In our newsletter, Microscopy Today, we attempt to publish monthly an on-going summary of ALL microscopy events (conferences/meetings, schools, training sessions, etc.) in the U.S. - plus major international events. A no-charge listing includes: Dates Title Sponsor/Organizer Location Contact/Tel/Fax We will consider longer write-ups and ask for an article contribution to the newsletter, rather than money, for the effort. The newsletter is mailed only to those who have requested copies - at no charge currently to some 8,000 microscopists in the U.S., Canada and the U.K. Unfortunately we must charge a modest subscription for other international readers. We invite your no charge event listing - or request for a subscription. Regards, Don Grimes, Editor
We would be obliged if you informed us about the specifications (including ISBN codes) of those books/atlases/workbooks which were published for students in the field of ULTRASTRUCTURAL HUMAN PATHOLOGY in the last ten years. Could you recommend for us some? Thank you in advance,sincerely yours
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)
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)
Dear Szabolcs and Evelin, I have several recommendations for you (though I'm sorry I don't have the ISBN codes handy). I would recommend the Journal: Ultrastructural Pathology. I would also recommend becoming a member of the Ultrastrucural Pathology Society. This society has among its members some of the "giants" in this field. To join, or inquire for more information please contact:
Claire M. Payne, Ph.D. Secretary, Ultrastructural Pathology Society University of Arizona Dept. of Microbiology & Immunology 1501 N. Campbell Tucson, AZ 85724 USA
Books:
Diagnostic Ultrastructure of Non-Neoplastic Diseases (1992), Papadimitriiou, Henderson and Spagnolo, Churchill Livingstone, Pub.
Diagnostic Electron Microscopy: A Text/Atlas (1988), Dickerson, Igaku-Shion, Pub.
} We would be obliged if you informed us about the specifications } (including ISBN codes) of those books/atlases/workbooks which were } published for students in the field of ULTRASTRUCTURAL HUMAN PATHOLOGY in } the last ten years. } Could you recommend for us some? } Thank you in advance,sincerely yours } } Szabolcs Viragh and Evelin Orso } }
Douglas W. Cromey, M.S. Cell Biology and Anatomy Arizona Health Sciences Center 1501 N. Campbell Ave. Tucson, AZ 85724 (520)626-2824 dcromey-at-ccit.arizona.edu
I see several folks trying to subscribe to the Fatfree list using the wrong address. This is the subscription address: FATFREE-REQUEST-at-HUSTLE.RAHUL.NET
To join, send e-mail to the subscription address using one of the following subjects: ADD to join as a regular member ADD DIGEST to join as a digest member
Hope this helps.
At 12:01 PM 4/3/95, JONES-at-KRDC.INT.ALCAN.CA wrote: } Sandra F. Zane, EM Tech. Biol. Dept. UNCC Charlotte, NC 28223 sfzane-at-unccvm.uncc.edu Fax (704) 547-3128
Dear Laurie, We have an old Ladd and a couple of home-made evaporators. The Ladd has been here longer than I (15 years), gets heavy use, and still works just fine (we just use it for coating grids, etc., so your mileage may vary). We use the 1/8" rods narrowed to ~1 mm (reduced section). I have coated objects from barely visible to very dark in one shot. BTW, a piece of paper folded so that one part shadows the other allows the color of the carbon to be fol- lowed relatively easily once one gets the hang of it. Any bell jar can, of course, be combined with the evaporating system (power supply, rod holder, etc.), so if you have the pumps available, you might save some $ by buying parts. Good luck. Yours, Bill Tivol
Dear Xiaogang, We have used both new and rebuilt W filaments, and have found the re- built ones to be at least as good as the new ones--in some ways better. The rebuilt ones we bought were from EBTEC (I am only a customer & have no finan- cial interest). The advantages of EBTEC's designs are 1) the connection of the hairpin to the posts is better than on some other designs, and 2) they have "regular" and "sharp" tips. The regular tips give a nice bright beam, and the sharp ones give a smaller source size. The intensity per unit area for the sharp tips is about the same as that for the regular tips, so the total inten- sity is less; however, the coherence is better, so for diffraction with radia- tion-sensitive specimens, the sharp tips are very good. We routinely preheat our filaments before mounting them in the Wehnelt cylinders. I don't know what the thin film you find sticking to your filaments is, but maybe preheating will help. Good luck. Yours, Bill Tivol
With 20 years of experience in servicing DV502's the most common problems I have encountered with carbon evaporation are:
1. Early units did not have a braided conduction wire between the moveable collet and the frame for sure current conduction. The fix for this is get the braid and drill/tap holes for fastening. If this is a problem, contact Denton to send it in for upgrade.
2. All versions work well with reduced section rods (approx .7mm diameter), pointed rods vary in conduction from start to finish and when the size nears the full 2mm the current can well go over 50 amps. I find reduced section rods will evaporate between 20-30 amps.
3. If the fuse is blown, often the fuse is replaced with a standard fuse. The correct fuse would be a ceramic type ABC fuse which is more tolerant and safer when it blows.
4. With reduced section rods if evaporation is continued past the reduced section you get the same overcurrent as with pointed rods.
5. Unfortunately there is a lot of variance in the way carbon is made into rods. You will see a variance of color, brittleness, ash formation (from some filler!). Only buy small quantities of any type until you find one that is good.
Obviously constant pressure on the rods during evaporation is essential. The leaf spring used for pressure can be easily abused by pulling it straight back to put in carbon rods. A better way of displacing it is to push it down for loading the carbon rod. If there is poor tension you really have to remove the spring and reform it to put more pressure on the rod.
7. If all of this doesn/t help, call the company and GET HELP, there is no reason to put up with poor performance, all parts and advice are available. The phone at Denton is 609-439-9100.
For any more detailed discussions you may contact me at Integrated Microsystems, Inc. 708-698-4210, fax 708-696-2541 or email.
I don't want to sound like I'm beating down the Egyptian meeting. But we've had multiple postings of that announcement for the last few weeks. I think we've had enough. Unfortunately, they are from different sources every time. So I cannot touch base with one person/organization and ask them to stop. So as a general announcement
Please cease posting this announcement until there is a significant change!
I remember seeing a request in the last few weeks about diamond knives. We are about to purchase one and are looking for suppliers and any thoughts on who makes the best diamond knives. I seem to remember the last time I bought one that the best were from Dupont or Diatome(?) but can't remember exactly which. Is that still true.? Any help greatly appreciated.
I've had a similar problem with this list that I haven't had with others. My postings get to the list, but somewhere out there multiple copies bounce off servers and come back over a period of days. Not being a UNIX guru, I have no idea why this is.
------------------------------------------------------------------ Heinz Hemken Departamento de Biologia Celular, CINVESTAV-IPN http://cell.cinvestav.mx/bchh.html
On Mon, 3 Apr 1995, Nestor J. Zaluzec-Argonne Nat. Lab. wrote:
} Colleagues... } } I don't want to sound like I'm beating down } the Egyptian meeting. But we've had multiple } postings of that announcement for the last few weeks. } I think we've had enough. Unfortunately, they } are from different sources every time. So I cannot } touch base with one person/organization and } ask them to stop. So as a general announcement } } Please cease posting this announcement until } there is a significant change! } } Thanks... } } Nestor } Your Friendly Neighborhood SysOp. } } } }
X-Mailer: InterCon TCP/Connect II 2.1.2 Mime-Version: 1.0 Message-Id: {9504032201.AA06079-at-usd14716.interramp.com}
I am working with sea urchin eggs and would like to have pictures of untreated eggs plus treated eggs ( with DTT and alpha-amylase) with SEM and TEM. Iam not sure what fixatives are appropriate for SEM and TEM . Please help me.
I did a considerable amount of TEM and SEM on urchin eggs a number of years ago. As I recall, the fixative giving the best results was something like 2% Glut. in seawater followed by osmium post-fixation. For references, see Byrd and Eppel, and Belisle and Byrd from the period of 1974 to 1979. Sorry I can't be more specific.
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
Good Morning Microscopists, Yesterday morning after observing several folks trying to subscribe to this list using the wrong address and some folks attempting to do the same with another list, I thought I would be helpful. Well....I somehow got my wires crossed and it seems that a great many of you now know how to join that other list. Please accept my apologies and I promise that I shall never again attempt to be helpful on a Monday morning. (Hopefully, I provided a chuckle for some of you.) Sandra Sandra F. Zane, EM Tech. Biol. Dept. UNCC Charlotte, NC 28223 sfzane-at-unccvm.uncc.edu Fax (704) 547-3128
Mark, I have had good luck, good quality and quick turn around from Micro Engineering (409)291-6891. My purchases were only resharpennings of old DuPont knives, but the price was good and so was the quality. Some of the folks I work with now will only buy Diatome diamond knives. Hopefully that means that almost any major manufaturer will have some high quality knives available. Doug
Douglas W. Cromey, M.S. Cell Biology and Anatomy Arizona Health Sciences Center 1501 N. Campbell Ave. Tucson, AZ 85724 (520)626-2824 dcromey-at-ccit.arizona.edu
The Electron Microscopy/Imaging Center at Colorado State University will be offering a 5 day Short Course in Freeze-Fracture Techniques this summer. In addition to comprehensive lectures, state-of-the-art instruments will be available for hands-on training in the laboratory.
For more detailed information, please visit our World Wide Web site at URL: http://www.vetmed.colostate.edu/anatomy/emic/homepage.html, or contact me directly (not to this list) for an email copy of the announcement.
John chandler-at-lamar.ColoState.EDU http://www.vetmed.colostate.edu/anatomy/faculty/chandler.html
Mark Elliott asked about diamond knives. Du Pont sold their diamond knife business to Delaware Diamond Knives, Inc. years ago. I don't know the further development. We have several users here; all use Diatome knives and all are satisfied with their knives. One of the users sent a Diatome diamond knife, which had been resharpened by a competitor, back to the Diatome factory for resharpening several years ago. Diatome would not touch it because it was not one of theirs, although the boat was. The diamond was of lower quality. This indicates their high standard of quality. There may be other good diamond knives in the marketplace, but you can't go wrong with a Diatome knife. BTW I have nothing to do with the Diatome except using their knives.
For the past 12 years, I have been dealing exclusively with MicroEngineering Inc. who make the Microstar diamond knife. I have no interest in this company other than the fact that they make a high quality, affordable product. In the rare cases when I have received a bad one, I have had a replacement by the next day or 2. Pricewise, I would have difficulty justifying any other brand to my procurement department. Again, this is no sales pitch, but just the way its been with my lab which buys or resharpens 1 or 2 knives a year.
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
} From: {MELLIOTT-at-prl.pulmonary.ubc.ca} } Organization: UBC Pulmonary Research Lab } To: microscopy-at-aaem.amc.anl.gov } Date: Mon, 3 Apr 1995 15:18:39 +0800PST } Subject: diamond knives } Priority: normal
} I remember seeing a request in the last few weeks about diamond } knives. We are about to purchase one and are looking for suppliers } and any thoughts on who makes the best diamond knives. I seem to } remember the last time I bought one that the best were from Dupont or } Diatome(?) but can't remember exactly which. Is that still true.? } Any help greatly appreciated. } } Yours } Mark Elliott PhD }
Mark,
I have always used Diatome diamond knives, as did the lab where I was trained. They are of superior quality and can be sharpened as often as needed, for life. Other companies sometimes limit the number of sharpenings that they will guarantee. I also have always had exceptional help and service from Stacey Kirsch who heads the US division of Diatome. She can be reached at 800-523-5874 for current pricing and additional information.
S. sesack-at-bns.pitt.edu Susan R. Sesack Dept. Neuroscience University of Pittsburgh Pittsburgh, PA 15260
CALIFORNIA ASSOCIATION OF CRIMINALISTS MEETING ANNOUNCEMENT
The California Association of Criminalists (CAC) is holding its 85th Semi-Annual meeting on May 10-13, 1995, at the Walnut Creek Marriott in Walnut Creek, California. The Contra Costa County Sheriff's Department Criminalistics Laboratory will be hosting this meeting.
The program is shaping up fast. Some of the subjects planned include: the Integrated Ballistics Identification System (IBIS), computer animation in casework, retrieving secured data in cases of computer crime, and bite mark analysis in the case of a mountain lion attack. This is also our last call for papers so this is your chance to share your latest project or most interesting case. Both your technical notes and innovative research are welcome. For an abstract form contact Rich Schorr, voice (510)646-2455 or fax (510)646-2913.
Workshops planned include: A Computer Workshop for Cyber- phobes (Everything You Ever Wanted To Do But Were Afraid To Admit You Didn't Know How), Practical Applications of GCMS In a Crime Laboratory Environment, a Glock Armorers Course, DNA Users Group Meeting and a Polaroid Film Product Workshop.
For more information or to receive a registration package contact Karen Sheldon, Contra Costa County, Sheriff-Coroner's Department, 1122 Escobar St., Martinez, CA 94553, voice (510) 646-2455 or fax (510)646-2913.
Dear Cryo Electronmicroscopists, I have a student who wants to compare the quantity of extracellular polysaccharide material produced by bacterial cells grown on agar plates (we don't want to use cells from broth) by TEM. The bacteria are involved in plant nodule formation. We would like to loose as little of the material as possible during processing. I think plunge freezing with a KF80, cryosubstituting the cells using perhaps 2% OsO4 in methanol would be good, followed by conventional dehydration and embedding in epoxy. Conventional room temperature fix methods use glutaraldehyde/ruthenium red/OsO4 in various concentrations to fix and stain the extracellular material but from the pictures I've seen I suspect much of the material has still been lost.
Is cryoprotection of the sample necessary? If I lightly fix then infiltrate in sucrose or some other protectant I'm a bit worried we'll start washing the polysaccharide away, if I don't then the cells (which will be a small scraping of colonies off the plate surface) could be ice damaged. If I don't use ruth. red as a stain will the material still be visible - even if present? I'd be grateful to hear from anyone who has had experiance of similar samples.
Regards, Richard E
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
Dear John, You asked about specific improvements in capabilities vs reduced life- times for pointed filaments in use with radiation-sensitive specimens. First, let me say that we experienced no reduction in lifetime. Since, with proper design, a pointed filament can be saturated at a lower current, this is not too surprising. In my work--as opposed to nearly everyone else I know--I often try to get rid of electrons. For EDX and crystallography I operate at the low- est setting for wehnelt bias, a maximally excited first condenser lens, and a small-bore condenser aperture. If I use a more intense beam for EDX, the dead- time of my system and my beam spot both get larger. For crystallography, I scan the specimen using low beam currents to avoid damage during the time I am not recording (damage during recording is inevitable). Furthermore, when I scan in diffraction mode, I want the beam size to be the same as the selected area, so that, again, every electron incident on the specimen is of some use. Bearing these facts in mind, the smaller beam size from a pointed fil- ament reduces the brehmsstrahlung radiation from the condenser aperture and other sources within the column and increases the signal-to-noise ratio and spatial resolution for microanalysis (not that my spatial resolution is too good; with a high-voltage EM and thick sections, the analysis volume will never be small). Exposure of unexamined areas of a specimen is crystallography is harder to characterize; however, given that scanning the specimen, tweaking the position and adjusting the beam, etc. take only a few percent of the expo- sure recorded on film, and that often several ED patterns can be taken from the same selected area, there would seem to be little effect on the data. It must be remembered that the ultimate, irreducible limit to the data obtainable is set by damage to the specimen and that the higher-resolution reflections are the most sensitive to this damage. Once again, it is the smaller size of the beam which is beneficial. Additionally, the ED pattern is better when the ob- jective lens is focussed (although, in the HVEM, this is not too critical), and focussing can be accomplished very rapidly using minimum contrast, which works better with a more coherent beam. I have not done quantitative comparisons between regular and pointed filaments for either case, so I can't give you specific figures of merit, but I can tell you that the beam is visibly smaller and the interference fringes are noticably more prominant with the pointed filament. I also have no idea whether any of these considerations are specific to 1.2 MV electrons, although I wouldn't think so. Another occasional consideration is that heating and charging effects are minimized at low beam currents, so the pointed filaments are more specimen- friendly in this way also. Yours, Bill Tivol
I mentioned in my last message about DIATOME would not touch the knife which had been resharpened by a competitor. I should have made it clear that "the diamond was of lower quality" was a direct quote from a sales rep.
I POSTED THIS MESSAGE SEVERAL WEEKS AGO AND RECEIVED NO RESPONSE. I THOUGHT I'D GIVE IT ANOTHER TRY JUST IN CASE THERE WAS A DELIVERY PROBLEM.
RE: Copy of: Polaron Polabed 812
Dear Microscopists-
I was wondering if anyone out there knows what happened to the Polaron line of products. I am particularly curious about some of their embedding resins such as the Polabed 812.
Thanks for your help!
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
} Dear Cryo Electronmicroscopists, } I have a student who wants to compare the quantity of extracellular } polysaccharide material produced by bacterial cells grown on agar plates } (we don't want to use cells from broth) by TEM. The bacteria are involved } in plant nodule formation. } We would like to loose as little of the material as possible during } processing.
Cryofixation would be a good way to go. Why not freeze sub in osmium/acetone, embed in epoxy, as normal, and then do a PAS stain on sections? The osmium and normal poststains are unlikely to stain all the polysaccharides. It would be interesting to find out if freeze substitution in ruthenium red/acetone would work (i.e., would the RR penetrate?).
R. Howard Berg, Ph.D. Biology Department University of Memphis Memphis, TN, 38152 E mail: bergrh-at-cc.memphis.edu phone: 901-678-4449 fax: 901-678-4457
Continuing the diamond knife theme, I have been using Diatome knives for many years, and have been impressed with their quality and reliabity. But in the commercial exibits at the Cell Biology meetings in San Francisco last December, I became aware of diamond knives being made in the Netherlands by Drukker International, B.V., an old dutch diamond firm (dating from 1906). The descriptions sounded quite impressive, including "superb wetting behavior". A 3 mm blade knife costs $2500. Or you can use their "exchange program" = if you send them an old diamond knife (any manufacturer, any condition), you will receive a new 3 mm Drukker knife for $1750 (if the new knife isn't shipped within two weeks, then you get it free). The U.S. distributor is Harris Diamond Corporation, 100 Stierli Court, suite 106, Mount Arlington, N.J. 07856, tel (201) 770-1520, fax (201)770-1549. I don't know anything about the company other than spending a few minutes at their booth at the meetings and receiving a nice brochure. If I had plenty of money and were buying a diamond knife today, I would probably buy Diatome, but I thought EM people would want to be aware of the Drukker knife, which could be very good.
To listserver people, Many thanks for the replies to my problem. We found that putting the Microcapsules between 2 squares of double sided tape and tearing them apart, left a good idea of the internal structure. Pretty simple really! This was adequate for what the user was wanting and they were pleased with the results. So pleased, they have decided to bring back about 80 samples!
rich.
***************************************************************** * Richard Lander * * South Campus Electron Microscope Unit * * c/- Pathology Department * * Otago Medical School * * P.O. Box 913 * * Dunedin * * N.Z. * * * * Tel. National 03 479 7301 International 64 3 479 7301 * * Fax. National 03 479 7254 International 64 3 479 7254 * *****************************************************************
} From: "W.L. Steffens" {STEFFENS.B-at-calc.vet.uga.edu} } Organization: College of Vet. Med } To: Microscopy-at-aaem.amc.anl.gov } Date sent: Tue, 4 Apr 1995 09:29:07 EST } Subject: diamond knives } Priority: normal
} To Mark Elliot: } } For the past 12 years, I have been dealing exclusively with } MicroEngineering Inc. who make the Microstar diamond knife. I have no } interest in this company other than the fact that they make a high } quality, affordable product. In the rare cases when I have received a bad } one, I have had a replacement by the next day or 2. Pricewise, I would } have difficulty justifying any other brand to my procurement department. } Again, this is no sales pitch, but just the way its been with my lab which } buys or resharpens 1 or 2 knives a year. } } -=W.L. Steffens=- } College of Veterinary Medicine } University of Georgia } MicroEngineering is now known as Micro Star Technologies. Contact: Cathy Zimmerman (800)533-2509, E-mail US3SNQ7N-at-IBMMAIL.COM *************************************** David Garrett "DGARRETT-at-GAB.UNT.EDU" University of North Texas Dept. Biological Sciences (817)565-3964 Fax (817)565-4136 ***************************************
Although, its been a while since I've polished aluminum. My initial impression is you have way too many steps. I'd go from 600 grit to 5 micron, to 1 micron, perhaps the .25 micron and then the colloidal. I also remember that cerium oxide paste gave better results. (probably not the greatest idea from a saftey and envirnomental standpoint) THe key to fine polishing of aluminium is to keep the steps to a minimum and the length of time at each step to a minimum. There may be a sure bet way of polishing aluminium....but I found it a real pain.
On Tue, 4 Apr 1995 IE09-at-VM.ACS.UNT.EDU wrote:
} We are currently trying to polish aluminum samples (7075). Before we etch } them, we are trying to get a 0.05 micron finish. The grinding/polishing } sequence is: 600 grit, 800 grit, 1200 grit, 3 micron diamond paste, 1 micron } diamond paste, 0.25 micron paste, 0.05 colloidal silica. Everytime we go from } the 1 micron step to the 0.25 micron step scratches appear.... } } We have been using Allied High Tech Spec-Cloth polishing pads. Doeas anyone } have suggestions to get to the final two steps without introducing scratches? } } I do not really understand why they get scratched so easily when the finer } pastes are used. } } Should these samples be stored in a dessicator to prevent oxidation? I have } never worked with Al, and therefore would appreciate any suggestions. } } Patrick Diehl } Center for Materials Characterization } University of North Texas
} Date: 05 Apr 95 12:18:19 EDT } From: South Bay Technology {73531.1344-at-compuserve.com} } To: Microscopy ListServer {MICROSCOPY-at-aaem.amc.anl.gov} } Subject: Copy of: Polaron Polabed 812
} I POSTED THIS MESSAGE SEVERAL WEEKS AGO AND RECEIVED NO RESPONSE. I THOUGHT I'D } GIVE IT ANOTHER TRY JUST IN CASE THERE WAS A DELIVERY PROBLEM. } } RE: Copy of: Polaron Polabed 812 } } Dear Microscopists- } } I was wondering if anyone out there knows what happened to the Polaron line of } products. I am particularly curious about some of their embedding resins such } as the Polabed 812. } } Thanks for your help! } } David Henriks } South Bay Technology, Inc. } 1120 Via Callejon } San Clemente, CA 92673 } } TEL: 714-492-2600 } FAX: 714-492-1499
David
I am not sure about specific products but the fate of Polaron goes Polaron was sold to BioRad who sold it to Fisons. By contacting a US rep of Fisons you may get more info, although the unit may have been sold to Thermo Instruments as part of the Fisons scientific instrument division sale.
