epithelial tissues, liver structure, etc. We would like to have the students prepare a hypertext program for each topic. They would integrate the hypertext with captured images from their light microscope observations as well as scanned diagrams and electron micrographs which they could obtain from texts or photographs. Students would be very much engaged in their own productions and could study lessons prepared by other students. These programs could be used and improved in subsequent years by other students. They could also be made available for other institutions.
If you prefer, you may respond to me personally at Greggk-at-Winthrop.edu
We are seriously interested in purchasing a setup for our Nikon Diaphot that could be used for the following: o quantitation of very low level fluorescence-- sensitive and linear o but also useful for critical Nomarski applications such as microtubule motility assays o automation of filter wheels for multiple probes or fluorescence / phase contrast image collection o automation of stage moving, for instance for making mosaics automatically or being able to move back to a specific field o maybe Z motor and deconvolution, although the are not our main concerns o ESSENTIAL: we are a multi-user facility so we need ease of operation
We are considering a Photometrics or Princeton Instruments cooled CCD camera. We have looked at Oncor Imaging, I.P. Lab and have spoken to other companies. For hardware, we have called companies that offer individual parts (e.g. just the filter wheels) and, of course, have checked out Ludl. We love NIH-Image, but want something does not require a large amount of macro or other programming to get the hardware to work; basically, we want plug and play. Any suggestions or "stay away from this product at all costs" comments on parts of systems or whole systems would be appreciated. Thanks- Michael Cammer cammer-at-aecom.yu.edu
Hi there, Could you help me with the problem of delineation of titanium in bone and soft tissue.Any information and/or references would be a great help to me. Study will involve the use of both SEM and EPMA, possibly FTIR as well. Thank you all, Wis Jablonski , Email W.Jablonski-at-csl.utas.edu.au
Place: Convene at Forensic Science Associates in Richmond, Conclude at Oakland PD. (See contact information below for reservations, maps, car pooling, and further details.)
Time: Convene by 10:00 am, Adjourn by 5:00 pm
Summary: This promises to be a most informative and interesting meeting as we hear from some of the Bay Areas foremost forensic microscopists, see some of the latest technology in microscopy, and hear about some interesting forensic applications of microscopy.
Agenda:
Forensic Science Associates, 10:00 am. Stephen A. Shaffer of MicroDataware will introduce the topic of forensic science in general and forensic microscopy in particular. Peter D. Barnett of Forensic Science Associates will then discuss two interesting cases involving forensic microscopy, and also the role of the consulting (i.e., non-government) microscopist in forensic cases. Pete will discuss cases involving questions of ballot marking and alleged surgical mistakes revealed by fibers in the nose! We'll conclude at FSA by 11:30 or 11:45.
Bureau of Alcohol, Tobacco, and Firearms, 1:00 pm. After a brief break for lunch at a Deli in Walnut Creek, we'll re-convene at the ATF Laboratory where we will divide into two groups to cover separate topics with two speakers. John Murdock will explain and demonstrate the use of comparison microscopy in firearms identification. John is the former Director of the Contra Costa County Crime Laboratory and also teaches criminalistics locally. He is an excellent speaker and promises a fascinating discussion. Also at ATF, we will meet with Robert Thompson who will demonstrate a new technology for automated comparison microscopy. This technology may revolutionize firearms work in criminalistics laboratories by screening cases and suggesting most promising comparisons for the human microscopist to perform. We will adjourn from ATF by 2:30 to proceed to the Oakland Police Department.
Oakland Police Department, 3:00 pm. At OPD, Diane Bowman will discuss the use of microchemical testing procedures for the identification of illicit drugs and narcotics, showing both the utility and the beauty of these procedures for the rapid identification of controlled substances. Mary Gibbons, Director of the OPD Lab, will discuss a fascinating case where microscopic particles of spray paint, identified and compared by microscopy, were instrumental in unraveling a suspect's story, leading to solution of a homicide. We will aduourn from OPD by 5:00 pm.
At both ATF and OPD, each of our sub-groups will take turns with each of the speakers so we all get to hear all of the topics. Each topic will be covered in a presentation of approximately 45 minutes duration.
Reservations Required!!! Because of the limited space available within the microscopy labs we will visit, it may be necessary to limit the number of people who attend. No more than 20 can be accommodated so call Steve Shaffer ASAP to reserve your space, receive maps to the locations, and to coordinate with others on car pooling to the three sites we will visit.
Contact: Stephen A. Shaffer at the numbers below. Don't delay, call right away!
************************************************************ * Stephen A. Shaffer * Publishers of The Particle Atlas * * MicroDataware * on CD-ROM. Developers of custom * * 2894 Tribune Avenue * image and database software for * * Hayward CA 94542-1637 * laboratories, specializing in * * 1-510-582-6624 voice * light and electron microscopy. * * 1-510-582-6624 fax * Email inquiries invited. * * sshaffer-at-microdataware.com * ************************************************************
Plant material can also be embedded in butyl methyl methacrylate and sectioned up to 20 um. Stick the sections down to glass slides with polyethylenimine. The resin is removed with acetone before antibody staining, but you don't need to remove it if staining with something small and water-soluble like aniline blue.
____________________________________________________________ Rosemary White __ / Department of Ecology _/ \__/ \ and Evolutionary Biology / \ Monash University / Australia \ Clayton, Victoria 3168 \ ____ / phone 61-3-905 5670 \_/ \_*_/ fax 61-3-905 5613 __ email r.g.white-at-sci.monash.edu.au \/
Message-Id: {1995Mar21.151709.2878643829-at-dmcmail.ucsf.edu} To: microscopy-at-aaem.amc.anl.gov (microscopy)
Subject: Time: 2:10 PM OFFICE MEMO Usage "Tracking" Programs Date: 3/21/95 Hi everybody! Does anyone have any usage tracking programs, or know where they might be obtained? I am especially interested in a DOS program that is compatible with Meridian software. Does anyone have any experience using tracking programs? I am also interested in programs that are compatible with HP and Macintosh acquisition/analysis software for flow cytometers (primarily Becton Dickinson.) Any suggestions or comments would be greatly appreciated. Thank you very much!
__________________________________________________________ Kris Kavanau Internet: kavanau-at-dmc.ucsf.edu Manager, Lab for Cell Analysis Telephone: 415-476-2631 Division of Molecular Cytometry FAX: 415-476-8218 University of California San Francisco, CA 94143 __________________________________________________________
-- [ From: Charles A. Garber, PH. D. * EMC.Ver #2.10P ] --
SPI Supplies
For TEM examination, we using the following procedure:
1] Pick up the unstained sections on a silicon dioxide coated TEM grid,
2] Gently etch (e.g. 10 seconds) the now supported section in a barrel geometry reactive etcher (for example, the SPI Model Plasma Prep II) using oxygen as the etching gas, which will immediately etch away all traces of the organic matrix, leaving the inorganics dispersed, in their original locations, on the silicon dioxide support film. Usually these residues form a ghost type of outline of original cell shapes. I don't know what bone might look like if etched this way.
The reason for the silicon dioxide support film is that it will not be etched by the oxygen plasma, where as any kind of organic or carbonaceous support film would otherwise be etched away.
The reason for the etching to remove the organic part of the sample is to reduce to almost zero the Bremstrahlung radiation, thereby increasing greatly the lower detection limits.
3] We have always found the TEM to be better for doing this kind of analysis than SEM/EDS, mainly because of the higher resolution possibilities.
Let me know if you would need further information. The making of the silicon dioxide coated grids requires a bit of art but they are not all that difficult to make, so long as you are patient.
Charles A. Garber, Ph. D. PRESIDENT SPI SUPPLIES PO Box 656 West Chester, PA 19381-0656 USA
The following position is available at the Johns Hopkins U. School of Medicine:
Electron Micrscopy Technician/19 hrs/wk PART-TIME:
Position will be responsible for semi- and ultra-thin sectioning on glass and diamond knives. Standard grid prep and staining of sections; maintenance of electron gun parts (polishing cleaning etc.). Replenising of photo- and lab chemical stocks, care of peripheral equipment and other related duties. College degree, BS or equivalent and minimum two years experience. Special skills/knowledge of diamond knive use and maintenance, routine EM lab operations.
All interested candidates should contact: Lou Cote (410) 955-0519 Job # M.3531.94
Kris et al, I use DIRECT ACCESS from Fifth Generation Systems. It keeps track of the time, reqires a pssword and has the option of also requiring a project name. This forces the issue with people who think it's better for me to chase down their grant number then for them to remember, and makes billing much easier. We also keep a log book for people to record problems and i modify billings accordingly. When I came here this was already installed on the XRD computer so I did no comparative shopping, & I don't know what else is out there. Their address is FIFTH GENERATION SYSTEMS INC 10049 n. Reiger Rd Baton Rouge LA 70809 (504) 291-7221
Maggy Piranian Memorial U of Newfoundland
On Tue, 21 Mar 1995 Kris_Kavanau-at-dmcmail.ucsf.edu wrote:
} Subject: Time: 2:10 PM } OFFICE MEMO Usage "Tracking" Programs Date: 3/21/95 } Hi everybody! } Does anyone have any usage tracking programs, or know where they might be } obtained? I am especially interested in a DOS program that is compatible } with Meridian software. Does anyone have any experience using tracking } programs? I am also interested in programs that are compatible with HP } and Macintosh acquisition/analysis software for flow cytometers (primarily } Becton Dickinson.) } Any suggestions or comments would be greatly appreciated. Thank you very } much! } } __________________________________________________________ } Kris Kavanau Internet: } kavanau-at-dmc.ucsf.edu } Manager, Lab for Cell Analysis Telephone: 415-476-2631 } Division of Molecular Cytometry FAX: 415-476-8218 } University of California } San Francisco, CA 94143 } __________________________________________________________ } } } } }
We regrettably learned Monday, of the untimely death this last Saturday of Mort Maser an individual whose efforts and contributions to the Microscopy Community here in the US are well known and valued by many of us.
Mort will be certainly missed and may I speak for the entire MSA family in expressing our condolences to his family and friends. For those of you who are interested I have gotten some details concerning a memorial for Mort and detail those below.
In memory of Mort, his family is establishing the Morton D Maser Scholarship Fund. Contributions can be sent to: Larry Maser, P.O. Box EM, Woods Hole, MA 02543. Please designate your gift to the Morton D Maser Scholarship Fund.
Memorial services celebrating Mort's life are being planned for the summer months. Details will be announced as they are finalized.
Morton D "Mort" Maser, President of Woods Hole Educational Associates, and Executive Secretary to Council of both the Microscopy Society of America and the Histochemical Society, died on Saturday, March 18, 1995 at the age of 60.
He earned his A.B. from the University of Pennsylvania in 1955, and his Ph.D. in biophysics from the University of Pittsburgh in 1962. He held research and faculty positions at the Mellon Institute, Harvard University, the Millard Fillmore Hospital, Northeastern University, Creighton University, and the Marine Biological Laboratory.
He developed roughly 100 short courses in electron microscopy and related fields for scientists and technicians, and made more than 50 contributions to peer-reviewed publications on electron microsopy and related topics.
He was a member of several professional societies including serving as Chair of EMSA's Education Committee; past President, Director of the New England Society for Electron Microscopy; the American Association for the Advancement of Science; American Society of Cell Biology; and the International Society for Stereology.
His research has spanned the sciences from electron microscopical techniques, to computer sciences, to software systems.
He is survived by his wife, Naomi, children Larry of Forestdale, Massachusetts and Jill, of Philadelphia, and grandson, Benjamin.
I was following the discussion on flatbed scanners with great interest, but I need to find some vendors to request additional information. The following units were mentioned: Umax PowerLink AGFA Arcus Perkin-Elmer and Optronics
Does any one have address, phone number or E-mail address for these or other vendors?
In message Mon, 20 Mar 95 14:23:11 EST, forbes-at-cip.org.ec writes: We would greatly appreciate } suggestions on other ways to clear and fix this tissue that do not } involve lactophenol or other products which require special } disposal facilities, which currently do not exist in Quito. *******
We routinely use 1 M NaOH to clear flowers to check in vivo pollen growth after Aniline Blue staining (for fluorescence). You might also want to try a clearing protocol specifically designed for fungal infected leaf tissue such as the one in the following publication:
AU HIGNIGHT-K-W. MUILENBURG-G-A. VAN-WIJK-A-J-P.
TI A CLEARING TECHNIQUE FOR DETECTING THE FUNGAL ENDOPHYTE ACREMONIUM- SP
IN GRASSES
SO BIOTECH HISTOCHEM
68 (2). 1993. 87-90.
JT BIOTECHNIC & HISTOCHEMISTRY.
KW FESTUCA-ARUNDINACEA FESTUCA-OVINA LOLIUM-PERENNE BRIGHT FIELD
MICROSCOPY ANALYTICAL METHOD
AB Leaf tissue of tall fescue Festuca arundinacea Schreb., hard fescue
Festuca ovina L., red fescue Festuca rubra L. and Perennial ryegrass
Lolium perenne L. was stained with rose Bengal or aniline blue to
detect the presence of the fungal endophyte Acremonium sp.. Specimens
were cleared using methyl salicylate, an optical clearing agent, and
viewed using bright field microscopy. Tissue was preserved as dried
tissue or stored in 70% aqueous ethyl alcohol before staining and
clearing. Tissue was observed at 2, 4 and 12 weeks following clearing
to check for stain retention. Staining with rose Bengal was inferior to
aniline blue when followed by the clearing agent methyl salicylate.
Fungal mycelia stained lighter with rose Bengal and were more difficult
to detect than mycelia stained with aniline blue. The results
illustrate the usefulness of combining staining and methyl salicylate
clearing for detecting fungal endophytes.
___________
Good Luck!
************************************************************************* M.V. Parthasarathy Professor of Plant Biology Section of Plant Biology 228 Plant Science Building Cornell University, Ithaca, NY 14850 Telephone: (607) 255-1734 Fax: (607) 255-5407 E-mail: mvp2-at-cornell.edu ***********************************************************************
Have you tried calling Universal Imaging (you mentioned some of the other competitive companies). They are at 610-344-9410. Their new metamorph system will do just about everything you requested...and works well with a cooled CCD. Nina Allen
} Date sent: Mon, 27 Feb 1995 09:30:43 -0500 (EST) } From: YANGA-at-NCCCOT.AGR.CA } Subject: RE:35mm film in TEM } To: microscopy-at-aaem.amc.anl.gov
} We use Eastman motion picture film (5302 fine grain release } positive film) in Philips EM300 in the past and currently in } Zeiss EM902 without any problem. } } Ann Fook Yang
We use copex PET10 in our CM12 and 301. It doesn't outgase much, is very thin so is easy to handle and has not sprocket hole to interfere with the images (depending onthe format).
I was following the discussion on flatbed scanners with great interest, but I need to find some vendors to request additional information. The following units were mentioned: Umax PowerLink AGFA Arcus Perkin-Elmer and Optronics
Does any one have address, phone number or E-mail address for these or other vendors?