As far as Polabed goes a number of suppliers have similar alternative to EPON 812.
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 (DuPont CR&D Analytical) have hosted girls in our lab for the past 2 years } on Take Your Daughters to Work Day and have set up several demonstrations using } materials we thought would be familiar to the girls. Lots of parents bend the } rules and bring quite small girls as young as 6, so be prepared. } My High School teacher wife read this over my shoulder and asked me to remind everyone who has these types of event to have them in the summer so that the students won't miss school. Teachers who try to do a good job for classrooms of 25 to 35 students, cannot tailor their lessons to meet the demands of students who take days off for going to work with mom or dad, and similar things can can just as easily be done during school vacation periods.
It is difficult to pinpoint the exact cause for the scratches, but there are a few things you may want to check.
1) What size are the scratches that you see? Are they significantly larger than the 1u scratches you are trying to remove? Perhaps you are getting an agglomeration of .25u particles if they are not properly wetted and dispersed in the paste. Have you used this same .25u paste before without trouble? Perhaps you should try a diamond suspension that has more uniformly dispersed particles.
2) Is your sample mounted in epoxy? If so, perhaps particles are emedding in the epoxy from a previous polishing step.
3) If your sample is not embedded in epoxy, perhaps the particles are embedded in the mounting wax or trapped inside the polishing fixture.
I don't think your cloth would be a problem. What you are using is the same as our Fine Rayon cloth and that is what I would recommend for what you are doing. I assume that you are using a fresh cloth when you reach this stage. Of course, oxidation could be the problem, but I have never had that problem so it doesn't seem likely to me.
Something else you may want to consider is trying Colloidal Alumina for the final polishing stage instead of Colloidal Silica. I understand that it can produce a better finish on softer metals. Of course, I understand you are having the problem before the colloidal silica, but it may be something to consider when you get to that point.
Of course, I would be pleased to supply you with a sample of the diamond suspension or the colloidal alumina if you would like to give that a try. I would also be interested to hear any suggestions you get from other people and ultimately your determination of the problem. I wish you good luck in figuring it out and I encourage you to contact me if I can be of any help.
Best regards-
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
Further continuation of the diamond knife theme::
1] I have heard essentially good things about the Drukker company. The only negatives I have ever heard came from Drukker competitors.
2] The Drukker people at the ICEM 94 in Paris and at MSA in 94 had in their exhibit both a video demonstration showing some purported advantages of their knife, because of the hydrophilic nature of the knife, over other knives. The advantage supposedly was that the sections come off more smoothly and with less vibration, e.g. slipping and sliding down the edge of the blade. At the ICEM 94 in Paris, the Drukker booth attracted some of the largest crowds.
Can anyone comment as to whether they have actually seen the hydrophilic edge to be a unique advantage, above and beyond diamond knives generally and therefore worth spending extra money?
Further with regard to Drukker, it is my understanding that they more or less "own" the world wide business for diamond scalpel blades used for radial keratotomy procedures being done by ophthamologists. They are now trying to apply their expertise gained from the surgical instruments end of the business to ultramicrotomy.
3] The "talk" seems to center around foreign made diamond knives. There are some excellent US made knives (e.g. MicroStar and DDK) and again, standing in our exhibit booths at the various meetings, I don't detect any unhappiness among their customers either. As a group, they do seem to be "happy campers". But as the US dollar stays weak relative to European currencies, those campers are going to have another reason to be "happy": They are going to be saving money.
Charles A. Garber PRESIDENT SPI SUPPLIES/STRUCTURE PROBE, INC. PO BOX 656 West Chester, PA 19380 USA
} Dear Cryo Electronmicroscopists, } I have a student who wants to compare the quantity of extracellular } polysaccharide material produced by bacterial cells grown on agar plates } (we don't want to use cells from broth) by TEM. The bacteria are involved } in plant nodule formation. } We would like to loose as little of the material as possible during } processing. I think plunge freezing with a KF80, cryosubstituting the cells } using perhaps 2% OsO4 in methanol would be good, followed by conventional } dehydration and embedding in epoxy. Conventional room temperature fix } methods use glutaraldehyde/ruthenium red/OsO4 in various concentrations to } fix and stain the extracellular material but from the pictures I've seen I } suspect much of the material has still been lost. } } Is cryoprotection of the sample necessary? If I lightly fix then infiltrate } in sucrose or some other protectant I'm a bit worried we'll start washing } the polysaccharide away, if I don't then the cells (which will be a small } scraping of colonies off the plate surface) could be ice damaged. } If I don't use ruth. red as a stain will the material still be visible - } even if present? } I'd be grateful to hear from anyone who has had experiance of similar samples. } } Regards, } Richard E } } Richard Easingwood } South Campus Electron Microscope Unit } Otago Medical School } PO Box 913 } Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } } "The southernmost electron microscope unit in the world"
You can try the method devoloped in Mueller's Lab, ETH, Switerland by using a capillary tube. "High-pressure freezing of cell suspensions in cellulose capillary tubes", J. Microscopy 175, 34-43 (1994).
Ya Chen
========================================================================= \ / Assistant Researcher-Cryo/SEM Coordinator TEL : 608-263-8481 \ / __ Integrated Microscopy Resource (IMR) FAX : 608-265-4076 \/ / / University of Wisconsin-Madison / / / 1675 Observatory Drive #167 Email:YChen-at-macc.wisc.edu / /__/_ Madison, WI 53706 Email:chen-at-calshp.cals.wisc.edu =========================================================================
I am not familiar with the Allied Spec-Cloth polishing cloths you are using, nor have I polished 7075 Al (to my knowledge), but here is some suggestions for ya to do with as ya see fit:
Using Buehler products, grind on grits 320, 400, 600, and 800; polish on: 1. 6 micron diamond paste [water-based] on Texmet, 2. 1 micron diamond paste [water-based] on either Texmet or Metcloth, 3. either (a) 1 micron Cerium Oxide solution on Mastercloth -or- (b) .5 micron Chrome Oxide solution on Mastercloth, then 4. .05 micron Mastermet [colloidal Silica] on Mastercloth.
Few hints: - make sure new cloths are used, and that before their use, you "beat" down the fibers on the Mastercloth by using a dummy/blank sample or such; - if using a ultrasonic to help in cleaning, this may dislodge particles that will then add scratches in later steps; - use very warm water and -much- soap to clean the samples between each step. I use my thumb to very gently rub and clean the sample surface; - use of a dessicator is highly recommened to help slow oxidation and to keep samples cleaner for further examination; - takes much time and is -very- frustrating, but experiment with the above and other recipes to get what works best for you; - have not done much myself, but don't overlook electropolishing if such can be used in your case; - for me, water-based diamond pastes seem to "cut" better for rough polishing, whereas oil-based seem to produce a smoother, cleaner finish; - if get frustrated, quit if can for awhile, and attack the sample(s) later on - have found myself that a recipe that works wonders one day seems to suck later, but then seems to work great again! The joys of hand-polishing!! oh boy...
Hope this is somewhat helpful. Good luck, -Rob disclaimer: [once again] am not affiliated nor have any financial interest in any products mentioned or used ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, | Rob Tayloe | MSM Spelunkers Club /\v/\ | | Metallographic Lab. | Missouri Speleological Survey /\v/\ | | Rolla Research Center | Bat Conservation International /\v/\ | | U.S. Bureau of Mines | Missouri Cave & Karst Conservancy | | tayloe-at-ptbma.usbm.gov | National Speleological Society #32993 /\v/\ | | (314) 364-3169 x247 | American Cave Conservation Association | ''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''
} You can try the method devoloped in Mueller's Lab, ETH, Switerland by using } a capillary tube. } "High-pressure freezing of cell suspensions in cellulose capillary tubes", } J. Microscopy 175, 34-43 (1994). } } Ya Chen } Does anyone know of a US Supplier of dialysis tubing in the dimensions specified in Mueller's paper (about the diameter of a capillary tube)?
R. Howard Berg, Ph.D. Biology Department University of Memphis Memphis, TN, 38152 E mail: bergrh-at-cc.memphis.edu phone: 901-678-4449 fax: 901-678-4457
In article {3lsfh6$51e-at-newsbf02.news.aol.com} , campbell36-at-aol.com (Campbell36) wrote:
} Is the Miscroscopy listserv still being echoed to this newgroup? I would } appreciate any info regarding this or the usenet address and instructions } on how to subscribe. } } } Thanks...Jim Campbell
OK, here is the scoop. The output of the Microscopy Mailserver at Argonne National Laboratory is no longer being automatically forwarded to the Newsgroup Sci.Techniques.Microscopy. The reason for this is that DEC closed down the gateway I was using to forward the mail to the newsgroup. I do not know where there is an alternative gateway and would like to know. So, PLEASE, PLEASE, IF YOU KNOW HOW TO FORWARD A MAILING LIST TO A NEWSGROUP THEN LET ME KNOW.
Also, if you know how to forward the newsgroup content to a mailing list then please also let me know. I know that we would have to parse the messages to prevent things being sent in infinite loops, but it should be doable.
Please do not send messages to Nestor Zaluzec asking him how to do this, he does know either!
Many thanks to whoever can help us.
-- John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
At the Environmental Scanning Electron Microscopy Users Group Meeting in Monterey CA this last weekend, it was suggested that we keep a list of Tips, Hacks and Frequently Asked Questions about ESEM. We have started to compile the list and I am soliciting input from the rest of the user community. Please send anything you think might be applicable to me at the address below (email only please).
By The Way: When we say ESEM people think mostly of the ElectroScan instrument. We are not talking exclusively about ElectroScan. We know that Topcon, Hitachi and JEOL (at the very least) make elevated pressure SEMs and all users of those instruments are encouraged to reply too.
In fact since Hitachi calll their instruments "Variable Pressure SEMs", Topcon call theirs "Wet SEMs" and JEOL call theirs "Low Pressure SEMs", I propose that we call the generic instruments "Lousy Vacuum SEMs". Trouble is, LVSEM is JEOL's accronym! If anyone has a better idea let me 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 or John.F.Mansfield-at-umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
to: Richard Easingwood subj: cryosubstitution of bacteria
Richard- We have had some success with preserving the polysaccharide layer in bacteria without the use of ruthenium red. A post-doc here custom-constructed a growth chamber which allowed the growth of the bacteria on a filter substrate. The idea was to examine the compaction of the bacteria under different osmotic stresses without actually being immersed in the culture media. A small paper disc of cigarette paper was placed on the supporting filter substrate and the bacteria were allowed to grow on top of it. The paper was then removed and propane-jet frozen in an RMC MF 7200 device. After a freeze-substitution regime using 1% osmium/0.1% UA in acetone, the sample was allowed to slowly warm to RT, then embedded in Epon/Araldite 502 epoxy and sectioned on a diamond knife. Overall preservation was excellent. I am aware of the preservative nature of RR on the polysaccharide layer but we were surprised to see a great deal of extracellular matrix between the cells without it. I am not sure what the mechanism of staining was in this case, but I think it is significant to consider the possible mechanical loss of the matrix due to processing/handling. We have not repeated the experiment with the addition of RR to the freeze-sub media so I do not yet have a comparison.
Regards,
Doug Davis EML, 26 Giannini Hall UC Berkeley 94720 (510) 642-2085 doug_davis-at-maillink.berkeley.edu
I'm studying lymphocytes in the bovine skin and I need some information.
Please, does anyone have information about the identification of T-lymphocytes subsets (T-helper and T-cytotoxic) in paraffin sections? I mean, it is possible or it only can be done in cryostat sections.
I will be very grateful for any help. ============================================================================= Francisco Javier Hernandez Blazquez | Universidade de Sao Paulo - Faculdade de | e-mail fjhblazq-at-fox.cce.usp.br Zootecnia e Engenharia de Alimentos | Voice: 55 195 616122 r. 265 Departamento de Ciencias Basicas/Histologia| r. 268 Av. Duque de Caxias Norte, 225 CP 23 | Fax: 55 195 611689 CEP 13630-000 Pirassununga (Sao Paulo) | BRAZIL | ==============================================================================
Dear John, I don't know how to forward mailservers to newsgroups or vice versa, but I'll bet Dave Kristoferson (sp?), who moderates the bionet newsgroups, does. Good luck; sorting this out will save a lot of people a lot of mailbox room. Yours, Bill Tivol
I HAVE BEEN ASKED TO DO SOME VIRAL PARTICLE COUNTING. THE INTERESTED PARTIES ARE CONCERNED ABOUT THE VIRAL CONCENTRATION FOR CALIBTRATION OF OTHER ASSAYS SUCH AS 260/280 RATIOS AND HPLC.
I HAVE DONE SOME READING ON THE SUBJECT BUT WOULD APPRECIATE ANY ADVICE ON WHICH TECHNIQUES ARE BETTER.
THANK YOU.
BARBARA HARTMAN SCHERING-PLOUGH RESEARCH LAFAYETTE, NJ 201-579-4343 201-579-4211 (FAX) E-MAIL: BARBARA.HARTMAN-at-1773.220.SCHERING.PLOUGH.SPRINT.COM
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IN REPLY TO ANTHONY GARRATT-REED, I DON'T DISAGREE WITH ALL OF YOUR COMMENTS, BUT MARK ELLIOTT WAS ASKING FOR ADVICE FROM ANYBODY WHO HAD MORE KNOWLEDGE THAN HIMSELF ON THE SUBJECT OF DIAMOND KNIFE PURCHASES. TRUE, THE RESPONSES WERE OVERWHELMINGLY IN FAVOR OF ONE PARTICULAR COMPANY, BUT ISN'T THAT THE INFORMATION THAT HE ASKED FOR? I WOULD PREFER TO PURCHASE A KNIFE FROM A COMPANY THAT MANY PEOPLE THOUGHT CONSISTANTLY SELLS GOOD QUALITY KNIVES AND HAS GREAT SERVICE THEN TO RISK BUYING A KNIFE FROM A COMPANY THAT DIDN'T GET THE RAVE REVIEWS. I CONSIDER DIAMOND KNIVES AN EXPENSIVE PURCHASE AND VALUE ANY WISDOM THAT MY CO-ELECTRON MICROSCOPISTS HAVE TO OFFER.
BARBARA HARTMAN SCHERING-PLOUGH RESEARCH LAFAYETTE, NJ 07848 (201) 579-4343 (201) 579-4211 (FAX) E-MAIL:BARBARA.HARTMAN-at-1773.220.SCHERING-PLOUGH.SPRINT.COM
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Hello....... If anybody out there knows anything about SEM or TEM studies of surface interactions between sea urchin eggs and beads( germ wheat, sugar beads) , please let me know. I will be very glad to know what fixative are you using and what is the proportional amount of eggs compare to beads? thanks
Dear colleagues of the AAEM: To whom should I contact so that I can place informations regarding 3rd Interamerican Congress on Electron Microscopy to be held from September 2 to 6, 1995, in Caxambu,MG, Brazil? Dr.Barbara Reine, from Univ.Washington,Seattle,who belongs to AMS is the representantive from US in the organizing committe. Would you plese contact me? Thanks. Kitajima. -- Kitajima, Elliot Watanabe (kitajima-at-guarany.cpd.unb.br) Departamento de Biologia Celular - Universidade de Brasilia 70919-970 - Brasilia - DF - Brasil tel. +55-61-348-2424/+55-61-340-9094 fax +55-61-274-1065
I have encountered your problem with fine scratches on aluminum and there are a couple of things you might want to try. The first option is polish as you have been, but skip the 1um and 0.25um diamond steps. I find that these smaller particles tend to embed in the polishing cloth which act as your scratch makers. And I recommend that you use 0.05 alumina suspension instead of silica. I think silica is too agressive for fine polishing the softer metals. If you feel this doesn't give you a fine enough finish you can go to 0.03 alumina.
Option two is instead of diamond paste try cubic boron nitride. The particles have rounded edges so they don't embed as easily. For soft metals like Al I use 2-4um CBN then 0.05 alumina and I get a very good finish. I get my CBN from Kay Industrial Diamond [1-800-327-8982]. Other than these are the only people I know of who manufactures CBN pastes, for a resonable price, I have no commercial interests.
In both options I try to keep actual polishing time to as little as I can get away with, because the longer you polish you get a statistically better chance of introducing some sort of scratch making contaminant. I hope this was helpful.
Respectfully,
Michael W. Odum Spec. Prep. Tech. Gatan, Inc. 6678 Owens Dr. Pleasanton, CA 94588 Tel: 510-463-0200 Fax: 510-463-0204
} We are currently trying to polish aluminum samples (7075). Before we etch } them, we are trying to get a 0.05 micron finish. The grinding/polishing } sequence is: 600 grit, 800 grit, 1200 grit, 3 micron diamond paste, 1 micron } diamond paste, 0.25 micron paste, 0.05 colloidal silica. Everytime we go from } the 1 micron step to the 0.25 micron step scratches appear.... } } We have been using Allied High Tech Spec-Cloth polishing pads. Doeas anyone } have suggestions to get to the final two steps without introducing scratches? } } I do not really understand why they get scratched so easily when the finer } pastes are used. } } Should these samples be stored in a dessicator to prevent oxidation? I have } never worked with Al, and therefore would appreciate any suggestions. } } Patrick Diehl } Center for Materials Characterization } University of North Texas
Mr-Received: by mta RANDB; Relayed; Fri, 07 Apr 1995 09:54:41 -0600 Mr-Received: by mta MCM$RAND; Relayed; Fri, 07 Apr 1995 09:54:46 -0600 Mr-Received: by mta RANDD; Relayed; Fri, 07 Apr 1995 09:55:02 -0600 Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
I tried to stain lymphocyte subsets in paraffin embedded sections of bird lung fixed with a wide variety of fixatives - alcoholic, aldehydes, PLP, imidates, etc. etc. - with no luck. I ended up using frozens. However, I recently read a paper in which glycol methacrylate sections were successfully stained for T cell subsets. The success was partly due to using protease inhibitors in fixatives. I haven't tried this method, but here is the reference:
Britten KM, Howarth PH, and Roche WR. 1993. Immunohistochemistry of resin sections: a comparison of resin embedding techniques for small mucosal biopsies. Biotechnic and Histochemistry 68(5):271-280.
The only other alternative is to use antibodies that recognize epitopes after formalin fixation and paraffin embedding. I don't know if these are available for bovine tissues. From past experience with veterinary tissues, I'd be surprised if they were!
Good luck!
Jane A. Fagerland, Ph.D. Dept. Cellular and Microscopic Research Abbott Laboratories D45M/AP31 Abbot Park IL, 60064 USA
I have been using an old Balzers BA360 freeze-fracture instrument for teaching for the past several years. I neeed to replace the stainless-steel bellows that is part of the LN2 plumbing for cooling the microtome knife. Spare parts for BA360 is not available any more from Balzers/Bal-Tec. If you have a BA360 that is scrapped and would like to sell or give away parts including the bellows, kindly get in touch with me directly through e-mail. Thanks!
************************************************************************* M.V. Parthasarathy Professor of Plant Biology & Director, EM Facility 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 ***********************************************************************
Someone was asking for information about POLABED 812. I know of Polybed 812 from Polysciences. In late 1970's Shell Chemicals decided to get out of Epon production. They sold the stock to every distributor by ration, just to be fair. I stocked up Epon myself and have some left. Two years later, one company came up with its name prefix to "bed 812". Then many companies followed with their own xxxbed 812 to replace Epon. The companies I dealt with recommended same formula for mixing the components and claimed that it behaved like Epon. Are these xxxbed 812 not the old Epon?
Your are correct in that there is some amount of selection of what are sometimes called the "Epon 812 substitutes".
With regard to these substitutes, they are most definitely NOT all the same. Our own SPI Pon 812 resin is indistinguishable from the original Epon 812 when it was being manufactured. I make this point because some of the Epon 812 that has been in the kind of long term storage described, has in fact started to change, since the shelf life is not unlimited, so if you were to compare one of the "substitutes"made today, with what you have had in your storage, indeed you might find that there are differences in the characteristics. Generally when the changes are occurring they are undesirable and are likely to erode the possibility of obtaining the best possible results.
Charles A. Garber SPI Supplies PO Box 656 West Chester, PA 19380 USA
Ph: 1-(610)-436-5400 1-(800)-242-4774
FAX:1-(610)-436-5755 e-mail: sp-supp-at-cerf.com or GVKM07A-at-prodigy.com ------- FORWARD, Original message follows -------
} Date: Saturday, 08-Apr-95 07:54 PM } } From: YANG, Ann-fook \ Internet: (yanga-at-em.agr.ca) } To: Charles A. Garber, Ph. D. \ PRODIGY: (GVKM07A) } } Subject: Re:Polabed 812 } } Someone was asking for information about POLABED 812. } I know of Polybed 812 from Polysciences. In late 1970's Shell Chemicals } decided to get out of Epon production. They sold the stock to every } distributor by ration, just to be fair. I stocked up Epon myself and have } some left. Two years later, one company came up with its name prefix to } "bed 812". Then many companies followed with their own xxxbed 812 to } replace Epon. The companies I dealt with recommended same formula for } mixing the components and claimed that it behaved like Epon. Are these } xxxbed 812 not the old Epon? } } Ann Fook Yang } } }
Reply to J.H. Mansfield's comments on ESEM type SEMs:
In addition to the ESEM, the Variable Pressure SEM of Hitachi and the Low Vacuum SEM of Jeol, there is now the new ECO-SEM (Environment controlled Sanning EM) introduced by AMRAY at Pittcon '95. How about coining the acronym SAPSEM--SubAtmospheric Pressure Scanning Electron Microscope--when referring to this class of SEMs?