} To: microscopy-at-aaem.amc.anl.gov } From: delannoy-at-welchlink.welch.jhu.edu (Michael Delannoy) } Subject: EM TECH POSITION } } The following position is available at the Johns Hopkins U. School of Medicine: } } Electron Micrscopy Technician/19 hrs/wk PART-TIME: } } Position will be responsible for semi- and ultra-thin sectioning on } glass and diamond knives. Standard grid prep and staining of sections; maintenance } of electron gun parts (polishing cleaning etc.). Replenising of photo- and lab } chemical stocks, care of peripheral equipment and other related duties. } College degree, BS or equivalent and minimum two years experience. Special } skills/knowledge of diamond knive use and maintenance, routine EM lab operations. } } All interested candidates should contact: } Lou Cote } (410) 955-0519 } Job # M.3531.94 } } or } } Mike Delannoy } (410) 955-1365 }
Dear Friends, As you all may remember, I was having crossing over problem between FITC & Rhodamine. I followed the suggestion that some of you gave me and used Cy5 (1.4 mg/ml) 1:100 and 1:400 and Cy3 (1.4 mg/ml) same dilution. I did a control and I got a lot background staining. Any suggestion? My next step will be to check my filters as some of you suggested. Nichole Dutton suggested that before applying primaries antibodies to block for nonspecifics with 20% horse serum solution, and use 1% bovine serum albumin in my diluents. Nichole, I'd like further information on this. Dave, also suggested using different blocking reagent (BSA, milk, gelatin, serum.) I'd like to learn more about this possibility, Dave. Thanks, Ciprian
__________________________________________________________ Ciprian Almonte Medical College of Pennsylvania Dept. of Anatomy and Neurobiology E-mail: almonte-at-medcolpa.edu 3200 Henry Ave. Voice: (215) 842-4081 Philadelphia, PA 19129 Fax: (215) 843-9082 __________________________________________________________
Dear Frank, There was a used Perkin-Elmer scanner available, which I heard about from the microscopy list. I don't remember the company name, since we didn't have the money to get the scanner. They sent me a write-up with specs, but I don't know whether I still have it. Perhaps you or Nestor can search the ar- chives for the posting. Good luck. Yours, Bill Tivol
REGARDING Position Available for FIB Engineer Position available for a FIB engineer to join a team working on process development and failure analysis of ULSI circuits in Motorola, Advanced Products Research and Development Laboratory, Austin, Texas. The engineer will be responsible for the setup and operation of a focused-ion-beam facility. The candidate must have hands-on experience on FIB. Qualified candidates please send resumes to
ra2169-at-email.mot.com
or mail to
David Hwang Motorola APRDL, K-10 3501 Ed Bluestein Boulevard Austin, TX 78721
MR-Received: by mta GATEV3; Relayed; Thu, 23 Mar 1995 12:19:38 -0600 (CST) Alternate-recipient: prohibited Disclose-recipients: prohibited
Hi, I am a TEM sample prep technician and is currently preparing samples by self supportive dimpling, tripod and acid etch(mainly for planview)techniques. I am interested in new methods out there being developed and used successfully. Is anyone out there preparing samples using FIB(Focus Ion Beam) as a tool.We have used it once in a while to thin our samples prepared with the wedge technique in the past, but would like to know more.We own a FEI800, but is currently used extensively for SEMs. Also any tips,and/or problems to avoid, associated with the sample prep(especially in the milling dept.) are most welcome. Most of my contamination problem occurs in the ion-milling process. Thanks, Lata Prabhu
Hello all- I am looking for recommendations for evaporation material used in rotary shadowing of proteins. I am referencing the text "Electron Microscopy in Molecular Biology", a practical approach; ed. by Sommerville and Scheer. The method described uses a Balzers electron beam gun to evaporate tungsten/tantalum. We do not have such a gun but plan to use a standard thermal evaporation. Anyone out there have experience/recommendations for evaporation metal, e.g., platinum, tungsten, palladium, carbon/platinum for finer grains?
Thanks, Doug Davis EML Berkeley (510) 642-2085 doug_davis-at-maillink.berkeley.edu
Job opening: Staff Research Associate II Electron Microscope Laboratory University of California at Berkeley $29,200 per annum, 80-100% appointment Job listing #03-345-30/SL closing date: 4-14-95 Contact the Office of Employment Room 7-G 2200 University Ave., Berkeley, CA 94720 (510) 642-1011 general info
Operate and maintain an SEM. Train users in SEM technique, including specimen preparation. Learn operation of JEOL 9000 freeze-fracture machine and train and supervise others in its use. Prepare samples for immunoelectron microscopy (TEM) and occasionally supervise TEM users. Operate cryofixation equipment for EM sample preparation . Train users in darkrom technique and supervise use of lab computers. Qualifications: Experience in cryofixation and immunocytochemical methods. General laboratory skills (preparation of buffer solutions, operating pH meters etc.). Familiarity with using computer programs (word processing, spreadsheets). Familiarity with electronic equipment and its routine maintenance. Darkroom experience. Ability to work independently.
Personnel will not send out job applications. If the applicants cannot come to the personnel office to pick up an application they can send a:
1. Cover Letter referencing the job number (03-345-30) 2. Resume
The cover letter and resume should be mailed to:
University of California Berkeley Berkeley Campus Employment Office Room 7-G (Ground Floor) 2200 University Ave. Berkeley, Ca. 94720
A message forwarded from my colleague Paul Sutherland
Has anyone got a good fixation method for insect gut tissue which has a thick peritrophic membrane. At present I am using 2% paraformaldehyde, 2.5% glutaraldehyde in 0.1M phosphate buffer pH 7.2 and a post fix in 1% osmium tetroxide. With this fix the microvilli are not well preserved.
Ian Hallett
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660
We are trying to localise enzyme sites (ATP) on the plasmalemma of salt glands in halophytic grasses. Has anyone out there got a method(s) for ATP-ase at the LM level for glut. fixed tissue already embedded in historesin. Also, methods for ATP-ase in samples for TEM.
Dear Doug, I suggest you look up articles by R.P. Apkarian. He has a Cr coating method which produces extremely fine grains. He told me of a detailed paper submitted to Scan. Microsc. Intl. which should have come out in 1994. I have a few of his papers, but not that one. Good luck. Yours, Bill Tivol
First Intarnational Conference on Electron Microscopy and Advances in Research in Differcent Fields of Sciance September 1995 Ismailia - Etap
Sponsored by Electron Microscopy Center Suez Canal University Ismailia - Egypt
Special Topics of the conference: 1- Role of EM in diagnostic virology. 2- Role of EM in diagnosis of tumors cylulogy and urinary atones. 3- Role of EM in ultrastructure pathology of the lung (non neoplastic conditions). 4- X-ray microanalysis: Applications particularly metalllurgical, mineralogical, and biological. 5- Scanning EM of plants, animal, ineccts, and mineral material. 6- Study of biological macromolecules from their characteristic electron diffraction patterns. 7- Skin pathology by EM. 8- Morphological ldentification of antigens by EM. 9- Different low temperature methods for biological EM. 10- Safety measures and maintenance needed for EM.