************************************************************************* M.V. Parthasarathy Professor of Plant Biology & Director, EM Facility 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 returned from the MSA Program Production Meeting this morning and found the Listserver looping a series of messages. I believe I have found the source and have temporairly shutdown the list until I clear up the mess (and also delete the addresses causing the problems). In the interim, we may loose a few postings. Hopefully they will be retrievable. Please wait until you receive a back on-line message from me before you repost your messages to the list.
Sorry, but as Murphy has it, the problem ocurred while I was out of town for a few days.
Well I've sorted out what I think is the source of the bounce. If your curious read on. Otherwise trash this. I'm going to leave the node down for the rest of the evening just to try make sure all of the bouncing/reflections have finished.
---------------------------------------------------- Here's what happened..
1.) A user turned on an automatic reply in his Email program. This automatic reply sent the message "Thank you for Remembering...." back to anyone that sent him a message. 2.) When a message that originated at the Microscopy list was sent to this user, his program sent a reply back to the list, which (by design) then sent this message out to everyone on the list. This of course included the user with the automatic reply. The automatic reply then kicked in (again) and the cycle started the FIRST infinite loop. 3.)After many of these cycles, a different user in Australia had their mailbox fillup. Their program next generated an error message which was sent back to the list, which went to everyone, including our "Thank-you" user. This started a SECOND infinite loop, since now everytime the message went out their mailbox was still full and the error message was sent again. 4.)At least 2 more users (one in Hungary and another in Slovenia) had their mailboxes fill.... 5.)The cycle just gets worse from there.
What made matters worse is that I was out of town from Friday AM, through Tuesday AM. Although I did a quick check on Friday PM I did not see anything that was grossly out of line, but by Tuesday AM there was a queue of some 100,000+ messages waiting to be delivered!
I had no intention of trying to sort throught that lot to see which were real or which were the echo so I shut down the whole system and started trashing things.
We will restart probably tomorrow, when I'm pretty sure that things have cleared up.
I've managed to save a some of the last few posting. I'll try to resend them as tests.
In our laboratory we have a old Edax 9100 detector with Be window. In future we wish replace this Be window with Moxtek window. I'm looking information, how is window mount mechanicaly mounted to the end cup. Has anyone had experience with replacing Be window in the EDS detector. Thank you.
Just a general chemistry question on OsO4 degradation. When OsO4 breaks down in aqueous solution and turns brown, we generally consider it no good. And quite often it is refered to as having been "oxidized'. But if you expose Osmium metal to ambient air it will oxidize to form OsO4 (Merck Index). I am just currious does anybody know what happens to the aqueous OsO4 when it turns brown?
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
As a non specialist in EDS I have a question. Is it possible that our EDS detector (UTW from EDAX) attached to SEM does not "see" nitrogen peak at 0.91 keV (even in AlN and GaN it is hardly visible), though at the same time it detects easily very close C and O peaks? 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 *****************************************************************
Does anyone out there have anything more specific on the following: 5th Intl Conference on Image Processing & its Applications, Details: S Griffiths, IEE, Savoy Place, London WC2R OBL? I am looking for an email address, tentative proceedings, etc. Thanks for your help
L E Porter Phone (216) 796-1620 Head of Microscopy Fax (216) 796-3304 The Goodyear Tire & Rubber Company EMail LPORTER-at-GOODYEAR.COM Dept 415A 142 Goodyear Blvd Akron, OH 44305 USA
{As a non specialist in EDS I have a question. {Is it possible that our EDS detector (UTW from EDAX) attached to SEM does not {"see" nitrogen peak at 0.91 keV (even in AlN and GaN it is hardly visible), {though at the same time it detects easily very close C and O peaks?
Sorry for the mistake. The considered nitrogen peak is of course at 0.39 keV. As you see I am really non-specialist.
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 *****************************************************************
Does anybody know a method of mounting sapphire to make cross- sections for TEM ?
We have used the GATAN G1 EPOXY glue whitout success.
We know a lot of methods to prepare cross-section specimens. We are able to prepare samples in plane without difficulty and cross-sections with different materials but sapphire.
It seems it would be the adhesion of the glue on sapphire which is the problem with our cross-section samples. Any idea on the type of glue that should be used.
The Listserver is up and running, but I will for the short term be checking/monitoring each contribution for a reflection/bounce before I turn it back into a free-for-all system as it was in the past. I'll not moderate, just intercept all messages and look at them first before passing them along. My guess is that in another day all the bounces will have finally cleared up. I saw a few from last week late last night. Because of this checking (manual) postings will appear abit behind what your used to, as I will be doing it late in the evening..
I've also had multiple requests for information about meeting schedules. I've now added a section to the Microscopy&Microanalysis WWW site which has dates/locations/contacts etc.. for different meetings that I know about. As I receive updates or see items of interest I will update the lists.
You may access this information at:
http://www.amc.anl.gov
Your Friendly (& lately blurry eyed) Neighborhood SysOp
The window mount is attached to the detector snout with epoxy. I should point out that it is difficult to predict the results of putting a light-element window on an old detector. You do not know the dead layer (or what is sometimes called the "detector window") thickness, or the attenuation from the front electrode. I also would like to hear of any experiences others have had doing this. regards mark
I am now sure that it weould be a good idea to forward the newsgroup stuff to the listserver........ the messages on this mail list outomatically get sent to all subscribers, many of whom are on commercial links and therefore have to pay by the message
Although this would not affect me cost-wise, it would vastly increase my volume & I am unsure about whether I would benefit from that -
Please make a concerted effot to check out all the ramifications of such an action before doing something that may change the character of this activity and try to get a concensus as to its desirability
thanks
marcelle gillott uwm
ps - I am not trying to be a spoilsport - more like a devil's advocate!
Ten days ago, I posted a message to ask advises about (a). new W filament vs. rebuilt one; (b). better sources to order filament; (c). a problem about a bad batch of filaments. A few people provided their experiences and more people asked for a summary. THANKS FOR ALL OF YOU!
1. New W filament vs. Rebuilt one (a). Price: A box of 12-piece JOEL K-type filament is about $500-$600 for the new ones from manufacturer, and $120-$180 for the rebuilt ones. (b).Pros: three people believe that the rebuilt one is as good as the new one, and two of them have better experience with the rebuilt one. (c). Cons: however, I also got "boo" about the rebuilt one from two people. One of them pointed out that W hairpin bend and distortion is a common problem with the rebuilt one.
2. Better vendor source for W filament No conclusion. Five different sources are mentioned in these response messages. Most people do not have preference in any specific vendor except two of them told me that they enjoy SPI and one prefers EBTEC for rebuilt filament.
3. Other tips A suggestion for achieving better filament performance that I got is to preheat the filament before mounting it.
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: {MAILQUEUE-101.950410165329.480-at-vanlab.paprican.ca} To: "DAVID VOWLES" {VOWLES-at-rsbs-central.anu.ed
Dear David, I received a very helpful responce from Dave Williams, and yes the (his?) book has been published. It is available from Plenum Press, NY. Their phone number is (212) 620-8000. It's ISBN 0306-448-580 and lists for $79.50 US. I'm looking forward to it. Laurie
} Dear Laurie, } } Recently you posted an enquiry to the Microscopy Listserver: } "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?" } } Did you receive any answer to this question? I am interested in } obtaining a copy of the above book by Williams and Newbury and } would be grateful if you could forward any details which you may } have. } } With thanks, } } David Vowles } Electron Microscopy Unit } Australian National University } Email: David.Vowles-at-anu.edu.au }
______________________________________________________________________ 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
Dear Colleges There is a full-time contract position in Electron Microscopy coming up at Trent University which is located in Peterborough Ontario. The position is available from May 15 to December 22 1995 and is in the dept. of biology. Applicants interested please reply and a job description can be faxed to you.
Subject: Time: 10:59 AM OFFICE MEMO Freeze-sub w/Ruthenium Red Date: 4/13/95
Anyone have any experience with Ruthenium Red in freeze-substitution? Seems to me that RR is not soluble in 100% acetone. Thanks.... doug_davis-at-maillink.berkeley.edu
Recently, I had seen a book entitled "MICROCOSMOS" BY Jeremy Burgess , Michael Marten and Rosemary Taylor . The book is now out of print and I'm interested in purchasing a second hand book. I work at Trent University and often do high school tours for the area. I thought this type of book would peek the interests of those students as well as lower year university students into the field of microscopy. If anyone would like to part with their copy or know where I can purchase a copy please reply. Or if there is a title of a more recent edition to the book recently in print please forward . THANKS DEBBIE
Does anyone out there have anything more specific on the following: 5th Intl Conference on Image Processing & its Applications, Details: S Griffiths, IEE, Savoy Place, London WC2R OBL? I am looking for an email address, tentative proceedings, etc. Thanks for your help
L E Porter Phone (216) 796-1620 Head of Microscopy Fax (216) 796-3304 The Goodyear Tire & Rubber Company EMail LPORTER-at-GOODYEAR.COM Dept 415A 142 Goodyear Blvd Akron, OH 44305 USA
Is there nayone out there who is using a UNIX based system, SGI Indy in particular to grab images from a Hitachi S-4000 or later FESEM. I would like to know how you configured your system to do this, e.g., do you get the digital signal or tv output? do you you go directly to the SGI video in or do you have something between etc.
Thanks much ****************************************************** * 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 ** * ******************************************************************
I'd like some recommendation on what kind of filter blocks and lines to use to view Cy5. I'm using a MRC 600. I haven't been able to view any fluorescence with Cy5 using filter block "RHS" line 647 DF10, Neutral Density set at 1. I have tried different dilutions: 1:100, 1:200, 1:400, 1:600 for Cy5 1.4 mg/ml. None of these dilutions have given me good results. Any recommendation would be appreciated. Thanks in advance, Ciprian
__________________________________________________________ Ciprian Almonte * * :) * Medical College of Pennsylvania Dept. of Anatomy and Neurobiology * * *
Just a general chemistry question on OsO4 degradation. When OsO4 breaks down in aqueous solution and turns brown, we generally consider it no good. And quite often it is refered to as having been "oxidized'. But if you expose Osmium metal to ambient air it will oxidize to form OsO4 (Merck Index). I am just currious does anybody know what happens to the aqueous OsO4 when it turns brown?
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
As a non specialist in EDS I have a question. Is it possible that our EDS detector (UTW from EDAX) attached to SEM does not "see" nitrogen peak at 0.91 keV (even in AlN and GaN it is hardly visible), though at the same time it detects easily very close C and O peaks? 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 *****************************************************************
It's not unusual to have relatively poor sensitivity for nitrogen in UTW or ATW detectors. Nitrogen x-rays are strongly absorbed by carbon that may be present on the sample surface(s), on the detector window, or--in some detectors--as a detector window material. Carbon and oxygen x-rays are less strongly absorbed by carbon films. You could also get high differential absorption of nitrogen x- rays by specific elements in bulk samples, but not by Al or Ga.
What can you do about it? 1. To enhance the detection of light elements in a SEM, try operating at low KV (~2-5 KV) to increase the yield of soft x-rays relative to that of higher energy x-rays. Increase the beam spot size to obtain the beam current needed to maintain an optimum counting rate.
2. Be sure the sample surface is clean. (A 10-20 nm thick anticharge coating of carbon on the sample won't affect the soft x-ray absorption significantly).
3. If the UTW window is dirty, it may require replacement (a factory job in most cases, but can be done by the user if the detector has a turret-type window assembly.
4. If the detector crystal is dirty, contact the manufacturer about cleaning or replacing it (usually very expensive).
5. For an SEM with a high rate of hydrocarbon contamination (also not unusual), consider one of the hydrocarbon cleaning systems now on the market (or make your own).
6. Probably most important: Try calculating what the x-ray spectra from your detector should look like, to find out what the nitrogen detectability should be for different experimental conditions. The DTSA program for Macintosh computers, from NIH/NIST, performs this function.
This message is on behalf of Professor Nemat-Nasser in the Applied Mechanics and Engineering Sciences Department at the University of CA, San Diego. Prof. Nemat-Nasser has an opening for a post-doctoral researcher with expertise in electron microscopy of deformation mechanisms and damage evolution. The candidate would be working closely with researchers in the area of mechanics of materials, and is expected to collaborate of several projects involving deformation and fracture of both ductile metals and brittle solids. Some of this research will be focused on understanding deformation behaviors at high strain rates, an area where there is currently considerable research in our group here at UCSD.
Interested candidates should sent a resume, list of publications, and three references directly to Professor Nemat-Nasser -at- Dept. of AMES, MC-0411, Univ. of CA, San Diego, La Jolla, CA 92093.
E-mail files may be send to: Sia-at-CEAM.UCSD.EDU, but should be followed up with a hard copy sent by regular mail. The position will be open until an suitable candidate is found to fill the position.
Please do not send any responses to the author of this message, but direct all future correspondences to Prof. Nemat-Nasser.
} email: kepinski-at-highscreen.int.pan.wroc.pl } ***************************************************************** } } Yes, it is quite possible that your window has poor Nitrogen transmission. For instance, Noran used to market a window material called ZMAX ( a diamond film?). ZMAX had very poor Nitrogen transmission. You should press EDAX for more information regarding your Window type and transmission properties.
I have one that is still under vacuum (was last used about 2 years ago for a demonstration) - we have lots of extra parts, including some specialized stages that were obtained when someone else's 11B was decommissioned. ... The space it is currently occupying is now needed for another activity -
Please contact me directly if you are interested -
Marcelle Gillott University of Wisconsin-Milwaukee
I POSTED THIS QUESTION LAST WEEK AND DID'T GET A RESPONSE SO I THOUGHT I'D GIVE IT ANOTHER TRY.
I HAVE BEEN ASKED TO DO SOME VIRAL PARTICLE COUNTING BY TEM. THE INTERESTED PARTIES ARE CONCERNED ABOUT THE VIRAL CONCENTRATION FOR CALIBRATION OF OTHER ASSAYS SUCH AS 260/280 RATIOS AND HPLC.
I HAVE DONE SOME READING ON THE SUBJECT BUT WOULD APPRECIATE ANY ADVICE ON TECHNIQUES.
THANK YOU.
BARBARA HARTMAN SCHERING-PLOUGH RESEARCH LAFAYETTE, NJ 201-579-4343 201-579-4211 (FAX) E-MAIL: BARBARA.HARTMAN-at-1773.220.SCHERING.PLOUGH.SPRINT.COM
Does anyone out there know of a good method for stainning fatty acids for TEM? Both intra and extracellular fatty acids are of intrest in red skeletal muscle, if it makes any difference in the procedure.
Thanks in Advance for any help.
Dirk W. Domaschko Electron Microscopy Facility Marshall University School of Medicine Huntington, WV 25755-9350 Ph: 304-696-7393 E-Mail: Domaschk-at-musom01.mu.wvnet.edu
how about tetrahydrofuran. it is a great freeze sub alternative to acetone.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (314)-882-4712 (voice) (314)-882-0123 (fax)
The valence state of Os in the tetroxide is presumed to be +8.
As the valence state is reduced to lower valence states there is a whole series of color changes, and by the time it gets to +2, the color is black.
When one has a vial that has started to turn brown, more likely than not, it is not a unique valence state but a mixture of valence states, or some purists might say a distribution of valence states and continues to become more and more black in color as the extent of reduction proceeds further.
At that point, in the ampoule, the change is irreversible and there is nothing anyone can do to reverse that change (without breaking open the ampoule) and doing actual chemistry to reverse the reactions..
Here are some options to discarding the spoiled (for some applications) ampoules::
1] Some people could still make use of the aqueous osmium , for example, those doing vapor phase staining of rubber modified polymers. For such work, it does not really matter that much whether the solution is "100% active" or not. It is the vapors that count. This might not be true for everyone but at least it is one reason why you need not automatically think of sending it off to some kind of hazardous waste container. I would imagine there would be student uses for the "brown" osmium product as well.
2] If you absolutely have no use for it and if you have some quantity of ampoules, the osmium can be recycled. Or even if you don't have a whole lot, we can still combine it with other ampoules and recycle the reduced osmium. Contact me by e-mail if you wanted to explore that option. Osmium is a non-renewable resource and we should try to preserve it. The osmium you would send back might not have any real economic value in the sense that you would get anything back for it, but it is usually a lower cost alternative relative to the cost of disposing of it as a hazardous waste.
Charles A. Garber SPI Supplies PO Box 656 West Chester, PA 19380
Hi barbara, I tried to post you personally, the mail bounced back. Probable reason for some lack of response!!
We mixed a known concentration of latex spheres with a known volume of virus culture. The sample was ultracentrifuged and resusupended with a known volume of water. Then we mixed well and negative stained the samples. Determine the counts using a ratio of # latex particles to known concentration and using this ratio with the particle count.
Understanding that this is going on the assumption that latex spheres and virus particles will have the same affinity for the formvar grid. (Some problems will occur if the latex agglutinates.)
In this method, both a general ideal of concentration is reached, and the size can be determined by comparison with the latex particle. A back up to measuring the particle on a negative.
I've done this about 2X for researchers.
Good luck, Lou Ann
*************************** Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine University of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lmiller-at-ux1.cso.uiuc.edu
To whom it may concern, I am a research assistant professor in Department of Restorative Dentistry,School of Dentistry,Ege University in Izmir,Turkiye since 1987. I have received my Ph.D. degree in endodontics in February,1994. I have been running a JEOL JSM-5200 scanning electron microscope in our faculty for two years. I had also used a Philips SEM in Amsterdam,The Netherlands for six months during my doctoral studies. I would like to continue my postdoctoral studies in biomedical sciences by using SEM. Language will not be a problem for me since I had graduated from Gar-Field High School in Dale City,VA. Upon your request, I will be glad to send you my CV. My researches by using SEM are: - Anatomical approaches to dental pulp and periodontal ligament, - Observation of microflora in dental root canals and dentinal tubules, - Observation of root surfaces in teeth with periapical resorption, - Adaptation and penetration of root canal sealers into dentinal tubules, - Morphological changes in red blood cells of patients with diabetes mellitus, - Local effect of tetracycline on teeth with periodontitis, - SEM investigations on natal and neonatal teeth, - The effect of polishing on dental filling materials, - Interactions of yeasts with dentin, - The effect of titanium tetrafluoride on dental hard tissues, - Durability of titanium tetrafluoride on enamel after in vivo applications, - Resorption patterns of decidous teeth.
Hope to collaborate with you. Kind regards, Dr. Bilge Hakan Sen
Try using LocTite 460 glue. It seems to be the highest quality super glue on the market. You must buy it through a distributor. You can contact me for that information.
Does anyone know of the availability of a used microtome capable of cutting undecalcified bone- something similar to the Reichert Autocut, Polycut etc? Also, is there a server on the network for histochemical people? Jim Swafford School of Dentistry University of Missouri Kansas City, MO muchas gracias and many thanks!
You may notice that Microscopy Email appears to be coming from a new address called
MicroscopyListZaluzec-at-aaem.amc.anl.gov...
PLEASE PLEASE, DO NOT use this address when posting to the list. Continue to use
Microscopy-at-AAEM.AMC.ANL.GOV
It is a temporary forwarding address that I am using to help clear up the looping that happened last week. Over the weekend one person tried to post to that address and started another loop. I caught it, but it still took me the morning to clear up things. Just post as you always did in the past.
Hi barbara, I tried to post you personally, the mail bounced back. Probable reason for some lack of response!!
We mixed a known concentration of latex spheres with a known volume of virus culture. The sample was ultracentrifuged and resusupended with a known volume of water. Then we mixed well and negative stained the samples. Determine the counts using a ratio of # latex particles to known concentration and using this ratio with the particle count.
Understanding that this is going on the assumption that latex spheres and virus particles will have the same affinity for the formvar grid. (Some problems will occur if the latex agglutinates.)
In this method, both a general ideal of concentration is reached, and the size can be determined by comparison with the latex particle. A back up to measuring the particle on a negative.
I've done this about 2X for researchers.
Good luck, Lou Ann
*************************** Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine University of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lmiller-at-ux1.cso.uiuc.edu
Hi Debbie; Re: "MICROCOSMOS" BY Jeremy Burgess , Michael Marten and Rosemary Taylor. This was First published in 1987 by Cambridge University Press and in 1990 republished under the tile of "UNDER THE MICROSCOPE'.
I see that you live in Peterborough you should be able to find it at The Worlds Biggest Bookstore in Toronto, I saw it there some time last year.
Another source for the book where they have copies of both editions listed is:
SAVONA BOOKS 9 Wilton Road, Hornsea, North Humberside, HU18 - 1QU England UK
The current catalogue lists the 1987 edition at 27 pounds sterling, and the 1990 edition at 15 pounds sterling.
For general interest to all on the list, SAVONA BOOKS has an extensive catalogue, some fifty pages long with as many topic areas of rare, old, and new books on microscopy and related subjects. I have had a number of books from them and they all arrive safely on this side of the pond.
Mr-Received: by mta RANDB; Relayed; Mon, 17 Apr 1995 09:16:22 -0600 Mr-Received: by mta MCM$RAND; Relayed; Mon, 17 Apr 1995 09:16:25 -0600 Mr-Received: by mta RANDC; Relayed; Mon, 17 Apr 1995 09:16:40 -0600 Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
Barbara Hartman asked for a technique for viral particle counting in the TEM. We routinely use a direct sedimentation/negative staining method for doing particle counts (see Miller M.F. and Rdzoc E.J., Proc. 39th Ann. Mtg. EMSA, p. 404 (1981) and Miller M.F., Proc. 37th Ann. Mtg. EMSA, p. 404 (1979)). This technique uses a Beckman Airfuge and a specially designed rotor (Model EM90) which produces uniform sedimentation across the grid, a potential source of error with some rotor designs. It is relatively easy to do if you have an airfuge and rotor. Hope this helps.
Joe Neilly Abbott Laboratories North Chicago, IL 60064 709-938-5024 E-mail: Joseph.neilly-at-abbott.com
On two occasions TechBooks have produced copies of out-of-print books for me, and have done a truly high quality job of it. The phone number I have for them is 800-767-1518.
Subject: Time: 7:57 AM OFFICE MEMO Another old TEM available Date: 4/17/95
Philips 200 at UC Berkeley, has not run in two years.
Contact: Prof. Ed Sylvester at (510) 642-7353 or Ethel Nakamura in Wellman Hall at (510) 642-1603 for details. Some funds available for moving, Ask Ed.
} I would be interested in hearing experiences/suggestions folks have } with Residual Gas Analyzers on their electron microscopes (specifics } please). (I have a Cameca SX50/51 electron microprobe that I am thinking } about installing one on.)
We have a Spectra VacScan Model 100 RGA permanently attached to our Philips CM-20 that is used primarily for cryoTEM. The ability to measure the partial pressure of contaminants (especially water vapor) makes it easy to track down problems. We bought this through Philips when we bought the microscope. We chose this one because it is the model they send to their service engineers when they need one. Our service engineer really like having this as a diagnostic tool. Given the high price most of us pay for our microscopes, I can't understand why more labs don't have this accessory that costs less than $10k.
Best Regards, John
John R. Minter, Ph. D. Phone: (716) 722-3407 Eastman Kodak Company FAX: (716) 477-3029 Analytical Technology Division email: minter-at- kodak.com Rochester, NY 14562-3712
I have preformed many many viral particle counts, mainly with enteric virus. The few references I have concerning particle counting are as follows:
Kinetic Attachment Method:
1) Sharp, D.G., 1974 Procedings of 32nd Annual Meeting of EMSA. Clayton's Publishing Division, p. 264-265
This methods describes using Al coated grids to allow virus to settle out onto the grids.
Pseudoreplication Method:
2) Smith,K.O. and Benyesh-Melnick,M., 1961 Proc. Soc. Exptl. Biol. 107:409-413.
3) Smith,K.O. and Melnick,J.L., 1962 J. Immunol. 89:279-284.
4) Smith,K.O. and Melnick,J.L., 1962 Virology 17:480-490
5) McCombs,R.M., Benyesh-Melnick,M. and Brunschwig,J.P., 1966 J. Bacteriol 91:803-812.
6) Benyesh-Melnick,M., et al 1966 J. Bacteriol. 92:1555-1561
The first method works vewry well with highly purified preps while the second method is great for cell culture supernatants, lysates or stool specimens. The first method is much more accurate and precise while the second is for good ball park estimations.
The one comment that I saw on the microscopy listserver about adding latex spheres to the viral prep and counting virus/spheres in my opinion is not a good suggestion only because of the different diffusion/settling constants between virus and spheres.
One comment on the counts obtained. You will probably be counting all viral particles and not just infectious ones, sokeep this in mind.
Good Luck, Ed Calomeni Medical College of Ohio Toledo, OH emlab-at-opus.mco.edu
I think that you are having trouble with the G1 epoxy for one or two reasons. First the epoxy is not bonding because the sapphire is not clean. When I make cross-sections I always use a cotton swab and a little acetone to clean my wafers right before putting them into a stack. Even if I have cleaned them previously.
The second reason may be that the G1 is being mixed wrong. It needs to be measured by weight in a 10:1, epoxy-to-hardener ratio. The 10 drops/1 drop instructions on the lid of the cross-sectioning kit have misled a lot of people. It is something that should have been fixed in the packaging long ago, and I am sorry if it has confused you.
If you are still having problems after this I would suggest getting a new batch of epoxy. It is possible that the epoxy is too old, or has been stored in an enviroment not suitable for long-term sitting.
Please feel free to contact me or, Dr. Alani, if you have an other questions. Thanx.
Michael W. Odum Gatan, Inc. 6678 Owens Dr. Pleasanton, CA 94588 Tel: 510-463-0200 Fax: 510-463-0204 E-Mail: modum-at-gatan.com
Reza Alani E-Mail: ralani-at-gatan.com
} Does anybody know a method of mounting sapphire to make cross- } sections for TEM ? } } We have used the GATAN G1 EPOXY glue whitout success. } } We know a lot of methods to prepare cross-section specimens. We are } able to prepare samples in plane without difficulty and cross-sections with } different materials but sapphire. } } It seems it would be the adhesion of the glue on sapphire which is the } problem with our cross-section samples. Any idea on the type of glue that } should be used. } } Thank you. } Georges Veilleux } Researcher } INRS-Energie et materiaux } C.P. 1020 } Varennes, Qc } Canada } J3X 1S2 } Tel.: (514) 929-8110 } Fax: (514) 929-8102
} From: SMTP%"GVKM07A-at-mail.prodigy.com" 14-APR-1995 03:25:23.55 } To: MICROSCOPY } CC: } Subj: Fwd: OsO4 'oxidation' } } I would like to respond as follows: } } The valence state of Os in the tetroxide is presumed to be +8. } } As the valence state is reduced to lower valence states there is a } whole series of color changes, and by the time it gets to +2, the color } is black. } } When one has a vial that has started to turn brown, more likely than } not, it is not a unique valence state but a mixture of valence states, } or some purists might say a distribution of valence states and } continues to become more and more black in color as the extent of } reduction proceeds further. } } At that point, in the ampoule, the change is irreversible and there is } nothing anyone can do to reverse that change (without breaking open the } ampoule) and doing actual chemistry to reverse the reactions.. } } Here are some options to discarding the spoiled (for some applications) } ampoules:: } } 1] Some people could still make use of the aqueous osmium , for } example, those doing vapor phase staining of rubber modified polymers. } For such work, it does not really matter that much whether the solution } is "100% active" or not. It is the vapors that count. This might not } be true for everyone but at least it is one reason why you need not } automatically think of sending it off to some kind of hazardous waste } container. I would imagine there would be student uses for the "brown" } osmium product as well. } } 2] If you absolutely have no use for it and if you have some quantity } of ampoules, the osmium can be recycled. Or even if you don't have a } whole lot, we can still combine it with other ampoules and recycle the } reduced osmium. Contact me by e-mail if you wanted to explore that } option. Osmium is a non-renewable resource and we should try to } preserve it. The osmium you would send back might not have any real } economic value in the sense that you would get anything back for it, } but it is usually a lower cost alternative relative to the cost of } disposing of it as a hazardous waste. } } Charles A. Garber } SPI Supplies } PO Box 656 } West Chester, PA 19380 } } Ph: 1-(610) 436-5400 FAX:1-(610) 436-5755 } } e-mail: GVKM07A-at-prodigy.com.
Dr. Garber,
I had no idea that you could recycle osmium or that is was a non-renewable resource. I suppose I could have figured this out but never thought about it. We collect the osmium that we use to fix tissue sections in waste containers and have them picked up when they're full. We also occasionally discard osmium that we've diluted with phosphate buffer once it gets beyond a certain age. Are you only interested in recycling un-opened ampoules, or are these other forms also of value? The jug of waste osmium eventuall turns completely to a black solid in a clear liquid. I don't care about getting any money back. My only monitary concern is the cost of hazardous chemical shipment. Please let me know whether it would be worth my initiating a recycling policy in my lab. Thanks.
S. sesack-at-bns.pitt.edu Susan R. Sesack Dept. Neuroscience University of Pittsburgh Pittsburgh, PA 15260
Try using LocTite 460 glue. It seems to be the highest quality super glue on the market. You must buy it through a distributor. You can contact me for that information.
Message-Id: {9504181500.AA02970-at-unlinfo.unl.edu} X-Sender: tvoiles-at-129.93.1.11 X-Mailer: Windows Eudora Version 1.4.3 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Hi,
I'd just like to inform all those persons from the U of Nebraska that happen to be on this list that the EM Lab is starting a "newsletter" of sorts and an electronic scheduling system. Please send me a note - tvoiles-at-unlinfo.unl.edu- if you wish further information.
Center for Materials Research and Analysis Central Facility for Electron Microscopy University of Nebraska at Lincoln
When I was finishing up my book on Vacuum Methods I made a survey of the sources of RGAs and found more than a dozen companies sell them (probably not all are manufacturers, however). These are listed in Appendix 2, p. 468, and their addresses are given in Appendix 3. There is also a discussion of the functioning of RGAs and some interesting examples of their application to problems in EM in Section 3.4.1, p. 114.
Residual Gas Analysis - I missed the original John Fournelle message but caught the John Minter reply.
I spent some time in the mid-80's using an Anavac-2 mass spectrometer from VG Gas Analysis, Cheshire, UK. This was mounted on both a JEOL 35C SEM and a JEOL 200CX STEM (obviously not simultaneously!).
The system was amazingly sensitive and showed rise and fall of hydrocarbons as the pumping system liquid nitrogen traps were filled and later emptied, or when specimen-area anti-contaminators were cooled or warmed. We actually wanted to monitor water vapour in cryo-setups, and in this regard it was again very useful.
A small paper was published in the now defunct Austrian joournal Mikroskopie (Wien) 42: 196-205 by KP Ryan, DH Purse & JW Wood titled: Cryo-stage performance and observation of freeze-drying in a scanning electron microscope. It shows a few graphs of water vaour partial pressure in various ways. Those were the days!
The instrument was very useful also for tracing vacuum leaks, tune it for helium (by letting some in via the airlock) and then probe around the column with a fine jet from a hypodermic needle or something similar. Mail again if I can be of further help - although it is now receding in the past! Good luck. Keith Ryan
While I do not know of any microtomes such as what you mentioned, we do produce a line of cutting and polishing equipment that is ideally suited for undecalcified bone. We produce diamond wheel saws and diamond band saws that are used extensively for this purpose.
If you would like more detailed information, please contact me either by e-mail or at:
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: 800-728-2233 (toll free in USA) 714-492-2600 FAX: 714-492-1499
} Date: Thu, 13 Apr 1995 15:06:11 -0400 (EDT) } From: DLIETZ-at-TrentU.ca } Subject: books for sale } To: microscopy-at-aaem.amc.anl.gov } Message-id: {01HPALSGL9E400DE1O-at-TRENTU.CA} } X-VMS-To: IN%"microscopy-at-aaem.amc.anl.gov" } MIME-version: 1.0 } Content-type: TEXT/PLAIN; CHARSET=3DUS-ASCII } Content-transfer-encoding: 7BIT } } Recently, I had seen a book entitled "MICROCOSMOS" BY Jeremy Burgess , } Michael Marten and Rosemary Taylor . The book is now out of print and I'm } interested in purchasing a second hand book. I work at Trent University } and often do high school tours for the area. I thought this type of book } would peek the interests of those students as well as lower year } university students into the field of microscopy. If anyone would like to } part with their copy or know where I can purchase a copy please reply. Or } if there is a title of a more recent edition to the book recently in print } please forward . } THANKS DEBBIE
Sir! I have this book in my library. My specimen was bought at the university book store here called "Akademica" , fax no. 22 85 30 53 , p.o. Box 84- 0314 Oslo,Norway. As they keep lots of books, there still may be a copy left. The book is ISBN 0-521-30433-4. and was published in 1987. That s not so old. We have both the original Hooke s "Micrographia" and Leeuvwenhookes "Achana Naturae" in two volumes here (1695) and most of what have been published ever. Sincerely yours Morten M.Laane , Professor of Biology, University of Oslo.
Hi, Would someone please send me a quick message, I'm trying to test this new subscription. Is there a special procedure for Macintosh cpu's through welchlink?
I heard about your news group from a friend. I am working with TEM, SEM and microanalysis on biological materials. I think I should have interesting informations with your list. How can I subscribe ?
Sincerely
Laurence Peru - Perul-at-ERE.UMontreal.Ca
Laboratory for Electron Microscopy Faculty of Dental Medicine Universite de Montreal 2900 Edouard Montpetit Montreal, QC, Canada - H3C 3J7
Yes I have seen all your questions as to the fact your not receiving mail. I once again shut things down. This time it was due to a major change in the Email router here at ANL. The new configuration completely swamped my host here, because, unlike the old router, when a message was delayed for more than 4 hours the router started sending messages back to the originator.
I started getting ~ 100 warning messages each day saying that the mail message to XXXX.YYYY.ZZZZ was not sent for 4 hours and a new attempt will be made in another 4 hours..
I'm going to try to restart today and see if the message problem is cured.
Keep posting your messages to Microscopy-at-aaem.amc.anl.gov I will intercept each and repost it after a short delay.
Doug, Ok got your message and I see the problem with DMSO although it would have been nicer to work with. Please let me know if lunch helped generate any other ideas. Have you tried the THF yet? Please let me know. I am still searching for other alternatives and will let you know if something comes up. Stacie
JEOL 1200exII Conventional TEM (straight TEM with no add-ons). This instrument is less than five years old, has been under service contract the entire time, and is in mint condition. It has had limited users and all were closely supervised. As a result, this machine has had no down time. For more information, call Larry Hawkey (919-6841-6425) or E-Mail (hawkey-at-neuro.duke.edu)
While I do not know of any microtomes such as what you mentioned, we do produce a line of cutting and polishing equipment that is ideally suited for undecalcified bone. We produce diamond wheel saws and diamond band saws that are used extensively for this purpose.
If you would like more detailed information, please contact me either by e-mail or at:
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: 800-728-2233 (toll free in USA) 714-492-2600 FAX: 714-492-1499
} To: Microscopy } From: delannoy-at-welchlink.welch.jhu.edu (Michael Delannoy) } Subject: EM Salaries } } Hello, } I'm trying to get some information on salary ranges for } electron microscopists and EM core facility managers. Is there a } source I can check? Thank you. } } M. Delannoy }
As we discussed today, there is a pretty interesting paper which is out that deals with your recent correspondance on the above subject. I have sent you a reprint of the article entitled "Pro's for a multiple/repeated use and disposal with recovery of osmium tet. solutions in routine e.m.. This paper was presented at the 1992 Boston EMSA meeting. It makes for very interesting reading and gives you a whole lot of alternatives of what to do with the Osmium. If you have any questions just let me know for I am in contact constantly with the gentlemen that did the work. If anyone else is interested in a copy of this protocol please contact me and I will be more then happy to send you one. Stacie Kirsch Electron Microscopy Sciences 215-646-1566
A staff member here would like to critical point dry millipore filters containing bacteria and zooplankton for SEM. She is concerned about losing the fine material, and was wondering if the filters could be mounted in some special holder in the CPD. Has anyone tried doing this, and if so, what sort of setup would you need to hold the filters?
Thanks for any suggestions, Thor
Dr Thor Bostrom Analytical EM Facility Queensland University of Technology (QUT) GPO Box 2434, Brisbane, QLD 4001, Australia Ph: 61-7-8642351 FAX: 61-7-8645100
I am looking for any information I can find concerning Tripod Polishing of Sapphire. I have spoken to a few people who have had some success, but even they would like to hear other suggestions. If anyone out there has any information, please respond to me directly. I'll summarize the responses for the listserver.
Thank you!
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: 800-728-2233 (toll free in USA) 714-492-2600 FAX: 714-492-1499
Hello. I am currently having difficulty getting cryo sections of mouse bone marrow macrophages to stick on grids.
I am using conventional techniques with a dry diamond knife, picking the sections up with sucrose and floating the grids on buffer before further prossing with immunogold. I have been using this method for some time with different types of cells and intracellular parisites with no difficulty. My problem seems to be with just this one cell type...
Any advise from someone in the know?
Thank you.
Jaime A. Dant Washington University School of Medicine St. Louis jaime-at-borcim.wustl.edu
My machine serves as an internet Gateway for 'space.com'. My machine has been receiving email with an incorrectly specified Fully Qualified Domain Name (FQDN).
We produce a wide range of "disc grinders" for TEM and other applications. Our lapping and polishing fixtures are ideally suited for TEM sample preparation. In fact, we have a few polishing fixtures that will allow you to transfer the sample mount from the polishing fixture, to any of our saws and also to our dimpling instrument.
Our polishing fixtures which are designed specifically for TEM are:
Model 140 $ 785.00 Model 141 395.00
Either of these will allow you to transfer the sample holder as described above. We also have larger fixtures for holding samples up to 2" in diameter. The price range on these fixtures is from $345 - $995.
If you would like additional information, please contact me at:
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: 800-728-2233 (toll free in USA) 714-492-2600 FAX: 714-492-1499
} } As a non specialist in EDS I have a question. } Is it possible that our EDS detector (UTW from EDAX) attached to SEM does not } "see" nitrogen peak at 0.91 keV (even in AlN and GaN it is hardly visible), } though at the same time it detects easily very close C and O peaks? } Leszek Kepinski } Leszek,
I don't know where I copied it from, but I wrote in my PhD thesis: "When operating with the thin window in place, it is not possible to analyse for nitrogen, as the carbon in the window heavily absorbs the nitrogen x-rays", meaning the nitrogen x-rays are the right sort of energy to interact with the carbon in your thin window, and only a few get through to be detected. I haven't bothered figuring out more precisely what is happening. Keith
MR-Received: by mta VULCAN.MUAS; Relayed; Thu, 20 Apr 1995 12:04:57 +1000 MR-Received: by mta VULCAN; Relayed; Thu, 20 Apr 1995 12:05:10 +1000 Alternate-recipient: prohibited Disclose-recipients: prohibited Content-return: prohibited
Has anyone ever used NIH Image to calculate the shape factor of a sessile drop, and if so, how. Calculation of the shape factor allows the surface tension of the droplet to be determined and requires the accurate measurement of the radius of curvature on many points of the drop surface.
Thanks in advance.
Leo Frawley B.Sc Electron Microscopist BHP Research - Melbourne Labs AARNet " frawley-at-a1.resmel.bhp.com.au "
Residual Gas Analysis - I missed the original John Fournelle message but caught the John Minter reply.
I spent some time in the mid-80's using an Anavac-2 mass spectrometer from VG Gas Analysis, Cheshire, UK. This was mounted on both a JEOL 35C SEM and a JEOL 200CX STEM (obviously not simultaneously!).
The system was amazingly sensitive and showed rise and fall of hydrocarbons as the pumping system liquid nitrogen traps were filled and later emptied, or when specimen-area anti-contaminators were cooled or warmed. We actually wanted to monitor water vapour in cryo-setups, and in this regard it was again very useful.
A small paper was published in the now defunct Austrian joournal Mikroskopie (Wien) 42: 196-205 by KP Ryan, DH Purse & JW Wood titled: Cryo-stage performance and observation of freeze-drying in a scanning electron microscope. It shows a few graphs of water vaour partial pressure in various ways. Those were the days!
The instrument was very useful also for tracing vacuum leaks, tune it for helium (by letting some in via the airlock) and then probe around the column with a fine jet from a hypodermic needle or something similar. Mail again if I can be of further help - although it is now receding in the past! Good luck. Keith Ryan
IOn Thu, 20 Apr 1995, Nestor J. Zaluzec-Argonne Nat. Lab. wrote:
} From: SMTP%"delannoy-at-welchlink.welch.jhu.edu" 18-APR-1995 15:19:23.95 } To: MICROSCOPY } CC: } Subj: EM Salaries } } Date: Tue, 18 Apr 1995 14:28:44 +0500 } Message-Id: {9504181828.AA21435-at-welchlink.welch.jhu.edu} } X-Sender: delannoy-at-welchlink.welch.jhu.edu } Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } To: Microscopy-at-anlemc.msd.anl.gov } From: delannoy-at-welchlink.welch.jhu.edu (Michael Delannoy) } Subject: EM Salaries } X-Mailer: {Windows Eudora Version 1.4.2b16} } content-length: 291 } } } To: Microscopy } } From: delannoy-at-welchlink.welch.jhu.edu (Michael Delannoy) } } Subject: EM Salaries } } } } Hello, } } I'm trying to get some information on salary ranges for } } electron microscopists and EM core facility managers. Is there a } } source I can check? Thank you. } } } } M. Delannoy } } } Hi, Statistics were taken by EMSA several years ago. I can tell you that salaries are pathetic for techs, at least in my experience. I just sent a man with 12 years of experience in histo and EM pathology & research, a recent BS, and MSA Certification , to Northwestern University for a position in downtown Chicago, and he was offered $21,000 per year.
I've just finished reworking the mailing list. I have now seperated out each group of users by country code. This is the LAST message you will receive using the original mailing list. As soon as I'm sure that all copies of this message have been sent out I will be disabling that old list of subscribers and restarting a new list.
If by Monday AM you DO NOT receive another message from me called TEST OF NEW LIST, then I have accidently left your subscription off the reconfigured list.
Please contact me at that time, at:
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so that we can get you back on the system.
The mailing address of the listserver has not changed it remains:
Microscopy-at-aaem.amc.anl.gov
You may continue to post all your messages to that address. I will continue to intercept and redirect the messages to the temporairy lists until I'm sure things are back to some sense of normal (i.e. minimum errors) operation.
Message-Id: {9504211955.AA2171-at-pho018.sb.com} To: MICROSCOPYLISTZALUZEC {MICROSCOPYLISTZALUZEC-at-AAEM.AMC.ANL.GOV}
Millipore filters can be successfully CPD'd in envelopes made from folded filter paper, closed with a staple at the open edge. I use Whatman 541 4.25cm papers routinely with 12mm filters. The sample code can be written on the paper with pencil, and this mark can then be used to determine the "up" side of the millipore filter - sometimes the sample is invisible to the eye.
Anyone have good web sites for microscopy or image processing? The address and a few words about it - topic, scope, usefulness - would be great.