There will be an equipment exblbition in conjunction with this meeting. Registration for foreigners will be US $ 150 inclusive of full board during the time of the meeting. For further information, contact the organizar: Prof, Dr. Khalifa Ibrahim Khalifa Electron Microscvpe Center Suez Canal University Ismailia - Egypt Fax: (20) 64- 329478 ( phone and Fax number) Fax: (20) 64- 333318 ( Phone and Fax number)
Microwave irradiation has been shown to enhance the penetration of aldehyde into insect and plant tissues and to improve fixation of these specimens for LM and EM. Five references follow:
1. Lindley VA: A new procedure for handling impervious biological specimens. Microsc Res Tech 21:355, 1992
2. Smid HM, Schooneveld H, Meerloo T: Microwave fixation of water-cooled insect tissues for immunohistochemistry. Histochem J 22:313, 1990
3. Heumann HG: Microwave-stimulated glutaraldehyde and osmium tetroxide fixation of plant tissue: ultrastructural preservation in seconds. Histochem 97:341, 1992
4. Login GR, Dvorak AM: Methods of microwave fixation for microscopy. A review of research and clinical applications: 1970-1992. Prog Histochem Cytochem 27/4:1, 1994
5. Login GR, Dvorak AM: The Microwave Toolbook. A Practical Guide for Microscopists. Boston, Beth Israel Hospital Press, 1994, 184
In message {7247D4029F-at-marc.cri.nz} "IAN HALLETT" writes: } A message forwarded from my colleague Paul Sutherland } } Has anyone got a good fixation method for insect gut tissue which has } a thick peritrophic membrane. At present I am using 2% } paraformaldehyde, 2.5% glutaraldehyde in 0.1M phosphate buffer pH } 7.2 and a post fix in 1% osmium tetroxide. With this fix the } microvilli are not well preserved. } } } Ian Hallett
GRL glogin-at- bih.harvard.edu Beth Israel Hospital - Department of Pathology Telephone: 617-667-2034 Fax: 617-667-8676
Dear Listserver Audience: The tektronix image recording crt (Current # CRTC818P4S) on our Hitachi S-500 scanning EM is no longer functional. We have temporarily replaced it with a high resolution monitor, however this is not satisfactory due to the smaller image. Effort at expanding the image have resulted in some edge distortion and a loss of resolution. Does anyone out there have a replacement tube that they wish to sell? If so, please contact me at:
Dear Microscopy Listserver Audience: The tektronix image recording crt (#CRTC 818 P4S) on our Hitachi S-500 scanning EM is no longer functional. We have temporarily replaced it with a high res. monitor, however this is not satisfactory due to problems associated with the small image produced by the shorter yoke on this crt. Attempts at expanding the image electronically have resulted in some edge distortion and an unaccept- able level of resolution loss. Does anyone out there have a replacement crt of this type? If so, please E-mail me at: Bray-at-hg.uleth.ca
We have an 18 years old EDAX EDS system. The MCA and computer is slow and unreliable. Does anybody has information on upgrading old EDS systems by replacing the multichannel analyser and computer with an IBM PC or MAC? We would like to keep it as cheap as possible.
Thanks!
Kris
Kristof Kovacs Central Laboratory University of Veszprem Veszprem, P.O.Box 158 H-8201 Hungary Phone: +36-(88)-421684 Fax: +36-(88)-426016 e-mail: kris-at-miat0.vein.hu
Yes, you can upgrade old EDS systems. There is a program called WinEDS. Coupled with a plug in board for your PC, your detector and pulse processor you get a Windows based quantitative system.
Sorry I do not have the address of the person who wrote/design the system, however I will try and track him down (he is in Australia).
This may take a few weeks.
Keith Moulding ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ HKUST MCPC, Clear Water Bay, Kowloon Hong Kong.
I would suggest a call to 4-pi analysis. Their product line emphasizes upgrading existing EDS systems and works with NIH Image and DTSA (and others)
4-pi analysis 3500 Westgate Dr. Suite 403 Durham, NC 27707-2534 USA
phone: Mike Sczyz (919) 489-1757
fax: (919) 489-1487
electronic mail: go4pi-at-applelink.apple.com
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
The WinEDS system described by Keith Moulding is designed and marketed by Paul Thomson: Thomson Scientific Instruments 216 Drummond Street, Carlton, 3053, Victoria, Australia phone (03) 663 2738. fax (03) 663 3680
I had the opportunity to demo this system at the New Orleans MAS/MSA. It is an impressive package. This is not an endorsement, just a personal, scientific opinion.
This is a test. I tried to resend a message to this address and it was retu returned by the Rockefeller mail director. I couldn't resend it. Let's see if this gets out.
Seems to be a great disparity between the other packages I've looked at :
DTSA $800 and quirky Flame $7000 and Fast
Anybody know of any pakages somewhere in the middle? Center for Materials Research and Analysis Central Facility for Electron Microscopy University of Nebraska at Lincoln
I am trying to do fluorescent in situ hybridization with bacteria directly on Nuclepore polycarbonate membrane filters, and need to omit the alcohol series that is usually included in these protocols. Does anyone have any F.I.S.H. protocol that does not include the alcohol series? It doesn't need to be a membrane filter method. I am, however, interested in any F.I.S.H. membrane filter protocols. What types of membranes work best?
Thanks, Barry Pyle, Montana State University - Bozeman.
I don't know how thick you mean for the peritrophic membrane, but I've had good results by injecting fixative into the gut lumen and letting it work awhile before dissecting the gut out in fixative solution. This works well for lep guts. The fixative etc is described in Ryerse et al, Tissue and Cell, 24:751-771 1992. See discussion of preventing midgut cell surface blebbing on p 768 by injecting fix directly into the gut lumen. Good luck. Jan Ryerse, Pathology, St. Louis University Med School, St. Louis, MO
Message-Id: {MAILQUEUE-101.950327093814.384-at-vanlab.paprican.ca} To: microscopy-at-aaem.amc.anl.gov
Hi, For an economic EDX system set up to utilize your existing detector, I suggest you contact the following:
Dapple Systems, Sunnyvale CA , (408) 733-3283 They supply a PC or Mac based system which I believe is passive only( ie. does not take control of the beam)
4pi Analysis, Inc. Durham NC , (919) 489- 1757 They supply a Mac or Power Mac based system (with beam control?)
I have only read their brochures not used the systems, but I think they are worth a look.
Good luck, Laurie
Kristsf Kovacs wrote: } } We have an 18 years old EDAX EDS system. The MCA and computer is slow and } unreliable. Does anybody has information on upgrading old EDS systems by } replacing the multichannel analyser and computer with an IBM PC or MAC? We } would like to keep it as cheap as possible. } } Thanks! } } Kris } } Kristof Kovacs } Central Laboratory } University of Veszprem } Veszprem, P.O.Box 158 } H-8201 Hungary } Phone: +36-(88)-421684 } Fax: +36-(88)-426016 } e-mail: kris-at-miat0.vein.hu } }
______________________________________________________________________ Laurie Frederick, A.SC.T. PAPRICAN Corrosion Control Group 3800 Wesbrook Mall The Pulp and Paper Research Vancouver, B.C. Institute of Canada Canada V6S 2L9
A fully-funded position is available in my laboratory for a postdoctoral fellow or an advanced EM technician who can work independently. I am looking for someone with experience in thin sectioning of mammalian tissue culture cells and tissues. Specifically, this project involves immunoEM analysis by post-embedding procedures (e.g. Lowicryl or L.R. White) and/or ultrathin frozen sectioning. I am an Associate Professor at The Rockefeller University in New York. For those of you who may not know, Rockefeller is located in one of the best and safest areas of New York City. With neighboring Cornell Medical School and Sloan-Kettering Cancer Center, this is one of the most scientifically exciting communities to work in. Subsidized housing is available across the street, and child care is available on campus. My laboratory is studying endothelial adhesion molecules and their role in inflammation. We are taking a multidisciplinary approach using techniques of cell biology, immunology, molecular biology, and biochemistry. We have identified and cloned two unique adhesion molecules concentrated in the inter- cellular borders between endothelial cells. These are PECAM-1 (CD31) and the endothelial-specific cadherin, cad5 (VE-cadherin). Our lab has shown that PECAM is required for the migration of leukocytes through the endothelial junctions during inflammation. The project involves determining the ultrastructural distribution of these proteins along the endothelial junctions in vitro and in vivo. (We have also cloned the corresponding mouse PECAM and cad5.) We believe that part of the function of these molecules is determined by their distribution along the membrane. This would be an excellent oppportunity for someone trained in electron microscopy to further his/her skills as well as to learn new techniques of molecular biology and immunology. Our group interacts well and often, so that everyone can benefit from the others' knowledge and skills.
Interested parties should send a letter along with a C.V. to the address below. For specific questions or more details, please fax (212) 327-8875. I look forward to hearing from you.
Sincerely,
William A. Muller, MD, PhD Laboratory of Cellular Physiology and Immunology Box 176 The Rockefeller University 11230 York Avenue New York, NY 10021-6399
P.S. Thanks to Chris Jeffries and Jeanne Barker for helping me connect to this listserver.
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I have problems in embedding two types of material, one is developing oil seed rape pods in order to look at changes leading to pod shatter. Even after several days in 100% Spurr resin, the centre of the section, which often comtains the developing seed, remains soft and falls out when sectioning. Similarly, in a study on the infection of rubber leaves by colletotricum spp., when sectioning infected material the sections split along the leaf surface. Any ideas on overcoming these problems? I routinely use Spurr hard resin and go through propylene oxide after ethanol dehydration. I have also tried holding the specimens at 45C in the moulds for 24h before raising the temperature to 65C for full polymerisation. I have tried LRWhite with no improvement
Richard Pring mentioned: } I have problems in embedding two types of material, one is developing oil } seed rape pods in order to look at changes leading to pod shatter. Even } after several days in 100% Spurr resin, the centre of the section, which } often comtains the developing seed, remains soft and falls out when } sectioning. Similarly, in a study on the infection of rubber leaves by } colletotricum spp., when sectioning infected material the sections split } along the leaf surface. Any ideas on overcoming these problems? I routinely } use Spurr hard resin and go through propylene oxide after ethanol } dehydration. I have also tried holding the specimens at 45C in the moulds } for 24h before raising the temperature to 65C for full polymerisation. I } have tried LRWhite with no improvement
Very often the failure to infiltrate material adequately with resin is due to inadequate fixation and not to inadequate resin infiltration. Especially when processing oily samples it is necessary to fix {very} well to destroy all membrane semi-permeability and to stabilise the lipids. Going to extremes during the fixation may lead to substandard structural preservation, but may be the only way to adequately infiltrate the sample. One such overkill schedule is: Fix material 24h at 40deg C in 2.5% glutaraldehyde, then postfix for another 24h in several changes of osmium, also at 40deg C (take due care during this step). Dehydrate in acetone, use a few prop. oxide rinses and embed in your standard resin. Other methods (adding Malachite Green to aldehydes in DMSO) are also known and may very well work. All these methods have in common that they are not optimal as far as structural preservation is concerned, but sometimes they may be the only ones that will result in thin sections. Richard's second problem, that of the adhesion of the resin to the leaf surface is a result of the wax layer on the leaf surface forming a parting layer between the leaf and the resin. One solution is to use an epoxy formulation that is not as easily parted from cuticular waxes - Quetol may be a possibility.
Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
I have a an old Kevex EDS system hooked up to an "old" Cameca MBX microprobe....both about 15 years old. I've recently replaced the multi- channel analyzer with a 4pi processing system and a Mac computer. The 4 pi system costs around $6,000 and is easy to incorporate into many different EDS systems.
The 4 pi is relatively cheap, it's efficient and easy to operate and is capable of interfacing with many different EDS systems. Not only can it generate very nice EDS spectra through DTSA, it does an excellent job imaging elementally specific X-ray transitions. The quality of these EDS maps is close to that of the digital WDS maps that we generate on our SX-50 microprobe. It's really quite impressive.
If you are interested, I would be happy to FAX you some information on the 4 pi system as well as some EDS images we have generated using that system. Just let me know....
or here's 4 pi info:
4 Pi Analysis, Inc. 3500 Westgate Dr., Suite 403 Durham, NC 27707-2534 Tel: 919-489-1757 FAX: 919-489-1487
Has anyone had experience with long term storage of images from a thermal printer (Codonics, Seikosha, etc.)? I have noticed that the images start to fade away after exposure (months) to fluorescent lights. I have been told that this will happen even when they are protected from exposure to light. Any suggestions for storage or other experience would be appreciated.
John Giles Honeywell Space Systems (813) 539-2270 (813) 539-3630 Fax e-mail: jegiles-at-space.honeywell.com
We've had a Seikosha thermal printer for 4 years and still have some of the first images printed with it saved. The images have faded slightly from gray to brown, but you can only tell if you hold next to a new print. The best storage for them is in a binder or folder that is stored in a closet or file cabinet. As long as they aren't exposed to drastic variations in heat they should store for long periods of time with minimal fading.
------------------------------------------------------------------ Heinz Hemken Departamento de Biologia Celular, CINVESTAV-IPN http://cell.cinvestav.mx/bchh.html
Energy Dispersive Spectroscopy, which is probably more correctly called X-ray Energy Dispersive Spectroscopy (XEDS) but various other rearrangements of the letters exist in the literature (EDXS,...)
It is an analytical methodology in which a solid state semi-conducting detector (either Si or Ge based) is used to measure the energy distribution of X-rays emitted from a specimen. The X-ray's may be excited by some sort of probe (usually a focussed beam of electrons, ions, or photons). The "Spectrum" is then displayed on a graphical output device (read computer screen) and the relative intensity of the measured characteristic X-ray peaks, analyzed to determine the relative elemental composition of the material.
Any good text book on Scanning Electron Microscopy or Electron Probe Microanalysis (published within the last 10 years or so will have a chapter or two on the subject).
An article about 4pi system for EDS was published in SCANNING Vol. 14, 233-240 (1992). The title is:
Mac X-ray: The NIST 'DTSA' program and the 4pi analysis board.
one of the authors is: David C. Joy EM lab U. of Tennessee Knoxville, TN 37996-0810
Our lab also installed both the 4pi spectral engine boards and the 4pi scanning interface box. We have been enjoying NIH Image and several other programs in playing the SEI, BEI, and X-ray mapping.
Xiaogang
**************************************** * Xiaogang Xie * * Department of Geology and Geophysics * * Louisiana State University * * Baton Rouge, LA 70803 * * Office (504)388-2240 * * Fax (504)388-2302 * ****************************************
As promised, following is the summary of responses received from my question, "How many of you have your curing ovens vented to a hood?".
From a total of 11 responses, one containing knowledge of two others, there were only two who were not curing their resins in, or venting their ovens to a hood. Both are making provisions to do so. Nine actually place their ovens in the hood while two are vented to the hood. There was only one reference sited...that being an article in the Proceedings of the RMS Vol. 16, pp. 265-270, 1981. Most of us seem to subscribe to the "just to be sure" school of reason. I attempted to thank each of you who responded individually, but I think that I may have lost one of you. So let me say now....thank you very much.
Sandra Zane Sandra F. Zane, EM Tech. Biol. Dept. UNCC Charlotte, NC 28223 sfzane-at-unccvm.uncc.edu Fax (704) 547-3128
The original paper on PLP was McLean and Nakane 1974, J Histochem Cytochem 22:1077-1083. Also see Pino 1984, Stain Technol. 59:307.
Some of the most impressive uses of PLP came out of Marilyn Farquhar's lab. If I remember correctly the ICC that Farquhar/Brown did utilized PLP, and would be a good source of method. For example, see: Cell 36:295(1984). PNAS 81:5135 (1984). JCB 106:1863 (1988). Meth Cell Biol 31:553 (1989).
Good luck.
A. Kent Christensen Department of Anatomy and Cell Biology University of Michigan Medical School Ann Arbor, MI 48109-0616 {akc-at-umich.edu}
---------------------------------
On Mon, 27 Mar 1995, Michel Deschuyteneer wrote:
} I would greatly appreciate the reference and/or the recipe for the } Paraformaldehyde - Lysine - Periodate (PLP) fixative. } Thank you very much in advance. } } Regards, } Michel. } Michel Deschuyteneer 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: +32-2-6568113 } } } }
I have a an old Kevex EDS system hooked up to an "old" Cameca MBX microprobe....both about 15 years old. I've recently replaced the multi- channel analyzer with a 4pi processing system and a Mac computer. The 4 pi system costs around $6,000 and is easy to incorporate into many different EDS systems.
The 4 pi is relatively cheap, it's efficient and easy to operate and is capable of interfacing with many different EDS systems. Not only can it generate very nice EDS spectra through DTSA, it does an excellent job imaging elementally specific X-ray transitions. The quality of these EDS maps is close to that of the digital WDS maps that we generate on our SX-50 microprobe. It's really quite impressive.