Thanks Dave
------------------------------------------------------------- David S. Bright dbright-at-nist.gov Microanalysis Research Group Bldg. 222 (Chem.) A113 National Institute of Standards & Technology (NIST, formerly NBS) Gaithersburg, MD 20899-0001 / USA 301-975-3911 (voice), 301-216-1134 (fax) "You must have accurate and honest weights and measures, so that you may live long in the land the Lord your God is giving you.", Deuteronomy 25:15
Just as I send out this posting the main DNS system here at ANL crashed. About 1/2 of the sites did not receive this message. So I apologize if you got it twice, but for some of you (~ 800-1000) this will be your first receipt.
Nestor
G'day Subscribers;
I've just finished reworking the mailing list. I have now seperated out each group of users by country code. This is the LAST message you will receive using the original mailing list. As soon as I'm sure that all copies of this message have been sent out I will be disabling that old list of subscribers and restarting a new list.
If by Monday AM you DO NOT receive another message from me called TEST OF NEW LIST, then I have accidently left your subscription off the reconfigured list.
Please contact me at that time, at:
Zaluzec-at-aaem.amc.anl.gov
so that we can get you back on the system.
The mailing address of the listserver has not changed it remains:
Microscopy-at-aaem.amc.anl.gov
You may continue to post all your messages to that address. I will continue to intercept and redirect the messages to the temporairy lists until I'm sure things are back to some sense of normal (i.e. minimum errors) operation.
} } Superglue is unstable under the electron beam!!!
Would you explain a little bit what you mean by unstable? Do you mean drifting specimen, or damage in the beam with release of sludge into the column? I can learn to deal with some specimen drift for certain specimens, but I don't want contamination.
Thanks for any input from anyone on the list.
John chandler-at-lamar.ColoState.EDU http://www.vetmed.colostate.edu/anatomy/faculty/chandler.html
Dear Dr. Robertson, I saw your message on the e-mail regarding the above mentioned subject. From what we understand you are currently using glass and yes I would agree that over a short period of time they would begin to crack. With our microwave and in conjunction with polyethylene coplin jars we have had much success. The high density polyethylene does not crack nor rupture over many uses. Our part # is 70319 if you have an interest. If you have any further questions please do not hesitate to contact us. Sincerely, Stacie Kirsch Electron Microscopy Sciences P.O.Box 251 Fort Washington, Pa, 19034 Tel:215-646-1566 Fax:215-646-8931
I shouldn't think there are any problems. However, it might be advisable to freeze dry part of the samples to compare the bacterial / plankton population. Filters can be cut up and prepared like thin plant leaves, marking the top might be advisable [unless you want to mount them vertically anyhow]. Yours sincerely
Dr Stephan Helfer, SSO Mycologist
Royal Botanic Garden Edinburgh, Inverleith Row, EDINBURGH EH3 5LR, Scotland UK
To all that has requested a copy of the protocol that I mentioned in my E-Mail message regarding the recycling of osmium please note that I have mailed to each of you a copy of the complete protocol. If anyone has questions please do not hesitate to contact me. Sincerely Stacie Kirsch Electron Microscopy Sciences P.O.Box 251 Fort washington, Pa 19034 Tel: 215-646-1566 Fax:215-646-8931
T. Robertson was asking about microwaving coplin jars. The answer is not to use glass coplin jars as they do indeed tend to break. Use plastic coplin jars available from a variety of scources. Any reference I have seen says to use the plastic ones. We obtained ours from either Fischer Scientific, or from Baxter-Canlab (now VWR-Canlab ) and have had no problems with them.
Yours, Mark Elliott, PhD UBC-Pulmonary Research Lab Vancouver, Canada
Krivanek and Mooney (Ultramicroscopy 49:95-108, 1993) state "However, whereas each pixel of the SSC (slow scan CCD) is capable of faithfully capturing an intensity between 0 and 2(to the 14th power), each pixel of film can capture only a few gray levels (roughly equivalent to the number of silver grains that can fit into the area of one pixel without overlapping)". Buonaquisti (Microscopy Today 95(1):12-13, 1995) states film "is an anlog process and, as such, one can argue that sheet film has an infinite gray scale resolution". Can anyone comment on this? I have always thought more along Krivanek and Mooney's reasoning. Does anyone know the number of grey levels film can capture? Thanks for any comments.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (314)-882-4712 (voice) (314)-882-0123 (fax)
This is in response to posting from Dr. Thor Bostrom on April 20:
There is an easy solution:
We have been making for some years a product called "Microporous Specimen Capsules". The original product had a pore size range of 120- 200 um, and is our part SPI #13215 and has overall dimensions of 10 mm x 10 mm. However, you will have to cut out a small piece of the membrane filter prior to insertion of the piece into the capsule.
We have a second product, exactly like the first, but with a pore size of 78um which should be used if the features on the membrane at that small.
We expect to have available yet a third product, just like the other two, but with pore size on the order of 30 um.
Charles A. Garber PRESIDENT SPI SUPPLIES PO BOX 656 WEST CHESTER, PA 19380
Dear Karen, I saw your message and I do have quite a bit of experience of uses with BDMA and I have spoken for quite a few years with Audrey and alot of my customers now do use BDMA in place of DMP-30 with great success. Please note the following: BDMA is less viscous than DMP-30,has a longer shelf life, and offers better penetration of the tissue. In order to achieve the optimum results my customers have all told me that when they use DMp the percentage is at 1.5-2% final volume and with the BDMA it needs to be 2.5-3%. No greater than 3 however. One should warm the resin and the anhydride prior to mixing and as well it would be beneficial to warm the stirring rod and the containers which will be used for mixing.You should make up the lot fresh however many of my customers has informed me that they have been storing the mixture at 4 degrees C in a bottle with a well fitting tight cap for several weeks and have never experienced any difficulties and have achieved fine results. It should be noted that if this is the avenue you will take prior to use the mixture must be allowed to reach room temperature. If you would like to delve deeper into this subject please just let me know. I hope this gives you some info you were looking for. Stacie Kirsch Electron Microscopy Sciences P.O.Box 251 Fort Washington, Pa. 19034 Tel:215-646-1566 Fax:215-646-8931
I don't know about coplin jars, but PARR INSTRUMENT COMPANY, 211 53rd St., Moline, IL 61265-1770 (Tel: 309 762-7716; Fax: 309-762-9453) sells microwave acid digestion vessels that can be used for LM & EM mivrowave fixation. The vessels are designed to provide complete containment of volatiles and stand pressures up to 1200 psi and temperatures up to 250 C. Hope this helps.
************************************************************************* M.V. Parthasarathy Professor of Plant Biology & Director, EM Facility 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 ***********************************************************************
IOn Thu, 20 Apr 1995, Nestor J. Zaluzec-Argonne Nat. Lab. wrote:
} From: SMTP%"delannoy-at-welchlink.welch.jhu.edu" 18-APR-1995 15:19:23.95 } To: MICROSCOPY } CC: } Subj: EM Salaries } } Date: Tue, 18 Apr 1995 14:28:44 +0500 } Message-Id: {9504181828.AA21435-at-welchlink.welch.jhu.edu} } X-Sender: delannoy-at-welchlink.welch.jhu.edu } Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } To: Microscopy-at-anlemc.msd.anl.gov } From: delannoy-at-welchlink.welch.jhu.edu (Michael Delannoy) } Subject: EM Salaries } X-Mailer: {Windows Eudora Version 1.4.2b16} } content-length: 291 } } } To: Microscopy } } From: delannoy-at-welchlink.welch.jhu.edu (Michael Delannoy) } } Subject: EM Salaries } } } } Hello, } } I'm trying to get some information on salary ranges for } } electron microscopists and EM core facility managers. Is there a } } source I can check? Thank you. } } } } M. Delannoy } } } Hi, Statistics were taken by EMSA several years ago. I can tell you that salaries are pathetic for techs, at least in my experience. I just sent a man with 12 years of experience in histo and EM pathology & research, a recent BS, and MSA Certification , to Northwestern University for a position in downtown Chicago, and he was offered $21,000 per year.
Always be carefull with using stuff like Unicryl or Lowicryl. If possible use a fumehood. Otherwise (like we do) wear a full-face gas-mask with an apropriate filter. Use the filter only once (every experiment).
I tried Unicryl, but after lots of disapointments we skipped to Lowicryl HM20. (we used the plastic for immunno Electron Microscopy). Do you get good results with Unicryl? If so, please let me know.
Bye.
Luc Analbers
*************************************************************************** * Luc Analbers * E-mail: Analbers-at-med.ruu.nl * *************************************************************************** * Utrecht University * LLL * * Medical Faculty * LLL * * Dept. Medical Physiology & * LLL A * * Sportsmedicine * LLL AA AA * * PO-box 80043 * LLL AA AA * * Zip: 3508 TA * LLLLLAAALLLAAALLL * * Utrecht The Netherlands * LLLLLAAALLLAAALLLL * * Tel: 030 - 538911 * AAA AAA * * Fax: 030 - 539036 * AAA AAA * ***************************************************************************
Be very certain that your Millipore filters are of the polycarbonate type and NOT nitrocellulose, as these will not survive the CPD. The problem with the polycarbonate filters though, is that they tend to roll up into tight tubes when dried. To hold them in place during CPD, I cut them into grid-sized pieces and clamp them between a 50 mesh folding (oyster) grid. Several of these can then be mounted on a single stub.
Hope this helps.
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
Message-Id: {9504211955.AA2171-at-pho018.sb.com} To: MICROSCOPYLISTZALUZEC {MICROSCOPYLISTZALUZEC-at-AAEM.AMC.ANL.GOV}
Millipore filters can be successfully CPD'd in envelopes made from folded filter paper, closed with a staple at the open edge. I use Whatman 541 4.25cm papers routinely with 12mm filters. The sample code can be written on the paper with pencil, and this mark can then be used to determine the "up" side of the millipore filter - sometimes the sample is invisible to the eye.
We have 50 fifth graders from Hammond Indiana coming to visit in May. They have been in a special science program and all want to see viruses in the TEM. Unfortunately, I have never worked with viruses. Does anyone have any ideas I could use? I have a JEOL 1200 TEMSCAN which is capable of visualizing them, but have to do a prep of some sort. Fortunately I have a CRT & also a computer with a program so they can see on either CRT & don't have to rely on the green screen. Thanks from Joyce at Chicago State University. .
I posted a message earlier for information on polishing sapphire for TEM. I'd now like to expand that to get any information at all on polishing sapphire. Ideal information would be suggested sample load, wheel speed, abrasive type etc. I've received several bits of information - naturally, it all conflicts to some degree. Perhaps if I get enough information, I can make a reasonable conclusion.
Thanks for your help!
David Henriks TEL: 800-728-2233 South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 e-mail: 75431.1344-at-compuserve.com
I run the day to day operations of the confocal microscopy/ video microscopy/ computer image analysis lab. I am the lowest rank faculty. My salary began at $26.5k in 1992 and is up to $32.5 now. To keep the Facility going, I work, on average, 9 hour days. We are in New York City. My salary feels low for the area. In another part of the country, this might be more comfortable.
You could try writing to macaluso-at-aecom.yu.edu to find out about the EM facility here.
Please do not post on the bboard.
----------------------------------------------- } } I'm trying to get some information on salary ranges for } } electron microscopists and EM core facility managers. Is there a } } source I can check? Thank you.
} Millipore filters can be successfully CPD'd in envelopes made from folded } filter paper, closed with a staple at the open edge. I use Whatman 541 4.25cm } papers routinely with 12mm filters. The sample code can be written on the } paper } with pencil, and this mark can then be used to determine the "up" side of the } millipore filter - sometimes the sample is invisible to the eye. } } John_Warrack-1%notes-at-sb.com } } SmithKline Beecham UK
One way we handle specimens like this for CPD is to sandwich them between two ring magnets. The thickness of the magnets protects the specimen.
The magnets are 1/2 inch diameter, 1/8 inch thick, with a 1/4 inch center hole. They are in the Edmond Scientific catalog as stock number D41,990. A pack of 25 was $5 in 1993.
Hope this helps.
John chandler-at-lamar.ColoState.EDU http://www.vetmed.colostate.edu/anatomy/faculty/chandler.html
We are currently using Kodak D-19 developer and Kodak Electron Microscopy Film 4489. We typically develop 123 pieces of film before changing developer and due to the relatively large number of potential users (6 people) we burn through the stock solution of D-19. Mixing D-19 from powder is time consuming activity, and in this era of doing more with less I would like to find a developer sold as a liquid concentrate. This would speed up darkroom maintenance and let us spend more time developing and less time preparing to develop.
Can anyone recommend a liquid developer? And while I'm asking, is there a better or equivalent film than 4489?
This is in response to posting from Dr. Thor Bostrom on April 20:
There is an easy solution:
We have been making for some years a product called "Microporous Specimen Capsules". The original product had a pore size range of 120- 200 um, and is our part SPI #13215 and has overall dimensions of 10 mm x 10 mm. However, you will have to cut out a small piece of the membrane filter prior to insertion of the piece into the capsule.
We have a second product, exactly like the first, but with a pore size of 78um which should be used if the features on the membrane at that small.
We expect to have available yet a third product, just like the other two, but with pore size on the order of 30 um.
Charles A. Garber PRESIDENT SPI SUPPLIES PO BOX 656 WEST CHESTER, PA 19380
I would like to have this information please. -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
What we have done is to put two filter face to face and hold them together using the cap of a BEEM capsule that has had a large hole punched in it with a cork borer and a ring cut from the top of the capsule that will fit inside the cap.
SO it goes cap, then two filters, then the ring on top and push them together. Something like an embroidery hoop/ -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
Good Morning Dr. Bostrom, In reply to your question about handling millipore filters containing bacteria and zooplankton for SEM, we do this sort of thing by covering the filter containing the sample with a spacer (which comes packaged between the filters) and sandwich that between two washers which are held together with small alligator clamps. Be sure when you purchase the washers that they are made of a material which will not rust. I'm not sure what the ones I use are made of....perhaps stainless or aluminum. I took them to a trophy shop and had numbers engraved on them to help with sample identification. I hope this is helpful. Sandra Zane
If the curling of membrane filters is a problem, then try using our silver membrane filter product, exactly like the polymer membrane (e.g. MCE) product in structure but made out of pure silver. Any curling is very slight and the bonus is that what ever conductive layer is eventually applied can be much less (maybe no conductive coating at all in fact) because of the inherent conductivity of the substrate. The aluminum oxide membrane filters are even stiffer, but are not conductive, but the pores are the smallest available in any membrane filter product (e.g. 20 nm). These are on pages 53 and 54 of our SPI Supplies 1991 SourceBook (catalog) of products for the EM laboratory. E-mail me if you need a copy.
Charles A. Garber PRESIDENT SPI SUPPLIES PO BOX 656 WEST CHESTER, PA 19380
Ph: (800) 2424-SPI FAX:(610) 436-5755
e-mail: GVKM07A-at-prodigy.com or sp-supp-at-cerf.com [April 25 to May 3 when I am abroad]
I seem to recollect this as a kind of wondrous upper limit, How many charges fit in a charge bucket vs the noise floor? Is the underlying question.
Candlelight to direct sun ratio is about 10E6
} each pixel of } film can capture only a few gray levels } (roughly equivalent to the number of silver grains
THE GRAINS ARE NOT ALL THE SAME SIZE - - -
} that can fit into the area of one pixel without } overlapping)". Buonaquisti (Microscopy Today 95(1):12-13, 1995) states } film "is an anlog process and, as such, one can argue that sheet film has } an infinite gray scale resolution". Can anyone comment on this? I have } always thought more along Krivanek and Mooney's reasoning. Does anyone } know the number of grey levels film can capture? Thanks for any comments. } } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } 3 Tucker Hall } University of Missouri } Columbia, MO 65211 } (314)-882-4712 (voice) } (314)-882-0123 (fax)
Regards,
Ed M.
Ed Monberg, General Manager Laser Motion and Development Co. 3101 Whipple Road Union City, CA 94587-1216 510-429-1060, FAX 429-1065 em-at-mediacity.com
On Sun, 23 Apr 1995, Nestor J. Zaluzec-Argonne Nat. Lab. wrote:
} From: SMTP%"cammer-at-aecom.yu.edu" 21-APR-1995 09:19:28.20 } To: ZALUZEC } CC: } Subj: info on EM Salaries } } Date: Fri, 21 Apr 1995 11:04:41 -0400 (EDT) } From: Michael Cammer {cammer-at-aecom.yu.edu} } Subject: info on EM Salaries } To: delannoy-at-welchlink.welch.jhu.edu } Cc: ZALUZEC-at-AAEM.AMC.ANL.GOV } In-Reply-To: {950420095748.1cc-at-AAEM.AMC.ANL.GOV} } Message-Id: {Pine.3.07.9504211140.A20241-a100000-at-alsys1} } Mime-Version: 1.0 } Content-Type: TEXT/PLAIN; charset=US-ASCII } } To keep the } Facility going, I work, on average, 9 hour days.
As I sit here on a Sunday night noticing that my reply was posted here, I would like to clarify or revise. Nine hour days was a glib comment. This means coming in at 9:30 and leaving at 7:00 with a break merely for lunch if nobody walks in demanding help. And this does not include administrative paperwork which must be prepared on weekend evenings when interruptions are less likely. I'm not complaining and, in fact, I love being constantly challenged with interesting projects; I'm merely clarifying the pay scale.
} ----------------------------------------------- } } } I'm trying to get some information on salary ranges } for } } electron microscopists and EM core facility managers. Is there a } } } source I can check? Thank you.
On Sun, 23 Apr 1995, Nestor J. Zaluzec-Argonne Nat. Lab. wrote:
} From: SMTP%"cammer-at-aecom.yu.edu" 21-APR-1995 09:19:28.20 } To: ZALUZEC } CC: } Subj: info on EM Salaries } } Date: Fri, 21 Apr 1995 11:04:41 -0400 (EDT) } From: Michael Cammer {cammer-at-aecom.yu.edu} } Subject: info on EM Salaries } To: delannoy-at-welchlink.welch.jhu.edu } Cc: ZALUZEC-at-AAEM.AMC.ANL.GOV } In-Reply-To: {950420095748.1cc-at-AAEM.AMC.ANL.GOV} } Message-Id: {Pine.3.07.9504211140.A20241-a100000-at-alsys1} } Mime-Version: 1.0 } Content-Type: TEXT/PLAIN; charset=US-ASCII } } To keep the } Facility going, I work, on average, 9 hour days.
As I sit here on a Sunday night noticing that my reply was posted here, I would like to clarify or revise. Nine hour days was a glib comment. This means coming in at 9:30 and leaving at 7:00 with a break merely for lunch if nobody walks in demanding help. And this does not include administrative paperwork which must be prepared on weekend evenings when interruptions are less likely. I'm not complaining and, in fact, I love being constantly challenged with interesting projects; I'm merely clarifying the pay scale.
} ----------------------------------------------- } } } I'm trying to get some information on salary ranges } for } } electron microscopists and EM core facility managers. Is there a } } } source I can check? Thank you.
How about making up a LARGE volume of D19 (say 10 gallons or more) and storing them in brown bottles. It will be cheaper than buying liquid developer and the D19 will last a long time if the bottles are filled and capped.
Dear Ed, Here is The infor you requested. If you can not find it let me know for I will send you a copy: To all others that have requested a copy they have all been send and posted today. 50th anniversary Meeting of EMSA proceedings, Boston. Author: W.H. Muss Let me know if you need anything else. Sincerely Stacie Kirsch Electron Microscopy Sciences
I used to CPD millipore filters with diatoms and found that there was little loss if the filters were simply a sandwich of two filters. The filters become very brittle after CPDing. If you decide to freeze dry the samples be sure to so from water free of buffers or salt or you will find all sorts of strange crystals. Good Luck.
"We haven't tried that particular brand but cyanoacrylates (superglues) shouldn't be used where they can be illuminated by the electron beam in our experience. They tend to bubble and release sludge into the column when not cured rock hard. M-Bond 610, Gatan G-1, Epotec 353ND, and Devcon-2-ton epoxys are better choices.
Superglues are far better than wax for mounting parts on preparation fixtures. They dissolve in solvents nearly as fast as wax but without a waxy residue."
What solvent do you use to dissolve and remove super glue?
Thanks, George C. Ruben Dept Biological Sciences Dartmouth College Hanover, NH 03755 tel 603 646-2144 fax 603 646-1347
I routinely CPD bacteria on a millipore filter that I roll like a "hand made" cigarette, the specimens lying inside the roll, and I simply put that roll in the metal mesh basket of the CPD, it works very well and i do not loose material.
Diane Montpetit Food research center St-Hyacinthe, Quebec, Canada tel: 514 773 1105 fax: 514 773 8461
I meant that it decomposes under the beam, releasing unwanted hydrocarbons into the column. I generally use M-Bond 610 for making cross sections, as it doesn't have this problem-also, it won't dissolve in acetone like Epoxy resins will.
} We are currently using Kodak D-19 developer and Kodak Electron Microscopy } Film 4489. We typically develop 123 pieces of film before changing } developer and due to the relatively large number of potential users (6 } people) we burn through the stock solution of D-19. Mixing D-19 from } powder is time consuming activity, and in this era of doing more with less } I would like to find a developer sold as a liquid concentrate. This would } speed up darkroom maintenance and let us spend more time developing and } less time preparing to develop. } } Can anyone recommend a liquid developer? And while I'm asking, is there a } better or equivalent film than 4489?
I think you'll find that you can use D-19 for up to about 800 plates developed without any side effects, assuming you have a nitrogen gas burst system. SO-163 is the film of choice. Its faster than 4489. Rob Keyse
Hi, I was a lens-designer and all-around optical engineer in one of my previous lives. A few times we had problems with contamination on lenses in various military instruments that we made. The problem was a cheesy goo (when examined over the microscope) and was always solved by me walking around the production/assembly floor and looking for the super-glue. Often a tech or assembler would bring in a tube in order to stake wires or subassemblies to make the work easier. When the instrument was later evacuated for back-filling with inert gas the glue would out-gas and spray all this goo around. I did a couple of experiments in a vacuum oven to verify that this is what we saw.
Now, I have to admit that nothing was done to 'cure' this super-glue, it may be that the heating was necessary (during the pump-down there was a bake-out to try to eliminate water vapor) to cause the out-gassing, but I am sure that NASA won't let super-glued components into space.