If you are interested, I would be happy to FAX you some information on the 4 pi system as well as some EDS images we have generated using that system. Just let me know....
or here's 4 pi info:
4 Pi Analysis, Inc. 3500 Westgate Dr., Suite 403 Durham, NC 27707-2534 Tel: 919-489-1757 FAX: 919-489-1487
I have a an old Kevex EDS system hooked up to an "old" Cameca MBX microprobe....both about 15 years old. I've recently replaced the multi- channel analyzer with a 4pi processing system and a Mac computer. The 4 pi system costs around $6,000 and is easy to incorporate into many different EDS systems.
The 4 pi is relatively cheap, it's efficient and easy to operate and is capable of interfacing with many different EDS systems. Not only can it generate very nice EDS spectra through DTSA, it does an excellent job imaging elementally specific X-ray transitions. The quality of these EDS maps is close to that of the digital WDS maps that we generate on our SX-50 microprobe. It's really quite impressive.
If you are interested, I would be happy to FAX you some information on the 4 pi system as well as some EDS images we have generated using that system. Just let me know....
or here's 4 pi info:
4 Pi Analysis, Inc. 3500 Westgate Dr., Suite 403 Durham, NC 27707-2534 Tel: 919-489-1757 FAX: 919-489-1487
Return-Path: u096585-at-mdanl3.md.dow.com Received: by mdanl3.md.dow.com (UCX V2.0-15) Wed, 29 Mar 1995 08:22:41 -0500 Received: from aaem.amc.anl.gov by inet1.ma.dow.com with SMTP id AA10523 (InterLock SMTP Gateway 1.1); Wed, 29 Mar 1995 08:22:24 -0500
Thanks to all who have explained to me what EDS is. I got several enlightening and courteous replies. This is a great list and a great community!
Keep up the rich info stream!
------------------------------------------------------------------ Heinz Hemken Departamento de Biologia Celular, CINVESTAV-IPN http://cell.cinvestav.mx/bchh.html
He is using SEM for industrial applications (various analyses of Clay, paints, paper, etc). They are looking for a good short course on SEM in materials science. The dates of both the Lehigh and McCrone courses will not fit into their schedule. I know of a course at Univ Michigan which I told them about. Are there any others?- Thanks for any help you can render.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
I am trying to do in situ hybridization and immunohistochemistry on glycol methacrylate-embedded wheat pollen. The reason I am using that resin, despite its uglyness to work with, is that the infiltration of pollen is very good and the overall morphology seems to be well preserved. But to gain a better access to the target in the tissue I would like to remove the resin after sectionning. The resin was polymerized with benzoyl peroxide at 55C. Is there a way to remove it without affecting antigenicity of the tissue???
If I cannot remove the glycolmethacrylate, I am planning to use a mixture of butyl and methyl methacrylate with a polymerization under UV. According to litterature, I can remove it with acetone. Some people use benzoin, others use benzoin ethyl ether; what is the difference between the two? Can I use bezoyl peroxide instead? I know that UV polymerization can lead to inconsistancy in the polymerization, is the use of a 4W placed on top of the specimen at a distance of about 10 cm could be good?
Since I do not subscribe to this group could you send me directly any suggestions you may have.
Greetings: I'm interested in using fluorescent probes to clarify the relationship between certain cellular structures, specifically the ER and what I believe to be vacuoles. Some of my colleages have suggested that the vesicles I believe to be vacuolar in origin may be dilated ER. In looking for likely probes I've found refences in the Molecular Probes handbook to DiIC (a long-chain carbocyanine) and Rhodamine B as ER-specific dyes. However, the cited references are based on experiments on animals. Ideally, I would like to use two probes that will discriminate between ER and vacuoles. Does anyone have experience with these dyes or could someone point me towards some references? I'm working with leaf tissue, specifically cells with the minor veins. Many thanks in advance.
Dwight Beebe IRBV, Dept. de sciences biologiques beebed-at-ere.umontreal.ca Universite de Montreal Voice:514-872-4563 4101, rue Sherbrooke est FAX:514-872-9406 Montreal, PQ H1X 2B2 Canada
Dear friends, Thanks for your responses. As you all may remember I was having some crossing over problem between FITC and Rhodamine-that's what I thought... My problem seems to be autoflorescense, yeah that simple and basic, I just overlooked it. I just looked at the neurons with just plain solution, they flouresce beautifully under the rhodamine filter, and slightly under the FITC filter and no fluorescence under the Cy5 filter. Also, I had a control (no antibodies present) which I fixed with 4% paraformaldehyde and used 3% normal goat serum as blocking reagent. This control flouresce better under the FITC filter than under the Rhodamine filter, the opposite of the control without fixative. It does not flouresce under the Cy5 filter. Also, I did a control with CY5 and Cy3. Again the cells flouresces under the FITC and rhodamine Filters but not under the CY5 filter. So my solution seems to stay away from FITC, and rhodamine. Any suggestion on how I can better solve my autoflourescence problem will be appreciated. Thanks again for your responses. Your friend, Ciprian
__________________________________________________________ Ciprian Almonte Medical College of Pennsylvania Dept. of Anatomy and Neurobiology E-mail: almonte-at-medcolpa.edu 3200 Henry Ave. Voice: (215) 842-4081 Philadelphia, PA 19129 Fax: (215) 843-9082 __________________________________________________________
There is a very good SEM short course given by Botany department of university of washington in Seattle. This course covers various subject and the scheduel is flexable. The instructor of the course is Barbra Reine.
Tel # for Botany Department: (206) 543-1942
On Wed, 29 Mar 1995, Jay Jerome wrote:
} This request is for a colleague: } } He is using SEM for industrial applications (various analyses of Clay, } paints, paper, etc). They are looking for a good short course on SEM in } materials science. The dates of both the Lehigh and McCrone courses will } not fit into their schedule. I know of a course at Univ Michigan which I } told them about. Are there any others?- } Thanks for any help you can render. } } } Jay Jerome } ************************************************************** } * aka: W. Gray Jerome * } * Dept. of Pathology * } * Bowman Gray School of Medicine of Wake Forest University * } * Medical Center Blvd * } * Winston-Salem, NC 27157-1092 * } * 910-716-4972 * } * jjerome-at-isnet.is.wfu.edu * } ************************************************************** } }
If someone knows, please, write me, which type of NCAM antibody can work for post- embedding method? (Araldite 6005 or Epon resin is used and pre embedding I have made an enzyme histochemical reaction. Fixation : paraformaldehyde- glutaraldehyde in cacodylate buffer) Thanks! Agnes Kittel
Both my lab and another lab here in Hawaii have Denton DV-502 vacuum evaporators. My evaporator is perfectly fine for my needs; I can carbon coat Formvar grids or prepare pure carbon substrates for TEM. The other lab, however, wants to carbon coat for SEM, with and without EDS. Their problem is that they cannot get the carbon to evaorate long enough to get a thick enough coating, and neither can I on my instrument. We both have the same spring-steel (?) driven carbon rod holders (which differ from the gravity-fed system in the very old Denton in still another Hawaii lab and also from the coiled spring feed on the non-adjustable holders). The problem seems to be that there is enough resistance somewhere that everything heats up like crazy, the moveable parts expand enough to get stuck, and the whole feed process stops until the unit cools down or blows a fuse, whichever comes first. This has happened with old holders, brand-new ones, and everything else I have been able to devise. Has anyone else had and oversome this problem? Any hints and tips will be appreciated!
Mahalo and aloha, Tina Weatherby Carvalho Biological EM Facility University of Hawaii
We need information on where to get a light scrambler for homogeneous illumination -to couple an HBO light source to a Zeiss microscope for fluorescence work. Sources, FAX numbers, price ranges -any information would be highly appreciated. dd
Daniel Dagan, Dept. Biophysics and Physiology, Faculty of Medicine, Technion, Israel Institute of Technology, POBox 9697 Haifa 31096 ISRAEL
From your description, it sounds like you need quenching rather than more blocking. Blocking uses miscellaneous protein to block non-specific protein-binding sites in the tissue, but doesn't actually deal with autofluorescence. Quenching uses reducing agents such as ammonium chloride or sodium borohydride to reduce double bonds in the tissue, which are usually the main source of autofluorescence. Try the following: make up a fresh 1% solution of sodium borohydride in distilled water (careful, borohydride is toxic), and leave a few drops of it on your sections for about 10 minutes early in your immunocytochemical run (before the primary antibody step). This should reduce the inherent autofluorescence in your tissue.