} Hello. I am currently having difficulty getting cryo sections of mouse bone } marrow macrophages to stick on grids. } } I am using conventional techniques with a dry diamond knife, picking the sections up with sucrose and floating the grids on buffer before further prossing } with immunogold. I have been using this method for some time with different } types of cells and intracellular parisites with no difficulty. My problem } seems to be with just this one cell type... } } Any advise from someone in the know? } } Thank you. } } Jaime A. Dant } Washington University School of Medicine } St. Louis } jaime-at-borcim.wustl.edu
G'day Jaime, I haven't tried this type of tissue, we work with peripheral nerve. One technique that we have found useful for some nerve types is the electrostatic transfer method. The procedure was published by Tsuji et al. 1992, in the Arch. Histol. Cytol. vol 55, p. 423-428. Hope this helps, Regards, Gerald Little.
Dr Gerald J. Little | Ph (61 49) 215618 The Neuroscience Group | Discipline of Anatomy | Fax (61 49) 216903 Faculty of Medicine and | Health Sciences | The University of Newcastle | Email ANGJL-at-medicine.newcastle.edu.au Australia, 2308 |
} } We are currently using Kodak D-19 developer and Kodak Electron Microscopy } Film 4489. We typically develop 123 pieces of film before changing } developer and due to the relatively large number of potential users (6 } people) we burn through the stock solution of D-19. Mixing D-19 from } powder is time consuming activity, and in this era of doing more with less } I would like to find a developer sold as a liquid concentrate. This would } speed up darkroom maintenance and let us spend more time developing and } less time preparing to develop. } } Can anyone recommend a liquid developer? And while I'm asking, is there a } better or equivalent film than 4489?
Try making up batches of 5 gal. or more in large cube-tainers, and then mix D19 1:2 parts H20. We develop our negs. (Kodak SO-163) for 4 min. The S0-163 film can be pushed for longer development times if necessary, up to 1\2 hr. for some special experiments.
We typically dilute the D-19 mixed stock with water 1:1 and develope 700 plates between changings. I've never seen any degradation of the developed negatives up to that time and one may even be able to go longer between developer changes-in fact, we've gone over 1000 on several occasions when people were too lazy to make up new developer.
Dont know if it is still in print (pub in 79) but there is a nice SEM atalas from The Freman Publ Co (the ones who do Scientific American) and As I seem to recall, it was relatively inexpensive:
Tisssues & Organs: a text atlas of scanning electron microscopy
Thomas E. Phillips wants comments on grey levels possible in film vs. digital. Film response is a sigmoidal curve and one adjusts exposure to fit the light coming off the subject to the linear portion of the curve in the film's response. Black & white film provides a greater number of lens aperture stops (6-8) to capture greys compared to color film (about 4, depends on the film). So the answer to the question of grey values possible in film is that one can capture a great variety of greys in film, but not all in one image-only a relatively narrow range can be captured, with the appropriate adjustment of exposure. CCDs have a linear response over several orders of magnitude. Slow scan CCDs can capture a remarkable range of greys in each image, in which case one can choose which range of greys to display on the moniter.
R. Howard Berg, Ph.D. Biology Department University of Memphis Memphis, TN, 38152 E mail: bergrh-at-cc.memphis.edu phone: 901-678-4449 fax: 901-678-4457
Anyone have a good, reproduceable protocol for making holey carbon films for high resolution work? We have tried numerous methods including Formvar/glycerol mixtures, steaming, followed by opening of pseudoholes with solvents and carbonization of the film. Major problem has been obtaining sufficient numbers of holes (either too many or not enough are obtained using glycerol). Another problem seems to be getting the Formvar to separate from the slide. Evaporated Victawet helps, but this seems a bit constraining. Thanks mucho.
John J. Bozzola Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 Phone: 618-453-3730 Fax: -2665 Email: bozzola-at-siu.edu OR bozzola-at-qm.c-cem.siu.edu
} } We are currently using Kodak D-19 developer and Kodak Electron Microscopy } Film 4489. We typically develop 123 pieces of film before changing } developer and due to the relatively large number of potential users (6 } people) we burn through the stock solution of D-19. Mixing D-19 from } powder is time consuming activity, and in this era of doing more with less } I would like to find a developer sold as a liquid concentrate. This would } speed up darkroom maintenance and let us spend more time developing and } less time preparing to develop. } } Can anyone recommend a liquid developer? And while I'm asking, is there a } better or equivalent film than 4489?
Try making up batches of 5 gal. or more in large cube-tainers, and then mix D19 1:2 parts H20. We develop our negs. (Kodak SO-163) for 4 min. The S0-163 film can be pushed for longer development times if necessary, up to 1\2 hr. for some special experiments.
} From: fskarl-at-goodyear.com (Frank Karl) } Subject: Alternates to D-19 } } Hi everyone! } } We are currently using Kodak D-19 developer and Kodak Electron Microscopy } Film 4489. We typically develop 123 pieces of film before changing } developer and due to the relatively large number of potential users (6 } people) we burn through the stock solution of D-19. Mixing D-19 from } powder is time consuming activity, and in this era of doing more with less } I would like to find a developer sold as a liquid concentrate. This would } speed up darkroom maintenance and let us spend more time developing and } less time preparing to develop. } } Can anyone recommend a liquid developer? And while I'm asking, is there a } better or equivalent film than 4489? } } } Thanks for your input. } } } Frank Karl fskarl-at-goodyear.com } Goodyear Tire & Rubber Co. Voice 216.796.7818 } Analytical Services - Dept 415B Fax 216.796.3304 } 142 Goodyear Blvd } Akron, OH 44305 } U.S.A. } } We use Ilford PQ Universal developer with Kodak 4489 film, for the convenience of using a liquid developer. Over the years we have tried different strengths, and are currently using a 1:9 dilution, as per the standard instructions for film. We normally use a fresh solution every day. The mismatch came about because we originally used Ilford film, and when switching to Kodak it was natural to test with the same processing chemicals. - the results were fine. However if there is a real incompatibility lurking in the background, we would be interested to know!
} best wishes, Sally Stowe
} } ---------------------------------------------------------------------- Sally Stowe Australian National Univ. Facility Coordinator Canberra, AUSTRALIA ANU Electron Microscopy Unit Ph 61 6 249 2743 RSBS, Box 475 Email stowe-at-rsbs-central.anu.edu.au FAX 61 6 249 4891 ------------------------------------- -------------------------------- -
Another problem seems to be getting the Formvar } to separate from the slide.
} John J. Bozzola } Center for Electron Microscopy } Southern Illinois University } Carbondale, IL 62901-4402 } Phone: 618-453-3730 } Fax: -2665 } Email: bozzola-at-siu.edu OR bozzola-at-qm.c-cem.siu.edu
To separate the formvar from the slide, I take a razor blade, and holding it at a 45 degree angle relative to the edge of the slide, I scrape all 3 sides of the slide, then I cut a line several time near the top of the slide, so that all sides have been cut with the razor blade. This should really help separate the formvar from the slide.
I see that we are back on the subject of glues again. There are two publications by M J Witcomb (The suitability of various adhesives and mounting media for scanning electron microscopy :Journal of Microscopy Vol 121 March 1981 pp. 289-308; The suitability of various mounting media for scanning electron microscopy. II. General glues Journal of Microscopy, Vol 139 Pt 1, July 1985 pp 75-114)
I think this may clear things up a bit and set a standard of criteria for glues.
Stephan H Coetee Electron Microscope Unit Private Bag 3 Wits 2050 Stephan-at-Gecko.biol.WITS.ac.za
Subject: Time: 8:00 AM OFFICE MEMO RE} D-19.alt Date: 4/25/95
We use Kodak HRP (high resolution plate) developer, Kodak catalog # 140-1306 with 4489 film. Kodak EIFilm SO-163 is a reasonable alternative and is more sensitive than 4489, reducing the time required for exposure, and is also a fine grain emulsion. Kodak has a tech sheet on this; call 1-800-242-2424, ext. 50 for technical info. I think it is publication # JJ-7
Subject: Time: 7:57 AM OFFICE MEMO Another old TEM available Date: 4/17/95
Philips 200 at UC Berkeley, has not run in two years.
Contact: Prof. Ed Sylvester at (510) 642-7353 or Ethel Nakamura in Wellman Hall at (510) 642-1603 for details. Some funds available for moving, Ask Ed.
In response to your query, a very good source of viruses are your local neighbor's baby's. Ask your friends if one of their children have a good case of diarrea, collect some stool from them, make a 10% suspension of the stuff and preform a negative stain. It will be possible to have a number of enteric viruses (adeno, rota, HAV, picorna, etc., also bacterial phages) in the preps. Of course not usually more than one species at a time. Also there will be various pieces or whole bacteria in it. If the child is a little older, there will be various undigested bits of food (ex: collegen fibers) which are neat to look at. Another side note is that if you find an enteric virus in the stool, you can tell the parents to quit giving the child antibiotics, unless of course the pediatrician has had a stool culture preformed and knows that there is a pathologic bacteria present. PS Work in a fume hood. Not fun to smell.
Good Luck, Ed Calomeni Medical College of Ohio Toledo, OH emlab-at-opus.mco.edu
If my memory serves me correctly 4489 and Ilford film are roughly equivalent in speed and grain size, although the Ilford stuff has the edge (I think). S0163 is much faster that 4489 because of the large grain but you obviously pay the price in resolution. For very fast work I have colleagues that have used DuPont mammography film (I think 10 to 15 times faster than Ilford). Does any one have a listing of what the grain size and realtive speed of the the em films is? Seems to me that this would be a useful list to maintain. As far as I know there are the 2 Kodak films, The Ilford (Ciba-Geigy) film, Agfa EM Film and Fuji EM film. Are there any others?
This info could be part of the Microscopy Web Pages. I will compile the info if we can collect it.
Jfm.
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
After reading all of the comings and goings on the listserver concerning the gluing of sapphire for TEM, I have noticed that we seem to be running parallel conversations. The initial question that started all of this was from Georges Veilleux who wanted to bond pieces together in order to make a cross section sample for TEM. The bond used to make a cross-section sample is intended to be a permanent bond - therefore, any questions about solvents would become irrelevent.
The entire conversation got twisted around when Gary Liechty suggested using Loctite 460 super glue. Super glue is an ideal material to use when mounting a sample for polishing (such as with a Tripod Polisher), but is not suitable for bonding together the cross section. Super glues or cyanoacrylates are soluble in acetone - an ideal characteristic for temporary bonding, but a very undesirable characteristic for a permanent bond. Materials such as: M-Bond 610 Gatan G1, South Bay Technology Golly G1, and EpoTek 353ND are all permanent bonding materials and I am not aware of anything that will disolve them after they have cured.
In short:
M-Bond, SBT Golly G1, Gatan G1, EpoTek 353ND are used for permanent bonding of a TEM cross section and are stable under the electron beam.
Superglues and other cyanoacrylates such as Loctite 460 are OK for temporary bonding, are soluble in acetone and are not stable under the electron beam.
If anyone has other questions concerning the bonding of materials, I would be pleased to offer whatever assistance I can. Please contact me directly and I will respond as quickly as possible. I hope this information has helped and has clarified the bonding question!
Best regards-
David Henriks TEL: 800-728-2233 South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 e-mail: 75431.1344-at-compuserve.com
Mr-Received: by mta RANDB; Relayed; Tue, 25 Apr 1995 09:38:25 -0600 Mr-Received: by mta MCM$RAND; Relayed; Tue, 25 Apr 1995 09:38:29 -0600 Mr-Received: by mta RANDB; Relayed; Tue, 25 Apr 1995 09:39:04 -0600 Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
I have used a method described by Paul Elsner in the proceedings of the 29th EMSA meeting, page 460. The method produced holey grids that even my microscope service engineer coveted!
In a chamber or room with 50-70% relative humidity, clean glass slides are dipped in a Coplin jar of 0.2-0.4% Formvar in ethylene dichloride. The films are mounted onto grids and then treated with either acetone, methanol, or chloroform-methanol (1:9) to open up pseudoholes and widen holes. Varying the concentration of Formvar and relative humidity results in different hole diameters and numbers of holes. After solvent treatment, the grids are rinsed in 95% ethanol, 50% ethanol, and distilled water - about 30 dips in each - and carbon coated.
A few tips to help strip the films from the slides: Rub a clean finger over the slide a couple of times before you dip it into the Formvar. After the film is dried, run a razor blade along the edges of the slide and cut across the top edge of the film. Then breathe on the film until it's fogged, and dip it into your stripping bath quickly. Sounds hokey, but it's a method that has never failed me.
Hope this is useful,
Jane A. Fagerland Dept. Cellular and Microscopic Research Abbott Laboratories
We use a liquid developer from Dupont called DL24 I believe. It is a graphic arts developer and we dilute it 1:7 for use. You have to buy it at least 5 gals. at a type. It comes in a polyethelene 'cubitanier' which collapses as it is emptied. It works well for us.
Dan
On Sun, 23 Apr 1995, Nestor J. Zaluzec-Argonne Nat. Lab. wrote:
} Date: Sun, 23 Apr 1995 7:43:38 -0500 (CDT) } From: Nestor J. Zaluzec-Argonne Nat. Lab. {ZALUZEC-at-AAEM.AMC.ANL.GOV} } To: MICROSCOPYLISTZALUZEC-at-AAEM.AMC.ANL.GOV } Cc: ZALUZEC-at-AAEM.AMC.ANL.GOV } Subject: Alternates to D-19 } } From: SMTP%"fskarl-at-goodyear.com" 21-APR-1995 14:21:32.08 } To: MICROSCOPY } CC: } Subj: Alternates to D-19 } } X-Sender: t456b15-at-rds163 } Message-Id: {v01510102abbdbeec0db8-at-[163.243.13.93]} } Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } Date: Fri, 21 Apr 1995 15:01:27 -0500 } To: microscopy-at-aaem.amc.anl.gov } From: fskarl-at-goodyear.com (Frank Karl) } Subject: Alternates to D-19 } } Hi everyone! } } We are currently using Kodak D-19 developer and Kodak Electron Microscopy } Film 4489. We typically develop 123 pieces of film before changing } developer and due to the relatively large number of potential users (6 } people) we burn through the stock solution of D-19. Mixing D-19 from } powder is time consuming activity, and in this era of doing more with less } I would like to find a developer sold as a liquid concentrate. This would } speed up darkroom maintenance and let us spend more time developing and } less time preparing to develop. } } Can anyone recommend a liquid developer? And while I'm asking, is there a } better or equivalent film than 4489? } } } Thanks for your input. } } } Frank Karl fskarl-at-goodyear.com } Goodyear Tire & Rubber Co. Voice 216.796.7818 } Analytical Services - Dept 415B Fax 216.796.3304 } 142 Goodyear Blvd } Akron, OH 44305 } U.S.A. } } } } } }
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% % % % Daniel Possin Work: 206/ 543-7489 % % Dept. of Ophthalmology RJ-10 FAX: 206/ 543-4414 % % University of Washington Home: 206/ 778-1714 % % Seattle WA 98195 USA Email: oemlab-at-u.washington.edu % % % % "The chinese expression 'cheung meng ba sui, gong hey fat choy' is % % equilvalent to Vulcan expression 'live long and prosper'. It's a % % small universe and getting smaller everyday". % % % %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
} } Subject: grey levels - film vs digital } } } } } } Krivanek and Mooney (Ultramicroscopy 49:95-108, 1993) state "However, } } } whereas each pixel of the SSC (slow scan CCD) is capable of faithfully } } } capturing an intensity between 0 and 2(to the 14th power), } } } } = 8 x 10E3 THAT is a HOT CCD ! } } } } I seem to recollect this as a kind of wondrous } } upper limit, How many charges } } fit in a charge bucket vs the noise floor? } } The full-well capacity of a CCD can be as high as half a million e/pixel } (as in TEK1024 CCDs, with a pixel size of 24 microns), and the noise level } is only ~10 e/sec when cooled to -40C, so a dynamic range of 14 bits can be } readily achieved. But, the true dynamic range is shot-noise limited. Say } your image is formed with 1,000,000 primary electrons/pixel (which is very, } very high), the dynamic range is 1000. } } ... } } } each pixel of film can capture only a few gray levels } } } (roughly equivalent to the number of silver grains } } I agree. } } } } that can fit into the area of one pixel without } } } overlapping)". Buonaquisti (Microscopy Today 95(1):12-13, 1995) states } } } film "is an anlog process and, as such, one can argue that sheet film has } } } an infinite gray scale resolution". Can anyone comment on this? } } Infinite? I read that too. Keep in mind that artcile was not refereed. } } Gary Fan } UC San Diego
I agree with the first half of Gary Fang's answer. Indeed, the dynamic range of cooled slow-scan CCD's can attain values in excess of } = 40,000 (we have an EG&G camera with a Thompson chip which does that when cooled to -65C). The second half of the answer, I believe, confuses dynamic range with signal/noise and whether the image is dark noise limited or shot noise limited: this will depend on the level of the signal (in photoelectrons/pixel/sec) as compared to the dark noise (in electrons/pixel/sec) due to the thermally excited electrons generated in the silicon and to the added electronic noise in the digitization electronics and in the bias voltage applied to the A/D converter. The shot noise is a phenomenon of quantum mechanical origin and cannot be eliminated, only minimized by accumulating as much signal as possible! In the example given, the image would be of very high quality indeed (S/N=1000/1)!!! Signal levels in low-light level fluorescence microscopy, which is the application I am most familiar with, almost never exceed 10,000 photoelectrons/pixel/sec with a pixel size of 20x20 micron.
No film can come even close to the dynamic range of a modern CCD: as explained by Nestor, film can be used to capture signals of various intensities by adjusting the f-stops on the camera, but the intrascene dynamic range (the number of different intensity levels that can be captured in the same image) is very limited (certainly less than 100).
Best regards, Massimo Sassaroli
_________________________________ Massimo Sassaroli, D.Sc. Dept. of Physiology & Biophysics Box 1218 Mount Sinai School of Medicine 1 Gustave L. Levy Pl. New York, NY 10029-6574
To apply for job MSD/3280, submit a resume to Lawrence Berkeley Laboratory. Following our receipt of your resume, you will receive a letter of acknowledgment. If you do not have a resume, please complete an LBL application form: this will not affect your being considered for employment.
Resumes will be retained in our active employment file for six months. During this period, you may refer to a specific job number by including it in your one-page cover letter or by writing it on the back of your resume. Individuals interviewed will be notified of selection or nonselection for the position.
Submit your resume by mail to -- The Staffing Office, 1 Cyclotron Road, MS 938A, Berkeley, CA 94720, or, if you prefer, via e-mail to the LBL Recruitment Coordinator, RLBoucher-at-LBL.gov. Please do not send your resume directly to, or otherwise contact, the NCEM.
DUTIES: Essential -- Responsible for daily operation and maintenance of several transmission electron microscopes. Duties include daily start-up, alignment, trouble shooting, user training in basic operation, alignment, microanalysis, and high-resolution imaging. Effective interaction with users and close coordination with Center staff and with manufacturers' service representatives are essential. Flexibility to work hours assigned based on Center needs.
QUALIFICATIONS: Essential -- Demonstrated ability in electron microscopy with emphasis on physical sciences. Detailed knowledge of operating principles of transmission electron microscopes, with demonstrated ability to maintain, operate and troubleshoot advanced and support transmission electron microscopes. Demonstrated experience in microscope alignment procedures for phase contrast imaging. Working knowledge of diffractogram analysis for diagnosis of defocus, astigmatism, beam tilt, sample tilt, drift, damping, information transfer etc. Sufficient understanding of principles of electron beam imaging, diffraction and crystallography to permit operating of the JEOL ARM-1000 under the scientific guidance of a user. Working knowledge of vacuum systems, specimen holders, high voltage systems and control electronics found in modern electron microscopes. Ability to isolate malfunctions in an electron microscope and to rectify failures. Practical experience with thin foil preparation methods. Elementary knowledge of principles of materials science. Elementary knowledge of computers for digital acquisition of images and microanalytical spectra. Ability to learn new techniques of imaging, diffraction, microanalysis and computer image processing. Ability to work with a minimum of supervision. Good communication skills for clear and efficient interaction with users and NCEM scientific staff scientists. Marginal -- B.S. or B.A. in general sciences, physics, or electronics, and coursework in electron microscopy.
To apply for job MSD/3280, submit a resume to Lawrence Berkeley Laboratory. Following our receipt of your resume, you will receive a letter of acknowledgment. If you do not have a resume, please complete an LBL application form: this will not affect your being considered for employment.
Resumes will be retained in our active employment file for six months. During this period, you may refer to a specific job number by including it in your one-page cover letter or by writing it on the back of your resume. Individuals interviewed will be notified of selection or nonselection for the position.
Submit your resume by mail to -- The Staffing Office, 1 Cyclotron Road, MS 938A, Berkeley, CA 94720, or, if you prefer, via e-mail to the LBL Recruitment Coordinator, RLBoucher-at-LBL.gov. Please do not send your resume directly to, or otherwise contact, the NCEM.
DUTIES: Essential -- Responsible for daily operation and maintenance of several transmission electron microscopes. Duties include daily start-up, alignment, trouble shooting, user training in basic operation, alignment, microanalysis, and high-resolution imaging. Effective interaction with users and close coordination with Center staff and with manufacturers' service representatives are essential. Flexibility to work hours assigned based on Center needs.
QUALIFICATIONS: Essential -- Demonstrated ability in electron microscopy with emphasis on physical sciences. Detailed knowledge of operating principles of transmission electron microscopes, with demonstrated ability to maintain, operate and troubleshoot advanced and support transmission electron microscopes. Demonstrated experience in microscope alignment procedures for phase contrast imaging. Working knowledge of diffractogram analysis for diagnosis of defocus, astigmatism, beam tilt, sample tilt, drift, damping, information transfer etc. Sufficient understanding of principles of electron beam imaging, diffraction and crystallography to permit operating of the JEOL ARM-1000 under the scientific guidance of a user. Working knowledge of vacuum systems, specimen holders, high voltage systems and control electronics found in modern electron microscopes. Ability to isolate malfunctions in an electron microscope and to rectify failures. Practical experience with thin foil preparation methods. Elementary knowledge of principles of materials science. Elementary knowledge of computers for digital acquisition of images and microanalytical spectra. Ability to learn new techniques of imaging, diffraction, microanalysis and computer image processing. Ability to work with a minimum of supervision. Good communication skills for clear and efficient interaction with users and NCEM scientific staff scientists. Marginal -- B.S. or B.A. in general sciences, physics, or electronics, and coursework in electron microscopy.
To apply for job MSD/3280, submit a resume to Lawrence Berkeley Laboratory. Following our receipt of your resume, you will receive a letter of acknowledgment. If you do not have a resume, please complete an LBL application form: this will not affect your being considered for employment.
Resumes will be retained in our active employment file for six months. During this period, you may refer to a specific job number by including it in your one-page cover letter or by writing it on the back of your resume. Individuals interviewed will be notified of selection or nonselection for the position.
Submit your resume by mail to -- The Staffing Office, 1 Cyclotron Road, MS 938A, Berkeley, CA 94720, or, if you prefer, via e-mail to the LBL Recruitment Coordinator, RLBoucher-at-LBL.gov. Please do not send your resume directly to, or otherwise contact, the NCEM.
DUTIES: Essential -- Responsible for daily operation and maintenance of several transmission electron microscopes. Duties include daily start-up, alignment, trouble shooting, user training in basic operation, alignment, microanalysis, and high-resolution imaging. Effective interaction with users and close coordination with Center staff and with manufacturers' service representatives are essential. Flexibility to work hours assigned based on Center needs.
QUALIFICATIONS: Essential -- Demonstrated ability in electron microscopy with emphasis on physical sciences. Detailed knowledge of operating principles of transmission electron microscopes, with demonstrated ability to maintain, operate and troubleshoot advanced and support transmission electron microscopes. Demonstrated experience in microscope alignment procedures for phase contrast imaging. Working knowledge of diffractogram analysis for diagnosis of defocus, astigmatism, beam tilt, sample tilt, drift, damping, information transfer etc. Sufficient understanding of principles of electron beam imaging, diffraction and crystallography to permit operating of the JEOL ARM-1000 under the scientific guidance of a user. Working knowledge of vacuum systems, specimen holders, high voltage systems and control electronics found in modern electron microscopes. Ability to isolate malfunctions in an electron microscope and to rectify failures. Practical experience with thin foil preparation methods. Elementary knowledge of principles of materials science. Elementary knowledge of computers for digital acquisition of images and microanalytical spectra. Ability to learn new techniques of imaging, diffraction, microanalysis and computer image processing. Ability to work with a minimum of supervision. Good communication skills for clear and efficient interaction with users and NCEM scientific staff scientists. Marginal -- B.S. or B.A. in general sciences, physics, or electronics, and coursework in electron microscopy.
To apply for job MSD/3280, submit a resume to Lawrence Berkeley Laboratory. Following our receipt of your resume, you will receive a letter of acknowledgment. If you do not have a resume, please complete an LBL application form: this will not affect your being considered for employment.
Resumes will be retained in our active employment file for six months. During this period, you may refer to a specific job number by including it in your one-page cover letter or by writing it on the back of your resume. Individuals interviewed will be notified of selection or nonselection for the position.
Submit your resume by mail to -- The Staffing Office, 1 Cyclotron Road, MS 938A, Berkeley, CA 94720, or, if you prefer, via e-mail to the LBL Recruitment Coordinator, RLBoucher-at-LBL.gov. Please do not send your resume directly to, or otherwise contact, the NCEM.
DESCRIPTION (Job MSD/3280) --
Research Technician, Senior
1.Job MSD/3280 2.Materials Sciences Division 3.National Center for Electron Microscopy 4.$12.10-$20.53/Hr.
DUTIES: Essential -- Responsible for daily operation and maintenance of several transmission electron microscopes. Duties include daily start-up, alignment, trouble shooting, user training in basic operation, alignment, microanalysis, and high-resolution imaging. Effective interaction with users and close coordination with Center staff and with manufacturers' service representatives are essential. Flexibility to work hours assigned based on Center needs. QUALIFICATIONS: Essential -- Demonstrated ability in electron microscopy with emphasis on physical sciences. Detailed knowledge of operating principles of transmission electron microscopes, with demonstrated ability to maintain, operate and troubleshoot advanced and support transmission electron microscopes. Demonstrated experience in microscope alignment procedures for phase contrast imaging. Working knowledge of diffractogram analysis for diagnosis of defocus, astigmatism, beam tilt, sample tilt, drift, damping, information transfer etc. Sufficient understanding of principles of electron beam imaging, diffraction and crystallography to permit operating of the JEOL ARM-1000 under the scientific guidance of a user. Working knowledge of vacuum systems, specimen holders, high voltage systems and control electronics found in modern electron microscopes. Ability to isolate malfunctions in an electron microscope and to rectify failures. Practical experience with thin foil preparation methods. Elementary knowledge of principles of materials science. Elementary knowledge of computers for digital acquisition of images and microanalytical spectra. Ability to learn new techniques of imaging, diffraction, microanalysis and computer image processing. Ability to work with a minimum of supervision. Good communication skills for clear and efficient interaction with users and NCEM scientific staff scientists. Marginal -- B.S. or B.A. in general sciences, physics, or electronics, and coursework in electron microscopy. CLOSING DATE: Open until filled.
Never, never submit a posting to the temporary address:
MicroscopyListZaluzec-at-aaem.amc.anl.gov
always submit your messages to the correct address:
Microscopy-at-aaem.amc.anl.gov.
The temporary address "MicroscopyListZaluzec-at-aaem.amc.anl.gov" was set up so that I can monitor and remove any problems which were created by the series of bouncing messages, DNS/Router crashes and several other things that went wrong last week. I have been manually checking each message which comes through and trashing the various error messages etc... and only forwarding the "real" mail. This minimizes your Email traffic and slows things down, and complicates my life, since I now check each message before it goes out. (BTW that is one reason for the longer delays now between the time you send a message and you see it back as a posting to the list).
If you post to the temporary address you WILL likely start another loop (Yes a person who will remain nameless did this today). I think I caught it before it got too far, but I'm sure several iterations went out. I have shut down the server and trashed any messages in the queue. This means that some things will likely be lost.
Please don't try to second guess the system. Just use the old address and thing will be fine, albeit abit slow for the next few more days....
Your Frustrated and Temporarily Unfriendly Neighborhood SysOp.
Bob Josephs suggests making up your D-19 stco and then storing it in brown bottles. What about making up your stock and freezing aliquots? You then freeze the quantity most suited to the final dilution you will use.
I have some 150 grams of dried-up colloidal silver paste that is not worth trying to get back into solution, partly because it costs more to ship the solvent to Hawaii than to purchase new paste. Is it worth trying to recycle somehow? What all is in it besides silver? Is it pretty pure? I'm not even sure there's anyone in Hawaii who recycles silver from darkroom chemicals.
Besides the osmium, what else should us environmentally-concerned microscopists be trying to conserve/recycle?
A belated Happy Earth Day to you all!
Mahalo, Tina Weatherby Carvalho Biological EM Facility University of Hawaii
I would like to get and exchange information about the Gatan Imaging Filter. We will receive ours at the end of the month, so we are absolute beginners. What I like to see are hints about the little things you just don't know because you have not enough experience.
Every help, sugestions and ideas is highly welcomed.
Re filters for cpd, I have made and used for years an aluminium holder about 15 mm on a side, ~cubic, consisting of folded thin alum sheet which switches back and forth in a multiple 'S' fashion so that individual slots about 1.5 mm wide by 15 * 15 mm are made. I have about 8 or 10 slots per holder. The multiple s is held together with a wide 'U' shape that holds the side and bottom of the holder. Individual filters are slipped down into the slots (I do this in a 20 or 30 mm beaker filled with enough alcohol that the filter stays emersed as it is fed into the slot, the filter is quickly transfered from the previous container to avoid drying by evaporation) By the way, I much prefer nucleopore filters to millipore, better background for SEM. I do not believe that material falls off the filter unless too much matrerial is applied. A good rule is to put about 500,000 cells on a 12 mm dia filter. Call me if you can't figure out the design of the holder. It took me 10 minutes to make with simple tools.
Alan S. Pooley ,PhD Bivalve shell SEM & shape analysis SEM/Morphometrics lab Marine & Coastal Sciences Rutgers University 908 932 8959 ext 225 Pooley-at-ahab.rutgers.edu
Would anyone out there have any experience with a resin called "Immuno-Bed" Embedding Resin (Polysciences?). It is in the EM Sciences catalogue and is claimed to be "A low viscosity media for light microscopy and immuno histochemistry techniques". I would like to know whether it would be suitable for staining of animal tissue using routine histo staining techniques, specifically H&E, PAS, Masson's Trichrome, Verhoeff's Elastic Stain and Periodic Acid Silver Haematoxylin Methamine. Comment regarding its staining reproducibility, ease of cutting etc are also welcome. Any suggestions of alternatives to the "Immuno-Bed" would also be appreciated.
Please correspond with me directly. Thanks in advance.
John//////// ////////////
-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--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- John F. Putterill -at- -at- Electron Microscopy Unit Tel: (Int) 27-12-529-9174 -at- -at- Pathology Section Fax: (Int) 27-12-529-9165 -at- -at- Onderstepoort Veterinary Institute E-mail: john-at-moon.ovi.ac.za -at- -at- Private Bag X05 -at- -at- Onderstepoort 0110 -at- -at- South Africa -at- -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--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-
X400-Received: by mta merit in /PRMD=internet/ADMD=telemail/C=us/; Relayed; Thu, 27 Apr 1995 10:56:16 -0400 X400-Received: by /ADMD=TELEMAIL/C=US/; Relayed; Thu, 27 Apr 1995 10:46:14 -0400 X400-Received: by /PRMD=SCHERING-PLOUGH/ADMD=TELEMAIL/C=US/; Relayed; Thu, 27 Apr 1995 10:44:20 -0400
Greetings,
I have to validate an EDAX DX-4. I am in the pharmaceutical industry and validation for the FDA on our equipment is top priority. Most of the advice that I have been receiving is more calibration than validation. What do others do to validate such a piece of equipment?
Thank you!!
Barbara Hartman Schering-Plough Research Lafayette, NJ 07848 (201) 579-4343 (201) 579-4211 FAX E-Mail: Barbara.Hartman-at-1773.220.Schering-Plough.sprint.com
In answer to John Mansfield's questions, there was another EM film in 1990, but I haven't looked lately to see whether it is still available: Mitsubishi MEM.
Let me refer you to a paper by Y. Kamiya and T. Arii, J. Electron Microscoy 39: 351-355 (1990). It shows that Kodak 4489 and Fuji FG are very similar. Mean sizes of developed grains in the Fuji film were measured at 0.24 and 0.33 micron for development times of 4 and 10 minutes, respectively. The Mitsubishi film was shown to be 5 times as sensitive as the Kodak or Fuji films at a development time of 4 minutes. Each film used a different developer.
Several years ago, we learned of a developing technique which uses a 2-part developer (Daniel Morse at Los Alamos National Laboratory). He (and we) used a particular product called Diafine, but there are other 2-part developers available. This procedure holds back development in areas of high exposure and increases development in areas of low exposure. The result is a negative which can be printed easily compared to a negative developed in a single-solution developer. Diafine also increases the effective film speed.
I have some data sheets from Kodak which compare 4489 and SO-163. I do not have rms granularity numbers for the films, but Kodak did provide relative electron speeds (sensitivities) for electron energies between 50 and 100 keV. These sensitivities were measured at a density of 1.0 above gross fog and are the reciprocal of the exposure measured in electrons per square micron.
Film Electron Speed Developer Devel. Time
SO-163 2.2 D19, full strength 12 min.
SO-163 0.8 1 part D19 to 2 parts 4 min. of water
SO-163 0.15 (1985) D19, full strength, 2 min. or 0.4 (1990) plus 1 gm/l of Kodak Anti-Fog #1
4489 0.4 1 part D19 to 2 parts 4 min. of water
By the way, the Kodak technical sheet for SO-163 recommends maintaining the relative humidity above 25% to mitigate against static build-up (and discharges) on the film.
Russell E. Cook Electron Microscopy Center for Materials Research Argonne National Laboratory
The Wisconsin Center 702 Langdon Street Madison, WI 53706
organized by the
Integrated Microscopy Resource University of Wisconsin-Madison
******************************************************************************** Presentations in this symposium will focus on biological problems for which a combination of microscopies (i.e. integrated microscopy) has been used. In this way the speakers will demonstrate by example the power, potential and limitations of various microscopical techniques. The techniques which will be discussed include: DIC, confocal, 2-photon excitation imaging, SEM, TEM, cryo-specimen preparation, and AFM. ********************************************************************************
FRIDAY (Evening), September 29, 1995 6:00 - 7:55 RECEPTION 8:00 - 8:45 Ken Jacobson, Univ. of N. Carolina-Chapel Hill 8:50 - 9:35 John Sedat, Univ. of California-San Francisco 9:40 - 10:25 Edward Salmon, Univ. of N.Carolina-Chapel Hill SATURDAY, September 30, 1995 8:30 - 9:15 D. Lansing Taylor, Carnegie-Mellon University 9:20 - 10:05 Jeff Lichtman, Washington University 10:10 - 10:40 COFFEE BREAK 10:45 - 11:30 John White, University of Wisconsin-Madison 11:35 - 12:20 Steven Smith, Stanford University 12:25 - 1:25 LUNCH (on your own) 1:30 - 2:15 Gary Borisy, University of Wisconsin-Madison 2:20 - 3:05 Ralph Albrecht, Univ. of Wisconsin-Madison 3:10 - 3:55 Paul Bridgman, Washington University-Madison 5:00 - 6:00 EXHIBITOR'S SOCIAL 6:00 - 8:00 BUFFET DINNER SUNDAY (Morning), October 1, 1995 9:00 - 9:45 Kent McDonald, Univ. of California-Berkley 9:50 - 10:35 Stanley Erlandsen, University of Minnesota 10:40 - 11:10 COFFEE BREAK 11:15 - 12:00 Hans Ris, University of Wisconsin-Madison 12:05 - 12:50 Eric Henderson, Iowa State University
Fees: General Registration $100.00 (Late fee: $130.00) (Includes: Fri. Reception, Sat. Social & Dinner, Coffee Breaks and Materials) Local Registration $ 80.00 (Late fee: $110.00) (Includes: Fri. Reception, Sat. Social, Coffee Breaks and Materials) Guest Ticket - Saturday Dinner $ 20.00 (On-site Unavailable)
Interested in attending?
Consult our Web site for additional information:
http://www.bocklabs.wisc.edu/imr/imr.html
To receive a brochure and registration form write to: IMR University of Wisconsin-Madison 1675 Observatory Drive Madison, WI 53706
or email: imradmin-at-calshp.cals.wisc.edu
Following the symposium, there will be a two-day workshop at the IMR. We will be presenting lectures and provide "hands-on" experience for the following techniques: * 2-photon excitation imaging * 4D DIC imaging * cryo-SEM * high pressure freezing * reversible embedment for SEM and TEM
Workshop attendence will be limited to 25 students. A letter of application is required. Fee: $150.00.
Does anyone know if there will be a Polaroid International Micrograph Contest this year? I've not been able to get hold of anyone at Polaroid who knows...
Aloha, Tina Carvalho Biological EM Facility University of Hawaii
Peggy: Thanks for inquiring about my vacuum book. The following is the information you requested: Title: Vacuum Methods in Electron Microscopy Author: Wilbur C. Bigelow Series: Practical Methods in Electron Microscopy, Vol. 15, Audrey M. Glauert, Editor Soft cover: ISBN 1-85578-053-4, 492 pp/116 illus/1994/$80 Hard cover: ISBN 1-85578-052-6, 492 pp/116 illus/1994/$175 In US and Canada, order from: Portland Press, Ashgate Publishing Co., Old Post Road, Brookfield, VT, 05036-9704. U.S.A. Ph: 802-276-3162; Fx: 802-276-3837 In Europe, order from: Portland Press Ltd., Commerce Way, Colchester CO2 8HP, UK Ph: 0206-796351; Fx: 0206-799331 I apologize for the fact that the prices are a bit higher than one might wish them to be; however, the book is nearly 500 pages long, and was a high quality production by Cambridge Univ. Press. It is printed on high quality paper, and the figures are of very high quality. In addition, I think you will find it to be filled with practically useful operational information. The two reviews I have seen so far have been pleasingly favourable. A quote from one for the Italian EM Society: "It is a (sic) almost perfect book, practical and convincing. The Series title is masterly (sic) fulfilled by practical hints, numerical data, references to everyday life, operation procedures, and suggestions, which, if followed, will save trouble and time for microscopists and will optimise their results". If you buy it, I hope you like it and find it useful. Wil Bigelow (bigelow-at-umich.edu)
} Subject: d-19 storage } } From: SMTP%"MAARTENP-at-biology.und.ac.za" 26-APR-1995 09:27:38.06 } To: ZALUZEC } CC: } Subj: d-19 srorage } } Message-ID: {MAILQUEUE-101.950426080958.523-at-biology.und.ac.za} } From: Mrs Priscilla Maartens {MAARTENP-at-biology.und.ac.za} } To: zaluzec-at-aaem.amc.anl.gov } Date: Wed, 26 Apr 1995 08:09:58 +0200 (SAST) } Subject: d-19 srorage } X-pmrqc: 1 } Priority: normal } X-mailer: PMail v3.0 (R1a) } } Bob Josephs suggests making up your D-19 stco and then storing it in } brown bottles. } What about making up your stock and freezing aliquots? You then } freeze the quantity most suited to the final dilution you will use. } I don't think it will work. The salts in the developer (sodium sulfite, etc.) will come out of solution and need to be redissolved before use, which maybe difficult (or impossible) depending on the other developer constituents.
Sorry-
Dan
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% % % % Daniel Possin Work: 206/ 543-7489 % % Dept. of Ophthalmology RJ-10 FAX: 206/ 543-4414 % % University of Washington Home: 206/ 778-1714 % % Seattle WA 98195 USA Email: oemlab-at-u.washington.edu % % % % "The chinese expression 'cheung meng ba sui, gong hey fat choy' is % % equilvalent to Vulcan expression 'live long and prosper'. It's a % % small universe and getting smaller everyday". % % % %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
} Can anyone tell me the origin of the term (lbs.) as it relates to the unit of } measure pounds? } } Thanks } } Jim Campbell
It comes from the latin word libra which was the original term and is still used in French, Spanish and other latin languages. The word 'pound' is the germanic equivalent also used in Dutch.
Doug
****************************************** Dr Douglas Arrell Mechanical Properties and Joining Division Institute for Advanced Materials Joint Research Centre European Commission Westerduinweg 2 1755 ZG Petten The Netherlands
we have a B&L stereozoom 7 in great shape. magnification varies from 10X to 70X and comes with a supplemental .5X lens to extend the magnification range from 5X to 35X. The asking price is US $1500.
I am interested in hearing from anyone who has worked with a JWS 7500E SEM for defect analysis on silicon wafers. Any information anyone can offer would be a great help!
Many thanks!
Steve McGarvey Photolithograhy Process Tech Fujitsu Microelectronics, Inc. Gresham, Oregon 97030
I will appreciate any trick (besides immersion in xylene) for removing coverslips from previously mounted 10 microns thick paraffin sections. In addition, any comments on antigenicity left over on those sections, which were previously stained with H&E?
************************************************************ *Cesar D. Fermin, Ph.D \|*|/ Fax (504) 587-7389 * *Tulane Medical School /|*|\ Answ. Mach.(504) 584-2618 * *Pathology/SL79 \|*|/ Secretary (504) 584-2436 * *New Orleans, La 70 112 /|*|\ Lab (504) 5841 * *Fermin-at-TMC.Tulane.edu -} Director of Morphological Services* ************************************************************
In response to Frank Scheltens query about the limits of the Mac OS to keep track of files, our local computer systems administrator/guru, Paul Domagala, said that the OS is limited to 65,000 per partition. That is, each partition of a drive can have up to 65K files. He is unaware of any other limitations. In other words, Frank, it appears that you won't have a file-limitation problem, especially if most of the files are multi-megabyte images. If you think you'll have too many files, just partition the drives.
Russell E. Cook Electron Microscopy Center for Materials Research Argonne National Laboratory
A short message for users of (or people interested in) the freeware version of 'S-SIMPLY' (SHRLI-SIMulation and diSPLaY of TEM & HRTEM images, for PCs), available on FTP.UNIV-LYON1.FR (/pub/dos/HRTEM) has been significantly re-updated, and new features are available (graphical displays, a "Make Interface" tool, "Beams" / Pendellosung plot....).
P.S. : S-SIMPLY will be available for use at the NCEM SummerSchool 1995 "Computer Simulation and Processing of HRTEM Images" to be held June 26-30, 1995, at NCEM, Berkeley, CA. (for information : Michael A. O'Keefe, MAOK-at-LBL.GOV)
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
In response to your question about the Polaroid Photomicrography Contest, it is no longer held. Polaroid called the event off in 1994, I believe for economic reasons. So we'll all have to live with the wonderful images it produced during its life span.