A. Kent Christensen, Department of Anatomy and Cell Biology, University of Michigan Medical School, {akc-at-umich.edu}
------------------------------------
On Wed, 29 Mar 1995 almonte-at-medcolpa.edu wrote:
} Dear friends, } Thanks for your responses. As you all may remember I was having } some crossing over problem between FITC and Rhodamine-that's what I } thought... My problem seems to be autoflorescense, yeah that simple and } basic, I just overlooked it. } I just looked at the neurons with just plain solution, they } flouresce beautifully under the rhodamine filter, and slightly under the } FITC filter and no fluorescence under the Cy5 filter. Also, I had a control } (no antibodies present) which I fixed with 4% paraformaldehyde and used 3% } normal goat serum as blocking reagent. This control flouresce better under } the FITC filter than under the Rhodamine filter, the opposite of the } control without fixative. It does not flouresce under the Cy5 filter. } Also, I did a control with CY5 and Cy3. Again the cells flouresces } under the FITC and rhodamine Filters but not under the CY5 filter. } So my solution seems to stay away from FITC, and rhodamine. Any } suggestion on how I can better solve my autoflourescence problem will be } appreciated. Thanks again for your responses. } Your friend, } Ciprian } } __________________________________________________________ } Ciprian Almonte } Medical College of Pennsylvania } Dept. of Anatomy and Neurobiology E-mail: almonte-at-medcolpa.edu } 3200 Henry Ave. Voice: (215) 842-4081 } Philadelphia, PA 19129 Fax: (215) 843-9082 } __________________________________________________________
A message about an article regarding the 4pi system sent out couple of days ago was bounced back. The paper was published in SCANNING Vol. 14, 233-240 (1992). The title is:
Mac X-ray: The NIST 'DTSA' program and the 4pi analysis board.
One of the authors is: David C. Joy EM lab U. of Tennessee Knoxville, TN 37996-0810
Our lab has installed both the 4pi spectral engine boards and the 4pi scanning interface unit. We have also been enjoying NIH Image and several other programs in playing the SEI, BEI, and X-ray mapping.
Xiaogang
**************************************** * Xiaogang Xie * * Department of Geology and Geophysics * * Louisiana State University * * Baton Rouge, LA 70803 * * Office (504)388-2240 * * Fax (504)388-2302 * ****************************************
} Does anyone know the mechanism of geode crystal formation in the midwest? And } why do some geodes contain oil?
According our resident geologist, geodes form when percolating waters precipitate into hollow cavities. Whatever happens to be carried with the water is deposited in layers into the cavity (this includes oil).
1. Has anyone made comparison about the performance, life time, and potential contamination between new filament and rebuilt one. The prices are four times difference. 2. Which company supplies better batch of filaments. This information may mail to me directly if you feel it may offend some one. 3. Last year, we had a batch of new filaments that tend to form thin film of metal sticking on the tip of the filament, though the heating temperature was low (with slightly large distance between filament and the cap, and the heating current is limited by a stopper). Is this a problem due to impurity of filament? Any idea?
Thanks for any input.
Xiaogang
**************************************** * Xiaogang Xie * * Department of Geology and Geophysics * * Louisiana State University * * Baton Rouge, LA 70803 * * Office (504)388-2240 * * Fax (504)388-2302 * ****************************************
Tina: In our lab, the DV-502 is used for C coating SEM and Microprobe samples for EDS and WDS works.
1. The length and size of the reduced C rod are important factors for the coating thickness. The coarser C rod does not only supply more C but also support longer reduced C rod. However, coarser C rod needs higher current to reach the evaporation temperature, hence it might hit some limits of the machine, like burn the fuses. This has to be balanced. 2. You also need to properly control the heating current during evaporating. Specially, you need quickly bring the current back to some position where evaporation can last longer at the very beginning when the evaporating starts.
Xiaogang
**************************************** * Xiaogang Xie * * Department of Geology and Geophysics * * Louisiana State University * * Baton Rouge, LA 70803 * * Office (504)388-2240 * * Fax (504)388-2302 * ****************************************
Message-Id: {m0ruVG4-0007AdC-at-stjohns.ohsu.edu} Message-Version: 2 } To: microscopy-at-aaem.amc.anl.gov
I have colleague that needs help setting up an automatic computer based method to count gold particles on TEM micrographs. He has NIH image, a good Macintosh and can import digital images but is having difficulties getting accurate counts of the gold particles. If anyone has experience with this please let me know.
Subject: Time: 4:13 PM OFFICE MEMO RE} C evap for EDS Date: 3/30/95
Tina- check your ceramic insulators through which the rod holder slides. If the ceramic gets enough carbon or metal on it, it creates a conductive pathway which corrupts your normal electrical flow and sends current through the metal parts of the electrode instead of the carbon rod. Could explain the hot metal and blown fuses from over-amping the system. This can even melt some of the smaller metal parts. Cleaning the ceramic is tough because of the porous surface and chances are the ceramic will crumble from the excessive heating when you dissassble the holder. Suggest you order some new ones from Denton and keep some spares. Have you tried the carbon yarn yet? Works well, never sparks or arcs, heavier thicknesses can be produced by reducing the source to target distance. Last time I checked, Denton had the best price. -Doug
We have done a comparison study of rods from different sources, and found some to be unworkable for SEM/EDS coating in our 502. Rather than belittle the ones that don't work, I can recomment those from Ladd. Hope this helps.
Subject: Time: 9:28 AM OFFICE MEMO Denton evap Date: 3/30/95
Tina- We have a 502A with turbo. One thing to check immediatly is the ceramic insulators which the electrode holder slides through. In the past, repeated applications of heavy coatings will eventually create a conductive surface coating on these parts and current through the carbon rod will be reduced as it flows along this alternate pathway. In extreme cases, carbon will not evaporate and metal parts of the electrode will actually begin to melt. Good thing your fuses are blowing rather than your top. The insulators are tough to clean because of the porous surface and you may find that they crumble to dust when you remove them due to the heating they have been subjected to. It's good to have a few extras around as this is a chronic problem. Have you tried the carbon yarn yet in the Denton? It is impossible to blowout/arc as with the rods and is much more controllable, just evaporate until it burns through. Heavier coatings can be acquired by reducing the distance from the source. Look out for metal contamination of the yarn if you evaporate shadowing metals; it shows up as tiny electron-dense spots on you substrates. Use the shutter to protect the unused yarn on the bobbin. You can get the yarn directly from Denton for the best price; e-mail me if you need info. -Doug
Message-Id: {MAILQUEUE-101.950330104455.320-at-vanlab.paprican.ca} To: microscopy-at-aaem.amc.anl.gov
Hello, We are intending to purchase a carbon evaporation unit for coating SEM/EDX samples . We have received quotes for 6 systems but have narrowed the choises to the following on the basis of cost: Denton 502A Ladd Edwards 306 The price range for these units is $24,000-$35,000 in Canadian dollars. We are very tight for money (of course) but do not want to buy a unit which will give us continuous headaches with overheating, jamming of the carbon rod feed mechanism or having to evacuate several times in order to recoat because enough carbon could not be applied the first time. If you have experience with any of these units, either positive or negative, I would appreciate hearing your comments. If you have any experience with using 1/4" rods vs. 1/8" rods or "reduced section" versus conically tipped rods, please pass it along.