Regards,
Ellie Solit Director of Publication The Microscope Book
On Thu, 27 Apr 1995, Nestor J. Zaluzec-Argonne Nat. Lab. wrote:
} From: SMTP%"tina-at-wana.pbrc.Hawaii.Edu" 27-APR-1995 14:43:51.17 } To: MICROSCOPY } CC: } Subj: Polaroid Micrograph Contest??? } } Date: Thu, 27 Apr 1995 10:37:40 -1000 (HST) } From: Tina Carvalho {tina-at-ahi.pbrc.Hawaii.Edu} } Subject: Polaroid Micrograph Contest??? } To: Microscopy Newsgroup {microscopy-at-aaem.amc.anl.gov} } Message-Id: {Pine.3.89.9504271047.A3193-0100000-at-ahi} } Mime-Version: 1.0 } Content-Type: TEXT/PLAIN; charset=US-ASCII } } Does anyone know if there will be a Polaroid International Micrograph } Contest this year? I've not been able to get hold of anyone at Polaroid } who knows... } } Aloha, } Tina Carvalho } Biological EM Facility } University of Hawaii }
} Fellow Microscopists, } } In the process of designing an image archive system (Mac based/huge external } HD/CD-ROM writer/CD-ROM Jukebox), I have heard rumblings concerning the } limitations of the Mac OS with regard to the number of individual files and } total external SCSI HD capacity that can be handled by the system. This } could be a big problem; we are planning on handling thousand of MB size } images on muli GB external drives before writing to the CD-ROM. } } Has anybody out there experienced problems of this type? } } Advise from anyone operating a similar system would be greatly appreciated! } } } Cheers, } Frank
MacOS up to version 7.1 was only capable of holding 2Gb per disk partition. I don't think that you could easily produce multiple partitions either, but I may be wrong here. System 7.5 allows disks of up to 4Gb. To use larger disks you _may_ need to purchase some disk partition software. We use 'Formatter Five' from Software Architects which allows us to put multiple partitions (mixed Mac and PC) on the same disk so that we can use the same removable disks on Macs and PCs as transferring a few hundred Mb at a time is very slow on our network.
Doug
****************************************** Dr Douglas Arrell Mechanical Properties and Joining Division Institute for Advanced Materials Joint Research Centre European Commission Westerduinweg 2 1755 ZG Petten The Netherlands
} } From: SMTP%"tina-at-wana.pbrc.Hawaii.Edu" 26-APR-1995 14:44:05.69 } To: MICROSCOPY } CC: } Subj: recycle silver paste? } } Date: Wed, 26 Apr 1995 10:26:53 -1000 (HST) } From: Tina Carvalho {tina-at-ahi.pbrc.Hawaii.Edu} } Subject: recycle silver paste? } To: Microscopy Newsgroup {microscopy-at-aaem.amc.anl.gov} } Message-Id: {Pine.3.89.9504261052.A29751-0100000-at-ahi} } Mime-Version: 1.0 } Content-Type: TEXT/PLAIN; charset=US-ASCII } } Aloha, all- } } I have some 150 grams of dried-up colloidal silver paste that is not worth } trying to get back into solution, partly because it costs more to ship } the solvent to Hawaii than to purchase new paste. Is it worth trying to } recycle somehow? What all is in it besides silver? Is it pretty pure? } I'm not even sure there's anyone in Hawaii who recycles silver from darkroom } chemicals. } } Besides the osmium, what else should us environmentally-concerned } microscopists be trying to conserve/recycle? } } A belated Happy Earth Day to you all! } } Mahalo, } Tina Weatherby Carvalho } Biological EM Facility } University of Hawaii
Tina, Mechanical vacuum pump oil to gas stations if nowhere else. Any large film developer should have a system for recycling fixer (which contains the Ag removed from the film). We bought containers to store several gallons in until we bring to a store down the street. There is a small charge for that service. Unfor tunately, the campus life safety group doesn't pick this up like they do other chemicals for proper disposal. We have them pick up used acetone and alcohol etc.} I know Hawaii is different in unexpected ways; i.e. the price of milk. Good Luck. I still remember the first Earth Day when I went to Herald Square, and collected bumper stickers etc. John Hunt
From: SMTP%"kris-at-miat0.vein.hu" 28-APR-1995 01:47:53.25 To: MICROSCOPY CC: Subj: Validation of EDS Equipment
Date: Fri, 28 Apr 1995 09:43:21 -0500 Message-Id: {9504281443.AA09249-at-miat0.vein.hu} X-Sender: kris-at-miat0.vein.hu X-Mailer: Windows Eudora Version 1.4.4 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" To: Microscopy-at-aaem.amc.anl.gov From: kris-at-miat0.vein.hu (Kovacs Kristof) Subject: Validation of EDS Equipment
Dear Barbara:
I don't know too much about the FDA validation process but if I were you I would do the following:
Have some good, approved standards e.g. from SPI Supplies in West Chester, PA (e-mail:sp-supp-at-cerf.com) and do several quantitative analysis runs on composite standards. Compare the results with the specified and approved standard values and report the results of comparison.
My other suggestions are connected to the operation of the instrument. In order to do this you have to have a good textbook dealing with EDS analysis, or some laboratory workbook such as Scanning Electron Microscopy, X-Ray Microanalysis, and Analytical Electron Microscopy by Charles E. Lyman et al.
1. Check the incoming vs. processed count rate. For a good instrument there should not be much difference up to about several thousand incoming counts.
2. Check the resolution of the detector with with a pure metal standard (not necessarily Mn), calculate it to Mn K alpha and see if it is in accordance with the specification.
3. Check the window constant of the detector. It is important to know the detector efficiency especially for light elements.
That's all, if you have any further question just send me a message.
From: SMTP%"S.Helfer-at-rbge.org.uk" 28-APR-1995 02:28:33.15 To: ZALUZEC CC: Subj: Re: Term lbs.
To: "Nestor J. Zaluzec-Argonne Nat. Lab." {ZALUZEC-at-aaem.amc.anl.gov} From: "STEPHAN HELFER" {S.Helfer-at-rbge.org.uk} Organization: Royal Botanic Garden Edinburgh Date: Fri, 28 Apr 1995 09:24:46 BST Subject: Re: Term lbs. Priority: normal X-mailer: WinPMail v1.0 (R2) Message-ID: {F09E282F0C-at-rbg-3.rbge.org.uk}
From: SMTP%"Campbell36-at-aol.com" 27-APR-1995 20:06:46.60 To: MICROSCOPY CC: Subj: Term lbs.
Can anyone tell me the origin of the term (lbs.) as it relates to the unit of measure pounds?
Thanks
Jim Campbell
Chambers Everyday Dictionary: Pound: unit of mass of varying value [hence not metric] long used in Western and Central Europe, more or less answering to the Roman LIBRA, whose symbol lb is used for pound.. ... British pound = 0.54359237kg...
Incidentally the Pound Sterling used to have the value of 0.543...kg of silver; comme ca change
Yours sincerely
Dr Stephan Helfer, SSO Mycologist
Royal Botanic Garden Edinburgh, Inverleith Row, EDINBURGH EH3 5LR, Scotland UK
Instead of all this discussion on the dynamic range of CCDs, how about considering the point spread function of scintillator/CCD systems?. If you want to measure intensities, it seems to me that you should be taking this into account.
Jfm
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
Two very quick questions: Is a short course (1 week)in biological TEM available in the US? Is Dr. Bendayan offering his course on colloidal gold labeling this year? Thanks Marge
Marge Hukee EM Core Facility hukee.margaret-at-mayo.edu Mayo Foundation (507) 284-3148 ---------------------------------------------------------------------------- --
If you are interested in this TEM please contact Dr. Fred Sack directly.
M.V. Parthasarathy ------------------------------
Partha - we have a Zeiss 10 TEM in good shape that the College wants to dispose of since they won't pay a service contract and moving charges (I have my own but can't swing another one..)
Zeiss will take it back for free and we could strip it, but would be happy to GIVE it away for anyone willing to pay for shipping. Do you know of anyone interested??
Fred Sack phone: 614-292-0896 Plant Biology fax: 614-292-6345 Ohio State University e-mail: sack.1-at-osu.edu 1735 Neil Avenue alternate e-mail address:fsack-at-magnus.acs.ohio-state.edu Columbus Ohio 43210
} From: "Fermin, Cesar" {fermin-at-tmc.tulane.edu} } Subject: FW: Coverslip removal } To: "Zaluzec, Nestor" {ZALUZEC-at-AAEM.AMC.ANL.GOV} } X-Mailer: Mail*Link SMTP-MS 3.0.2 } } } } I will appreciate any trick (besides immersion in xylene) for removing } coverslips from previously mounted 10 microns thick paraffin sections. In } addition, any comments on antigenicity left over on those sections, which were } previously stained with H&E? }
Try freezing. I have had success by freeezing from an inverted can of dust-off or similar compressed air product that uses a compressed gas as a propellant. Direct the spray right onto the cover slip, the pop off with a razor blade or scalpel blade.
Peter D. Barnett - Forensic Science Associates - Richmond CA pbarnett-at-crl.com VOICE: 510-222-8883 FAX: 510-222-8887
Can anyone with experience point me in the right direction toward chosing a high resolution video capture board. We are about to buy a fuji pictography 3000 color printer at $26,000. (Thanks to all who helped me earier with advice about this and other printers). We are trying to capture the highest possible spatial and color resolution from Betacam SP tape via a Sony BVW 75 tape reader, which gives COMPONENT signal (intensity Y, red-Y, Blue-Y). We hear various 'line' resolutions from this, ranging from 450 to 750 analog lines of video resolution. Does anyone know what the actual is? How does this translate into pixels? Do we need mo Do we need more than 640 * 480 * 8 bits/color channel? Some of the boards support 1024 * 1024 and various color resolution depths. Where do you see a differnce? We are willing to go either powermac or pentium and want to find the capture card with the best. We also want to be able to capture 5-10 images 'on the fly' as the tape plays, rather than slowwing it down or freezing the particular frame we want to capture. The subject matter is color video of tube worm etc fields growing rapidly at the hydrothermal vents as captured on betacam sp for national geographic tv last november by Rich Lutz. Our purpose in digitizing is both for measurement and for high quality image hard copy to the fuji printer via photoshop on the powermac. (Does photoshop come on the PC?)
I would appreciate any help you can give me.
Alan Pooley Marine Science SEM & Morphometrics lab, Rutgers univ 908 932 8959 ext 225 pooley-at-ahab.rutgers.edu
In your message dated Thursday 27, April 1995 you wrote :
} Can anyone tell me the origin of the term (lbs.) as it relates to the unit of } measure pounds?
This is quite an interesting one, actually.
In Roman times the Latin work for a pair of scales or a balance was 'libra', this word is still used for the astronomical constellation Libra, the scales. Over time the word for scales became associated with a specific mass, what we call a pound today.
The lb stands for 'libra', lbs is really a sort of false plural, adding an Anglo-Saxon ending to a Latin word! 'Libras' is nonsense, it should have been 'librae' I suppose. That explains the lbs, but leaves the new question, 'Why did we start to call the mass a 'pound', and when? Anyone know?
Oh, by the way. The English *monetary* value, the pound, was originally the value of one pound in weight of metallic silver. Inflation has long since made nonsense of that! But that explains the pound sign, it's an old-fashioned copperplate capital 'L', for 'libra' again.
Kind regards,
Chris --------------------------------------------------------------------------- | Chris Jefferies E-Mail at home - chris-at-stowey.demon.co.uk | | at work - chris.jefferies-at-bbsrc.ac.uk | | Microscopy page {URL: http://metro.turnpike.net/jefferie/index.html} | ---------------------------------------------------------------------------
external=20 } HD/CD-ROM writer/CD-ROM Jukebox), I have heard rumblings concerning the=20 } limitations of the Mac OS with regard to the number of individual files and= =20 } total external SCSI HD capacity that can be handled by the system. This=20 } could be a big problem; we are planning on handling thousand of MB size=20 } images on muli GB external drives before writing to the CD-ROM. } } Has anybody out there experienced problems of this type? } } Advise from anyone operating a similar system would be greatly appreciated!= =20 } } } Cheers, } Frank Dear Frank, This is a problem that we are also facing. At the moment it appears= =20 that we are collecting about 1.5 Gb of confocal images a month. On top of=20 this our network has also to deal with a large volume of images from other= =20 sources such as laser scaning densitometers. Our network consists of a SUN= =20 based PCNFS network with ~80 PCs and 10 MACs all acessing files held=20 centrally on our unix machine. All our file should ideally be available to= =20 all machines and we also need to be sure that they are all properly backed= =20 up in case of accidents. I am currently talking to a number of computer suppliers regarding= =20 data storage. The current favourite option appears to be to have a large=20 disk farm, about 27 Gb for online storage, then as this becomes fully to=20 move of data onto a read/write optical juke box then finally as this becomes= =20 full to write CD roms. The price for this system complete with backup=20 software appears to be about =A320-30 sterling.=20 What I was reallly wondering is what do you other confocal=20 microscopist do? I am a microscopist and I don't want to spend all day being= =20 a network manager so I am looking for a simple answer.
Peter
Peter McHardy, Technology Services Manager, Dept. Medical Oncology, Glasgow University, Alexander Stone Building, Garscube Estate, Switchback Road, Bearsden, Glasgow G61 1BD Tel No. 041-330 4811 Fax No. 041-330 4127 e-mail:gpma44-at-udcf.gla.ac.uk
This message got accidently confused by me and was incorrectly titled. My apologizes
Nestor.
---------------------
Meeting Announcement
Symposium on
"INTEGRATED MICROSCOPY"
September 29 to October 1, 1995
at
The Wisconsin Center 702 Langdon Street Madison, WI 53706
organized by the
Integrated Microscopy Resource University of Wisconsin-Madison
******************************************************************************** Presentations in this symposium will focus on biological problems for which a combination of microscopies (i.e. integrated microscopy) has been used. In this way the speakers will demonstrate by example the power, potential and limitations of various microscopical techniques. The techniques which will be discussed include: DIC, confocal, 2-photon excitation imaging, SEM, TEM, cryo-specimen preparation, and AFM. ********************************************************************************
FRIDAY (Evening), September 29, 1995 6:00 - 7:55 RECEPTION 8:00 - 8:45 Ken Jacobson, Univ. of N. Carolina-Chapel Hill 8:50 - 9:35 John Sedat, Univ. of California-San Francisco 9:40 - 10:25 Edward Salmon, Univ. of N.Carolina-Chapel Hill SATURDAY, September 30, 1995 8:30 - 9:15 D. Lansing Taylor, Carnegie-Mellon University 9:20 - 10:05 Jeff Lichtman, Washington University 10:10 - 10:40 COFFEE BREAK 10:45 - 11:30 John White, University of Wisconsin-Madison 11:35 - 12:20 Steven Smith, Stanford University 12:25 - 1:25 LUNCH (on your own) 1:30 - 2:15 Gary Borisy, University of Wisconsin-Madison 2:20 - 3:05 Ralph Albrecht, Univ. of Wisconsin-Madison 3:10 - 3:55 Paul Bridgman, Washington University-Madison 5:00 - 6:00 EXHIBITOR'S SOCIAL 6:00 - 8:00 BUFFET DINNER SUNDAY (Morning), October 1, 1995 9:00 - 9:45 Kent McDonald, Univ. of California-Berkley 9:50 - 10:35 Stanley Erlandsen, University of Minnesota 10:40 - 11:10 COFFEE BREAK 11:15 - 12:00 Hans Ris, University of Wisconsin-Madison 12:05 - 12:50 Eric Henderson, Iowa State University
Fees: General Registration $100.00 (Late fee: $130.00) (Includes: Fri. Reception, Sat. Social & Dinner, Coffee Breaks and Materials) Local Registration $ 80.00 (Late fee: $110.00) (Includes: Fri. Reception, Sat. Social, Coffee Breaks and Materials) Guest Ticket - Saturday Dinner $ 20.00 (On-site Unavailable)
Interested in attending?
Consult our Web site for additional information:
http://www.bocklabs.wisc.edu/imr/imr.html
To receive a brochure and registration form write to: IMR University of Wisconsin-Madison 1675 Observatory Drive Madison, WI 53706
or email: imradmin-at-calshp.cals.wisc.edu
Following the symposium, there will be a two-day workshop at the IMR. We will be presenting lectures and provide "hands-on" experience for the following techniques: * 2-photon excitation imaging * 4D DIC imaging * cryo-SEM * high pressure freezing * reversible embedment for SEM and TEM
Workshop attendence will be limited to 25 students. A letter of application is required. Fee: $150.00.
Sorry to be late in seeing this discussion but, looking over the points made so far, I think that there seems to be a misunderstanding.
Yes, the CCD part of the Gatan (or Teitz etc.) digital TEM image recording system may have a dynamic range of 14 to 16 bits, however, there is not a 1-to-1 relation between electrons-in-the-beam and electrons-in-the-CCD. The beam electrons are first converted to photons in some sort of phosphor or scintillator. Although the exact number of photons produced/incident-beam -electron varies with the energy of the beam (less at higher kV) and with the thickness and type of phosphor, 500 might be a good number.
Of these, only a small fraction make it to the CCD while others are lost because they are going in the wrong direction to start off with and never get out of the phosphor layer (because of total internal reflection, scattering or absorbtion in the phosphor, either YAG-Ce or P-20) or because they never get into the fiber-optical coupling (or lens system), as they do not strike the lens or fiber-optic plate at the correct angle. As a result, 50-100 photons may reach the CCD, producing 25-50 electrons/incident-beam-electron. For this reason the dynamic range of the entire detector is 25-50 times less for electron-optical use than for use with light. One can use a thinner phosphor or a FO-plate with a lower NA to reduce this factor but then you lose the single-electron sensitivity that is also a good feature of this detector.
As mentioned by others, one of the other good features is the linearity of response of this detector. However, one should remember that the response of film to direct kV electron exposure is far less non-linear than is its response to light. This is because it is generally believed that two-photons are needed to expose a silver grain (and therefore the blackening is proportional to intensity squared) whereas only a single kV-electron is needed to expose a grain (because this single electron is likely to deposit far more energy with each interaction.).
The dynamic range of film exposed to electrons is likewise less than the linear exposure range (or the range of your digitizer) because each 100 kV beam electron (though perhaps not each 1000 keV etectron) is likely to expose (many) more than one grain (depending on grain size, emulsion thickness and silver density). As a result, although one may be able to digitize a brightness ratio or 10,000:1 from the developed film, this may actually only represent an electron intensity range of 1,000:1 or less.
Because of its very low "fog" level, the Fugi imaging plate is (extremely) linear over a larger range but the resolution is also low so one must generally operate at a higher magnification (and hence a higher dose rate to the specimen).
An excellent reference on the response (resolution and "speed") of a wide variety of photographic emulsions to 100 keV electrons is given in K.H. Downing and D.A. Grano, Ultramicroscopy 7:381 (1982) } } Thomas E. Phillips wants comments on grey levels possible in film vs. } digital. Film response is a sigmoidal curve and one adjusts exposure to } fit the light coming off the subject to the linear portion of the curve in } the film's response. Black & white film provides a greater number of lens } aperture stops (6-8) to capture greys compared to color film (about 4, } depends on the film). So the answer to the question of grey values } possible in film is that one can capture a great variety of greys in film, } but not all in one image-only a relatively narrow range can be captured, } with the appropriate adjustment of exposure. CCDs have a linear response } over several orders of magnitude. Slow scan CCDs can capture a remarkable } range of greys in each image, in which case one can choose which range of } greys to display on the moniter. } } } } } R. Howard Berg, Ph.D. } Biology Department } University of Memphis } Memphis, TN, 38152 } E mail: bergrh-at-cc.memphis.edu } phone: 901-678-4449 fax: 901-678-4457 }
***************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
I am going to attempt a restart of the automatic delivery mode tonight. Over the weekend we had only slightly more than normal "error" which generally were due to network connection failures, computers shutdown, disk full problems. Basically, nothing that looked like a loop, or bouncing messages. Remember if a delivery error occurs, the originator of the message will get the report and I will only see it if the originator's node is down. We continue to have problems with some nodes shutting down & or crashing. so bewarned that some delivery error messages will virtually always appear. Generally much less than 1% of the mail that comes thorough this system is not delivered. Just remember that for every single message that comes into the host over 2000 copies leave.
I am going to attempt a restart of the automatic delivery mode tonight. Over the weekend we had only slightly more than normal "error" which generally were due to network connection failures, computers shutdown, disk full problems. Basically, nothing that looked like a loop, or bouncing messages. Remember if a delivery error occurs, the originator of the message will get the report and I will only see it if the originator's node is down. We continue to have problems with some nodes shutting down & or crashing. so bewarned that some delivery error messages will virtually always appear. Generally much less than 1% of the mail that comes thorough this system is not delivered. Just remember that for every single message that comes into the host over 2000 copies leave.
Indeed Mansfields remark is correct. The point spread function is the definitive description of the performance of any detector. The psf of CCD cameras is described by Kujawa and Krahl in Ultramicroscopy 46, 395-403 (1992) and by Daberkow et al Ibid 38 215-223 (1991).
Instead of compressed gases, we routinely use liquid nitrogen. Just hold the slide in the 'fumes' until the coverslip goes opaque (you can try submersing for 10 s). The coverslip will pop off with gently prying from a single edge safety razor blade.
As for antigenicity on H&E it is very much an individual trial an error. Start by de-coloring followed by a 1 hour soak in buffer. Then proceed as normal.
Peter O. Steele, Ph.D. All Children's Hospital St. Petersburg, FL
Osmium solutions that have reduced (and changed color) can be partially or wholly restored by reoxidation using hydrogen peroxide. Just add dilute peroxide dropwise until the colour lightens. If the change is not too advanced, the original oxidation state can be completely restored.
We saved up discarded osmium from around australia for years then sent it to Johnson Matthey. But the concentration was so low the value of the product only just covered the cost of reclamation and refining. We didn't make money but I guess we got a warm fuzzy from saving a non renewable product
FS: NEW EYEPIECES 30mm BARRELS "10x/20" (fit Zeiss, Leitz, Wild)
I have a few sets of some new, modern design microscope eyepieces with a quite wide field of view, and fit ZEISS and LEITZ and WILD WF binocular eyetubes. Also work quite well with astronomical optics.
"WF10x/20" TUBE DIAMETER 30mm/1.18" MFG B&L MODEL 31-15-77 ORIGINAL COST $264/set CONDITION New in original packaging
My Price $89/pair
Ed Monberg em-at-mediacity.com
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