I was all set to make a purchase before I read the recent comments on the listserver. Thanks very much for any assistance. Laurie
______________________________________________________________________ Laurie Frederick, A.SC.T. PAPRICAN Corrosion Control Group 3800 Wesbrook Mall The Pulp and Paper Research Vancouver, B.C. Institute of Canada Canada V6S 2L9
Message-Id: {MAILQUEUE-101.950330143551.320-at-vanlab.paprican.ca} To: microscopy-at-aaem.amc.anl.gov
Hi, I am trying to wrap up a summary of referances on EDX and require some information: 1) Has the book from MAS entitled "X-ray Spectrometry in Electron Beam Instruments" by Dave Williams and Dale Newbury actually been published yet and if so, is there a phone number for orders?
2) Is anyone familiar with an EDX upgrade system from "Aptec"? I believe it is located somewhere in Ontario, Canada but I am not sure if the name I have is a company or product name. Contact # ?
3) When and where is the next International Congress on Electron Microscopy (ICEM) and what is a contact number for information?
4) When and where is the next International Conference on X-ray Optics and Microanalysis (ICXOM) and what is a contact # for info?
I will be making a summary of the information I have gathered on EDX available shortly. Thanks for any assistance. Laurie
______________________________________________________________________ Laurie Frederick, A.SC.T. PAPRICAN Corrosion Control Group 3800 Wesbrook Mall The Pulp and Paper Research Vancouver, B.C. Institute of Canada Canada V6S 2L9
Reply to: RE} TEM: Gold particle counting Hi Bob, Your colleague may be suffering from low contrast images. Try acquiring or scanning in the images with greater contrast range. The contrast of the images should be such that the black gold particles should be black (an extreme gray level) and everything else in the image should be a different gray value (mid-gray to white). For the images that are already digitized try doing a "Sharpen" or an "Unsharp Masking" operation on the image.
Sincerely,
George McNamara Universal Imaging Corporation --------------------------------------
I have colleague that needs help setting up an automatic computer based method to count gold particles on TEM micrographs. He has NIH image, a good Macintosh and can import digital images but is having difficulties getting accurate counts of the gold particles. If anyone has experience with this please let me know.
Message-Id: {9503311445.AA01865-at-dnagel.bio.fc.ul.pt} X-Sender: bjfeijo-at-skull.cc.fc.ul.pt X-Mailer: Windows Eudora Version 1.4.3 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I there: One of our users has plans to look at entire styles and to do some immuno labeling of the stylar channel proteins. He wants to get rid of embedding and/or freeze-sectioning so he figured confocal would be the answer. Among others, I antecipate some problem with self fluorescence coming either from chlorophyls and/or lignins. One way around these would be to use the red line of the Kr-Ar (we have a MRC-600). So, does anyone has experience on working with secondary antibodies (in this case against chicken) labeled with red excitable fluorophores in plant tissues? What may the major drawbacks in this kind of preparation? Any help would be highly appreciated.
___________________________________________________________ Jose A. Feijo Dept. Biologia Vegetal, Fac. Ciencias Lisboa Ed. C2, Campo Grande, P-1700 LISBOA, PORTUGAL
The Carnegie Mellon Research Institute (a division of Carnegie Mellon University) is trying to find a new home for a 13 year old ISI SS-40 SEM. If anyone out there is interested please send mail to me at bz0c+-at-andrew.cmu.edu
} From the Characterization Facility at the University of Minnesota
MASTER CLASS
Cryo-TEM of Colloidal Materials
=A5 Specimen Preparation
=A5 Cryo-Transfer to the Electron Microscope
=A5 Low-Electron Dose Image and Diffraction Pattern Recording on Film
=A5 Image-Processing and Analysis of Digital Cryo-TEM Images
May 18-20, 1995
Intended Audience
This Master Class is intended for any materials scientists or process eng= ineers =
working with biological or colloidal materials. This class will explore t= he use =
of the technique of cryo-transmission electron microscopy to probe the =
microstructure of these samples in the vitrified, hydrated state at resol= utions =
around 1 nm. Previous TEM experience, although welcome, is not required.=
Description
The lectures will introduce the basics of cryo-transmission electron micr= oscopy, including sample preparation and microscopy techniques. Applications to s= pecific biological and colloidal materials=D5 samples will be demonstrated, and =
complementary characterization techniques will be described.
The details of sample preparation, focussing on vitrification parameters = and =
transfer to the microscope, will be described and demonstrated. Imaging =
techniques and interpretation, artifact identification, and quantitative =
analysis will also be covered.
Supervised hands-on laboratories demonstrating the techniques that were =
introduced in the lectures will be interspersed throughout the class. Po= ssible =
topics include: low dose imaging of soft latex particles; low dose imagi= ng of =
phospholipid vesicles; image processing of cryo-TEM images using NIH Imag= e and =
Digital Micrograph; CEVS sample preparation; and video-enhanced light mic= roscopy of colloidal materials.
Upon completion of the class, participants will be able to select an appr= opriate cryogen, prepare vitrified thin films of colloidal specimens, transfer th= ese =
specimens to the microscope, and record images free from artifacts caused= by =
electron beam damage.
Registration
For more information, reply to Beth Trend, trend-at-cems.umn.edu
Instructors
Frank Booy is a Senior Staff Fellow, National Institute of Arthritis =
Musculoskeletal and Skin Diseases at the National Institutes of Health. = As a =
special expert in electron microscopy, he has worked in the field of =
cryo-electron microscopy of biological materials since 1978, at the CNRS = in =
Grenoble, France, the European Molecular Biological Laboratory in Heidelb= erg, =
Germany, and the Rijksuniversiteit Groningen in the Netherlands. Dr. Boo= y is =
particularly involved in the use of cryo-electron microscopy and 3-D imag= e =
reconstruction techniques to localize the proteins that make up the =
nucleocapsids of herpes simplex virus.
John Minter, a Research Associate in the Analytical Technology Division o= f the =
Eastman Kodak Company, is the technical group leader of the Quantitative = Colloid Microscopy Group. Minter=D5s group applies cryo-TEM and electron crystall= ography =
to the study of photographic colloids. During 1990, he was an Industrial = Fellow =
at CIE collaborating with Professors H. Ted Davis and L. E. Scriven to st= udy =
surfactant morphology and crystal precipitation by cryo-TEM. He is curren= tly an =
Adjunct Professor at CIE and Industrial Co-chair of the CIE Characterizat= ion =
Facility Advisory Committee.
David P. Siegel is a Senior Scientist in the Research & Development Dept.= , =
Procter & Gamble Co. His general interest areas are membrane biophysics = and the physical chemistry of biologically relevant lipids. Dr. Siegel is partic= ularly =
interested in the mechanisms of biomembrane fusion and of lamellar-to-inv= erted =
phase transitions in phospholipids. Since 1991, he has used cryo-TEM and= =
time-resolved cryo-TEM to image intermediates in these processes.
Schedule
Thursday May 18 =
8:00 a.m. Registration =
8:30 Lecture I: Introduction to Cryo-TEM =A5 Basics steps in cryo-TEM =A5 Biological and colloidal problems that can be solved using c= ryo-TEM =
=
10:15 Lecture II: Sample Preparation, Vitrification, Storage, and Trans= fer to =
the Microscope =
Lunch =
1:00 p.m. Lab Session - Shepherd Labs
Dinner
7:00-10:00 Lab Session continues as needed =
Friday, May 19
8:00 Coffee =
8:30 Lecture III: Imaging in Cryo-TEM =A5 Operation of Electron Microscope =A5 Recognizing the good samples to examine =
10:15 Lecture IV: Image interpretation, processing, and analysis =A5 Artifacts - a rogue's gallery =A5 Quantitative analysis
____________________________________________________________________= ___ Beth Trend trend-at-cems.umn.edu (faster) or btrend-at-maroon.tc.= 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=
The Carnegie Mellon Research Institute (a division of Carnegie Mellon University) is trying to find a new home for a 13 year old ISI SS-40 SEM. If anyone out there is interested please respond to me at bz0c+-at-andrew .cmu.edu
Thanks, Brian
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